title: Demineralized bone promotes chondrocyte or osteoblast differentiation of human marrow stromal cells cultured in collagen sponges.
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-beta, 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-beta/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.
authors.affiliation: Department of Orthopedic Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Shuanhu
authors.initials: S
authors.lastname: Zhou
authors.forename: Karen E
authors.initials: KE
authors.lastname: Yates
authors.forename: Karim
authors.initials: K
authors.lastname: Eid
authors.forename: Julie
authors.initials: J
authors.lastname: Glowacki
mesh_terms: D053674:Aggrecans; D018775:Bone Demineralization Technique; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D020219:Chondrogenesis / Q000187:drug effects; D003094:Collagen / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D037181:Lectins, C-Type; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D011208:Powders / Q000494:pharmacology; D011509:Proteoglycans / Q000235:genetics; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053773:Transforming Growth Factor beta1
chemical_list: D053674:Aggrecans; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011208:Powders; D011509:Proteoglycans; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D003094:Collagen
references.citation: J Cell Biochem. 2001;82(4):583-90
references.pmid: 11500936
references.citation: Clin Orthop Relat Res. 1996 Mar;(324):47-54
references.pmid: 8595776
references.citation: Biomaterials. 2003 Jul;24(15):2593-603
references.pmid: 12726713
references.citation: J Bone Joint Surg Am. 2003;85-A Suppl 3:19-28
references.pmid: 12925605
references.citation: Connect Tissue Res. 2003;44 Suppl 1:305-11
references.pmid: 12952214
references.citation: J Orthop Sci. 2003;8(5):740-8
references.pmid: 14557946
references.citation: Clin Orthop Relat Res. 2003 Dec;(417):183-94
references.pmid: 14646716
references.citation: Cells Tissues Organs. 2004;176(1-3):109-19
references.pmid: 14745240
references.citation: J Bone Miner Res. 2004 Mar;19(3):463-70
references.pmid: 15040835
references.citation: Calcif Tissue Int. 2004 May;74(5):458-68
references.pmid: 14961210
references.citation: J Bone Miner Res. 2004 Oct;19(10):1732-41
references.pmid: 15355569
references.citation: Science. 1965 Nov 12;150(3698):893-9
references.pmid: 5319761
references.citation: Proc Natl Acad Sci U S A. 1972 Jun;69(6):1601-5
references.pmid: 4504376
references.citation: Lancet. 1981 May 2;1(8227):959-62
references.pmid: 6112384
references.citation: Clin Plast Surg. 1985 Apr;12(2):233-41
references.pmid: 3886260
references.citation: Clin Orthop Relat Res. 1985 Nov;(200):159-64
references.pmid: 3905107
references.citation: Endocr Rev. 1985 Fall;6(4):544-51
references.pmid: 3000756
references.citation: J Periodontol. 1985 Aug;56(8):470-9
references.pmid: 3915013
references.citation: Proc Natl Acad Sci U S A. 1988 Dec;85(24):9484-8
references.pmid: 3200834
references.citation: Calcif Tissue Int. 1988 Dec;43(6):346-51
references.pmid: 3146423
references.citation: Clin Orthop Relat Res. 1991 Feb;(263):30-48
references.pmid: 1993386
references.citation: J Hand Surg Am. 1992 Jul;17(4):704-13
references.pmid: 1629553
references.citation: Am J Orthop (Belle Mead NJ). 1995 Oct;24(10):752-61
references.pmid: 8593557
references.citation: Endocr Rev. 1994 Feb;15(1):27-39
references.pmid: 8156937
references.citation: Exp Cell Res. 1996 Aug 25;227(1):89-97
references.pmid: 8806455
references.citation: Biomaterials. 1996 Sep;17(18):1819-25
references.pmid: 8879522
references.citation: Int Orthop. 1998;22(5):335-42
references.pmid: 9914941
references.citation: Science. 1999 Apr 2;284(5411):143-7
references.pmid: 10102814
references.citation: Calcif Tissue Int. 1999 May;64(5):394-401
references.pmid: 10203416
references.citation: Clin Orthop Relat Res. 1999 Jul;(364):61-9
references.pmid: 10416393
references.citation: Bone. 2000 Aug;27(2):177-84
references.pmid: 10913909
references.citation: Exp Cell Res. 2001 May 1;265(2):203-11
references.pmid: 11302685
references.citation: Cytokine Growth Factor Rev. 2002 Jun;13(3):241-57
references.pmid: 12486877