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Demineralized bone promotes chondrocyte or osteoblast differentiation of human marrow stromal cells cultured in collagen sponges

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Abstract

Demineralized bone implants have been used for many types of craniomaxillofacial, orthopedic, periodontal, and hand reconstruction procedures. In previous studies, we showed that demineralized bone powder (DBP) induces chondrogenesis of human dermal fibroblasts in a DBP/collagen sponge system that optimized interactions between particles of DBP and target cells in cell culture. In this study, we test the hypothesis that DBP promotes chondrogenesis or osteogenesis of human marrow stromal cells (hMSCs) in 3-D collagen sponge culture, depending upon the culture conditions. We first confirmed that hMSCs have chondrogenic potential when treated with TGF-β, either in 2-D monolayer cultures or in 3-D porous collagen sponges. Second, we found that DBP markedly enhanced chondrogenesis in hMSCs in 3-D sponges, as assessed by metachromasia and expression of chondrocyte-specific genes AGGRECAN, COL II, and COL X. Human dermal fibroblasts (hDFs) were used to define mechanisms of chondroinduction because unlike hMSCs they have no inherent chondrogenic potential. In situ hybridization revealed that hDFs vicinal to DBPs express chondrocyte-specific genes AGGRECAN or COL II. Macroarray analysis showed that DBP activates TGF-β/BMP signaling pathway genes in hDFs. Finally, DBP induced hMSCs to express the osteoblast phenotype when cultured with osteogenic supplements. These studies show how culture conditions can influence the differentiation pathway that human marrow stromal cells follow when stimulated by DBP. These results support the potential to engineer cartilage or bone in vitro by using human bone marrow stromal cells and DBP/collagen scaffolds.

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Abbreviations

3-D:

three-dimensional

DBP:

Demineralized Bone Powder

DMEM:

Dulbecco’s Modifed Eagle’s Medium

FBS-HI:

Fetal Bovine Serum–Heat Inactivated

hDFs:

human dermal fibroblasts

hMSCs:

human marrow stromal cells

IRB:

Institutional Review Board

References

  • H. Breitschopf G. Suchanek et al. (1996) Detection of mRNA in tissue using DIG-labeled RNA and oligonucleotide probes S. Grunewald-Janho (Eds) Nonradioactive In Situ Hybridization Application Manual EditionNumber2 Roche Molecular Biochemicals, Boehringer Mannheim GmbH Germany 126–135

    Google Scholar 

  • E. Canalis T. McCarthy M. Centrella (1988) ArticleTitleIsolation of growth factors from adult bovine bone Calcif. Tissue Intl. 43 346–351 Occurrence Handle1:CAS:528:DyaL1MXhtlWmtLs%3D

    CAS  Google Scholar 

  • M. Centrella E. Canalis (1985) ArticleTitleLocal regulators of skeletal growth: a perspective Endocr. Rev. 6 544–551 Occurrence Handle10.1210/edrv-6-4-544 Occurrence Handle3000756 Occurrence Handle1:CAS:528:DyaL28Xmt1Wmug%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • M. Centrella M.C. Horowitz J.M. Wozney T.L. McCarthy (1994) ArticleTitleTransforming growth factor-β gene family members and bone Endocr. Rev. 15 27–39 Occurrence Handle8156937 Occurrence Handle1:CAS:528:DyaK2cXivFShsLw%3D

    PubMed  CAS  Google Scholar 

  • D.L. Diefenderfer A.M. Osyczka G.C. Reilly P.S. Leboy (2003a) ArticleTitleB.M.P. responsiveness in human mesenchymal stem cells Connect. Tissue Res. 44 305–311 Occurrence Handle1:CAS:528:DC%2BD3sXjtlanu7s%3D

    CAS  Google Scholar 

  • D.L. Diefenderfer A.M. Osyczka J.P. Garino P.S. Leboy (2003b) ArticleTitleRegulation of BMP-induced transcription in cultured human bone marrow stromal cells J. Bone Joint Surgery. Am. 85-A 19–28

    Google Scholar 

  • S.C. Gamradt J.R. Lieberman (2003) ArticleTitleBone graft for revision hip arthroplasty: biology and future applications Clin. Orthop. 417 183–194 Occurrence Handle14646716

    PubMed  Google Scholar 

  • J. Glowacki L.B. Kaban J.E. Murray J. Folkman J.B. Mulliken (1981) ArticleTitleApplication of the biological principle of induced osteogenesis for craniofacial defects Lancet 1 959–962 Occurrence Handle6112384 Occurrence Handle1:STN:280:Bi6C1MnitFI%3D

    PubMed  CAS  Google Scholar 

  • J. Glowacki J.B. Mulliken (1985) ArticleTitleDemineralized bone implants Clin. Plast Surg. 12 233–241 Occurrence Handle3886260 Occurrence Handle1:STN:280:BiqC2sfgt1E%3D

    PubMed  CAS  Google Scholar 

  • J. Glowacki (1996) ArticleTitleCellular reactions to bone-derived material Clin. Orthop. Relat. Res. 324 47–54 Occurrence Handle8595776

    PubMed  Google Scholar 

  • J. Glowacki K.E. Yates G. Little S. Mizuno (1998) ArticleTitleInduced chondroblastic differentiation of human fibroblasts by three-dimensional culture with demineralized bone matrix Mat. Sci. Eng. C 6 199–203

    Google Scholar 

  • E. Grimaud D. Heymann F. Redini (2002) ArticleTitleRecent advances in TGF-β effects on chondrocyte metabolism Potential therapeutic roles of TGF-β in cartilage disorders Cytokine Growth Factor Rev. 13 241–257 Occurrence Handle12486877 Occurrence Handle1:CAS:528:DC%2BD38Xktlylt78%3D

    PubMed  CAS  Google Scholar 

  • A.A. Kale P.E. Di Cesare (1995) ArticleTitleOsteoinductive agents. Basic science and clinical applications Am. J. Orthop. 24 752–761 Occurrence Handle8593557 Occurrence Handle1:STN:280:BymC3M3ktlA%3D

    PubMed  CAS  Google Scholar 

  • P. Kasten R. Luginbuhl M. van Griensven T. Barkhausen C. Krettek M. Bohner U. Bosch (2003) ArticleTitleComparison of human bone marrow stromal cells seeded on calcium-deficient hydroxyapatitebeta-tricalcium phosphate and demineralized bone matrix Biomaterials 24 2593–2603 Occurrence Handle10.1016/S0142-9612(03)00062-0 Occurrence Handle1:CAS:528:DC%2BD3sXjt1emu7o%3D Occurrence Handle12726713

    Article  CAS  PubMed  Google Scholar 

  • A. Lomri O. Fromigue M Hott P.J. Marie (1999) ArticleTitleGenomic insertion of the SV-40 large T oncogene in normal adult human trabecular osteoblastic cells induces cell growth without loss of the differentiated phenotype Calcif. Tissue Intl. 64 394–401 Occurrence Handle1:CAS:528:DyaK1MXislynsLw%3D

    CAS  Google Scholar 

  • Mauney J.R., Sjostorm S., Blumberg J., Horan R., O’Leary J.P., Vunjak-Novakovic G., Volloch V. and Kaplan D.L. 2004. Mechanical Stimulation Promotes Osteogenic Differentiation of Human Bone Marrow Stromal Cells on 3-D Partially Demineralized Bone Scaffolds In Vitro. Calcif. Tissue Intl. 74: 458–468.

    Google Scholar 

  • S. Mizuno J. Glowacki (1996a) ArticleTitleChondroinduction of human dermal fibroblasts by demineralized bone in three-dimensional culture Exp. Cell Res. 227 89–97 Occurrence Handle1:CAS:528:DyaK28Xltlymtb4%3D

    CAS  Google Scholar 

  • S. Mizuno J. Glowacki (1996b) ArticleTitleThree-dimensional composite of demineralized bone powder and collagen for in vitro analysis of chondroinduction of human dermal fibroblasts Biomaterials 17 1819–1825 Occurrence Handle1:CAS:528:DyaK28Xls1OmtbY%3D

    CAS  Google Scholar 

  • S. Mohan D.J. Baylink (1991) ArticleTitleBone growth factors Clin. Orthop. Relat. Res. 263 30–48 Occurrence Handle1993386

    PubMed  Google Scholar 

  • S.M. Mueller J. Glowacki (2001) ArticleTitleAge-related decline in the osteogenic potential of human bone marrow cells cultured in three-dimensional collagen sponges J. Cell Biochem. 82 583–590 Occurrence Handle11500936 Occurrence Handle1:CAS:528:DC%2BD3MXlvFKrt7w%3D

    PubMed  CAS  Google Scholar 

  • N. Muthukumaran A.H. Reddi (1985) ArticleTitleBone matrix-induced local bone induction Clin. Orthop. 200 159–164 Occurrence Handle3905107 Occurrence Handle1:CAS:528:DyaL28XkvFShtQ%3D%3D

    PubMed  CAS  Google Scholar 

  • M.E. Nuttall J.M. Gimble (2000) ArticleTitleIs there a therapeutic opportunity to either prevent or treat osteopenic disorders by inhibiting marrow adipogenesis? Bone 27 177–184 Occurrence Handle10913909 Occurrence Handle1:CAS:528:DC%2BD3cXltVGqsbw%3D

    PubMed  CAS  Google Scholar 

  • A.M. Osyczka D.L. Diefenderfer G. Bhargave P.S. Leboy (2004) ArticleTitleDifferent effects of BMP-2 on marrow stromal cells from human and rat bone Cells Tissue Organs 176 109–119 Occurrence Handle1:CAS:528:DC%2BD2cXptVCmug%3D%3D

    CAS  Google Scholar 

  • M.F. Pittenger A.M. Mackay S.C. Beck R.K. Jaiswal R. Douglas J.D. Mosca M.A. Moorman D.W. Simonetti S. Craig D.R. Marshak (1999) ArticleTitleMultilineage potential of adult human mesenchymal stem cells Science 284 143–147 Occurrence Handle10.1126/science.284.5411.143 Occurrence Handle1:CAS:528:DyaK1MXitlCnu7o%3D Occurrence Handle10102814

    Article  CAS  PubMed  Google Scholar 

  • A.H. Reddi C. Huggins (1972) ArticleTitleBiochemical sequences in the transformation of normal fibroblasts in adolescent rats Proc. Natl. Acad. Sci. USA 69 1601–1605 Occurrence Handle4504376 Occurrence Handle1:CAS:528:DyaE38XksF2ltL4%3D

    PubMed  CAS  Google Scholar 

  • B.A. Roelen P. Dijke (2003) ArticleTitleControlling mesenchymal stem cell differentiation by TGFβ family members J. Orthop. Sci. 8 740–748 Occurrence Handle14557946

    PubMed  Google Scholar 

  • R.K. Rosenthal J. Folkman J. Glowacki (1999) ArticleTitleDemineralized bone implants for non-union fractures, bone cysts, and fibrous bone lesions Clin. Orthop. Relat. Res. 364 61–69 Occurrence Handle10416393

    PubMed  Google Scholar 

  • E. Solheim (1998) ArticleTitleOsteoinduction by demineralised bone Intl. Orthop. 22 335–342 Occurrence Handle1:STN:280:DyaK1M7htleksg%3D%3D

    CAS  Google Scholar 

  • S.T. Sonis R.C. Williams M.K. Jeffcoat R. Black G. Shklar (1985) ArticleTitleHealing of spontaneous periodontal defects in dogs treated with xenogeneic demineralized bone J. Periodontol. 56 470–479 Occurrence Handle3915013 Occurrence Handle1:STN:280:BimB38npt1E%3D

    PubMed  CAS  Google Scholar 

  • J. Upton J. Glowacki (1992) ArticleTitleHand reconstruction with allograft demineralized bone: twenty-six implants in twelve patients J. Hand. Surg. [Am] 17 704–713 Occurrence Handle1:STN:280:By2A3s3ps10%3D

    CAS  Google Scholar 

  • M.R. Urist (1965) ArticleTitleBone: formation by autoinduction Science 150 893–899 Occurrence Handle1:STN:280:CCmD3Mbjsl0%3D Occurrence Handle5319761

    CAS  PubMed  Google Scholar 

  • E.A. Wang V. Rosen P. Cordes R.M. Hewick M.J. Kriz D.P. Luxenberg B.S. Sibley J.M. Wozney (1988) ArticleTitlePurification and characterization of other distinct bone-inducing factors Proc. Natl. Acad. Sci. USA 85 9484–9488 Occurrence Handle3200834 Occurrence Handle1:CAS:528:DyaL1MXmvF2qsQ%3D%3D

    PubMed  CAS  Google Scholar 

  • K.E. Yates S. Mizuno J. Glowacki (2001) ArticleTitleEarly shifts in gene expression during chondroinduction of human dermal fibroblasts Exp. Cell Res. 265 203–211 Occurrence Handle10.1006/excr.2001.5192 Occurrence Handle11302685 Occurrence Handle1:CAS:528:DC%2BD3MXis1ertLk%3D

    Article  PubMed  CAS  Google Scholar 

  • S. Zhou K. Eid J. Glowacki (2004a) ArticleTitleCooperation between TGF-β and Wnt pathways during chondrocyte and adipocyte differentiation of human marrow stromal cells J. Bone Miner. Res. 19 463–470 Occurrence Handle1:CAS:528:DC%2BD2cXisFCrt74%3D

    CAS  Google Scholar 

  • S. Zhou J. Glowacki K.E. Yates (2004b) ArticleTitleComparison of TGF-β/BMP pathways signaled by demineralized bone powder and BMP-2 in human dermal fibroblasts J. Bone Miner. Res. 19 1732–1741 Occurrence Handle1:CAS:528:DC%2BD2cXovFWmtL8%3D

    CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Orthopedic Surgery, Brigham and Women’s Hospital, Harvard Medical School, 02115, Boston, MA, USA

    Shuanhu Zhou, Karen E. Yates, Karim Eid & Julie Glowacki

Authors
  1. Shuanhu Zhou
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  2. Karen E. Yates
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  3. Karim Eid
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  4. Julie Glowacki
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Correspondence to Julie Glowacki.

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Zhou, S., Yates, K.E., Eid, K. et al. Demineralized bone promotes chondrocyte or osteoblast differentiation of human marrow stromal cells cultured in collagen sponges. Cell Tissue Banking 6, 33–44 (2005). https://doi.org/10.1007/s10561-005-4253-y

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  • DOI: https://doi.org/10.1007/s10561-005-4253-y

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