alpha-smooth muscle actin and contractile behavior of bovine meniscus cells seeded in type I and type II collagen-GAG matrices.
alpha-smooth muscle actin and contractile behavior of bovine meniscus cells seeded in type I and type II collagen-GAG matrices.
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DOI：
10.1002/(sici)1097-4636(19990605)45:3
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发表时间：
1999-06
期刊：
影响因子：
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通讯作者：
Stefan M. Mueller;T. O. Schneider;Sonya Shortkroff;H. Breinan;Myron Spector;Myron Spector
中科院分区：
文献类型：
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作者：
Stefan M. Mueller;T. O. Schneider;Sonya Shortkroff;H. Breinan;Myron Spector;Myron Spector
膝关节半月板的许多类型的损伤导致不愈合的缺陷，导致疼痛和功能障碍。一些正在进行的研究正在开发多孔可吸收基质，可单独使用或与培养的细胞接种，以促进该组织的再生。本研究的目的是在体外评估种植在I型和II型胶原基质中的胰岛细胞的收缩行为。在许多结缔组织中，已经发现具有收缩表型的成纤维细胞（肌成纤维细胞）在愈合和病理状况中起重要作用。这种表型，如果表达的大肠癌细胞，可能会影响他们的行为，在细胞接种基质开发的组织工程。在这项研究中，存在的收缩肌动蛋白亚型，α-平滑肌（α-SM）肌动蛋白，通过免疫组织化学在正常小牛直肠组织和直肠细胞在2-和3-维培养进行了评估。将小牛半月板细胞接种在I型和II型胶原-糖胺聚糖（GAG）基质中。每2-3天测量基质的直径。在1、3和7天后对α-SM肌动蛋白的二维培养物进行免疫组织化学染色，并在1、7、14和21天对接种的基质进行染色。对选定的样品进行透射电子显微镜（TEM）。3周后，接种的I型基质显示出几乎50%的显著收缩，而II型基质和两种类型的未接种对照在同一时间段内几乎没有收缩。在10%的正常组织细胞中观察到α-SM肌动蛋白表型的阳性染色，但在单层接种的所有细胞和两种类型的基质中均存在。代表性细胞接种基质的TEM显示约7 nm厚的微丝，与肌成纤维细胞表型一致。这是第一次报告的α-SM肌动蛋白含有细胞在膝关节半月板。在某些条件下，牙周细胞可以表达肌成纤维细胞表型，这一发现保证了研究它们在牙周愈合中的作用和对植入物的组织反应，以促进组织再生。
Many types of injuries to the meniscus of the knee joint result in defects that do not heal, leading to pain and dysfunction. Several ongoing investigations are developing porous absorbable matrices to be used alone or seeded with cultured cells to facilitate regeneration of this tissue. The objective of this study was to evaluate in vitro the contractile behavior of meniscal cells seeded in type I and type II collagen matrices. In many connective tissues, fibroblasts that have assumed a contractile phenotype (myofibroblasts) have been found to play an important role in healing and in pathological conditions. This phenotype, if expressed by meniscal cells, could affect their behavior in cell-seeded matrices developed for tissue engineering. In this study, the presence of a contractile actin isoform, alpha-smooth muscle (alpha-SM) actin, was assessed by immunohistochemistry in normal calf meniscal tissue and in meniscal cells in 2- and 3-dimensional culture. Calf meniscus cells were seeded in type I and type II collagen-glycosaminoglycan (GAG) matrices. The diameter of the matrices was measured every 2-3 days. Immunohistochemical staining of the 2-dimensional cultures for alpha-SM actin was performed after 1, 3, and 7 days and the staining of the seeded matrices was at 1, 7, 14, and 21 days. Transmission electron microscopy (TEM) was performed on selected samples. After 3 weeks the seeded type I matrices displayed a significant shrinkage of almost 50% whereas the type II matrix and both types of unseeded controls showed almost no contraction over the same time period. Positive staining for the alpha-SM actin phenotype was seen in 10% of the cells of the normal tissue but was present in all cells seeded in monolayer and in both types of matrices. TEM of representative cell-seeded matrices showed microfilaments approximately 7 nm thick, consistent with the myofibroblast phenotype. This is the first report of alpha-SM actin containing cells in the knee meniscus. The finding that, under certain conditions, meniscal cells can express the myofibroblast phenotype warrants study of their role in meniscal healing and the tissue response to implants to facilitate tissue regeneration.