Gene expression by fibroblasts seeded on small intestinal submucosa and subjected to cyclic stretching
Gene expression by fibroblasts seeded on small intestinal submucosa and subjected to cyclic stretching
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DOI：
10.1089/ten.2006.0318
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发表时间：
2007-06-01
影响因子：
--
通讯作者：
Woo, Savio L-Y.
中科院分区：
文献类型：
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作者：
Gilbert, Thomas W.;Stewart-Akers, Ann M.;Woo, Savio L-Y.
来源于猪小肠粘膜下层的细胞外基质支架（SIS-ECM）在许多临床前动物研究中已被证明可促进部位特异性组织的形成。然而，这种建设性的重塑过程需要支架经受特定场地的机械环境。机械负荷对SIS-ECM上成纤维细胞基因表达模式的具体定量影响尚不清楚，但在组织重塑过程中非常重要。本研究的目的是评估I型胶原（Col I）、III型胶原（Col III）、平滑肌肌动蛋白（SMA）、腱素- c （TN-C）、基质金属蛋白酶-2 （MMP-2）、基质金属蛋白酶-9 （MMP-9）、转化生长因子- β 1 （tgf - β 1）和转化生长因子- β 3 （tgf - β 3）在不同强度（0%、5%、10%和15%）和频率（0.1 Hz、0.3 Hz和0.5 Hz）拉伸下的表达情况。建立了一种新的循环拉伸组织培养（CSTC）体系。该系统由8个独立控制的培养室组成，可以在无菌培养箱中操作。每个腔室包括一个测压元件，以便监测每个支架中的载荷。研究发现，不同的拉伸方案导致成纤维细胞在SIS-ECM上复杂而独特的基因表达模式。一般来说，随着拉伸频率的增加，成纤维细胞Col I的表达增加了5倍，Col III的表达减少了5倍。此外，成纤维细胞表现出收缩表型，SMA、TN-C和tgf - β 1的表达增加。这些发现支持了一个概念，即重塑ECM支架的机械环境可能对支架内细胞的行为有实质性影响，并有助于在体内研究中观察到的部位特异性组织重塑。
Extracellular matrix scaffolds derived from porcine small intestinal submucosa (SIS-ECM) have been shown to promote the formation of site-specific tissue in a number of preclinical animal studies. However, this constructive remodeling process requires that the scaffold be subjected to a site-specific mechanical environment. The specific quantitative effects of mechanical loading on the gene expression patterns of fibroblasts seeded on SIS-ECM are unknown and yet very important in the tissue remodeling process. The objective of the present study was to evaluate the expression of collagen type I (Col I), collagen type III (Col III), smooth muscle actin (SMA), tenascin-C (TN-C), matrix metalloprotease-2 (MMP-2), matrix metalloprotease-9 (MMP-9), transforming growth factor-beta 1 (TGF-beta 1), and transforming growth factor-beta 3 (TGF-beta 3) by fibroblasts subjected to various magnitudes (0%, 5%, 10%, and 15%) and frequencies (0.1 Hz, 0.3 Hz, and 0.5 Hz) of stretch. A new cyclic-stretching tissue culture (CSTC) system was developed. This system consists of eight independently controlled culture chambers that can be operated in a sterile incubator. Each chamber includes a load cell so that the load in each scaffold can be monitored. It was found that different stretching regimens led to complex and distinctive patterns of gene expression by fibroblasts seeded onto SIS-ECM. In general, the fibroblasts increased expression of Col I up to 5-fold and decreased that of Col III with increased frequency of stretch. In addition, the fibroblasts exhibited a contractile phenotype with increased expression of SMA, TN-C, and TGF-beta 1. These findings support the concept that the mechanical environment of a remodeling ECM scaffold may have substantial effects on the behavior of cells within the scaffold and contribute to the site-specific tissue remodeling that has been observed in in vivo studies.