GEORG-SCHMORL-PRIZE OF THE GERMAN SPINE SOCIETY (DWG) 2016: Comparison of in vitro osteogenic potential of iliac crest and degenerative facet joint bone autografts for intervertebral fusion in lumbar spinal stenosis
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The promotion of spinal fusion using bone autografts is largely mediated by the osteoinductive potential of progenitors/mesenchymal stem cells (MSC) that reside in the marrow spaces of cancellous bone. Iliac crest is the common autograft donor site, but its use presents an increased risk for donor site pain, morbidity and infection. Degenerative bone samples harvested during facetectomy might provide an alternative viable source of osteoinductive autografts. In this study, we conducted an intra-individual comparison of the osteogenic potential of isolated low passage MSC from both sources.
Iliac crest and degenerative facet joints were harvested from eight consecutive patients undergoing transforaminal lumbar interspinal fusion due to lumbar spinal stenosis. MSC were isolated by collagenase digestion, selected by plastic adherence and minimally expanded for downstream assays. Clonogenic and osteogenic potential was evaluated by colony formation assays in control and osteogenic culture medium. Osteogenic properties, including alkaline phosphatase (ALP) induction, matrix mineralization and type I collagen mRNA and protein expression were characterized using quantitative histochemical staining and reverse transcription PCR. Spontaneous adipogenesis was analysed by adipocyte enumeration and gene expression analysis of adipogenic markers.
Average colony-forming efficiency in osteogenic medium was equal between iliac crest (38 ± 12%) and facet joint (36 ± 11%). Osteogenic potential at the clonal level was 55 ± 26 and 68 ± 17% for iliac crest and facet joint MSC, respectively. Clonogenic and osteogenic potential were significantly negatively associated with donor age. Osteogenic differentiation led to significant induction of ALP activity in iliac crest (sixfold) and facet joint (eightfold) MSC. Matrix mineralization quantified by Alizarin red staining was increased by osteogenic differentiation, yet similar between both MSC sources. Protein expression of type I collagen was enhanced during osteogenesis and significantly greater in iliac crest MSC. Correspondingly, COL1A2 mRNA expression was higher in osteogenically differentiated MSC from iliac crest. Adipocyte numbers showed significant differences between iliac crest (63 ± 60) and facet joint (18 ± 15) MSC under osteogenic conditions. Negative (GREM1) and positive (FABP4) adipogenic markers were not differentially expressed between sources.
MSC from iliac crest and degenerative facet joints largely display similar clonogenic and osteogenic properties in vitro. Differences at the molecular level are not likely to impair the osteoinductive capacity of facet joint MSC. Bone autografts from facetectomy would be viable alternatives as bone autografts for intervertebral spinal fusion in lumbar spinal stenosis.
KeywordsSpinal fusion Bone graft Facetectomy Mesenchymal stem cell Osteogenesis
Compliance with ethical standards
Conflict of interest
None of the authors has any potential conflict of interest.
This research was supported by a Grant from the Gottfried and Julia Bangerter-Rhyner Foundation to J.G. and C.N.
- 1.Niu CC, Tsai TT, Fu TS, Lai PL, Chen LH, Chen WJ (2009) A comparison of posterolateral lumbar fusion comparing autograft, autogenous laminectomy bone with bone marrow aspirate, and calcium sulphate with bone marrow aspirate: a prospective randomized study. Spine (Phila Pa 1976) 34:2715–2719. doi: 10.1097/BRS.0b013e3181b47232 CrossRefGoogle Scholar
- 2.Ito Z, Imagama S, Kanemura T, Hachiya Y, Miura Y, Kamiya M, Yukawa Y, Sakai Y, Katayama Y, Wakao N, Matsuyama Y, Ishiguro N (2013) Bone union rate with autologous iliac bone versus local bone graft in posterior lumbar interbody fusion (PLIF): a multicenter study. Eur Spine J 22:1158–1163. doi: 10.1007/s00586-012-2593-4 CrossRefPubMedPubMedCentralGoogle Scholar
- 3.Fu TS, Wang IC, Lu ML, Hsieh MK, Chen LH, Chen WJ (2016) The fusion rate of demineralized bone matrix compared with autogenous iliac bone graft for long multi-segment posterolateral spinal fusion. BMC Musculoskelet Disord 17:3. doi: 10.1186/s12891-015-0861-2 CrossRefPubMedPubMedCentralGoogle Scholar
- 5.Summers BN, Eisenstein SM (1989) Donor site pain from the ilium. A complication of lumbar spine fusion. J Bone Jt Surg Br 71:677–680Google Scholar
- 8.Radcliff K, Hwang R, Hilibrand A, Smith HE, Gruskay J, Lurie JD, Zhao W, Albert T, Weinstein J (2012) The effect of iliac crest autograft on the outcome of fusion in the setting of degenerative spondylolisthesis: a subgroup analysis of the Spine Patient Outcomes Research Trial (SPORT). J Bone Jt Surg Am 94:1685–1692. doi: 10.2106/JBJS.K.00952 CrossRefGoogle Scholar
- 11.Ito Z, Matsuyama Y, Sakai Y, Imagama S, Wakao N, Ando K, Hirano K, Tauchi R, Muramoto A, Matsui H, Matsumoto T, Kanemura T, Yoshida G, Ishikawa Y, Ishiguro N (2010) Bone union rate with autologous iliac bone versus local bone graft in posterior lumbar interbody fusion. Spine (Phila Pa 1976) 35:E1101–E1105. doi: 10.1097/BRS.0b013e3181de4f2e CrossRefGoogle Scholar
- 12.Eder C, Chavanne A, Meissner J, Bretschneider W, Tuschel A, Becker P, Ogon M (2011) Autografts for spinal fusion: osteogenic potential of laminectomy bone chips and bone shavings collected via high speed drill. Eur Spine J 20:1791–1795. doi: 10.1007/s00586-011-1736-3 CrossRefPubMedPubMedCentralGoogle Scholar
- 15.Niu CC, Lin SS, Yuan LJ, Chen LH, Pan TL, Yang CY, Lai PL, Chen WJ (2014) Identification of mesenchymal stem cells and osteogenic factors in bone marrow aspirate and peripheral blood for spinal fusion by flow cytometry and proteomic analysis. J Orthop Surg Res 9:32. doi: 10.1186/1749-799X-9-32 CrossRefPubMedPubMedCentralGoogle Scholar
- 16.Harada Y, Furukawa K, Asari T, Chin S, Ono A, Tanaka T, Mizukami H, Murakami M, Yagihashi S, Motomura S, Ishibashi Y (2014) Osteogenic lineage commitment of mesenchymal stem cells from patients with ossification of the posterior longitudinal ligament. Biochem Biophys Res Commun 443:1014–1020. doi: 10.1016/j.bbrc.2013.12.080 CrossRefPubMedGoogle Scholar
- 17.Kristjansson B, Limthongkul W, Yingsakmongkol W, Thantiworasit P, Jirathanathornnukul N, Honsawek S (2016) Isolation and characterization of human mesenchymal stem cells from facet joints and interspinous ligaments. Spine (Phila Pa 1976) 41:E1–E7. doi: 10.1097/BRS.0000000000001178 CrossRefGoogle Scholar
- 20.Appel H, Maier R, Loddenkemper C, Kayser R, Meier O, Hempfing A, Sieper J (2010) Immunohistochemical analysis of osteoblasts in zygapophyseal joints of patients with ankylosing spondylitis reveal repair mechanisms similar to osteoarthritis. J Rheumatol 37:823–828. doi: 10.3899/jrheum.090986 CrossRefPubMedGoogle Scholar
- 22.Zhen G, Wen C, Jia X, Li Y, Crane JL, Mears SC, Askin FB, Frassica FJ, Chang W, Yao J, Carrino JA, Cosgarea A, Artemov D, Chen Q, Zhao Z, Zhou X, Riley L, Sponseller P, Wan M, Lu WW, Cao X (2013) Inhibition of TGF-beta signaling in mesenchymal stem cells of subchondral bone attenuates osteoarthritis. Nat Med 19:704–712. doi: 10.1038/nm.3143 CrossRefPubMedPubMedCentralGoogle Scholar
- 24.Hunter DJ, Gerstenfeld L, Bishop G, Davis AD, Mason ZD, Einhorn TA, Maciewicz RA, Newham P, Foster M, Jackson S, Morgan EF (2009) Bone marrow lesions from osteoarthritis knees are characterized by sclerotic bone that is less well mineralized. Arthritis Res Ther 11:R11. doi: 10.1186/ar2601 CrossRefPubMedPubMedCentralGoogle Scholar
- 30.Ghali O, Broux O, Falgayrac G, Haren N, van Leeuwen JP, Penel G, Hardouin P, Chauveau C (2015) Dexamethasone in osteogenic medium strongly induces adipocyte differentiation of mouse bone marrow stromal cells and increases osteoblast differentiation. BMC Cell Biol 16:9. doi: 10.1186/s12860-015-0056-6 CrossRefPubMedPubMedCentralGoogle Scholar
- 31.Worthley DL, Churchill M, Compton JT, Tailor Y, Rao M, Si Y, Levin D, Schwartz MG, Uygur A, Hayakawa Y, Gross S, Renz BW, Setlik W, Martinez AN, Chen X, Nizami S, Lee HG, Kang HP, Caldwell JM, Asfaha S, Westphalen CB, Graham T, Jin G, Nagar K, Wang H, Kheirbek MA, Kolhe A, Carpenter J, Glaire M, Nair A, Renders S, Manieri N, Muthupalani S, Fox JG, Reichert M, Giraud AS, Schwabe RF, Pradere JP, Walton K, Prakash A, Gumucio D, Rustgi AK, Stappenbeck TS, Friedman RA, Gershon MD, Sims P, Grikscheit T, Lee FY, Karsenty G, Mukherjee S, Wang TC (2015) Gremlin 1 identifies a skeletal stem cell with bone, cartilage, and reticular stromal potential. Cell 160:269–284. doi: 10.1016/j.cell.2014.11.042 CrossRefPubMedPubMedCentralGoogle Scholar
- 32.Kim JS, Ali MH, Wydra F, Li X, Hamilton JL, An HS, Cs-Szabo G, Andrews S, Moric M, Xiao G, Wang JH, Chen D, Cavanaugh JM, Im HJ (2015) Characterization of degenerative human facet joints and facet joint capsular tissues. Osteoarthr Cartil 23:2242–2251. doi: 10.1016/j.joca.2015.06.009 CrossRefPubMedPubMedCentralGoogle Scholar
- 33.Baboolal TG, Boxall SA, El-Sherbiny YM, Moseley TA, Cuthbert RJ, Giannoudis PV, McGonagle D, Jones E (2014) Multipotential stromal cell abundance in cellular bone allograft: comparison with fresh age-matched iliac crest bone and bone marrow aspirate. Regen Med 9:593–607. doi: 10.2217/rme.14.17 CrossRefPubMedPubMedCentralGoogle Scholar
- 35.Bianco D, Todorov A Jr, Čengić T, Pagenstert GI, Hügle T, Forster-Horvath C, Martin I, Schären S, Geurts J (2016) Subchondral bone mesenchymal stromal cells from osteoarthritic lesions give rise to aberrant in vitro and in vivo mineralization. Osteoarthr Cartil 24:S133. doi: 10.1016/j.joca.2016.01.259 CrossRefGoogle Scholar
- 36.Paul J, Barg A, Kretzschmar M, Pagenstert G, Studler U, Hugle T, Wegner NJ, Valderrabano V, Geurts J (2015) Increased Osseous (99m)Tc-DPD uptake in end-stage ankle osteoarthritis: correlation between SPECT-CT imaging and histologic findings. Foot Ankle Int 36:1438–1447. doi: 10.1177/1071100715596745 CrossRefPubMedGoogle Scholar