Research ArticleGenetic Polymorphisms of Interleukin-1 Alpha andthe Vitamin D Receptor in Mexican Mestizo Patients withIntervertebral Disc Degeneration

Salvador Cervin Serrano1 Dalia Gonzaacutelez Villareal2

Maribel Aguilar-Medina2 Jose Guillermo Romero-Navarro2

Jose Geovanni Romero Quintana2 Eliakym Araacutembula Meraz2

Ignacio Osuna Ramiacuterez2 Veronica Picos-Caacuterdenas3

Julio Granados4 Iris Estrada-Garciacutea5

Guzman Saacutenchez-Schmitz6 and Rosaliacuteo Ramos-Payaacuten27

1 Division of Pain ISSSTE Hospital 80200 Culiacan SIN Mexico2 Faculty of Biological and Chemical Sciences Doctoral Programs in Biotechnology and Biomedical SciencesAutonomous University of Sinaloa 80010 Culiacan SIN Mexico

3Human Genetics Laboratory of the Research Unit Faculty of Medicine Autonomous University of Sinaloa80019 Culiacan SIN Mexico

4Division of Immunogenetics National Institute of Medical Sciences and Nutrition 14000 Mexico City DF Mexico5 Department of Immunology National School of Biological Science National Polytechnic Institute 11340 Mexico City DF Mexico6Division of Infectious Diseases Boston Childrenrsquos Hospital and Harvard Medical School Harvard University Boston MA 02115 USA7 Laboratory of Immunology Faculty of Biological and Chemical Sciences Autonomous University of SinaloaAvenida de las Americas y Universitarios sn Ciudad Universitaria 80010 Culiacan SIN Mexico

Correspondence should be addressed to Rosalıo Ramos-Payan ramospayanyahoocommx

Received 11 September 2014 Accepted 12 November 2014 Published 20 November 2014

Academic Editor Mohamed Salem

Copyright copy 2014 Salvador Cervin Serrano et alThis is an open access article distributed under theCreativeCommonsAttributionLicense which permits unrestricted use distribution and reproduction in anymedium provided the originalwork is properly cited

Intervertebral disc degeneration (IDD) is themost common diagnosis in patients with back pain a leading cause ofmusculoskeletaldisability worldwide Several conditions such as occupational activities gender age and obesity have been associated with IDDHowever the development of this disease has strong genetic determinants In this study we explore the possible association betweenrs1800587 (c-949CgtT) of interleukin-1 alpha (IL1A) and rs2228570 (c2TgtV) and rs731236 (c1056TgtC) of vitamin D receptor(VDR) gene polymorphisms and the development of IDD in northwestern Mexican Mestizo population Gene polymorphismswere analyzed by polymerase chain reaction followed by restriction fragment length polymorphism in two groups matched byage and gender patients with symptomatic lumbar IDD (119899 = 100) and subjects with normal lumbar-spine MRI-scans (119899 = 100)Distribution of the mutated alleles in patients and controls was 270 versus 280 (119875 = 0455) for T of rs1800587 (IL1A) 530versus 580 (119875 = 0183) for V of rs2228570 (VDR) and 180 versus 210 (119875 = 0262) for C of rs731236 (VDR) Our resultsshowed no association between the studied polymorphisms and IDD in this populationThis is the first report on the contributionof gene polymorphisms on IDD in a Mexican population

1 Introduction

Intervertebral disc degeneration (IDD) is one of the mostcommon musculoskeletal disorders IDD is often diag-nosed in patients suffering back pain and is an important

contributor to workforce-absence with staggering economicimpact worldwide [1 2]

Spinal degeneration includes both osteoarthriticchanges of the facet joint and IDD Pathophysiology of discdegeneration is not well understood and is thought to

Hindawi Publishing CorporationInternational Journal of GenomicsVolume 2014 Article ID 302568 7 pageshttpdxdoiorg1011552014302568

2 International Journal of Genomics

occur gradually with aging Recently a study in Finnishpostmenopausal women showed that higher lumbar bonemineral density (BMD) and 119885-score from T2-weightedmagnetic resonance imaging (MRI) were associated withmore severe lumbar disc degeneration at L1ndashL4 levels [3] andexperimental animal models have found that osteoporosiscould have a role in IDD progression [4]

Decreased production of extracellular matrix (ECM)increased production of degrading enzymes and increasedexpression of inflammatory cytokines in the intervertebraldisc are usually observed in IDD These findings may leadto several alterations including reduced strength of thenucleus pulposus and annulus fibrosus dehydration heightreduction trabecular fissures end-plate changes Schmorlnodes osteophyte formation spinal stenosis disc bulgingherniation and discogenic pain [5 6] However IDD oftenoccurs without associated symptoms as demonstrated by thehigh incidence of degenerative changes in the asymptomaticpopulation [7ndash9]

Environmental behavioral and anthropometric factorssuch as gender obesity height occupational activities andsmoking have been associated with a higher risk for devel-oping IDD In contrast to the relatively minor contributionof these risk factors [10ndash12] twin-pair studies have found astrong familial aggregation and heritability for IDD [13ndash16]In fact genetic factors account for up to 75 of individualsusceptibility to IDD [12 16ndash19]

Association studies of genes encoding for structural andfunctional components of intervertebral disc have high-lighted the participation of polymorphisms in IDD [18 20ndash22] including collagens I [23] IX [24] and XI [25] aggre-can [26] cartilage intermediate layer protein (CILP) [27]ECM-degrading enzymes such as MMP-3 [28] and MMP9thrombospondin-2 (THBS2) [29] inflammatory cytokinesinterleukin-1 alpha (IL-1120572) [30] IL-18 [21] IL-6 and tumornecrosis factor alpha (TNF-120572) [31] and vitamin D receptor(VDR) [32]

The hormonal form of vitamin D (calcitriol) plays akey role in mineralization of bone absorption of calciumfrom the gut control of calcium and phosphate homeosta-sis and regulation of parathyroid hormone secretion andmay affect intervertebral disc maintenance by altering thesulfation of glycosaminoglycans as well as other changesrelating to the nucleus pulposus ECM Upon activation byvitamin D VDR forms a heterodimer with the retinoid-Xreceptor and binds to hormone response elements on thegenome resulting in the expression of several genes involvedin mineral metabolism and other metabolic and immunepathways Therefore it seems possible that alterations in thefunction of the VDR could participate in the pathogenesisof IDD or other diseases where vitamin D plays a role onbone and cartilage maintenance In fact several studies havedemonstrated a strong association between polymorphismsof the VDR gene with osteoporosis [33] osteoarthritis andeven IDD [32 34] Allelic variants of FokI (c2TgtV) of VDRcode for structurally different receptors the product of Vallele (mutated) shows relatively increased function com-pared with the T allele (larger product) This functional SNPis not in linkage disequilibriumwith adjacent polymorphisms

and therefore could be considered independent and directlyassociated with pathological findings [35]

IL-1120572 is a proinflammatory cytokine involved in theregulation of immune responses inflammatory processeshematopoiesis and induction of apoptosis in response to cellinjury IL-1120572 produced mainly by activated macrophagesneutrophils and epithelial and endothelial cells is thoughtto be involved in the pathogenesis of disc degeneration byincreasing the production of ECM degradation enzymes andby inhibiting ECM synthesis [36ndash38] Some polymorphismsof the IL-1120572 gene (IL1A) have been associated with discdegeneration [30 39]

The contribution of genetic factors on IDD has beenstudied mainly in Finnish Spanish Chinese and Japanesepopulations but not in Mexico Therefore in this work weselected polymorphisms rs1800587 (c-949CgtT) of IL1A andrs2228570 (c2TgtV) and rs731236 (c1056TgtC) of VDR toevaluate their potential association with lumbar IDD in aMestizo population from Sinaloa a northwestern state fromMexico

2 Materials and Methods

21 Study Subjects The population studied consisted of twogroups matched by age gender and ethnicity consecutiveunrelated patients with symptomatic lumbar IDD (119899 = 100)and nonrelated control individuals without IDD (119899 = 100)

Patients were subjected to T2-weighted MRI of thelumbar spine (L1ndashL5) to assess disc degeneration by decreasedsignal intensity height reduction bulging annular tearsherniation and osteophytes Patients were diagnosed at theDivision of Pain ISSSTE Hospital over a period of fouryears and were classified according to Modic changes onMRI and the American Society of Neuroradiology (ASNR)criteria [5 40] Controls were subjects who attended otherdivisions of the same hospital with normal lumbar-spineMRI-scans and lifetime history of no chronic low back painaccidental back injuries or spine pathology For both groupscases and controls the inclusion criteria for age were from18 to 50 years both male and female weight range from 45to 90Kg and natives of the northwestern state of SinaloaMexico (for two previous generations) Exclusion criteriawere the presence or history of spondylopathologies spinaltumors infections traumatisms diabetes systemic lupuserythematosus rheumatoid arthritis osteoporosis and otherbone diseases fibromyalgia and collagen pathologies

Age gender weight height and body mass index (BMI)were registered In accordance with the World Health Orga-nizationrsquos categories subjects with BMI ge 25 kgm2 wereconsidered overweight and ge27 as class-I obese The Ethicaland Research Committee of the ISSSTE Hospital approvedthis study and written informed consent according to theHelsinki Declaration was obtained from all subjects beforetheir enrollment in the study

22 Blood Samples Peripheral bloodwas collected by a singlevenipuncture from subjects All blood samples were drawn inEDTA containing tubes according to guidelines approved by

International Journal of Genomics 3

Table 1 Conditions and products of PCR-RFLP

Gene SNPs Primers Tm RE bp Alleles(∘C) (bp)

IL1A rs1800587 51015840-GCATGCCATCACACCTAGTT-31015840 56 NcoI 193 C 174 and 1951015840-TTACATATGAGCCTTCCATG-31015840 T 193

VDRrs2228570 51015840-AGCTGGCCCTGGCACTGCTCTGCTCT-31015840 60 FokI 265 f 197 and 68

51015840-ATGGAAACACCTTGCTTCTTCTCCCTC-31015840 F 265

rs731236 51015840-CAGAGCATGGACAGGGAGCAAG-31015840 59 119879119886119902120572I 746 T 495 and 251

51015840-GCAACTCCTCATGGCTGAGGTCTCA-31015840 t 294 251 and 201Tm annealing temperature RE restriction enzyme

the Internal Review Committee of the ISSSTE and processedwithin 1 hour

23 DNAAmplification and Restriction Fragment Length Poly-morphism Genomic DNA was isolated from whole bloodusing the salt precipitation method [41] Gene polymor-phisms were analyzed by polymerase chain reaction followedby restriction fragment length polymorphism (PCR-RFLP)Reaction conditions primers and restriction fragments aresummarized in Table 1

A region of 193 bp spanning the rs1800587 polymorphicsite at the promoter region of IL1A gene in chromosome2q14 (formerly c-899CgtT or c-949CgtT) was amplified anddigested with NcoI (2U for 2 h at 37∘C) [31] Allele C (wild)has one restriction site giving two fragments of 174 and 19 bpwhile allele T has no recognition site

PCR product of 265 bp containing the single nucleotidepolymorphism (SNP) rs2228570 at the translation initiationcodon of exon 2 of VDR gene in chromosome 12q1311(formerly c2TgtC)was digested with FokI (1 U for 2 h at 37∘C)[31] Allele T (ancestral) has one restriction site giving twofragments of 197 and 68 bp while allele V (A C or G) hasno recognition site In the same way the 747 bp ampliconspanning the synonymous polymorphism rs731236 in exon9 of VDR gene (formerly c352TgtC or c1056TgtC) [32] wascleaved with TaqI (2U for 2 h at 37∘C) Allele T (ancestral)showed two fragments of 496 and 251 bp while allele Cgenerated an additional restriction site giving three fragmentsof 294 251 and 201 bp

24 Statistical Analysis Demographic and clinical variablesof patients and controls were presented as mean plusmn SD andfrequencies Power calculation showed that a significancelevel of 119875 le 005 one-tailed directional would yield a powerof 80 with a sample size of 96 individuals per group [42]Differences in allelic genotype and haplotype frequencieswere evaluated using Fisherrsquos exact test SNPs associationswere tested under dominant and recessive genetic modelsand odds ratios (OR)with 95 confidence intervals (CI) wereused as themeasure of association between specific alleles andgenotypes with IDD and its clinical subtypes [43] Significant119875 values were corrected with the Bonferroni test for multiplecomparisons [44] Hardy-Weinbergrsquos equilibrium was calcu-lated by 1205942 test for all genotype combinations of each SNPin patients and controls PASW v180 (SPSS Inc Chicago

IL USA) and Arlequin v3512 (Swiss National ScienceFoundation) software packages were used for analysis

3 Results and Discussion

IDD is a complex pathology and genetic predisposition couldimpact the physiological maintenance of intervertebral discsor the control of inflammation and pain [15 19 22 45] ulti-mately leading to the clinical manifestations of IDD Geneticstudies on IDD have been carried out mainly in populationof European (Finnish and Spanish) or Asian (Chinese andJapanese) ancestry but not in Mexican Mestizos a majorpopulation in North America Our work carried out in anorthwestern Mexican population represents the first reportof genetic polymorphisms and IDD in a population withmixed native Mexican-European ancestry also known asMestizos

Patients with IDD and control subjects with a mean ageof 3922 (plusmn688) versus 3913 (plusmn680) years (119875 = 0918) wereincluded in this study In both groups 890 of individualswere female and 110 were male BMI data between patientsand controls was 2599 (plusmn433) versus 2623 (plusmn372) kgm2(119875 = 0673) Both groups fell into the overweight WorldHealth Organization category for BMI Covariates followeda normal distribution according to the Kolmogorov-Smirnovtest with no statistically significant differences between thegroups Disc herniation was evident in 800 of the patientsand herniation with intervertebral osteochondrosis in 200The percentages of cases based on Modic changes of spineMRIwere type I (560) type II (340) and type III (100)Clinical symptomatology included 360with sciatica 220with lumbago and 420 with lumbosciatica

Our study on the rs1800587 polymorphism of the IL1Ashowed no significant association with IDD (Table 2) Thedistribution of T allele in patients and controls was 270versus 280 (119875 = 0455 OR = 095 95 CI = 061ndash147) Genotype frequencies between the groups (119875 = 0455)were 40 versus 100 for TT 450 versus 350 for CTand 510 versus 550 for CC respectively However otherreports have shown an association between the T allele andIDD A report on the Finnish population showed that T allelecarriers have a higher risk (OR = 24) of developing discbulges than those without it [38] Another study found thatT allele was associated (25-fold risk) with Modic changes[46 47] An analysis of occupational and genetic factors

4 International Journal of Genomics

Table 2 Allelic (af ) and genotype (gf ) frequencies of IL1A andVDRpolymorphisms in controls subjects and patients with IDD

SNPs Patients Controls(119899 = 100) (119899 = 100)

rs1800587 (IL1A)Alleles 119899 af 119899 afT 53 027 55 028C 147 074 145 073Genotypes 119899 gf 119899 gfTT 4 004 10 010CT 45 045 35 035CC 51 051 55 055rs2228570 (VDR)Alleles 119899 af 119899 afV 105 053 115 058T 95 048 85 043Genotypes 119899 gf 119899 gfVV 20 020 32 032TV 65 065 51 051TT 15 015 17 017rs731236 (VDR)Alleles 119899 af 119899 afC 35 018 41 021T 165 083 159 080Genotypes 119899 gf 119899 gfCC 4 004 3 003TC 27 027 35 035TT 69 069 62 062

demonstrated that T allele represented a significant risk factorfor the disc degeneration phenotype [48] A study of a Danishpopulation also found an association of this polymorphismwith disc degeneration [39] In contrast a case-control studyof a population of Spain foundno association betweenT alleleand symptomatic lumbar disc herniation [49]The reason forthis inconsistent association between the T allele and IDDmay be due to ethnic differences in the studied populationsor by different haplotypes in the promoter or enhancerregions Mexican Mestizo populations have a high degree ofgenetic heterogeneity [50] carrying Amerindian and a fewEuropean and African HLA haplotypes [51] Moreover thepopulation studied by us in Mexico has a particular geneticbackground since half of the most common haplotypesfound in this population have a proposed European originmost likely from Spanish origin [52] Not surprisingly thefrequency of the T allele in our population (280) wassimilar to the one observed in a Spanish population (355)[49] and alike our results was not associated with IDD

Genotype frequencies observed between patients andcontrols (119875 = 0161) for rs2228570 polymorphism of VDRwere 200 versus 320 for VV 650 versus 510 forTV and 150 versus 170 for TT respectively (Table 2)

Frequencies of V allele were 530 versus 580 (119875 = 0183OR = 081 95 CI = 055ndash121) In the case of rs731236allele C of VDR (Table 2) the distribution in patients andcontrols was 180 versus 210 (119875 = 0262 OR = 08295 CI = 049ndash135) Genotype frequencies (119875 = 0262)were 40 versus 30 for CC 270 versus 350 for TCand 690 versus 620 for TT respectively In accordancewith our allelic frequencies a recent study on the associationbetween VDR polymorphisms and BMD in postmenopausalMexican women allelic frequencies of 53 for C of FokI and27 for V of TaqI were reported in the control group [53]Other reports have shown an association between these VDRpolymorphisms and IDDThe report in a Finnish populationthat demonstrated for the first time that VDR polymor-phism was associated with IDD quantitatively assessed signalintensities of thoracic and lumbar discs finding those tobe 129 worse in men with the TaqI CC genotype and93 in the men with FokI VV genotype [32] Reports onJapanese [54] Chinese [55] Turkish [56] English [57] andAustralian [58] populations also found association betweenVDR polymorphisms and IDD

Therefore for all the studied polymorphisms (Table 2)there were no significant differences in the distribution ofalleles and genotypes between the groups (119875 gt 005) underthe genetic models tested The study of haplotypes showedno differences in frequencies between the groups for anycombination of rs2228570 and rs731236 SNPs of VDR Thestratified analysis of the data (patients by pathology symp-tomatology and Modic changes) also showed no significantdifferences (all 119875 ge 005) The genotypes in the groupswere not significantly different from the expected distributionfor a population in a Hardy-Weinberg equilibrium with theexception of rs2228570 (VDR) in patients (119875 = 0010)

Our work assessed for the first time the potential contri-bution of gene polymorphisms on IDD in aMexicanMestizopopulation with homogeneous genetic background sinceboth patients and controls were ethnically and geographi-cally matched The results showed that no association existsbetween the studied polymorphisms and IDD in this pop-ulation Further analysis of other relevant polymorphismsand more Mestizo populations may contribute to finding thegenetic determinants of this disease in our country

4 Conclusion

In conclusion this is the first report on the potentialcontribution of gene polymorphisms on IDD in a Mestizopopulation in Mexico Our results showed that there is noassociation of the IL1A (rs1800587) and VDR (rs2228570and rs731236) polymorphisms with IDD The study of moreMestizo populations and more candidate genes may providefurther insight into the etiology of the disease

Consent

Written consent was obtained from patients for publicationof this work

International Journal of Genomics 5

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Authorsrsquo Contribution

Salvador Cervin Serrano diagnosed and treated the patientsSalvador Cervin Serrano Dalia Gonzalez Villareal MaribelAguilar-Medina Jose Geovanni Romero Quintana Veron-ica Picos-Cardenas and Jose Guillermo Romero-Navarroperformed the research Julio Granados Eliakym ArambulaMeraz and Ignacio Osuna Ramırez analyzed the data Sal-vador Cervin Serrano and Rosalıo Ramos-Payan designedthe research Iris Estrada-Garcıa Guzman Sanchez-Schmitzand Rosalıo Ramos-Payan wrote the paper All authors readand approved the final paper

Acknowledgment

This work was supported by CONACYT (106152)

References

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[2] D Chou D Samartzis C Bellabarba et al ldquoDegenerativemagnetic resonance imaging changes in patients with chroniclow back pain a systematic reviewrdquo Spine vol 36 supplementno 21 pp S43ndashS53 2011

[3] S Salo V Leinonen T Rikkonen et al ldquoAssociation betweenbonemineral density and lumbar disc degenerationrdquoMaturitas2014

[4] L Wang W Cui J P Kalala T V Hoof and B G Liu ldquoToinvestigate the effect of osteoporosis and intervertebral disc deg-eneration on the endplate cartilage injury in ratsrdquo Asian PacificJournal of Tropical Medicine vol 7 no 10 pp 796ndash800 2014

[5] M T Modic P M Steinberg J S Ross T J Masaryk and JR Carter ldquoDegenerative disk disease assessment of changes invertebral body marrow with MR imagingrdquo Radiology vol 166no 1 part 1 pp 193ndash199 1988

[6] S Roberts H Evans J Trivedi and J Menage ldquoHistology andpathology of the human intervertebral discrdquoThe Journal of Boneamp Joint Surgery Series A vol 88 supplement 2 pp 10ndash14 2006

[7] R Quiroz-Moreno G Lezama-Suarez and C Gomez-JimenezldquoDisc alterations of lumbar spine onmagnetic resonance imagesin asymptomatic workersrdquo Revista Medica del Instituto Mexi-cano del Seguro Social vol 46 no 2 pp 185ndash190 2008

[8] S D Boden D O Davis T S Dina N J Patronas and SW Wiesel ldquoAbnormal magnetic-resonance scans of the lumbarspine in asymptomatic subjects A prospective investigationrdquoJournal of Bone and Joint Surgery A vol 72 no 3 pp 403ndash4081990

[9] K M C Cheung J Karppinen D Chan et al ldquoPrevalenceand pattern of lumbar magnetic resonance imaging changes ina population study of one thousand forty-three individualsrdquoSpine vol 34 no 9 pp 934ndash940 2009

[10] M B Joslashrgensen A Holtermann F Gyntelberg and P Suadi-cani ldquoPhysical fitness as a predictor of herniated lumbar discdiseasemdasha 33-year follow-up in the Copenhagen male studyrdquoBMCMusculoskeletal Disorders vol 14 article 86 2013

[11] J A Buckwalter ldquoAging and degeneration of the humanintervertebral discrdquo Spine vol 20 no 11 pp 1307ndash1314 1995

[12] L Ala-Kokko ldquoGenetic risk factors for lumbar disc diseaserdquoAnnals of Medicine vol 34 no 1 pp 42ndash47 2002

[13] M C Battie T Videman L E Gibbons L D Fisher HManni-nen and K Gill ldquoDeterminants of lumbar disc degenerationa study relating lifetime exposures and magnetic resonanceimaging findings in identical twinsrdquo Spine vol 20 no 24 pp2601ndash2612 1995

[14] M C Batite D R Haynor L D Fisher S K Gill L EGibbons and T Videman ldquoSimilarities in degenerative findingson magnetic resonance images of the lumbar spines of identicaltwinsrdquo Journal of Bone and Joint SurgerymdashSeries A vol 77 no11 pp 1662ndash1670 1995

[15] P N Sambrook A J MacGregor and T D Spector ldquoGeneticinfluences on cervical and lumbar disc degeneration amagneticresonance imaging study in twinsrdquo Arthritis amp Rheumatologyvol 42 no 2 pp 366ndash372 1999

[16] M C Battie T Videman J Kaprio et al ldquoThe Twin SpineStudy contributions to a changing view of disc degenerationrdquoThe Spine Journal vol 9 no 1 pp 47ndash59 2009

[17] C Bijkerk J J Houwing-Duistermaat H A Valkenburg etal ldquoHeritabilities of radiologic osteoarthritis in peripheraljoints and of disc degeneration of the spinerdquo Arthritis ampRheumatology vol 42 no 8 pp 1729ndash1735 1999

[18] C K Kepler R K Ponnappan C A Tannoury M V Risbudand D G Anderson ldquoThemolecular basis of intervertebral discdegenerationrdquo Spine Journal vol 13 no 3 pp 318ndash330 2013

[19] S Kalb N L Martirosyan M Y Kalani G G Broc and NTheodore ldquoGenetics of the degenerated intervertebral discrdquoWorld Neurosurgery vol 77 no 3-4 pp 491ndash501 2012

[20] A A Patel W R Spiker M Daubs D Brodke and L ACannon-Albright ldquoEvidence for an inherited predisposition tolumbar disc diseaserdquo Journal of Bone and Joint SurgerymdashSeriesA vol 93 no 3 pp 225ndash229 2011

[21] T Videman J Saarela J Kaprio et al ldquoAssociations of 25structural degradative and inflammatory candidate geneswith lumbar disc desiccation bulging and height narrowingrdquoArthritis amp Rheumatism vol 60 no 2 pp 470ndash481 2009

[22] J E Mayer J C Iatridis D Chan S A Qureshi O Gottesmanand A C Hecht ldquoGenetic polymorphisms associated withintervertebral disc degenerationrdquoThe Spine Journal vol 13 no3 pp 299ndash317 2013

[23] SM F Pluijm HW Van Essen N Bravenboer et al ldquoCollagentype I 1205721 Sp1 polymorphism osteoporosis and intervertebraldisc degeneration in older men and womenrdquo Annals of theRheumatic Diseases vol 63 no 1 pp 71ndash77 2004

[24] S Annunen P Paassilta J Lohiniva et al ldquoAn allele of COL9A2associated with intervertebral disc diseaserdquo Science vol 285 no5426 pp 409ndash412 1999

[25] F Mio K Chiba Y Hirose et al ldquoA functional polymorphismin COL11A1 which encodes the 1205721 chain of type XI collagenis associated with susceptibility to lumbar disc herniationrdquoAmerican Journal of Human Genetics vol 81 no 6 pp 1271ndash1277 2007

6 International Journal of Genomics

[26] Y Kawaguchi R Osada M Kanamori et al ldquoAssociationbetween an aggrecan gene polymorphism and lumbar discdegenerationrdquo Spine vol 24 no 23 pp 2456ndash2460 1999

[27] S Seki Y Kawaguchi K Chiba et al ldquoA functional SNP inCILPencoding cartilage intermediate layer protein is associated withsusceptibility to lumbar disc diseaserdquo Nature Genetics vol 37no 6 pp 607ndash612 2005

[28] M Takahashi H Haro Y Wakabayashi T Kawa-uchi HKomori and K Shinomiya ldquoThe association of degenerationof the intervertebral disc with 5a6a polymorphism in thepromoter of the human matrix metalloproteinase-3 generdquoJournal of Bone and Joint Surgery vol 83 no 4 pp 491ndash4952001

[29] Y Hirose K Chiba T Karasugi et al ldquoA functional polymor-phism inTHBS2 that affects alternative splicing andMMPbind-ing is associated with lumbar-disc herniationrdquo The AmericanJournal of Human Genetics vol 82 no 5 pp 1122ndash1129 2008

[30] S Solovieva J Lohiniva P Leino-Arjas et al ldquoIntervertebraldisc degeneration in relation to the COL9A3 and the IL-1120573 genepolymorphismsrdquo European Spine Journal vol 15 no 5 pp 613ndash619 2006

[31] N Noponen-Hietala I Virtanen R Karttunen et al ldquoGeneticvariations in IL6 associate with intervertebral disc diseasecharacterized by sciaticardquo Pain vol 114 no 1-2 pp 186ndash1942005

[32] T Videman J Leppavuori J Kaprio et al ldquoIntragenic poly-morphisms of the vitamin D receptor gene associated withintervertebral disc degenerationrdquo Spine vol 23 no 23 pp2477ndash2485 1998

[33] R JWood and J C Fleet ldquoThe genetics of osteoporosis vitaminD receptor polymorphismsrdquoAnnual Review of Nutrition vol 18pp 233ndash258 1998

[34] A Colombini S Cauci G Lombardi et al ldquoRelationshipbetween vitamin D receptor gene (VDR) polymorphismsvitamin D status osteoarthritis and intervertebral disc degen-erationrdquo The Journal of Steroid Biochemistry and MolecularBiology vol 138 pp 24ndash40 2013

[35] A G Uitterlinden Y Fang J B J van Meurs H A P Polsand J P T M van Leeuwen ldquoGenetics and biology of vitaminD receptor polymorphismsrdquo Gene vol 338 no 2 pp 143ndash1562004

[36] K L E Phillips N Jordan-Mahy M J H Nicklin and CL Le Maitre ldquoInterleukin-1 receptor antagonist deficient miceprovide insights into pathogenesis of human intervertebral discdegenerationrdquo Annals of the Rheumatic Diseases vol 72 no 11pp 1860ndash1867 2013

[37] W Ye D S Huang W J Chen et al ldquoAssociation of 86 bpvariable number tandem repeat polymorphism of interleukin-1receptor antagonist gene with lumbar disc diseaserdquoNan Fang YiKe Da Xue Xue Bao vol 27 no 10 pp 1485ndash1488 2007

[38] S Solovieva S Kouhia P Leino-Arjas et al ldquoInterleukin 1polymorphisms and intervertebral disc degenerationrdquo Epidemi-ology vol 15 no 5 pp 626ndash633 2004

[39] P J Eskola P Kjaer I M Daavittila et al ldquoGenetic risk factorsof disc degeneration among 12-14-year-old Danish children apopulation studyrdquo International Journal of Molecular Epidemi-ology and Genetics vol 1 no 2 pp 158ndash165 2010

[40] M T Modic T J Masaryk J S Ross and J R Carter ldquoImagingof degenerative disk diseaserdquo Radiology vol 168 no 1 pp 177ndash186 1988

[41] S Gustincich G Manfioletti G Del Sal C Schneider andP Carninci ldquoA fast method for high-quality genomic DNAextraction fromwhole human bloodrdquo BioTechniques vol 11 no3 pp 298ndash302 1991

[42] M Gail R Williams D P Byar and C Brown ldquoHow manycontrolsrdquo Journal of Chronic Diseases vol 29 no 11 pp 723ndash731 1976

[43] B Woolf ldquoOn estimating the relation between blood group anddiseaserdquo Annals of Human Genetics vol 19 no 4 pp 251ndash2531955

[44] T V Perneger ldquoWhatrsquos wrong with Bonferroni adjustmentsrdquoBritish Medical Journal vol 316 no 7139 pp 1236ndash1238 1998

[45] I Tegeder and J Lotsch ldquoCurrent evidence for a modulation oflow back pain by human genetic variantsrdquo Journal of Cellularand Molecular Medicine vol 13 no 8B pp 1605ndash1619 2009

[46] J Karppinen S Solovieva K Luoma R Raininko P Leino-Arjas and H Riihimaki ldquoModic changes and interleukin 1 genelocus polymorphisms in occupational cohort of middle-agedmenrdquo European Spine Journal vol 18 no 12 pp 1963ndash19702009

[47] J Karppinen I Daavittila S Solovieva et al ldquoGenetic factorsare associated with modic changes in endplates of lumbarvertebral bodiesrdquo Spine vol 33 no 11 pp 1236ndash1241 2008

[48] I M Virtanen J Karppinen S Taimela et al ldquoOccupationaland genetic risk factors associated with intervertebral discdiseaserdquo Spine vol 32 no 10 pp 1129ndash1134 2007

[49] J Paz Aparicio I Fernandez Bances E Lopez-AngladaFernandez et al ldquoThe IL-1120573 (+3953 TC) gene polymorphismassociates to symptomatic lumbar disc herniationrdquo EuropeanSpine Journal vol 20 no 3 pp 383ndash389 2011

[50] AMoreno-Estrada C R Gignoux J C Fernandez-Lopez et alldquoHuman genetics The genetics of Mexico recapitulates NativeAmerican substructure and affects biomedical traitsrdquo Sciencevol 344 no 6189 pp 1280ndash1285 2014

[51] R Barquera J Zuniga R Hernandez-Dıaz et al ldquoHLA class Iand class II haplotypes in admixed families from several regionsof MexicordquoMolecular Immunology vol 45 no 4 pp 1171ndash11782008

[52] J G Romero-Quintana L O Frıas-Castro E Arambula-Merazet al ldquoIdentification of novel mutation in cathepsin C genecausing Papillon-Lefevre Syndrome in Mexican patientsrdquo BMCMedical Genetics vol 14 no 1 article 7 2013

[53] A Gonzalez-Mercado J Y Sanchez-Lopez J A Regla-Nava etal ldquoAssociation analysis of vitamin D receptor gene polymor-phisms and bone mineral density in postmenopausal Mexican-Mestizo womenrdquo Genetics and Molecular Research vol 12 no3 pp 2755ndash2763 2013

[54] Y KawaguchiMKanamoriH Ishihara KOhmoriHMatsuiand T Kimura ldquoThe association of lumbar disc disease withvitamin-D receptor gene polymorphismrdquo Journal of Bone andJoint SurgerymdashSeries A vol 84 no 11 pp 2022ndash2028 2002

[55] K M C Cheung D Chan J Karppinen et al ldquoAssociation ofthe Taq I allele in vitamin D receptor with degenerative discdisease and disc bulge in a Chinese populationrdquo Spine vol 31no 10 pp 1143ndash1148 2006

[56] B Eser T Cora O Eser et al ldquoAssociation of the polymor-phisms of vitamin d receptor and aggrecan genes with degen-erative disc diseaserdquo Genetic Testing and Molecular Biomarkersvol 14 no 3 pp 313ndash317 2010

International Journal of Genomics 7

[57] A M Valdes G Hassett D J Hart and T D SpectorldquoRadiographic progression of lumbar spine disc degenerationis influenced by variation at inflammatory genes a candidateSNP association study in the Chingford cohortrdquo Spine vol 30no 21 pp 2445ndash2451 2005

[58] G Jones C White P Sambrook and J Eisman ldquoAllelicvariation in the vitamin D receptor lifestyle factors and lumbarspinal degenerative diseaserdquo Annals of the Rheumatic Diseasesvol 57 no 2 pp 94ndash99 1998

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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2 International Journal of Genomics

occur gradually with aging Recently a study in Finnishpostmenopausal women showed that higher lumbar bonemineral density (BMD) and 119885-score from T2-weightedmagnetic resonance imaging (MRI) were associated withmore severe lumbar disc degeneration at L1ndashL4 levels [3] andexperimental animal models have found that osteoporosiscould have a role in IDD progression [4]

Decreased production of extracellular matrix (ECM)increased production of degrading enzymes and increasedexpression of inflammatory cytokines in the intervertebraldisc are usually observed in IDD These findings may leadto several alterations including reduced strength of thenucleus pulposus and annulus fibrosus dehydration heightreduction trabecular fissures end-plate changes Schmorlnodes osteophyte formation spinal stenosis disc bulgingherniation and discogenic pain [5 6] However IDD oftenoccurs without associated symptoms as demonstrated by thehigh incidence of degenerative changes in the asymptomaticpopulation [7ndash9]

Environmental behavioral and anthropometric factorssuch as gender obesity height occupational activities andsmoking have been associated with a higher risk for devel-oping IDD In contrast to the relatively minor contributionof these risk factors [10ndash12] twin-pair studies have found astrong familial aggregation and heritability for IDD [13ndash16]In fact genetic factors account for up to 75 of individualsusceptibility to IDD [12 16ndash19]

Association studies of genes encoding for structural andfunctional components of intervertebral disc have high-lighted the participation of polymorphisms in IDD [18 20ndash22] including collagens I [23] IX [24] and XI [25] aggre-can [26] cartilage intermediate layer protein (CILP) [27]ECM-degrading enzymes such as MMP-3 [28] and MMP9thrombospondin-2 (THBS2) [29] inflammatory cytokinesinterleukin-1 alpha (IL-1120572) [30] IL-18 [21] IL-6 and tumornecrosis factor alpha (TNF-120572) [31] and vitamin D receptor(VDR) [32]

The hormonal form of vitamin D (calcitriol) plays akey role in mineralization of bone absorption of calciumfrom the gut control of calcium and phosphate homeosta-sis and regulation of parathyroid hormone secretion andmay affect intervertebral disc maintenance by altering thesulfation of glycosaminoglycans as well as other changesrelating to the nucleus pulposus ECM Upon activation byvitamin D VDR forms a heterodimer with the retinoid-Xreceptor and binds to hormone response elements on thegenome resulting in the expression of several genes involvedin mineral metabolism and other metabolic and immunepathways Therefore it seems possible that alterations in thefunction of the VDR could participate in the pathogenesisof IDD or other diseases where vitamin D plays a role onbone and cartilage maintenance In fact several studies havedemonstrated a strong association between polymorphismsof the VDR gene with osteoporosis [33] osteoarthritis andeven IDD [32 34] Allelic variants of FokI (c2TgtV) of VDRcode for structurally different receptors the product of Vallele (mutated) shows relatively increased function com-pared with the T allele (larger product) This functional SNPis not in linkage disequilibriumwith adjacent polymorphisms

and therefore could be considered independent and directlyassociated with pathological findings [35]

IL-1120572 is a proinflammatory cytokine involved in theregulation of immune responses inflammatory processeshematopoiesis and induction of apoptosis in response to cellinjury IL-1120572 produced mainly by activated macrophagesneutrophils and epithelial and endothelial cells is thoughtto be involved in the pathogenesis of disc degeneration byincreasing the production of ECM degradation enzymes andby inhibiting ECM synthesis [36ndash38] Some polymorphismsof the IL-1120572 gene (IL1A) have been associated with discdegeneration [30 39]

The contribution of genetic factors on IDD has beenstudied mainly in Finnish Spanish Chinese and Japanesepopulations but not in Mexico Therefore in this work weselected polymorphisms rs1800587 (c-949CgtT) of IL1A andrs2228570 (c2TgtV) and rs731236 (c1056TgtC) of VDR toevaluate their potential association with lumbar IDD in aMestizo population from Sinaloa a northwestern state fromMexico

2 Materials and Methods

21 Study Subjects The population studied consisted of twogroups matched by age gender and ethnicity consecutiveunrelated patients with symptomatic lumbar IDD (119899 = 100)and nonrelated control individuals without IDD (119899 = 100)

Patients were subjected to T2-weighted MRI of thelumbar spine (L1ndashL5) to assess disc degeneration by decreasedsignal intensity height reduction bulging annular tearsherniation and osteophytes Patients were diagnosed at theDivision of Pain ISSSTE Hospital over a period of fouryears and were classified according to Modic changes onMRI and the American Society of Neuroradiology (ASNR)criteria [5 40] Controls were subjects who attended otherdivisions of the same hospital with normal lumbar-spineMRI-scans and lifetime history of no chronic low back painaccidental back injuries or spine pathology For both groupscases and controls the inclusion criteria for age were from18 to 50 years both male and female weight range from 45to 90Kg and natives of the northwestern state of SinaloaMexico (for two previous generations) Exclusion criteriawere the presence or history of spondylopathologies spinaltumors infections traumatisms diabetes systemic lupuserythematosus rheumatoid arthritis osteoporosis and otherbone diseases fibromyalgia and collagen pathologies

Age gender weight height and body mass index (BMI)were registered In accordance with the World Health Orga-nizationrsquos categories subjects with BMI ge 25 kgm2 wereconsidered overweight and ge27 as class-I obese The Ethicaland Research Committee of the ISSSTE Hospital approvedthis study and written informed consent according to theHelsinki Declaration was obtained from all subjects beforetheir enrollment in the study

22 Blood Samples Peripheral bloodwas collected by a singlevenipuncture from subjects All blood samples were drawn inEDTA containing tubes according to guidelines approved by

International Journal of Genomics 3

Table 1 Conditions and products of PCR-RFLP

Gene SNPs Primers Tm RE bp Alleles(∘C) (bp)

IL1A rs1800587 51015840-GCATGCCATCACACCTAGTT-31015840 56 NcoI 193 C 174 and 1951015840-TTACATATGAGCCTTCCATG-31015840 T 193

VDRrs2228570 51015840-AGCTGGCCCTGGCACTGCTCTGCTCT-31015840 60 FokI 265 f 197 and 68

51015840-ATGGAAACACCTTGCTTCTTCTCCCTC-31015840 F 265

rs731236 51015840-CAGAGCATGGACAGGGAGCAAG-31015840 59 119879119886119902120572I 746 T 495 and 251

51015840-GCAACTCCTCATGGCTGAGGTCTCA-31015840 t 294 251 and 201Tm annealing temperature RE restriction enzyme

the Internal Review Committee of the ISSSTE and processedwithin 1 hour

23 DNAAmplification and Restriction Fragment Length Poly-morphism Genomic DNA was isolated from whole bloodusing the salt precipitation method [41] Gene polymor-phisms were analyzed by polymerase chain reaction followedby restriction fragment length polymorphism (PCR-RFLP)Reaction conditions primers and restriction fragments aresummarized in Table 1

A region of 193 bp spanning the rs1800587 polymorphicsite at the promoter region of IL1A gene in chromosome2q14 (formerly c-899CgtT or c-949CgtT) was amplified anddigested with NcoI (2U for 2 h at 37∘C) [31] Allele C (wild)has one restriction site giving two fragments of 174 and 19 bpwhile allele T has no recognition site

PCR product of 265 bp containing the single nucleotidepolymorphism (SNP) rs2228570 at the translation initiationcodon of exon 2 of VDR gene in chromosome 12q1311(formerly c2TgtC)was digested with FokI (1 U for 2 h at 37∘C)[31] Allele T (ancestral) has one restriction site giving twofragments of 197 and 68 bp while allele V (A C or G) hasno recognition site In the same way the 747 bp ampliconspanning the synonymous polymorphism rs731236 in exon9 of VDR gene (formerly c352TgtC or c1056TgtC) [32] wascleaved with TaqI (2U for 2 h at 37∘C) Allele T (ancestral)showed two fragments of 496 and 251 bp while allele Cgenerated an additional restriction site giving three fragmentsof 294 251 and 201 bp

24 Statistical Analysis Demographic and clinical variablesof patients and controls were presented as mean plusmn SD andfrequencies Power calculation showed that a significancelevel of 119875 le 005 one-tailed directional would yield a powerof 80 with a sample size of 96 individuals per group [42]Differences in allelic genotype and haplotype frequencieswere evaluated using Fisherrsquos exact test SNPs associationswere tested under dominant and recessive genetic modelsand odds ratios (OR)with 95 confidence intervals (CI) wereused as themeasure of association between specific alleles andgenotypes with IDD and its clinical subtypes [43] Significant119875 values were corrected with the Bonferroni test for multiplecomparisons [44] Hardy-Weinbergrsquos equilibrium was calcu-lated by 1205942 test for all genotype combinations of each SNPin patients and controls PASW v180 (SPSS Inc Chicago

IL USA) and Arlequin v3512 (Swiss National ScienceFoundation) software packages were used for analysis

3 Results and Discussion

IDD is a complex pathology and genetic predisposition couldimpact the physiological maintenance of intervertebral discsor the control of inflammation and pain [15 19 22 45] ulti-mately leading to the clinical manifestations of IDD Geneticstudies on IDD have been carried out mainly in populationof European (Finnish and Spanish) or Asian (Chinese andJapanese) ancestry but not in Mexican Mestizos a majorpopulation in North America Our work carried out in anorthwestern Mexican population represents the first reportof genetic polymorphisms and IDD in a population withmixed native Mexican-European ancestry also known asMestizos

Patients with IDD and control subjects with a mean ageof 3922 (plusmn688) versus 3913 (plusmn680) years (119875 = 0918) wereincluded in this study In both groups 890 of individualswere female and 110 were male BMI data between patientsand controls was 2599 (plusmn433) versus 2623 (plusmn372) kgm2(119875 = 0673) Both groups fell into the overweight WorldHealth Organization category for BMI Covariates followeda normal distribution according to the Kolmogorov-Smirnovtest with no statistically significant differences between thegroups Disc herniation was evident in 800 of the patientsand herniation with intervertebral osteochondrosis in 200The percentages of cases based on Modic changes of spineMRIwere type I (560) type II (340) and type III (100)Clinical symptomatology included 360with sciatica 220with lumbago and 420 with lumbosciatica

Our study on the rs1800587 polymorphism of the IL1Ashowed no significant association with IDD (Table 2) Thedistribution of T allele in patients and controls was 270versus 280 (119875 = 0455 OR = 095 95 CI = 061ndash147) Genotype frequencies between the groups (119875 = 0455)were 40 versus 100 for TT 450 versus 350 for CTand 510 versus 550 for CC respectively However otherreports have shown an association between the T allele andIDD A report on the Finnish population showed that T allelecarriers have a higher risk (OR = 24) of developing discbulges than those without it [38] Another study found thatT allele was associated (25-fold risk) with Modic changes[46 47] An analysis of occupational and genetic factors

4 International Journal of Genomics

Table 2 Allelic (af ) and genotype (gf ) frequencies of IL1A andVDRpolymorphisms in controls subjects and patients with IDD

SNPs Patients Controls(119899 = 100) (119899 = 100)

rs1800587 (IL1A)Alleles 119899 af 119899 afT 53 027 55 028C 147 074 145 073Genotypes 119899 gf 119899 gfTT 4 004 10 010CT 45 045 35 035CC 51 051 55 055rs2228570 (VDR)Alleles 119899 af 119899 afV 105 053 115 058T 95 048 85 043Genotypes 119899 gf 119899 gfVV 20 020 32 032TV 65 065 51 051TT 15 015 17 017rs731236 (VDR)Alleles 119899 af 119899 afC 35 018 41 021T 165 083 159 080Genotypes 119899 gf 119899 gfCC 4 004 3 003TC 27 027 35 035TT 69 069 62 062

demonstrated that T allele represented a significant risk factorfor the disc degeneration phenotype [48] A study of a Danishpopulation also found an association of this polymorphismwith disc degeneration [39] In contrast a case-control studyof a population of Spain foundno association betweenT alleleand symptomatic lumbar disc herniation [49]The reason forthis inconsistent association between the T allele and IDDmay be due to ethnic differences in the studied populationsor by different haplotypes in the promoter or enhancerregions Mexican Mestizo populations have a high degree ofgenetic heterogeneity [50] carrying Amerindian and a fewEuropean and African HLA haplotypes [51] Moreover thepopulation studied by us in Mexico has a particular geneticbackground since half of the most common haplotypesfound in this population have a proposed European originmost likely from Spanish origin [52] Not surprisingly thefrequency of the T allele in our population (280) wassimilar to the one observed in a Spanish population (355)[49] and alike our results was not associated with IDD

Genotype frequencies observed between patients andcontrols (119875 = 0161) for rs2228570 polymorphism of VDRwere 200 versus 320 for VV 650 versus 510 forTV and 150 versus 170 for TT respectively (Table 2)

Frequencies of V allele were 530 versus 580 (119875 = 0183OR = 081 95 CI = 055ndash121) In the case of rs731236allele C of VDR (Table 2) the distribution in patients andcontrols was 180 versus 210 (119875 = 0262 OR = 08295 CI = 049ndash135) Genotype frequencies (119875 = 0262)were 40 versus 30 for CC 270 versus 350 for TCand 690 versus 620 for TT respectively In accordancewith our allelic frequencies a recent study on the associationbetween VDR polymorphisms and BMD in postmenopausalMexican women allelic frequencies of 53 for C of FokI and27 for V of TaqI were reported in the control group [53]Other reports have shown an association between these VDRpolymorphisms and IDDThe report in a Finnish populationthat demonstrated for the first time that VDR polymor-phism was associated with IDD quantitatively assessed signalintensities of thoracic and lumbar discs finding those tobe 129 worse in men with the TaqI CC genotype and93 in the men with FokI VV genotype [32] Reports onJapanese [54] Chinese [55] Turkish [56] English [57] andAustralian [58] populations also found association betweenVDR polymorphisms and IDD

Therefore for all the studied polymorphisms (Table 2)there were no significant differences in the distribution ofalleles and genotypes between the groups (119875 gt 005) underthe genetic models tested The study of haplotypes showedno differences in frequencies between the groups for anycombination of rs2228570 and rs731236 SNPs of VDR Thestratified analysis of the data (patients by pathology symp-tomatology and Modic changes) also showed no significantdifferences (all 119875 ge 005) The genotypes in the groupswere not significantly different from the expected distributionfor a population in a Hardy-Weinberg equilibrium with theexception of rs2228570 (VDR) in patients (119875 = 0010)

Our work assessed for the first time the potential contri-bution of gene polymorphisms on IDD in aMexicanMestizopopulation with homogeneous genetic background sinceboth patients and controls were ethnically and geographi-cally matched The results showed that no association existsbetween the studied polymorphisms and IDD in this pop-ulation Further analysis of other relevant polymorphismsand more Mestizo populations may contribute to finding thegenetic determinants of this disease in our country

4 Conclusion

In conclusion this is the first report on the potentialcontribution of gene polymorphisms on IDD in a Mestizopopulation in Mexico Our results showed that there is noassociation of the IL1A (rs1800587) and VDR (rs2228570and rs731236) polymorphisms with IDD The study of moreMestizo populations and more candidate genes may providefurther insight into the etiology of the disease

Consent

Written consent was obtained from patients for publicationof this work

International Journal of Genomics 5

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Authorsrsquo Contribution

Salvador Cervin Serrano diagnosed and treated the patientsSalvador Cervin Serrano Dalia Gonzalez Villareal MaribelAguilar-Medina Jose Geovanni Romero Quintana Veron-ica Picos-Cardenas and Jose Guillermo Romero-Navarroperformed the research Julio Granados Eliakym ArambulaMeraz and Ignacio Osuna Ramırez analyzed the data Sal-vador Cervin Serrano and Rosalıo Ramos-Payan designedthe research Iris Estrada-Garcıa Guzman Sanchez-Schmitzand Rosalıo Ramos-Payan wrote the paper All authors readand approved the final paper

Acknowledgment

This work was supported by CONACYT (106152)

References

[1] USBJDUSBJI ldquoSpine low back and neck painrdquo in The Burdenof Musculoskeletal Diseases in the United States vol 1 pp 21ndash56American Academy of Orthopaedic Surgeons 2011

[2] D Chou D Samartzis C Bellabarba et al ldquoDegenerativemagnetic resonance imaging changes in patients with chroniclow back pain a systematic reviewrdquo Spine vol 36 supplementno 21 pp S43ndashS53 2011

[3] S Salo V Leinonen T Rikkonen et al ldquoAssociation betweenbonemineral density and lumbar disc degenerationrdquoMaturitas2014

[4] L Wang W Cui J P Kalala T V Hoof and B G Liu ldquoToinvestigate the effect of osteoporosis and intervertebral disc deg-eneration on the endplate cartilage injury in ratsrdquo Asian PacificJournal of Tropical Medicine vol 7 no 10 pp 796ndash800 2014

[5] M T Modic P M Steinberg J S Ross T J Masaryk and JR Carter ldquoDegenerative disk disease assessment of changes invertebral body marrow with MR imagingrdquo Radiology vol 166no 1 part 1 pp 193ndash199 1988

[6] S Roberts H Evans J Trivedi and J Menage ldquoHistology andpathology of the human intervertebral discrdquoThe Journal of Boneamp Joint Surgery Series A vol 88 supplement 2 pp 10ndash14 2006

[7] R Quiroz-Moreno G Lezama-Suarez and C Gomez-JimenezldquoDisc alterations of lumbar spine onmagnetic resonance imagesin asymptomatic workersrdquo Revista Medica del Instituto Mexi-cano del Seguro Social vol 46 no 2 pp 185ndash190 2008

[8] S D Boden D O Davis T S Dina N J Patronas and SW Wiesel ldquoAbnormal magnetic-resonance scans of the lumbarspine in asymptomatic subjects A prospective investigationrdquoJournal of Bone and Joint Surgery A vol 72 no 3 pp 403ndash4081990

[9] K M C Cheung J Karppinen D Chan et al ldquoPrevalenceand pattern of lumbar magnetic resonance imaging changes ina population study of one thousand forty-three individualsrdquoSpine vol 34 no 9 pp 934ndash940 2009

[10] M B Joslashrgensen A Holtermann F Gyntelberg and P Suadi-cani ldquoPhysical fitness as a predictor of herniated lumbar discdiseasemdasha 33-year follow-up in the Copenhagen male studyrdquoBMCMusculoskeletal Disorders vol 14 article 86 2013

[11] J A Buckwalter ldquoAging and degeneration of the humanintervertebral discrdquo Spine vol 20 no 11 pp 1307ndash1314 1995

[12] L Ala-Kokko ldquoGenetic risk factors for lumbar disc diseaserdquoAnnals of Medicine vol 34 no 1 pp 42ndash47 2002

[13] M C Battie T Videman L E Gibbons L D Fisher HManni-nen and K Gill ldquoDeterminants of lumbar disc degenerationa study relating lifetime exposures and magnetic resonanceimaging findings in identical twinsrdquo Spine vol 20 no 24 pp2601ndash2612 1995

[14] M C Batite D R Haynor L D Fisher S K Gill L EGibbons and T Videman ldquoSimilarities in degenerative findingson magnetic resonance images of the lumbar spines of identicaltwinsrdquo Journal of Bone and Joint SurgerymdashSeries A vol 77 no11 pp 1662ndash1670 1995

[15] P N Sambrook A J MacGregor and T D Spector ldquoGeneticinfluences on cervical and lumbar disc degeneration amagneticresonance imaging study in twinsrdquo Arthritis amp Rheumatologyvol 42 no 2 pp 366ndash372 1999

[16] M C Battie T Videman J Kaprio et al ldquoThe Twin SpineStudy contributions to a changing view of disc degenerationrdquoThe Spine Journal vol 9 no 1 pp 47ndash59 2009

[17] C Bijkerk J J Houwing-Duistermaat H A Valkenburg etal ldquoHeritabilities of radiologic osteoarthritis in peripheraljoints and of disc degeneration of the spinerdquo Arthritis ampRheumatology vol 42 no 8 pp 1729ndash1735 1999

[18] C K Kepler R K Ponnappan C A Tannoury M V Risbudand D G Anderson ldquoThemolecular basis of intervertebral discdegenerationrdquo Spine Journal vol 13 no 3 pp 318ndash330 2013

[19] S Kalb N L Martirosyan M Y Kalani G G Broc and NTheodore ldquoGenetics of the degenerated intervertebral discrdquoWorld Neurosurgery vol 77 no 3-4 pp 491ndash501 2012

[20] A A Patel W R Spiker M Daubs D Brodke and L ACannon-Albright ldquoEvidence for an inherited predisposition tolumbar disc diseaserdquo Journal of Bone and Joint SurgerymdashSeriesA vol 93 no 3 pp 225ndash229 2011

[21] T Videman J Saarela J Kaprio et al ldquoAssociations of 25structural degradative and inflammatory candidate geneswith lumbar disc desiccation bulging and height narrowingrdquoArthritis amp Rheumatism vol 60 no 2 pp 470ndash481 2009

[22] J E Mayer J C Iatridis D Chan S A Qureshi O Gottesmanand A C Hecht ldquoGenetic polymorphisms associated withintervertebral disc degenerationrdquoThe Spine Journal vol 13 no3 pp 299ndash317 2013

[23] SM F Pluijm HW Van Essen N Bravenboer et al ldquoCollagentype I 1205721 Sp1 polymorphism osteoporosis and intervertebraldisc degeneration in older men and womenrdquo Annals of theRheumatic Diseases vol 63 no 1 pp 71ndash77 2004

[24] S Annunen P Paassilta J Lohiniva et al ldquoAn allele of COL9A2associated with intervertebral disc diseaserdquo Science vol 285 no5426 pp 409ndash412 1999

[25] F Mio K Chiba Y Hirose et al ldquoA functional polymorphismin COL11A1 which encodes the 1205721 chain of type XI collagenis associated with susceptibility to lumbar disc herniationrdquoAmerican Journal of Human Genetics vol 81 no 6 pp 1271ndash1277 2007

6 International Journal of Genomics

[26] Y Kawaguchi R Osada M Kanamori et al ldquoAssociationbetween an aggrecan gene polymorphism and lumbar discdegenerationrdquo Spine vol 24 no 23 pp 2456ndash2460 1999

[27] S Seki Y Kawaguchi K Chiba et al ldquoA functional SNP inCILPencoding cartilage intermediate layer protein is associated withsusceptibility to lumbar disc diseaserdquo Nature Genetics vol 37no 6 pp 607ndash612 2005

[28] M Takahashi H Haro Y Wakabayashi T Kawa-uchi HKomori and K Shinomiya ldquoThe association of degenerationof the intervertebral disc with 5a6a polymorphism in thepromoter of the human matrix metalloproteinase-3 generdquoJournal of Bone and Joint Surgery vol 83 no 4 pp 491ndash4952001

[29] Y Hirose K Chiba T Karasugi et al ldquoA functional polymor-phism inTHBS2 that affects alternative splicing andMMPbind-ing is associated with lumbar-disc herniationrdquo The AmericanJournal of Human Genetics vol 82 no 5 pp 1122ndash1129 2008

[30] S Solovieva J Lohiniva P Leino-Arjas et al ldquoIntervertebraldisc degeneration in relation to the COL9A3 and the IL-1120573 genepolymorphismsrdquo European Spine Journal vol 15 no 5 pp 613ndash619 2006

[31] N Noponen-Hietala I Virtanen R Karttunen et al ldquoGeneticvariations in IL6 associate with intervertebral disc diseasecharacterized by sciaticardquo Pain vol 114 no 1-2 pp 186ndash1942005

[32] T Videman J Leppavuori J Kaprio et al ldquoIntragenic poly-morphisms of the vitamin D receptor gene associated withintervertebral disc degenerationrdquo Spine vol 23 no 23 pp2477ndash2485 1998

[33] R JWood and J C Fleet ldquoThe genetics of osteoporosis vitaminD receptor polymorphismsrdquoAnnual Review of Nutrition vol 18pp 233ndash258 1998

[34] A Colombini S Cauci G Lombardi et al ldquoRelationshipbetween vitamin D receptor gene (VDR) polymorphismsvitamin D status osteoarthritis and intervertebral disc degen-erationrdquo The Journal of Steroid Biochemistry and MolecularBiology vol 138 pp 24ndash40 2013

[35] A G Uitterlinden Y Fang J B J van Meurs H A P Polsand J P T M van Leeuwen ldquoGenetics and biology of vitaminD receptor polymorphismsrdquo Gene vol 338 no 2 pp 143ndash1562004

[36] K L E Phillips N Jordan-Mahy M J H Nicklin and CL Le Maitre ldquoInterleukin-1 receptor antagonist deficient miceprovide insights into pathogenesis of human intervertebral discdegenerationrdquo Annals of the Rheumatic Diseases vol 72 no 11pp 1860ndash1867 2013

[37] W Ye D S Huang W J Chen et al ldquoAssociation of 86 bpvariable number tandem repeat polymorphism of interleukin-1receptor antagonist gene with lumbar disc diseaserdquoNan Fang YiKe Da Xue Xue Bao vol 27 no 10 pp 1485ndash1488 2007

[38] S Solovieva S Kouhia P Leino-Arjas et al ldquoInterleukin 1polymorphisms and intervertebral disc degenerationrdquo Epidemi-ology vol 15 no 5 pp 626ndash633 2004

[39] P J Eskola P Kjaer I M Daavittila et al ldquoGenetic risk factorsof disc degeneration among 12-14-year-old Danish children apopulation studyrdquo International Journal of Molecular Epidemi-ology and Genetics vol 1 no 2 pp 158ndash165 2010

[40] M T Modic T J Masaryk J S Ross and J R Carter ldquoImagingof degenerative disk diseaserdquo Radiology vol 168 no 1 pp 177ndash186 1988

[41] S Gustincich G Manfioletti G Del Sal C Schneider andP Carninci ldquoA fast method for high-quality genomic DNAextraction fromwhole human bloodrdquo BioTechniques vol 11 no3 pp 298ndash302 1991

[42] M Gail R Williams D P Byar and C Brown ldquoHow manycontrolsrdquo Journal of Chronic Diseases vol 29 no 11 pp 723ndash731 1976

[43] B Woolf ldquoOn estimating the relation between blood group anddiseaserdquo Annals of Human Genetics vol 19 no 4 pp 251ndash2531955

[44] T V Perneger ldquoWhatrsquos wrong with Bonferroni adjustmentsrdquoBritish Medical Journal vol 316 no 7139 pp 1236ndash1238 1998

[45] I Tegeder and J Lotsch ldquoCurrent evidence for a modulation oflow back pain by human genetic variantsrdquo Journal of Cellularand Molecular Medicine vol 13 no 8B pp 1605ndash1619 2009

[46] J Karppinen S Solovieva K Luoma R Raininko P Leino-Arjas and H Riihimaki ldquoModic changes and interleukin 1 genelocus polymorphisms in occupational cohort of middle-agedmenrdquo European Spine Journal vol 18 no 12 pp 1963ndash19702009

[47] J Karppinen I Daavittila S Solovieva et al ldquoGenetic factorsare associated with modic changes in endplates of lumbarvertebral bodiesrdquo Spine vol 33 no 11 pp 1236ndash1241 2008

[48] I M Virtanen J Karppinen S Taimela et al ldquoOccupationaland genetic risk factors associated with intervertebral discdiseaserdquo Spine vol 32 no 10 pp 1129ndash1134 2007

[49] J Paz Aparicio I Fernandez Bances E Lopez-AngladaFernandez et al ldquoThe IL-1120573 (+3953 TC) gene polymorphismassociates to symptomatic lumbar disc herniationrdquo EuropeanSpine Journal vol 20 no 3 pp 383ndash389 2011

[50] AMoreno-Estrada C R Gignoux J C Fernandez-Lopez et alldquoHuman genetics The genetics of Mexico recapitulates NativeAmerican substructure and affects biomedical traitsrdquo Sciencevol 344 no 6189 pp 1280ndash1285 2014

[51] R Barquera J Zuniga R Hernandez-Dıaz et al ldquoHLA class Iand class II haplotypes in admixed families from several regionsof MexicordquoMolecular Immunology vol 45 no 4 pp 1171ndash11782008

[52] J G Romero-Quintana L O Frıas-Castro E Arambula-Merazet al ldquoIdentification of novel mutation in cathepsin C genecausing Papillon-Lefevre Syndrome in Mexican patientsrdquo BMCMedical Genetics vol 14 no 1 article 7 2013

[53] A Gonzalez-Mercado J Y Sanchez-Lopez J A Regla-Nava etal ldquoAssociation analysis of vitamin D receptor gene polymor-phisms and bone mineral density in postmenopausal Mexican-Mestizo womenrdquo Genetics and Molecular Research vol 12 no3 pp 2755ndash2763 2013

[54] Y KawaguchiMKanamoriH Ishihara KOhmoriHMatsuiand T Kimura ldquoThe association of lumbar disc disease withvitamin-D receptor gene polymorphismrdquo Journal of Bone andJoint SurgerymdashSeries A vol 84 no 11 pp 2022ndash2028 2002

[55] K M C Cheung D Chan J Karppinen et al ldquoAssociation ofthe Taq I allele in vitamin D receptor with degenerative discdisease and disc bulge in a Chinese populationrdquo Spine vol 31no 10 pp 1143ndash1148 2006

[56] B Eser T Cora O Eser et al ldquoAssociation of the polymor-phisms of vitamin d receptor and aggrecan genes with degen-erative disc diseaserdquo Genetic Testing and Molecular Biomarkersvol 14 no 3 pp 313ndash317 2010

International Journal of Genomics 7

[57] A M Valdes G Hassett D J Hart and T D SpectorldquoRadiographic progression of lumbar spine disc degenerationis influenced by variation at inflammatory genes a candidateSNP association study in the Chingford cohortrdquo Spine vol 30no 21 pp 2445ndash2451 2005

[58] G Jones C White P Sambrook and J Eisman ldquoAllelicvariation in the vitamin D receptor lifestyle factors and lumbarspinal degenerative diseaserdquo Annals of the Rheumatic Diseasesvol 57 no 2 pp 94ndash99 1998

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Molecular Biology International

GenomicsInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Signal TransductionJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

Evolutionary BiologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Genetics Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Enzyme Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Microbiology

International Journal of Genomics 3

Table 1 Conditions and products of PCR-RFLP

Gene SNPs Primers Tm RE bp Alleles(∘C) (bp)

IL1A rs1800587 51015840-GCATGCCATCACACCTAGTT-31015840 56 NcoI 193 C 174 and 1951015840-TTACATATGAGCCTTCCATG-31015840 T 193

VDRrs2228570 51015840-AGCTGGCCCTGGCACTGCTCTGCTCT-31015840 60 FokI 265 f 197 and 68

51015840-ATGGAAACACCTTGCTTCTTCTCCCTC-31015840 F 265

rs731236 51015840-CAGAGCATGGACAGGGAGCAAG-31015840 59 119879119886119902120572I 746 T 495 and 251

51015840-GCAACTCCTCATGGCTGAGGTCTCA-31015840 t 294 251 and 201Tm annealing temperature RE restriction enzyme

the Internal Review Committee of the ISSSTE and processedwithin 1 hour

23 DNAAmplification and Restriction Fragment Length Poly-morphism Genomic DNA was isolated from whole bloodusing the salt precipitation method [41] Gene polymor-phisms were analyzed by polymerase chain reaction followedby restriction fragment length polymorphism (PCR-RFLP)Reaction conditions primers and restriction fragments aresummarized in Table 1

A region of 193 bp spanning the rs1800587 polymorphicsite at the promoter region of IL1A gene in chromosome2q14 (formerly c-899CgtT or c-949CgtT) was amplified anddigested with NcoI (2U for 2 h at 37∘C) [31] Allele C (wild)has one restriction site giving two fragments of 174 and 19 bpwhile allele T has no recognition site

PCR product of 265 bp containing the single nucleotidepolymorphism (SNP) rs2228570 at the translation initiationcodon of exon 2 of VDR gene in chromosome 12q1311(formerly c2TgtC)was digested with FokI (1 U for 2 h at 37∘C)[31] Allele T (ancestral) has one restriction site giving twofragments of 197 and 68 bp while allele V (A C or G) hasno recognition site In the same way the 747 bp ampliconspanning the synonymous polymorphism rs731236 in exon9 of VDR gene (formerly c352TgtC or c1056TgtC) [32] wascleaved with TaqI (2U for 2 h at 37∘C) Allele T (ancestral)showed two fragments of 496 and 251 bp while allele Cgenerated an additional restriction site giving three fragmentsof 294 251 and 201 bp

24 Statistical Analysis Demographic and clinical variablesof patients and controls were presented as mean plusmn SD andfrequencies Power calculation showed that a significancelevel of 119875 le 005 one-tailed directional would yield a powerof 80 with a sample size of 96 individuals per group [42]Differences in allelic genotype and haplotype frequencieswere evaluated using Fisherrsquos exact test SNPs associationswere tested under dominant and recessive genetic modelsand odds ratios (OR)with 95 confidence intervals (CI) wereused as themeasure of association between specific alleles andgenotypes with IDD and its clinical subtypes [43] Significant119875 values were corrected with the Bonferroni test for multiplecomparisons [44] Hardy-Weinbergrsquos equilibrium was calcu-lated by 1205942 test for all genotype combinations of each SNPin patients and controls PASW v180 (SPSS Inc Chicago

IL USA) and Arlequin v3512 (Swiss National ScienceFoundation) software packages were used for analysis

3 Results and Discussion

IDD is a complex pathology and genetic predisposition couldimpact the physiological maintenance of intervertebral discsor the control of inflammation and pain [15 19 22 45] ulti-mately leading to the clinical manifestations of IDD Geneticstudies on IDD have been carried out mainly in populationof European (Finnish and Spanish) or Asian (Chinese andJapanese) ancestry but not in Mexican Mestizos a majorpopulation in North America Our work carried out in anorthwestern Mexican population represents the first reportof genetic polymorphisms and IDD in a population withmixed native Mexican-European ancestry also known asMestizos

Patients with IDD and control subjects with a mean ageof 3922 (plusmn688) versus 3913 (plusmn680) years (119875 = 0918) wereincluded in this study In both groups 890 of individualswere female and 110 were male BMI data between patientsand controls was 2599 (plusmn433) versus 2623 (plusmn372) kgm2(119875 = 0673) Both groups fell into the overweight WorldHealth Organization category for BMI Covariates followeda normal distribution according to the Kolmogorov-Smirnovtest with no statistically significant differences between thegroups Disc herniation was evident in 800 of the patientsand herniation with intervertebral osteochondrosis in 200The percentages of cases based on Modic changes of spineMRIwere type I (560) type II (340) and type III (100)Clinical symptomatology included 360with sciatica 220with lumbago and 420 with lumbosciatica

Our study on the rs1800587 polymorphism of the IL1Ashowed no significant association with IDD (Table 2) Thedistribution of T allele in patients and controls was 270versus 280 (119875 = 0455 OR = 095 95 CI = 061ndash147) Genotype frequencies between the groups (119875 = 0455)were 40 versus 100 for TT 450 versus 350 for CTand 510 versus 550 for CC respectively However otherreports have shown an association between the T allele andIDD A report on the Finnish population showed that T allelecarriers have a higher risk (OR = 24) of developing discbulges than those without it [38] Another study found thatT allele was associated (25-fold risk) with Modic changes[46 47] An analysis of occupational and genetic factors

4 International Journal of Genomics

Table 2 Allelic (af ) and genotype (gf ) frequencies of IL1A andVDRpolymorphisms in controls subjects and patients with IDD

SNPs Patients Controls(119899 = 100) (119899 = 100)

rs1800587 (IL1A)Alleles 119899 af 119899 afT 53 027 55 028C 147 074 145 073Genotypes 119899 gf 119899 gfTT 4 004 10 010CT 45 045 35 035CC 51 051 55 055rs2228570 (VDR)Alleles 119899 af 119899 afV 105 053 115 058T 95 048 85 043Genotypes 119899 gf 119899 gfVV 20 020 32 032TV 65 065 51 051TT 15 015 17 017rs731236 (VDR)Alleles 119899 af 119899 afC 35 018 41 021T 165 083 159 080Genotypes 119899 gf 119899 gfCC 4 004 3 003TC 27 027 35 035TT 69 069 62 062

demonstrated that T allele represented a significant risk factorfor the disc degeneration phenotype [48] A study of a Danishpopulation also found an association of this polymorphismwith disc degeneration [39] In contrast a case-control studyof a population of Spain foundno association betweenT alleleand symptomatic lumbar disc herniation [49]The reason forthis inconsistent association between the T allele and IDDmay be due to ethnic differences in the studied populationsor by different haplotypes in the promoter or enhancerregions Mexican Mestizo populations have a high degree ofgenetic heterogeneity [50] carrying Amerindian and a fewEuropean and African HLA haplotypes [51] Moreover thepopulation studied by us in Mexico has a particular geneticbackground since half of the most common haplotypesfound in this population have a proposed European originmost likely from Spanish origin [52] Not surprisingly thefrequency of the T allele in our population (280) wassimilar to the one observed in a Spanish population (355)[49] and alike our results was not associated with IDD

Genotype frequencies observed between patients andcontrols (119875 = 0161) for rs2228570 polymorphism of VDRwere 200 versus 320 for VV 650 versus 510 forTV and 150 versus 170 for TT respectively (Table 2)

Frequencies of V allele were 530 versus 580 (119875 = 0183OR = 081 95 CI = 055ndash121) In the case of rs731236allele C of VDR (Table 2) the distribution in patients andcontrols was 180 versus 210 (119875 = 0262 OR = 08295 CI = 049ndash135) Genotype frequencies (119875 = 0262)were 40 versus 30 for CC 270 versus 350 for TCand 690 versus 620 for TT respectively In accordancewith our allelic frequencies a recent study on the associationbetween VDR polymorphisms and BMD in postmenopausalMexican women allelic frequencies of 53 for C of FokI and27 for V of TaqI were reported in the control group [53]Other reports have shown an association between these VDRpolymorphisms and IDDThe report in a Finnish populationthat demonstrated for the first time that VDR polymor-phism was associated with IDD quantitatively assessed signalintensities of thoracic and lumbar discs finding those tobe 129 worse in men with the TaqI CC genotype and93 in the men with FokI VV genotype [32] Reports onJapanese [54] Chinese [55] Turkish [56] English [57] andAustralian [58] populations also found association betweenVDR polymorphisms and IDD

Therefore for all the studied polymorphisms (Table 2)there were no significant differences in the distribution ofalleles and genotypes between the groups (119875 gt 005) underthe genetic models tested The study of haplotypes showedno differences in frequencies between the groups for anycombination of rs2228570 and rs731236 SNPs of VDR Thestratified analysis of the data (patients by pathology symp-tomatology and Modic changes) also showed no significantdifferences (all 119875 ge 005) The genotypes in the groupswere not significantly different from the expected distributionfor a population in a Hardy-Weinberg equilibrium with theexception of rs2228570 (VDR) in patients (119875 = 0010)

Our work assessed for the first time the potential contri-bution of gene polymorphisms on IDD in aMexicanMestizopopulation with homogeneous genetic background sinceboth patients and controls were ethnically and geographi-cally matched The results showed that no association existsbetween the studied polymorphisms and IDD in this pop-ulation Further analysis of other relevant polymorphismsand more Mestizo populations may contribute to finding thegenetic determinants of this disease in our country

4 Conclusion

In conclusion this is the first report on the potentialcontribution of gene polymorphisms on IDD in a Mestizopopulation in Mexico Our results showed that there is noassociation of the IL1A (rs1800587) and VDR (rs2228570and rs731236) polymorphisms with IDD The study of moreMestizo populations and more candidate genes may providefurther insight into the etiology of the disease

Consent

Written consent was obtained from patients for publicationof this work

International Journal of Genomics 5

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Authorsrsquo Contribution

Salvador Cervin Serrano diagnosed and treated the patientsSalvador Cervin Serrano Dalia Gonzalez Villareal MaribelAguilar-Medina Jose Geovanni Romero Quintana Veron-ica Picos-Cardenas and Jose Guillermo Romero-Navarroperformed the research Julio Granados Eliakym ArambulaMeraz and Ignacio Osuna Ramırez analyzed the data Sal-vador Cervin Serrano and Rosalıo Ramos-Payan designedthe research Iris Estrada-Garcıa Guzman Sanchez-Schmitzand Rosalıo Ramos-Payan wrote the paper All authors readand approved the final paper

Acknowledgment

This work was supported by CONACYT (106152)

References

[1] USBJDUSBJI ldquoSpine low back and neck painrdquo in The Burdenof Musculoskeletal Diseases in the United States vol 1 pp 21ndash56American Academy of Orthopaedic Surgeons 2011

[2] D Chou D Samartzis C Bellabarba et al ldquoDegenerativemagnetic resonance imaging changes in patients with chroniclow back pain a systematic reviewrdquo Spine vol 36 supplementno 21 pp S43ndashS53 2011

[3] S Salo V Leinonen T Rikkonen et al ldquoAssociation betweenbonemineral density and lumbar disc degenerationrdquoMaturitas2014

[4] L Wang W Cui J P Kalala T V Hoof and B G Liu ldquoToinvestigate the effect of osteoporosis and intervertebral disc deg-eneration on the endplate cartilage injury in ratsrdquo Asian PacificJournal of Tropical Medicine vol 7 no 10 pp 796ndash800 2014

[5] M T Modic P M Steinberg J S Ross T J Masaryk and JR Carter ldquoDegenerative disk disease assessment of changes invertebral body marrow with MR imagingrdquo Radiology vol 166no 1 part 1 pp 193ndash199 1988

[6] S Roberts H Evans J Trivedi and J Menage ldquoHistology andpathology of the human intervertebral discrdquoThe Journal of Boneamp Joint Surgery Series A vol 88 supplement 2 pp 10ndash14 2006

[7] R Quiroz-Moreno G Lezama-Suarez and C Gomez-JimenezldquoDisc alterations of lumbar spine onmagnetic resonance imagesin asymptomatic workersrdquo Revista Medica del Instituto Mexi-cano del Seguro Social vol 46 no 2 pp 185ndash190 2008

[8] S D Boden D O Davis T S Dina N J Patronas and SW Wiesel ldquoAbnormal magnetic-resonance scans of the lumbarspine in asymptomatic subjects A prospective investigationrdquoJournal of Bone and Joint Surgery A vol 72 no 3 pp 403ndash4081990

[9] K M C Cheung J Karppinen D Chan et al ldquoPrevalenceand pattern of lumbar magnetic resonance imaging changes ina population study of one thousand forty-three individualsrdquoSpine vol 34 no 9 pp 934ndash940 2009

[10] M B Joslashrgensen A Holtermann F Gyntelberg and P Suadi-cani ldquoPhysical fitness as a predictor of herniated lumbar discdiseasemdasha 33-year follow-up in the Copenhagen male studyrdquoBMCMusculoskeletal Disorders vol 14 article 86 2013

[11] J A Buckwalter ldquoAging and degeneration of the humanintervertebral discrdquo Spine vol 20 no 11 pp 1307ndash1314 1995

[12] L Ala-Kokko ldquoGenetic risk factors for lumbar disc diseaserdquoAnnals of Medicine vol 34 no 1 pp 42ndash47 2002

[13] M C Battie T Videman L E Gibbons L D Fisher HManni-nen and K Gill ldquoDeterminants of lumbar disc degenerationa study relating lifetime exposures and magnetic resonanceimaging findings in identical twinsrdquo Spine vol 20 no 24 pp2601ndash2612 1995

[14] M C Batite D R Haynor L D Fisher S K Gill L EGibbons and T Videman ldquoSimilarities in degenerative findingson magnetic resonance images of the lumbar spines of identicaltwinsrdquo Journal of Bone and Joint SurgerymdashSeries A vol 77 no11 pp 1662ndash1670 1995

[15] P N Sambrook A J MacGregor and T D Spector ldquoGeneticinfluences on cervical and lumbar disc degeneration amagneticresonance imaging study in twinsrdquo Arthritis amp Rheumatologyvol 42 no 2 pp 366ndash372 1999

[16] M C Battie T Videman J Kaprio et al ldquoThe Twin SpineStudy contributions to a changing view of disc degenerationrdquoThe Spine Journal vol 9 no 1 pp 47ndash59 2009

[17] C Bijkerk J J Houwing-Duistermaat H A Valkenburg etal ldquoHeritabilities of radiologic osteoarthritis in peripheraljoints and of disc degeneration of the spinerdquo Arthritis ampRheumatology vol 42 no 8 pp 1729ndash1735 1999

[18] C K Kepler R K Ponnappan C A Tannoury M V Risbudand D G Anderson ldquoThemolecular basis of intervertebral discdegenerationrdquo Spine Journal vol 13 no 3 pp 318ndash330 2013

[19] S Kalb N L Martirosyan M Y Kalani G G Broc and NTheodore ldquoGenetics of the degenerated intervertebral discrdquoWorld Neurosurgery vol 77 no 3-4 pp 491ndash501 2012

[20] A A Patel W R Spiker M Daubs D Brodke and L ACannon-Albright ldquoEvidence for an inherited predisposition tolumbar disc diseaserdquo Journal of Bone and Joint SurgerymdashSeriesA vol 93 no 3 pp 225ndash229 2011

[21] T Videman J Saarela J Kaprio et al ldquoAssociations of 25structural degradative and inflammatory candidate geneswith lumbar disc desiccation bulging and height narrowingrdquoArthritis amp Rheumatism vol 60 no 2 pp 470ndash481 2009

[22] J E Mayer J C Iatridis D Chan S A Qureshi O Gottesmanand A C Hecht ldquoGenetic polymorphisms associated withintervertebral disc degenerationrdquoThe Spine Journal vol 13 no3 pp 299ndash317 2013

[23] SM F Pluijm HW Van Essen N Bravenboer et al ldquoCollagentype I 1205721 Sp1 polymorphism osteoporosis and intervertebraldisc degeneration in older men and womenrdquo Annals of theRheumatic Diseases vol 63 no 1 pp 71ndash77 2004

[24] S Annunen P Paassilta J Lohiniva et al ldquoAn allele of COL9A2associated with intervertebral disc diseaserdquo Science vol 285 no5426 pp 409ndash412 1999

[25] F Mio K Chiba Y Hirose et al ldquoA functional polymorphismin COL11A1 which encodes the 1205721 chain of type XI collagenis associated with susceptibility to lumbar disc herniationrdquoAmerican Journal of Human Genetics vol 81 no 6 pp 1271ndash1277 2007

6 International Journal of Genomics

[26] Y Kawaguchi R Osada M Kanamori et al ldquoAssociationbetween an aggrecan gene polymorphism and lumbar discdegenerationrdquo Spine vol 24 no 23 pp 2456ndash2460 1999

[27] S Seki Y Kawaguchi K Chiba et al ldquoA functional SNP inCILPencoding cartilage intermediate layer protein is associated withsusceptibility to lumbar disc diseaserdquo Nature Genetics vol 37no 6 pp 607ndash612 2005

[28] M Takahashi H Haro Y Wakabayashi T Kawa-uchi HKomori and K Shinomiya ldquoThe association of degenerationof the intervertebral disc with 5a6a polymorphism in thepromoter of the human matrix metalloproteinase-3 generdquoJournal of Bone and Joint Surgery vol 83 no 4 pp 491ndash4952001

[29] Y Hirose K Chiba T Karasugi et al ldquoA functional polymor-phism inTHBS2 that affects alternative splicing andMMPbind-ing is associated with lumbar-disc herniationrdquo The AmericanJournal of Human Genetics vol 82 no 5 pp 1122ndash1129 2008

[30] S Solovieva J Lohiniva P Leino-Arjas et al ldquoIntervertebraldisc degeneration in relation to the COL9A3 and the IL-1120573 genepolymorphismsrdquo European Spine Journal vol 15 no 5 pp 613ndash619 2006

[31] N Noponen-Hietala I Virtanen R Karttunen et al ldquoGeneticvariations in IL6 associate with intervertebral disc diseasecharacterized by sciaticardquo Pain vol 114 no 1-2 pp 186ndash1942005

[32] T Videman J Leppavuori J Kaprio et al ldquoIntragenic poly-morphisms of the vitamin D receptor gene associated withintervertebral disc degenerationrdquo Spine vol 23 no 23 pp2477ndash2485 1998

[33] R JWood and J C Fleet ldquoThe genetics of osteoporosis vitaminD receptor polymorphismsrdquoAnnual Review of Nutrition vol 18pp 233ndash258 1998

[34] A Colombini S Cauci G Lombardi et al ldquoRelationshipbetween vitamin D receptor gene (VDR) polymorphismsvitamin D status osteoarthritis and intervertebral disc degen-erationrdquo The Journal of Steroid Biochemistry and MolecularBiology vol 138 pp 24ndash40 2013

[35] A G Uitterlinden Y Fang J B J van Meurs H A P Polsand J P T M van Leeuwen ldquoGenetics and biology of vitaminD receptor polymorphismsrdquo Gene vol 338 no 2 pp 143ndash1562004

[36] K L E Phillips N Jordan-Mahy M J H Nicklin and CL Le Maitre ldquoInterleukin-1 receptor antagonist deficient miceprovide insights into pathogenesis of human intervertebral discdegenerationrdquo Annals of the Rheumatic Diseases vol 72 no 11pp 1860ndash1867 2013

[37] W Ye D S Huang W J Chen et al ldquoAssociation of 86 bpvariable number tandem repeat polymorphism of interleukin-1receptor antagonist gene with lumbar disc diseaserdquoNan Fang YiKe Da Xue Xue Bao vol 27 no 10 pp 1485ndash1488 2007

[38] S Solovieva S Kouhia P Leino-Arjas et al ldquoInterleukin 1polymorphisms and intervertebral disc degenerationrdquo Epidemi-ology vol 15 no 5 pp 626ndash633 2004

[39] P J Eskola P Kjaer I M Daavittila et al ldquoGenetic risk factorsof disc degeneration among 12-14-year-old Danish children apopulation studyrdquo International Journal of Molecular Epidemi-ology and Genetics vol 1 no 2 pp 158ndash165 2010

[40] M T Modic T J Masaryk J S Ross and J R Carter ldquoImagingof degenerative disk diseaserdquo Radiology vol 168 no 1 pp 177ndash186 1988

[41] S Gustincich G Manfioletti G Del Sal C Schneider andP Carninci ldquoA fast method for high-quality genomic DNAextraction fromwhole human bloodrdquo BioTechniques vol 11 no3 pp 298ndash302 1991

[42] M Gail R Williams D P Byar and C Brown ldquoHow manycontrolsrdquo Journal of Chronic Diseases vol 29 no 11 pp 723ndash731 1976

[43] B Woolf ldquoOn estimating the relation between blood group anddiseaserdquo Annals of Human Genetics vol 19 no 4 pp 251ndash2531955

[44] T V Perneger ldquoWhatrsquos wrong with Bonferroni adjustmentsrdquoBritish Medical Journal vol 316 no 7139 pp 1236ndash1238 1998

[45] I Tegeder and J Lotsch ldquoCurrent evidence for a modulation oflow back pain by human genetic variantsrdquo Journal of Cellularand Molecular Medicine vol 13 no 8B pp 1605ndash1619 2009

[46] J Karppinen S Solovieva K Luoma R Raininko P Leino-Arjas and H Riihimaki ldquoModic changes and interleukin 1 genelocus polymorphisms in occupational cohort of middle-agedmenrdquo European Spine Journal vol 18 no 12 pp 1963ndash19702009

[47] J Karppinen I Daavittila S Solovieva et al ldquoGenetic factorsare associated with modic changes in endplates of lumbarvertebral bodiesrdquo Spine vol 33 no 11 pp 1236ndash1241 2008

[48] I M Virtanen J Karppinen S Taimela et al ldquoOccupationaland genetic risk factors associated with intervertebral discdiseaserdquo Spine vol 32 no 10 pp 1129ndash1134 2007

[49] J Paz Aparicio I Fernandez Bances E Lopez-AngladaFernandez et al ldquoThe IL-1120573 (+3953 TC) gene polymorphismassociates to symptomatic lumbar disc herniationrdquo EuropeanSpine Journal vol 20 no 3 pp 383ndash389 2011

[50] AMoreno-Estrada C R Gignoux J C Fernandez-Lopez et alldquoHuman genetics The genetics of Mexico recapitulates NativeAmerican substructure and affects biomedical traitsrdquo Sciencevol 344 no 6189 pp 1280ndash1285 2014

[51] R Barquera J Zuniga R Hernandez-Dıaz et al ldquoHLA class Iand class II haplotypes in admixed families from several regionsof MexicordquoMolecular Immunology vol 45 no 4 pp 1171ndash11782008

[52] J G Romero-Quintana L O Frıas-Castro E Arambula-Merazet al ldquoIdentification of novel mutation in cathepsin C genecausing Papillon-Lefevre Syndrome in Mexican patientsrdquo BMCMedical Genetics vol 14 no 1 article 7 2013

[53] A Gonzalez-Mercado J Y Sanchez-Lopez J A Regla-Nava etal ldquoAssociation analysis of vitamin D receptor gene polymor-phisms and bone mineral density in postmenopausal Mexican-Mestizo womenrdquo Genetics and Molecular Research vol 12 no3 pp 2755ndash2763 2013

[54] Y KawaguchiMKanamoriH Ishihara KOhmoriHMatsuiand T Kimura ldquoThe association of lumbar disc disease withvitamin-D receptor gene polymorphismrdquo Journal of Bone andJoint SurgerymdashSeries A vol 84 no 11 pp 2022ndash2028 2002

[55] K M C Cheung D Chan J Karppinen et al ldquoAssociation ofthe Taq I allele in vitamin D receptor with degenerative discdisease and disc bulge in a Chinese populationrdquo Spine vol 31no 10 pp 1143ndash1148 2006

[56] B Eser T Cora O Eser et al ldquoAssociation of the polymor-phisms of vitamin d receptor and aggrecan genes with degen-erative disc diseaserdquo Genetic Testing and Molecular Biomarkersvol 14 no 3 pp 313ndash317 2010

International Journal of Genomics 7

[57] A M Valdes G Hassett D J Hart and T D SpectorldquoRadiographic progression of lumbar spine disc degenerationis influenced by variation at inflammatory genes a candidateSNP association study in the Chingford cohortrdquo Spine vol 30no 21 pp 2445ndash2451 2005

[58] G Jones C White P Sambrook and J Eisman ldquoAllelicvariation in the vitamin D receptor lifestyle factors and lumbarspinal degenerative diseaserdquo Annals of the Rheumatic Diseasesvol 57 no 2 pp 94ndash99 1998

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Molecular Biology International

GenomicsInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Signal TransductionJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

Evolutionary BiologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Genetics Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Enzyme Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Microbiology

4 International Journal of Genomics

Table 2 Allelic (af ) and genotype (gf ) frequencies of IL1A andVDRpolymorphisms in controls subjects and patients with IDD

SNPs Patients Controls(119899 = 100) (119899 = 100)

rs1800587 (IL1A)Alleles 119899 af 119899 afT 53 027 55 028C 147 074 145 073Genotypes 119899 gf 119899 gfTT 4 004 10 010CT 45 045 35 035CC 51 051 55 055rs2228570 (VDR)Alleles 119899 af 119899 afV 105 053 115 058T 95 048 85 043Genotypes 119899 gf 119899 gfVV 20 020 32 032TV 65 065 51 051TT 15 015 17 017rs731236 (VDR)Alleles 119899 af 119899 afC 35 018 41 021T 165 083 159 080Genotypes 119899 gf 119899 gfCC 4 004 3 003TC 27 027 35 035TT 69 069 62 062

demonstrated that T allele represented a significant risk factorfor the disc degeneration phenotype [48] A study of a Danishpopulation also found an association of this polymorphismwith disc degeneration [39] In contrast a case-control studyof a population of Spain foundno association betweenT alleleand symptomatic lumbar disc herniation [49]The reason forthis inconsistent association between the T allele and IDDmay be due to ethnic differences in the studied populationsor by different haplotypes in the promoter or enhancerregions Mexican Mestizo populations have a high degree ofgenetic heterogeneity [50] carrying Amerindian and a fewEuropean and African HLA haplotypes [51] Moreover thepopulation studied by us in Mexico has a particular geneticbackground since half of the most common haplotypesfound in this population have a proposed European originmost likely from Spanish origin [52] Not surprisingly thefrequency of the T allele in our population (280) wassimilar to the one observed in a Spanish population (355)[49] and alike our results was not associated with IDD

Genotype frequencies observed between patients andcontrols (119875 = 0161) for rs2228570 polymorphism of VDRwere 200 versus 320 for VV 650 versus 510 forTV and 150 versus 170 for TT respectively (Table 2)

Frequencies of V allele were 530 versus 580 (119875 = 0183OR = 081 95 CI = 055ndash121) In the case of rs731236allele C of VDR (Table 2) the distribution in patients andcontrols was 180 versus 210 (119875 = 0262 OR = 08295 CI = 049ndash135) Genotype frequencies (119875 = 0262)were 40 versus 30 for CC 270 versus 350 for TCand 690 versus 620 for TT respectively In accordancewith our allelic frequencies a recent study on the associationbetween VDR polymorphisms and BMD in postmenopausalMexican women allelic frequencies of 53 for C of FokI and27 for V of TaqI were reported in the control group [53]Other reports have shown an association between these VDRpolymorphisms and IDDThe report in a Finnish populationthat demonstrated for the first time that VDR polymor-phism was associated with IDD quantitatively assessed signalintensities of thoracic and lumbar discs finding those tobe 129 worse in men with the TaqI CC genotype and93 in the men with FokI VV genotype [32] Reports onJapanese [54] Chinese [55] Turkish [56] English [57] andAustralian [58] populations also found association betweenVDR polymorphisms and IDD

Therefore for all the studied polymorphisms (Table 2)there were no significant differences in the distribution ofalleles and genotypes between the groups (119875 gt 005) underthe genetic models tested The study of haplotypes showedno differences in frequencies between the groups for anycombination of rs2228570 and rs731236 SNPs of VDR Thestratified analysis of the data (patients by pathology symp-tomatology and Modic changes) also showed no significantdifferences (all 119875 ge 005) The genotypes in the groupswere not significantly different from the expected distributionfor a population in a Hardy-Weinberg equilibrium with theexception of rs2228570 (VDR) in patients (119875 = 0010)

Our work assessed for the first time the potential contri-bution of gene polymorphisms on IDD in aMexicanMestizopopulation with homogeneous genetic background sinceboth patients and controls were ethnically and geographi-cally matched The results showed that no association existsbetween the studied polymorphisms and IDD in this pop-ulation Further analysis of other relevant polymorphismsand more Mestizo populations may contribute to finding thegenetic determinants of this disease in our country

4 Conclusion

In conclusion this is the first report on the potentialcontribution of gene polymorphisms on IDD in a Mestizopopulation in Mexico Our results showed that there is noassociation of the IL1A (rs1800587) and VDR (rs2228570and rs731236) polymorphisms with IDD The study of moreMestizo populations and more candidate genes may providefurther insight into the etiology of the disease

Consent

Written consent was obtained from patients for publicationof this work

International Journal of Genomics 5

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Authorsrsquo Contribution

Salvador Cervin Serrano diagnosed and treated the patientsSalvador Cervin Serrano Dalia Gonzalez Villareal MaribelAguilar-Medina Jose Geovanni Romero Quintana Veron-ica Picos-Cardenas and Jose Guillermo Romero-Navarroperformed the research Julio Granados Eliakym ArambulaMeraz and Ignacio Osuna Ramırez analyzed the data Sal-vador Cervin Serrano and Rosalıo Ramos-Payan designedthe research Iris Estrada-Garcıa Guzman Sanchez-Schmitzand Rosalıo Ramos-Payan wrote the paper All authors readand approved the final paper

Acknowledgment

This work was supported by CONACYT (106152)

References

[1] USBJDUSBJI ldquoSpine low back and neck painrdquo in The Burdenof Musculoskeletal Diseases in the United States vol 1 pp 21ndash56American Academy of Orthopaedic Surgeons 2011

[2] D Chou D Samartzis C Bellabarba et al ldquoDegenerativemagnetic resonance imaging changes in patients with chroniclow back pain a systematic reviewrdquo Spine vol 36 supplementno 21 pp S43ndashS53 2011

[3] S Salo V Leinonen T Rikkonen et al ldquoAssociation betweenbonemineral density and lumbar disc degenerationrdquoMaturitas2014

[4] L Wang W Cui J P Kalala T V Hoof and B G Liu ldquoToinvestigate the effect of osteoporosis and intervertebral disc deg-eneration on the endplate cartilage injury in ratsrdquo Asian PacificJournal of Tropical Medicine vol 7 no 10 pp 796ndash800 2014

[5] M T Modic P M Steinberg J S Ross T J Masaryk and JR Carter ldquoDegenerative disk disease assessment of changes invertebral body marrow with MR imagingrdquo Radiology vol 166no 1 part 1 pp 193ndash199 1988

[6] S Roberts H Evans J Trivedi and J Menage ldquoHistology andpathology of the human intervertebral discrdquoThe Journal of Boneamp Joint Surgery Series A vol 88 supplement 2 pp 10ndash14 2006

[7] R Quiroz-Moreno G Lezama-Suarez and C Gomez-JimenezldquoDisc alterations of lumbar spine onmagnetic resonance imagesin asymptomatic workersrdquo Revista Medica del Instituto Mexi-cano del Seguro Social vol 46 no 2 pp 185ndash190 2008

[8] S D Boden D O Davis T S Dina N J Patronas and SW Wiesel ldquoAbnormal magnetic-resonance scans of the lumbarspine in asymptomatic subjects A prospective investigationrdquoJournal of Bone and Joint Surgery A vol 72 no 3 pp 403ndash4081990

[9] K M C Cheung J Karppinen D Chan et al ldquoPrevalenceand pattern of lumbar magnetic resonance imaging changes ina population study of one thousand forty-three individualsrdquoSpine vol 34 no 9 pp 934ndash940 2009

[10] M B Joslashrgensen A Holtermann F Gyntelberg and P Suadi-cani ldquoPhysical fitness as a predictor of herniated lumbar discdiseasemdasha 33-year follow-up in the Copenhagen male studyrdquoBMCMusculoskeletal Disorders vol 14 article 86 2013

[11] J A Buckwalter ldquoAging and degeneration of the humanintervertebral discrdquo Spine vol 20 no 11 pp 1307ndash1314 1995

[12] L Ala-Kokko ldquoGenetic risk factors for lumbar disc diseaserdquoAnnals of Medicine vol 34 no 1 pp 42ndash47 2002

[13] M C Battie T Videman L E Gibbons L D Fisher HManni-nen and K Gill ldquoDeterminants of lumbar disc degenerationa study relating lifetime exposures and magnetic resonanceimaging findings in identical twinsrdquo Spine vol 20 no 24 pp2601ndash2612 1995

[14] M C Batite D R Haynor L D Fisher S K Gill L EGibbons and T Videman ldquoSimilarities in degenerative findingson magnetic resonance images of the lumbar spines of identicaltwinsrdquo Journal of Bone and Joint SurgerymdashSeries A vol 77 no11 pp 1662ndash1670 1995

[15] P N Sambrook A J MacGregor and T D Spector ldquoGeneticinfluences on cervical and lumbar disc degeneration amagneticresonance imaging study in twinsrdquo Arthritis amp Rheumatologyvol 42 no 2 pp 366ndash372 1999

[16] M C Battie T Videman J Kaprio et al ldquoThe Twin SpineStudy contributions to a changing view of disc degenerationrdquoThe Spine Journal vol 9 no 1 pp 47ndash59 2009

[17] C Bijkerk J J Houwing-Duistermaat H A Valkenburg etal ldquoHeritabilities of radiologic osteoarthritis in peripheraljoints and of disc degeneration of the spinerdquo Arthritis ampRheumatology vol 42 no 8 pp 1729ndash1735 1999

[18] C K Kepler R K Ponnappan C A Tannoury M V Risbudand D G Anderson ldquoThemolecular basis of intervertebral discdegenerationrdquo Spine Journal vol 13 no 3 pp 318ndash330 2013

[19] S Kalb N L Martirosyan M Y Kalani G G Broc and NTheodore ldquoGenetics of the degenerated intervertebral discrdquoWorld Neurosurgery vol 77 no 3-4 pp 491ndash501 2012

[20] A A Patel W R Spiker M Daubs D Brodke and L ACannon-Albright ldquoEvidence for an inherited predisposition tolumbar disc diseaserdquo Journal of Bone and Joint SurgerymdashSeriesA vol 93 no 3 pp 225ndash229 2011

[21] T Videman J Saarela J Kaprio et al ldquoAssociations of 25structural degradative and inflammatory candidate geneswith lumbar disc desiccation bulging and height narrowingrdquoArthritis amp Rheumatism vol 60 no 2 pp 470ndash481 2009

[22] J E Mayer J C Iatridis D Chan S A Qureshi O Gottesmanand A C Hecht ldquoGenetic polymorphisms associated withintervertebral disc degenerationrdquoThe Spine Journal vol 13 no3 pp 299ndash317 2013

[23] SM F Pluijm HW Van Essen N Bravenboer et al ldquoCollagentype I 1205721 Sp1 polymorphism osteoporosis and intervertebraldisc degeneration in older men and womenrdquo Annals of theRheumatic Diseases vol 63 no 1 pp 71ndash77 2004

[24] S Annunen P Paassilta J Lohiniva et al ldquoAn allele of COL9A2associated with intervertebral disc diseaserdquo Science vol 285 no5426 pp 409ndash412 1999

[25] F Mio K Chiba Y Hirose et al ldquoA functional polymorphismin COL11A1 which encodes the 1205721 chain of type XI collagenis associated with susceptibility to lumbar disc herniationrdquoAmerican Journal of Human Genetics vol 81 no 6 pp 1271ndash1277 2007

6 International Journal of Genomics

[26] Y Kawaguchi R Osada M Kanamori et al ldquoAssociationbetween an aggrecan gene polymorphism and lumbar discdegenerationrdquo Spine vol 24 no 23 pp 2456ndash2460 1999

[27] S Seki Y Kawaguchi K Chiba et al ldquoA functional SNP inCILPencoding cartilage intermediate layer protein is associated withsusceptibility to lumbar disc diseaserdquo Nature Genetics vol 37no 6 pp 607ndash612 2005

[28] M Takahashi H Haro Y Wakabayashi T Kawa-uchi HKomori and K Shinomiya ldquoThe association of degenerationof the intervertebral disc with 5a6a polymorphism in thepromoter of the human matrix metalloproteinase-3 generdquoJournal of Bone and Joint Surgery vol 83 no 4 pp 491ndash4952001

[29] Y Hirose K Chiba T Karasugi et al ldquoA functional polymor-phism inTHBS2 that affects alternative splicing andMMPbind-ing is associated with lumbar-disc herniationrdquo The AmericanJournal of Human Genetics vol 82 no 5 pp 1122ndash1129 2008

[30] S Solovieva J Lohiniva P Leino-Arjas et al ldquoIntervertebraldisc degeneration in relation to the COL9A3 and the IL-1120573 genepolymorphismsrdquo European Spine Journal vol 15 no 5 pp 613ndash619 2006

[31] N Noponen-Hietala I Virtanen R Karttunen et al ldquoGeneticvariations in IL6 associate with intervertebral disc diseasecharacterized by sciaticardquo Pain vol 114 no 1-2 pp 186ndash1942005

[32] T Videman J Leppavuori J Kaprio et al ldquoIntragenic poly-morphisms of the vitamin D receptor gene associated withintervertebral disc degenerationrdquo Spine vol 23 no 23 pp2477ndash2485 1998

[33] R JWood and J C Fleet ldquoThe genetics of osteoporosis vitaminD receptor polymorphismsrdquoAnnual Review of Nutrition vol 18pp 233ndash258 1998

[34] A Colombini S Cauci G Lombardi et al ldquoRelationshipbetween vitamin D receptor gene (VDR) polymorphismsvitamin D status osteoarthritis and intervertebral disc degen-erationrdquo The Journal of Steroid Biochemistry and MolecularBiology vol 138 pp 24ndash40 2013

[35] A G Uitterlinden Y Fang J B J van Meurs H A P Polsand J P T M van Leeuwen ldquoGenetics and biology of vitaminD receptor polymorphismsrdquo Gene vol 338 no 2 pp 143ndash1562004

[36] K L E Phillips N Jordan-Mahy M J H Nicklin and CL Le Maitre ldquoInterleukin-1 receptor antagonist deficient miceprovide insights into pathogenesis of human intervertebral discdegenerationrdquo Annals of the Rheumatic Diseases vol 72 no 11pp 1860ndash1867 2013

[37] W Ye D S Huang W J Chen et al ldquoAssociation of 86 bpvariable number tandem repeat polymorphism of interleukin-1receptor antagonist gene with lumbar disc diseaserdquoNan Fang YiKe Da Xue Xue Bao vol 27 no 10 pp 1485ndash1488 2007

[38] S Solovieva S Kouhia P Leino-Arjas et al ldquoInterleukin 1polymorphisms and intervertebral disc degenerationrdquo Epidemi-ology vol 15 no 5 pp 626ndash633 2004

[39] P J Eskola P Kjaer I M Daavittila et al ldquoGenetic risk factorsof disc degeneration among 12-14-year-old Danish children apopulation studyrdquo International Journal of Molecular Epidemi-ology and Genetics vol 1 no 2 pp 158ndash165 2010

[40] M T Modic T J Masaryk J S Ross and J R Carter ldquoImagingof degenerative disk diseaserdquo Radiology vol 168 no 1 pp 177ndash186 1988

[41] S Gustincich G Manfioletti G Del Sal C Schneider andP Carninci ldquoA fast method for high-quality genomic DNAextraction fromwhole human bloodrdquo BioTechniques vol 11 no3 pp 298ndash302 1991

[42] M Gail R Williams D P Byar and C Brown ldquoHow manycontrolsrdquo Journal of Chronic Diseases vol 29 no 11 pp 723ndash731 1976

[43] B Woolf ldquoOn estimating the relation between blood group anddiseaserdquo Annals of Human Genetics vol 19 no 4 pp 251ndash2531955

[44] T V Perneger ldquoWhatrsquos wrong with Bonferroni adjustmentsrdquoBritish Medical Journal vol 316 no 7139 pp 1236ndash1238 1998

[45] I Tegeder and J Lotsch ldquoCurrent evidence for a modulation oflow back pain by human genetic variantsrdquo Journal of Cellularand Molecular Medicine vol 13 no 8B pp 1605ndash1619 2009

[46] J Karppinen S Solovieva K Luoma R Raininko P Leino-Arjas and H Riihimaki ldquoModic changes and interleukin 1 genelocus polymorphisms in occupational cohort of middle-agedmenrdquo European Spine Journal vol 18 no 12 pp 1963ndash19702009

[47] J Karppinen I Daavittila S Solovieva et al ldquoGenetic factorsare associated with modic changes in endplates of lumbarvertebral bodiesrdquo Spine vol 33 no 11 pp 1236ndash1241 2008

[48] I M Virtanen J Karppinen S Taimela et al ldquoOccupationaland genetic risk factors associated with intervertebral discdiseaserdquo Spine vol 32 no 10 pp 1129ndash1134 2007

[49] J Paz Aparicio I Fernandez Bances E Lopez-AngladaFernandez et al ldquoThe IL-1120573 (+3953 TC) gene polymorphismassociates to symptomatic lumbar disc herniationrdquo EuropeanSpine Journal vol 20 no 3 pp 383ndash389 2011

[50] AMoreno-Estrada C R Gignoux J C Fernandez-Lopez et alldquoHuman genetics The genetics of Mexico recapitulates NativeAmerican substructure and affects biomedical traitsrdquo Sciencevol 344 no 6189 pp 1280ndash1285 2014

[51] R Barquera J Zuniga R Hernandez-Dıaz et al ldquoHLA class Iand class II haplotypes in admixed families from several regionsof MexicordquoMolecular Immunology vol 45 no 4 pp 1171ndash11782008

[52] J G Romero-Quintana L O Frıas-Castro E Arambula-Merazet al ldquoIdentification of novel mutation in cathepsin C genecausing Papillon-Lefevre Syndrome in Mexican patientsrdquo BMCMedical Genetics vol 14 no 1 article 7 2013

[53] A Gonzalez-Mercado J Y Sanchez-Lopez J A Regla-Nava etal ldquoAssociation analysis of vitamin D receptor gene polymor-phisms and bone mineral density in postmenopausal Mexican-Mestizo womenrdquo Genetics and Molecular Research vol 12 no3 pp 2755ndash2763 2013

[54] Y KawaguchiMKanamoriH Ishihara KOhmoriHMatsuiand T Kimura ldquoThe association of lumbar disc disease withvitamin-D receptor gene polymorphismrdquo Journal of Bone andJoint SurgerymdashSeries A vol 84 no 11 pp 2022ndash2028 2002

[55] K M C Cheung D Chan J Karppinen et al ldquoAssociation ofthe Taq I allele in vitamin D receptor with degenerative discdisease and disc bulge in a Chinese populationrdquo Spine vol 31no 10 pp 1143ndash1148 2006

[56] B Eser T Cora O Eser et al ldquoAssociation of the polymor-phisms of vitamin d receptor and aggrecan genes with degen-erative disc diseaserdquo Genetic Testing and Molecular Biomarkersvol 14 no 3 pp 313ndash317 2010

International Journal of Genomics 7

[57] A M Valdes G Hassett D J Hart and T D SpectorldquoRadiographic progression of lumbar spine disc degenerationis influenced by variation at inflammatory genes a candidateSNP association study in the Chingford cohortrdquo Spine vol 30no 21 pp 2445ndash2451 2005

[58] G Jones C White P Sambrook and J Eisman ldquoAllelicvariation in the vitamin D receptor lifestyle factors and lumbarspinal degenerative diseaserdquo Annals of the Rheumatic Diseasesvol 57 no 2 pp 94ndash99 1998

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Molecular Biology International

GenomicsInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Signal TransductionJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

Evolutionary BiologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Genetics Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Enzyme Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Microbiology

International Journal of Genomics 5

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Authorsrsquo Contribution

Salvador Cervin Serrano diagnosed and treated the patientsSalvador Cervin Serrano Dalia Gonzalez Villareal MaribelAguilar-Medina Jose Geovanni Romero Quintana Veron-ica Picos-Cardenas and Jose Guillermo Romero-Navarroperformed the research Julio Granados Eliakym ArambulaMeraz and Ignacio Osuna Ramırez analyzed the data Sal-vador Cervin Serrano and Rosalıo Ramos-Payan designedthe research Iris Estrada-Garcıa Guzman Sanchez-Schmitzand Rosalıo Ramos-Payan wrote the paper All authors readand approved the final paper

Acknowledgment

This work was supported by CONACYT (106152)

References

[1] USBJDUSBJI ldquoSpine low back and neck painrdquo in The Burdenof Musculoskeletal Diseases in the United States vol 1 pp 21ndash56American Academy of Orthopaedic Surgeons 2011

[2] D Chou D Samartzis C Bellabarba et al ldquoDegenerativemagnetic resonance imaging changes in patients with chroniclow back pain a systematic reviewrdquo Spine vol 36 supplementno 21 pp S43ndashS53 2011

[3] S Salo V Leinonen T Rikkonen et al ldquoAssociation betweenbonemineral density and lumbar disc degenerationrdquoMaturitas2014

[4] L Wang W Cui J P Kalala T V Hoof and B G Liu ldquoToinvestigate the effect of osteoporosis and intervertebral disc deg-eneration on the endplate cartilage injury in ratsrdquo Asian PacificJournal of Tropical Medicine vol 7 no 10 pp 796ndash800 2014

[5] M T Modic P M Steinberg J S Ross T J Masaryk and JR Carter ldquoDegenerative disk disease assessment of changes invertebral body marrow with MR imagingrdquo Radiology vol 166no 1 part 1 pp 193ndash199 1988

[6] S Roberts H Evans J Trivedi and J Menage ldquoHistology andpathology of the human intervertebral discrdquoThe Journal of Boneamp Joint Surgery Series A vol 88 supplement 2 pp 10ndash14 2006

[7] R Quiroz-Moreno G Lezama-Suarez and C Gomez-JimenezldquoDisc alterations of lumbar spine onmagnetic resonance imagesin asymptomatic workersrdquo Revista Medica del Instituto Mexi-cano del Seguro Social vol 46 no 2 pp 185ndash190 2008

[8] S D Boden D O Davis T S Dina N J Patronas and SW Wiesel ldquoAbnormal magnetic-resonance scans of the lumbarspine in asymptomatic subjects A prospective investigationrdquoJournal of Bone and Joint Surgery A vol 72 no 3 pp 403ndash4081990

[9] K M C Cheung J Karppinen D Chan et al ldquoPrevalenceand pattern of lumbar magnetic resonance imaging changes ina population study of one thousand forty-three individualsrdquoSpine vol 34 no 9 pp 934ndash940 2009

[10] M B Joslashrgensen A Holtermann F Gyntelberg and P Suadi-cani ldquoPhysical fitness as a predictor of herniated lumbar discdiseasemdasha 33-year follow-up in the Copenhagen male studyrdquoBMCMusculoskeletal Disorders vol 14 article 86 2013

[11] J A Buckwalter ldquoAging and degeneration of the humanintervertebral discrdquo Spine vol 20 no 11 pp 1307ndash1314 1995

[12] L Ala-Kokko ldquoGenetic risk factors for lumbar disc diseaserdquoAnnals of Medicine vol 34 no 1 pp 42ndash47 2002

[13] M C Battie T Videman L E Gibbons L D Fisher HManni-nen and K Gill ldquoDeterminants of lumbar disc degenerationa study relating lifetime exposures and magnetic resonanceimaging findings in identical twinsrdquo Spine vol 20 no 24 pp2601ndash2612 1995

[14] M C Batite D R Haynor L D Fisher S K Gill L EGibbons and T Videman ldquoSimilarities in degenerative findingson magnetic resonance images of the lumbar spines of identicaltwinsrdquo Journal of Bone and Joint SurgerymdashSeries A vol 77 no11 pp 1662ndash1670 1995

[15] P N Sambrook A J MacGregor and T D Spector ldquoGeneticinfluences on cervical and lumbar disc degeneration amagneticresonance imaging study in twinsrdquo Arthritis amp Rheumatologyvol 42 no 2 pp 366ndash372 1999

[16] M C Battie T Videman J Kaprio et al ldquoThe Twin SpineStudy contributions to a changing view of disc degenerationrdquoThe Spine Journal vol 9 no 1 pp 47ndash59 2009

[17] C Bijkerk J J Houwing-Duistermaat H A Valkenburg etal ldquoHeritabilities of radiologic osteoarthritis in peripheraljoints and of disc degeneration of the spinerdquo Arthritis ampRheumatology vol 42 no 8 pp 1729ndash1735 1999

[18] C K Kepler R K Ponnappan C A Tannoury M V Risbudand D G Anderson ldquoThemolecular basis of intervertebral discdegenerationrdquo Spine Journal vol 13 no 3 pp 318ndash330 2013

[19] S Kalb N L Martirosyan M Y Kalani G G Broc and NTheodore ldquoGenetics of the degenerated intervertebral discrdquoWorld Neurosurgery vol 77 no 3-4 pp 491ndash501 2012

[20] A A Patel W R Spiker M Daubs D Brodke and L ACannon-Albright ldquoEvidence for an inherited predisposition tolumbar disc diseaserdquo Journal of Bone and Joint SurgerymdashSeriesA vol 93 no 3 pp 225ndash229 2011

[21] T Videman J Saarela J Kaprio et al ldquoAssociations of 25structural degradative and inflammatory candidate geneswith lumbar disc desiccation bulging and height narrowingrdquoArthritis amp Rheumatism vol 60 no 2 pp 470ndash481 2009

[22] J E Mayer J C Iatridis D Chan S A Qureshi O Gottesmanand A C Hecht ldquoGenetic polymorphisms associated withintervertebral disc degenerationrdquoThe Spine Journal vol 13 no3 pp 299ndash317 2013

[23] SM F Pluijm HW Van Essen N Bravenboer et al ldquoCollagentype I 1205721 Sp1 polymorphism osteoporosis and intervertebraldisc degeneration in older men and womenrdquo Annals of theRheumatic Diseases vol 63 no 1 pp 71ndash77 2004

[24] S Annunen P Paassilta J Lohiniva et al ldquoAn allele of COL9A2associated with intervertebral disc diseaserdquo Science vol 285 no5426 pp 409ndash412 1999

[25] F Mio K Chiba Y Hirose et al ldquoA functional polymorphismin COL11A1 which encodes the 1205721 chain of type XI collagenis associated with susceptibility to lumbar disc herniationrdquoAmerican Journal of Human Genetics vol 81 no 6 pp 1271ndash1277 2007

6 International Journal of Genomics

[26] Y Kawaguchi R Osada M Kanamori et al ldquoAssociationbetween an aggrecan gene polymorphism and lumbar discdegenerationrdquo Spine vol 24 no 23 pp 2456ndash2460 1999

[27] S Seki Y Kawaguchi K Chiba et al ldquoA functional SNP inCILPencoding cartilage intermediate layer protein is associated withsusceptibility to lumbar disc diseaserdquo Nature Genetics vol 37no 6 pp 607ndash612 2005

[28] M Takahashi H Haro Y Wakabayashi T Kawa-uchi HKomori and K Shinomiya ldquoThe association of degenerationof the intervertebral disc with 5a6a polymorphism in thepromoter of the human matrix metalloproteinase-3 generdquoJournal of Bone and Joint Surgery vol 83 no 4 pp 491ndash4952001

[29] Y Hirose K Chiba T Karasugi et al ldquoA functional polymor-phism inTHBS2 that affects alternative splicing andMMPbind-ing is associated with lumbar-disc herniationrdquo The AmericanJournal of Human Genetics vol 82 no 5 pp 1122ndash1129 2008

[30] S Solovieva J Lohiniva P Leino-Arjas et al ldquoIntervertebraldisc degeneration in relation to the COL9A3 and the IL-1120573 genepolymorphismsrdquo European Spine Journal vol 15 no 5 pp 613ndash619 2006

[31] N Noponen-Hietala I Virtanen R Karttunen et al ldquoGeneticvariations in IL6 associate with intervertebral disc diseasecharacterized by sciaticardquo Pain vol 114 no 1-2 pp 186ndash1942005

[32] T Videman J Leppavuori J Kaprio et al ldquoIntragenic poly-morphisms of the vitamin D receptor gene associated withintervertebral disc degenerationrdquo Spine vol 23 no 23 pp2477ndash2485 1998

[33] R JWood and J C Fleet ldquoThe genetics of osteoporosis vitaminD receptor polymorphismsrdquoAnnual Review of Nutrition vol 18pp 233ndash258 1998

[34] A Colombini S Cauci G Lombardi et al ldquoRelationshipbetween vitamin D receptor gene (VDR) polymorphismsvitamin D status osteoarthritis and intervertebral disc degen-erationrdquo The Journal of Steroid Biochemistry and MolecularBiology vol 138 pp 24ndash40 2013

[35] A G Uitterlinden Y Fang J B J van Meurs H A P Polsand J P T M van Leeuwen ldquoGenetics and biology of vitaminD receptor polymorphismsrdquo Gene vol 338 no 2 pp 143ndash1562004

[36] K L E Phillips N Jordan-Mahy M J H Nicklin and CL Le Maitre ldquoInterleukin-1 receptor antagonist deficient miceprovide insights into pathogenesis of human intervertebral discdegenerationrdquo Annals of the Rheumatic Diseases vol 72 no 11pp 1860ndash1867 2013

[37] W Ye D S Huang W J Chen et al ldquoAssociation of 86 bpvariable number tandem repeat polymorphism of interleukin-1receptor antagonist gene with lumbar disc diseaserdquoNan Fang YiKe Da Xue Xue Bao vol 27 no 10 pp 1485ndash1488 2007

[38] S Solovieva S Kouhia P Leino-Arjas et al ldquoInterleukin 1polymorphisms and intervertebral disc degenerationrdquo Epidemi-ology vol 15 no 5 pp 626ndash633 2004

[39] P J Eskola P Kjaer I M Daavittila et al ldquoGenetic risk factorsof disc degeneration among 12-14-year-old Danish children apopulation studyrdquo International Journal of Molecular Epidemi-ology and Genetics vol 1 no 2 pp 158ndash165 2010

[40] M T Modic T J Masaryk J S Ross and J R Carter ldquoImagingof degenerative disk diseaserdquo Radiology vol 168 no 1 pp 177ndash186 1988

[41] S Gustincich G Manfioletti G Del Sal C Schneider andP Carninci ldquoA fast method for high-quality genomic DNAextraction fromwhole human bloodrdquo BioTechniques vol 11 no3 pp 298ndash302 1991

[42] M Gail R Williams D P Byar and C Brown ldquoHow manycontrolsrdquo Journal of Chronic Diseases vol 29 no 11 pp 723ndash731 1976

[43] B Woolf ldquoOn estimating the relation between blood group anddiseaserdquo Annals of Human Genetics vol 19 no 4 pp 251ndash2531955

[44] T V Perneger ldquoWhatrsquos wrong with Bonferroni adjustmentsrdquoBritish Medical Journal vol 316 no 7139 pp 1236ndash1238 1998

[45] I Tegeder and J Lotsch ldquoCurrent evidence for a modulation oflow back pain by human genetic variantsrdquo Journal of Cellularand Molecular Medicine vol 13 no 8B pp 1605ndash1619 2009

[46] J Karppinen S Solovieva K Luoma R Raininko P Leino-Arjas and H Riihimaki ldquoModic changes and interleukin 1 genelocus polymorphisms in occupational cohort of middle-agedmenrdquo European Spine Journal vol 18 no 12 pp 1963ndash19702009

[47] J Karppinen I Daavittila S Solovieva et al ldquoGenetic factorsare associated with modic changes in endplates of lumbarvertebral bodiesrdquo Spine vol 33 no 11 pp 1236ndash1241 2008

[48] I M Virtanen J Karppinen S Taimela et al ldquoOccupationaland genetic risk factors associated with intervertebral discdiseaserdquo Spine vol 32 no 10 pp 1129ndash1134 2007

[49] J Paz Aparicio I Fernandez Bances E Lopez-AngladaFernandez et al ldquoThe IL-1120573 (+3953 TC) gene polymorphismassociates to symptomatic lumbar disc herniationrdquo EuropeanSpine Journal vol 20 no 3 pp 383ndash389 2011

[50] AMoreno-Estrada C R Gignoux J C Fernandez-Lopez et alldquoHuman genetics The genetics of Mexico recapitulates NativeAmerican substructure and affects biomedical traitsrdquo Sciencevol 344 no 6189 pp 1280ndash1285 2014

[51] R Barquera J Zuniga R Hernandez-Dıaz et al ldquoHLA class Iand class II haplotypes in admixed families from several regionsof MexicordquoMolecular Immunology vol 45 no 4 pp 1171ndash11782008

[52] J G Romero-Quintana L O Frıas-Castro E Arambula-Merazet al ldquoIdentification of novel mutation in cathepsin C genecausing Papillon-Lefevre Syndrome in Mexican patientsrdquo BMCMedical Genetics vol 14 no 1 article 7 2013

[53] A Gonzalez-Mercado J Y Sanchez-Lopez J A Regla-Nava etal ldquoAssociation analysis of vitamin D receptor gene polymor-phisms and bone mineral density in postmenopausal Mexican-Mestizo womenrdquo Genetics and Molecular Research vol 12 no3 pp 2755ndash2763 2013

[54] Y KawaguchiMKanamoriH Ishihara KOhmoriHMatsuiand T Kimura ldquoThe association of lumbar disc disease withvitamin-D receptor gene polymorphismrdquo Journal of Bone andJoint SurgerymdashSeries A vol 84 no 11 pp 2022ndash2028 2002

[55] K M C Cheung D Chan J Karppinen et al ldquoAssociation ofthe Taq I allele in vitamin D receptor with degenerative discdisease and disc bulge in a Chinese populationrdquo Spine vol 31no 10 pp 1143ndash1148 2006

[56] B Eser T Cora O Eser et al ldquoAssociation of the polymor-phisms of vitamin d receptor and aggrecan genes with degen-erative disc diseaserdquo Genetic Testing and Molecular Biomarkersvol 14 no 3 pp 313ndash317 2010

International Journal of Genomics 7

[57] A M Valdes G Hassett D J Hart and T D SpectorldquoRadiographic progression of lumbar spine disc degenerationis influenced by variation at inflammatory genes a candidateSNP association study in the Chingford cohortrdquo Spine vol 30no 21 pp 2445ndash2451 2005

[58] G Jones C White P Sambrook and J Eisman ldquoAllelicvariation in the vitamin D receptor lifestyle factors and lumbarspinal degenerative diseaserdquo Annals of the Rheumatic Diseasesvol 57 no 2 pp 94ndash99 1998

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Molecular Biology International

GenomicsInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Signal TransductionJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

Evolutionary BiologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Genetics Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Enzyme Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Microbiology

6 International Journal of Genomics

[26] Y Kawaguchi R Osada M Kanamori et al ldquoAssociationbetween an aggrecan gene polymorphism and lumbar discdegenerationrdquo Spine vol 24 no 23 pp 2456ndash2460 1999

[27] S Seki Y Kawaguchi K Chiba et al ldquoA functional SNP inCILPencoding cartilage intermediate layer protein is associated withsusceptibility to lumbar disc diseaserdquo Nature Genetics vol 37no 6 pp 607ndash612 2005

[28] M Takahashi H Haro Y Wakabayashi T Kawa-uchi HKomori and K Shinomiya ldquoThe association of degenerationof the intervertebral disc with 5a6a polymorphism in thepromoter of the human matrix metalloproteinase-3 generdquoJournal of Bone and Joint Surgery vol 83 no 4 pp 491ndash4952001

[29] Y Hirose K Chiba T Karasugi et al ldquoA functional polymor-phism inTHBS2 that affects alternative splicing andMMPbind-ing is associated with lumbar-disc herniationrdquo The AmericanJournal of Human Genetics vol 82 no 5 pp 1122ndash1129 2008

[30] S Solovieva J Lohiniva P Leino-Arjas et al ldquoIntervertebraldisc degeneration in relation to the COL9A3 and the IL-1120573 genepolymorphismsrdquo European Spine Journal vol 15 no 5 pp 613ndash619 2006

[31] N Noponen-Hietala I Virtanen R Karttunen et al ldquoGeneticvariations in IL6 associate with intervertebral disc diseasecharacterized by sciaticardquo Pain vol 114 no 1-2 pp 186ndash1942005

[32] T Videman J Leppavuori J Kaprio et al ldquoIntragenic poly-morphisms of the vitamin D receptor gene associated withintervertebral disc degenerationrdquo Spine vol 23 no 23 pp2477ndash2485 1998

[33] R JWood and J C Fleet ldquoThe genetics of osteoporosis vitaminD receptor polymorphismsrdquoAnnual Review of Nutrition vol 18pp 233ndash258 1998

[34] A Colombini S Cauci G Lombardi et al ldquoRelationshipbetween vitamin D receptor gene (VDR) polymorphismsvitamin D status osteoarthritis and intervertebral disc degen-erationrdquo The Journal of Steroid Biochemistry and MolecularBiology vol 138 pp 24ndash40 2013

[35] A G Uitterlinden Y Fang J B J van Meurs H A P Polsand J P T M van Leeuwen ldquoGenetics and biology of vitaminD receptor polymorphismsrdquo Gene vol 338 no 2 pp 143ndash1562004

[36] K L E Phillips N Jordan-Mahy M J H Nicklin and CL Le Maitre ldquoInterleukin-1 receptor antagonist deficient miceprovide insights into pathogenesis of human intervertebral discdegenerationrdquo Annals of the Rheumatic Diseases vol 72 no 11pp 1860ndash1867 2013

[37] W Ye D S Huang W J Chen et al ldquoAssociation of 86 bpvariable number tandem repeat polymorphism of interleukin-1receptor antagonist gene with lumbar disc diseaserdquoNan Fang YiKe Da Xue Xue Bao vol 27 no 10 pp 1485ndash1488 2007

[38] S Solovieva S Kouhia P Leino-Arjas et al ldquoInterleukin 1polymorphisms and intervertebral disc degenerationrdquo Epidemi-ology vol 15 no 5 pp 626ndash633 2004

[39] P J Eskola P Kjaer I M Daavittila et al ldquoGenetic risk factorsof disc degeneration among 12-14-year-old Danish children apopulation studyrdquo International Journal of Molecular Epidemi-ology and Genetics vol 1 no 2 pp 158ndash165 2010

[40] M T Modic T J Masaryk J S Ross and J R Carter ldquoImagingof degenerative disk diseaserdquo Radiology vol 168 no 1 pp 177ndash186 1988

[41] S Gustincich G Manfioletti G Del Sal C Schneider andP Carninci ldquoA fast method for high-quality genomic DNAextraction fromwhole human bloodrdquo BioTechniques vol 11 no3 pp 298ndash302 1991

[42] M Gail R Williams D P Byar and C Brown ldquoHow manycontrolsrdquo Journal of Chronic Diseases vol 29 no 11 pp 723ndash731 1976

[43] B Woolf ldquoOn estimating the relation between blood group anddiseaserdquo Annals of Human Genetics vol 19 no 4 pp 251ndash2531955

[44] T V Perneger ldquoWhatrsquos wrong with Bonferroni adjustmentsrdquoBritish Medical Journal vol 316 no 7139 pp 1236ndash1238 1998

[45] I Tegeder and J Lotsch ldquoCurrent evidence for a modulation oflow back pain by human genetic variantsrdquo Journal of Cellularand Molecular Medicine vol 13 no 8B pp 1605ndash1619 2009

[46] J Karppinen S Solovieva K Luoma R Raininko P Leino-Arjas and H Riihimaki ldquoModic changes and interleukin 1 genelocus polymorphisms in occupational cohort of middle-agedmenrdquo European Spine Journal vol 18 no 12 pp 1963ndash19702009

[47] J Karppinen I Daavittila S Solovieva et al ldquoGenetic factorsare associated with modic changes in endplates of lumbarvertebral bodiesrdquo Spine vol 33 no 11 pp 1236ndash1241 2008

[48] I M Virtanen J Karppinen S Taimela et al ldquoOccupationaland genetic risk factors associated with intervertebral discdiseaserdquo Spine vol 32 no 10 pp 1129ndash1134 2007

[49] J Paz Aparicio I Fernandez Bances E Lopez-AngladaFernandez et al ldquoThe IL-1120573 (+3953 TC) gene polymorphismassociates to symptomatic lumbar disc herniationrdquo EuropeanSpine Journal vol 20 no 3 pp 383ndash389 2011

[50] AMoreno-Estrada C R Gignoux J C Fernandez-Lopez et alldquoHuman genetics The genetics of Mexico recapitulates NativeAmerican substructure and affects biomedical traitsrdquo Sciencevol 344 no 6189 pp 1280ndash1285 2014

[51] R Barquera J Zuniga R Hernandez-Dıaz et al ldquoHLA class Iand class II haplotypes in admixed families from several regionsof MexicordquoMolecular Immunology vol 45 no 4 pp 1171ndash11782008

[52] J G Romero-Quintana L O Frıas-Castro E Arambula-Merazet al ldquoIdentification of novel mutation in cathepsin C genecausing Papillon-Lefevre Syndrome in Mexican patientsrdquo BMCMedical Genetics vol 14 no 1 article 7 2013

[53] A Gonzalez-Mercado J Y Sanchez-Lopez J A Regla-Nava etal ldquoAssociation analysis of vitamin D receptor gene polymor-phisms and bone mineral density in postmenopausal Mexican-Mestizo womenrdquo Genetics and Molecular Research vol 12 no3 pp 2755ndash2763 2013

[54] Y KawaguchiMKanamoriH Ishihara KOhmoriHMatsuiand T Kimura ldquoThe association of lumbar disc disease withvitamin-D receptor gene polymorphismrdquo Journal of Bone andJoint SurgerymdashSeries A vol 84 no 11 pp 2022ndash2028 2002

[55] K M C Cheung D Chan J Karppinen et al ldquoAssociation ofthe Taq I allele in vitamin D receptor with degenerative discdisease and disc bulge in a Chinese populationrdquo Spine vol 31no 10 pp 1143ndash1148 2006

[56] B Eser T Cora O Eser et al ldquoAssociation of the polymor-phisms of vitamin d receptor and aggrecan genes with degen-erative disc diseaserdquo Genetic Testing and Molecular Biomarkersvol 14 no 3 pp 313ndash317 2010

International Journal of Genomics 7

[57] A M Valdes G Hassett D J Hart and T D SpectorldquoRadiographic progression of lumbar spine disc degenerationis influenced by variation at inflammatory genes a candidateSNP association study in the Chingford cohortrdquo Spine vol 30no 21 pp 2445ndash2451 2005

[58] G Jones C White P Sambrook and J Eisman ldquoAllelicvariation in the vitamin D receptor lifestyle factors and lumbarspinal degenerative diseaserdquo Annals of the Rheumatic Diseasesvol 57 no 2 pp 94ndash99 1998

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Molecular Biology International

GenomicsInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Signal TransductionJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

Evolutionary BiologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Genetics Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Enzyme Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Microbiology

International Journal of Genomics 7

[57] A M Valdes G Hassett D J Hart and T D SpectorldquoRadiographic progression of lumbar spine disc degenerationis influenced by variation at inflammatory genes a candidateSNP association study in the Chingford cohortrdquo Spine vol 30no 21 pp 2445ndash2451 2005

[58] G Jones C White P Sambrook and J Eisman ldquoAllelicvariation in the vitamin D receptor lifestyle factors and lumbarspinal degenerative diseaserdquo Annals of the Rheumatic Diseasesvol 57 no 2 pp 94ndash99 1998

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Molecular Biology International

GenomicsInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Signal TransductionJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

Evolutionary BiologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Genetics Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Enzyme Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Microbiology

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Molecular Biology International

GenomicsInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Signal TransductionJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

Evolutionary BiologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Genetics Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Enzyme Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Microbiology