Smooth muscle actin expression by human articular chondrocytes and their contraction of a collagen-glycosaminoglycan matrix in vitro
Smooth muscle actin expression by human articular chondrocytes and their contraction of a collagen-glycosaminoglycan matrix in vitro
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DOI：
10.1016/s0736-0266(00)00081-4
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发表时间：
2001-03-01
影响因子：
2.8
通讯作者：
Spector, M
中科院分区：
文献类型：
--
作者：
Kinner, B;Spector, M
最近的研究表明，人关节软骨细胞可以原位表达一种收缩肌肌动蛋白的基因，即ct-平滑肌肌动蛋白（SMA）。这项工作的目的之一是利用Western blot分析评估分离的人关节软骨细胞的SMA含量，并将细胞中SMA的含量与传代数和培养天数相关联。第二个目的是确定在体外表达SMA基因的关节软骨源性细胞是否也继续表达II型胶原。当前研究的最终目的是确定含有sma的软骨来源细胞是否能够在体外收缩细胞外基质的胶原-糖胺聚糖类似物。从13例全关节置换术患者中分离关节软骨细胞。细胞依次传代至第7代。分配样本进行SMA的Western blot分析。单层培养的细胞也进行免疫组织化学染色，检测SMA和II型胶原。将第3代和第7代的细胞接种到多孔的1型胶原-糖胺聚糖基质中，每隔一天测量支架直径，共21天。关节软骨样本的免疫组化显示原位关节软骨细胞中存在SMA，且软骨浅层部分（60 +/- 1.2%；平均+/- SEM）的细胞染色阳性比例高于基底部分（28 +/- 1.3%）。单层培养细胞中SMA含量随传代数的增加而增加，且SMA含量与培养天数呈显著相关（线性回归分析，R-2 = 0.72）。SMA和II型胶原双染色显示，单层表达II型胶原的细胞也能表达SMA。发现含有SMA的细胞收缩胶原-糖胺聚糖基质，含有更多SMA的细胞（第7代细胞）比含有较少SMA的细胞（第3代细胞）表现出非基质收缩。结果表明，当使用关节软骨细胞，在单层培养中扩增，单独植入或在细胞种子基质中进行软骨修复手术时，控制SMA的表达可能是重要的。(C) 2001骨科研究学会。Elsevier Science Ltd.出版。版权所有。
Recent studies have demonstrated that human articular chondrocytes can express the gene for a contractile muscle actin, ct-smooth muscle actin (SMA), in situ. One objective of this work was to evaluate the SMA-content of isolated human articular chondrocytes using Western blot analysis and to correlate the amount of SMA in the cells with passage number and the number of days in culture. A second objective was to determine if articular cartilage-derived cells expressing the gene for SMA in vitro also continue to express type II collagen. A final aim of the current study was to determine if SMA-containing cartilage-derived cells were capable of contracting a collagen-glycosaminoglycan analog of extracellular matrix in vitro. Articular chondrocytes were isolated from 13 patients undergoing total joint arthroplasty. Cells were serially passaged through passage 7. Samples were allocated for Western blot analysis of SMA. Cells in monolayer culture were also stained immunohistochemically for SMA and type II collagen. Cells from passage 3 and 7 were seeded into a porous type 1 collagen-glycosaminoglycan matrix and the diameter of the scaffolds measured every other day for 21 days. Immunohistochemistry of the articular cartilage samples revealed SMA in the articular chondrocytes in situ with a greater percentage of cells staining positive in the superficial half (60 +/- 1.2%; mean +/- SEM) of the cartilage than in the basal half (28 +/- 1.3%). There was an increasing amount of SMA in the cells in monolayer culture with passage number and a meaningful correlation of the SMA content with the days in culture (linear regression analysis; R-2 = 0.72). Double staining for SMA and type II collagen showed that type II collagen-expressing cells in monolayer could also express SMA. SMA-containing cells were found to contract the collagen-glycosaminoglycan matrix, with the cells containing more SMA (passage 7 cells) displaying mon matrix contraction than those with a lesser amount of SMA (passage 3 cells). The results indicate that control of the expression of SMA may be important when employing articular chondrocytes, expanded in monolayer culture, for implantation alone or in a cell-seeded matrix for cartilage repair procedures. (C) 2001 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.