Differential Effect of ECM Molecules on Re-Expression of Cartilaginous Markers in Near Quiescent Human Chondrocytes
Differential Effect of ECM Molecules on Re-Expression of Cartilaginous Markers in Near Quiescent Human Chondrocytes
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DOI：
10.1002/jcp.22530
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发表时间：
2011-08-01
影响因子：
5.6
通讯作者：
Tsai, Yu-Hui
中科院分区：
文献类型：
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作者：
Chiu, Li-Hsuan;Chen, Shih-Ching;Tsai, Yu-Hui
健康的原代软骨细胞来源有限，限制了组织工程软骨修复的临床应用。因此，在体外扩增过程中维持或恢复软骨细胞表型的方法是必不可少的。本研究的目的是确定ECM分子对恢复广泛单层扩增后原代人软骨细胞中软骨标记物的再表达的有益作用。在软骨细胞连续扩增的过程中，连续传代的细胞中COL 2A 1、SOX 9和AGN mRNA表达水平以及GAG积累水平显著降低。外源性II型胶原剂量依赖性地升高GAG水平，并诱导P7软骨细胞中软骨标记物mRNA的重新表达。硫酸软骨素对P7软骨细胞无明显影响，而透明质酸抑制了SOX 9和AGN mRNA的表达。在与II型胶原蛋白，FAK，ERK 1/2，和JNK的治疗后，激活通过磷酸化在P7软骨细胞在15分钟内。此外，GFOGER整合素阻断肽，MEK抑制剂和JNK抑制剂，而不是p38抑制剂，显着降低II型胶原蛋白诱导的GAG沉积水平。最后，在TGF-β 1和IGF-I的存在下，在3D II型胶原基质中培养的P7软骨细胞表现出比在I型胶原基质中培养的那些细胞更好的软骨特征。总之，单独的II型胶原蛋白可以有效地恢复扩张的P7人软骨细胞的软骨特征。可能通过激活FAK-ERK 1/2和FAK-JNK信号通路介导。II型胶原蛋白的潜在应用在扩大稀缺的健康软骨细胞在体外进一步组织工程的牵连。J.细胞。226：1981-1988，2011。(C)2010 Wiley-Liss，Inc.
The limited source of healthy primary chondrocytes restricts the clinical application of tissue engineering for cartilage repair. Therefore, method to maintain or restore the chondrocyte phenotype during in vitro expansion is essential. The objective of this study is to establish the beneficial effect of ECM molecules on restoring the re-expression of cartilaginous markers in primary human chondrocytes after extensive monolayer expansion. During the course of chondrocyte serial expansion, COL2A1, SOX9, and AGN mRNA expression levels, and GAG accumulation level were reduced significantly in serially passaged cells. Exogenous type II collagen dose-dependently elevated GAG level and induced the re-expression of cartilaginous marker mRNAs in P7 chondrocytes. Chondroitin sulfate did not show significant effect on P7 chondrocytes, while hyaluronic acid inhibited the expression of SOX9 and AGN mRNAs. Upon treatment with type II collagen, FAK, ERK1/2, and JNK were activated via phosphorylation in P7 chondrocytes within 15 min. Furthermore, GFOGER integrin blocking peptide, MEK inhibitor and JNK inhibitor, not p38 inhibitor, significantly reduced the type II collagen-induced GAG deposition level. Finally, in the presence of TGF-beta 1 and IGF-I, P7 chondrocytes cultured in 3D type II collagen matrix exhibited better cartilaginous features than those cells cultured in the type I collagen matrix. In conclusion, type II collagen alone can effectively restore cartilaginous features of expanded P7 human chondrocytes. It is probably mediated via the activation of FAK-ERK1/2 and FAK-JNK signaling pathways. The potential application of type II collagen in expanding a scarcity of healthy chondrocytes in vitro for further tissue engineering is implicated. J. Cell. Physiol. 226: 1981-1988, 2011. (C) 2010 Wiley-Liss, Inc.