In vitro expansion affects the response of chondrocytes to mechanical stimulation
In vitro expansion affects the response of chondrocytes to mechanical stimulation
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DOI：
10.1016/j.joca.2007.07.014
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发表时间：
2008-03-01
影响因子：
7
通讯作者：
Weinans, H.
中科院分区：
文献类型：
--
作者：
Das, R. H. J.;Jahr, H.;Weinans, H.
目的：自体软骨细胞扩增是软骨缺损修复的常用步骤。随后的去分化可以改变细胞对机械负荷的反应，对细胞在再植后的体内行为具有重要影响。因此，我们研究的主要和扩大的人关节软骨细胞的机械loading.Method的反应：主要和扩大的软骨细胞被拉伸在0.5%或3.0%，0.5 Hz，每天2小时，为3天。将基质组分（聚集蛋白聚糖（AGC1）、润滑素（PRG4）、I型胶原（COL1）、II型胶原（COL2）和X型胶原（COL10））以及基质酶（基质金属蛋白酶1（MMP 1）、MMP 3、MMP 13）和SOX 9的基因表达水平与未拉伸对照进行比较。为了评估软骨形成环境对细胞响应拉伸的影响，再分化培养基中使用的扩展cells.Results：在原代软骨细胞，拉伸导致AGC1，COLI和COL10基因表达（最大的3.8倍）和PRG4（2.0倍）的上调轻微下降。在扩增的软骨细胞中，AGC1（高达21倍）、PRG4（高达5.0倍）、COO（10倍）和COL2（2.9倍）的表达下调。此外，MMP 1（20倍）和MMP 3（高达4倍）的表达上调，而MMP 13下调（2.8倍）。一个软骨形成的环境出现回火的影响stretch.Discussion：我们的研究结果表明，扩张改变了人类软骨细胞的反应，拉伸。扩张的软骨细胞大大降低了基质成分的基因表达，并增加了MMPs的表达，而原代软骨细胞几乎没有反应。本研究结果可作为软骨细胞再植入的细胞来源优化及扩增方案的参考。(C)2007年国际骨关节炎研究学会。由爱思唯尔有限公司出版。保留所有权利。
Objective: Expansion of autologous chondrocytes is a common step in procedures for cartilage defect repair. Subsequent dedifferentiation can alter cellular response to mechanical loading, having major consequences for the cell's behavior in vivo after reimplantation. Therefore, we examined the response of primary and expanded human articular chondrocytes to mechanical loading.Method: Primary and expanded chondrocytes were stretched at either 0.5% or 3.0% at 0.5 Hz, 2 h per day, for 3 days. Gene expression levels of matrix components (aggrecan (AGC1), lubricin (PRG4), collagen type I (COL1), type II (COL2) and type X (COL10)) as well as matrix enzymes (matrix metalloproteinase 1 (MMP1), MMP3, MMP13) and SOX9 were compared to unstretched controls. To evaluate the effect of a chondrogenic environment on cellular response to stretch, redifferentiation medium was used on expanded cells.Results: In primary chondrocytes, stretch led to mild decreases in AGC1, COLI and COL10 gene expression (maximum of 3.8-fold) and an up-regulation of PRG4 (2.0-fold). In expanded chondrocytes, expression was down-regulated for AGC1 (up to 21-fold), PRG4 (up to 5.0-fold), COO (10-fold) and COL2 (2.9-fold). Also, expression was up-regulated for MMP1 (20-fold) and MMP3 (up to 4-fold), while MMP13 was down-regulated (2.8-fold). A chondrogenic environment appeared to temper effects of stretch.Discussion: Our results show that expansion alters the response of human chondrocytes to stretch. Expanded chondrocytes greatly decrease gene expression of matrix constituents and increase expression of MMPs, whereas primary chondrocytes hardly respond. Our data could be a reference for optimization of cell sources or expansion protocols for reimplanted chondrocytes. (C) 2007 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.