Kisiday John D, Kopesky Paul W, Evans Christopher H, Grodzinsky Alan J, McIlwraith C Wayne, Frisbie David D
Orthopaedic Research Center, Department of Clinical Science, Colorado State University, 300 W. Drake Road, Fort Collins, Colorado 80523, USA.
J Orthop Res. 2008 Mar;26(3):322-31. doi: 10.1002/jor.20508.
Bone marrow mesenchymal stem cells (BM-MSCs) and adipose-derived progenitor cells (ADPCs) are potential alternatives to autologous chondrocytes for cartilage resurfacing strategies. In this study, the chondrogenic potentials of these cell types were compared by quantifying neo-tissue synthesis and assaying gene expression and accumulation of extracellular matrix (ECM) components of cartilage. Adult equine progenitor cells encapsulated in agarose or self-assembling peptide hydrogels were cultured in the presence or absence of TGFbeta1 for 3 weeks. In BM-MSCs-seeded hydrogels, TGFbeta1 stimulated ECM synthesis and accumulation 3-41-fold relative to TGFbeta1-free culture. In ADPC cultures, TGFbeta1 stimulated a significant increase in ECM synthesis and accumulation in peptide (18-29-fold) but not agarose hydrogels. Chromatographic analysis of BM-MSC-seeded agarose and peptide hydrogels cultured in TGFbeta1 medium showed extensive synthesis of aggrecan-like proteoglycan monomers. ADPCs seeded in peptide hydrogel also synthesized aggrecan-like proteoglycans, although to a lesser extent than seen in BM-MSC hydrogels, whereas aggrecan-like proteoglycan synthesis in ADPC-seeded agarose was minimal. RT-PCR analysis of TGFbeta1 cultures showed detectable levels of type II collagen gene expression in BM-MSC but not ADPC cultures. Histological analysis of TGFbeta1-cultured peptide hydrogels showed the deposition of a continuous proteoglycan- and type II collagen rich ECM for BM-MSCs but not ADPCs. Therefore, this study showed both protein and gene expression evidence of superior chondrogenesis of BM-MSCs relative to ADPCs.
骨髓间充质干细胞(BM-MSCs)和脂肪来源的祖细胞(ADPCs)是用于软骨表面修复策略的自体软骨细胞的潜在替代物。在本研究中,通过量化新组织合成以及检测软骨细胞外基质(ECM)成分的基因表达和积累,比较了这些细胞类型的软骨形成潜力。将成年马祖细胞封装在琼脂糖或自组装肽水凝胶中,在有或没有转化生长因子β1(TGFbeta1)的情况下培养3周。在接种BM-MSCs的水凝胶中,相对于无TGFbeta1的培养,TGFbeta1刺激ECM合成和积累增加3至41倍。在ADPC培养物中,TGFbeta1刺激肽水凝胶中ECM合成和积累显著增加(18至29倍),但在琼脂糖水凝胶中未出现这种情况。对在TGFbeta1培养基中培养的接种BM-MSCs的琼脂糖和肽水凝胶进行色谱分析,结果显示有大量类聚集蛋白聚糖蛋白多糖单体的合成。接种在肽水凝胶中的ADPCs也合成类聚集蛋白聚糖蛋白多糖,尽管程度低于在BM-MSCs水凝胶中的情况,而接种在ADPCs的琼脂糖中类聚集蛋白聚糖蛋白多糖的合成极少。对TGFbeta1培养物进行逆转录聚合酶链反应(RT-PCR)分析显示,在BM-MSC培养物中可检测到II型胶原基因表达水平,但在ADPC培养物中未检测到。对TGFbeta1培养的肽水凝胶进行组织学分析显示,BM-MSCs有富含蛋白聚糖和II型胶原的连续ECM沉积,而ADPCs则没有。因此,本研究显示了蛋白质和基因表达方面的证据,表明BM-MSCs相对于ADPCs具有更强的软骨形成能力。