TGFβ affects collagen cross-linking independent of chondrocyte phenotype but strongly depending on physical environment
TGFβ affects collagen cross-linking independent of chondrocyte phenotype but strongly depending on physical environment
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DOI：
10.1089/tea.2007.0345
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发表时间：
2008-06-01
影响因子：
4.1
通讯作者：
Van Osch, Gerjo J. V. M.
中科院分区：
文献类型：
--
作者：
Bastiaansen-Jenniskens, Yvonne Maria;Koevoet, Wendy;Van Osch, Gerjo J. V. M.
转化生长因子β（TGFβ）通常用于软骨组织工程，以增加具有各种表型的细胞的基质形成。然而，TGFβ 的副作用，例如培养的成纤维细胞中的广泛交联，也有报道。我们的目标是研究 TGF beta 对胶原交联的影响并评估细胞表型和物理环境的作用。因此，我们在两种截然不同的物理环境中使用了四种不同的细胞群：包埋在藻酸盐凝胶中并附着在组织培养塑料上的原代和扩增软骨细胞和成纤维细胞。在使用或不使用 2.5 ng/mL TGF beta 2 的情况下，在 4 周内分析基质生成、胶原交联和 α-平滑肌肌动蛋白 (α SMA)。TGF beta 2 不影响原代细胞的胶原沉积。在扩增的细胞中，TGF beta 2 增加了胶原沉积。单层中的软骨细胞和成纤维细胞产生更多与 TGF β 2 的胶原交联。在藻酸盐中，原代细胞和扩增细胞显示出与 TGF β 2 的胶原交联意外减少。藻酸盐培养物中不存在 aSMA，并且 TGF β 2 上调裸层。有组织的 α SMA 纤维存在于所有单层培养物中，并且在 TGF β 2 中变得更加明显。这项研究表明，由所用基质决定的物理环境共同决定了细胞对 TGF beta 的反应。通过 α SMA 染色确定的机械应力的存在可能是添加 TGF β 后胶原交联增加的原因。
Transforming growth factor beta (TGF beta) is often used in cartilage tissue engineering to increase matrix formation by cells with various phenotypes. However, adverse effects of TGF beta, such as extensive cross-linking in cultured fibroblasts, have also been reported. Our goal was to study effects of TGF beta on collagen cross-linking and evaluating the role of cellular phenotype and physical environment. We therefore used four different cell populations in two very different physical environments: primary and expanded chondrocytes and fibroblasts embedded in alginate gel and attached to tissue culture plastic. Matrix production, collagen cross-linking, and alpha-smooth muscle actin (alpha SMA) were analyzed during 4 weeks with or without 2.5 ng/mL TGF beta 2. TGF beta 2 did not affect collagen deposition by primary cells. In expanded cells, TGF beta 2 increased collagen deposition. Chondrocytes and fibroblasts in monolayer produced more collagen cross-links with TGF beta 2. In alginate, primary and expanded cells displayed an unexpected decrease in collagen cross-linking with TGF beta 2. aSMA was not present in alginate cultures and barelay upregulated by TGF beta 2. Organized alpha SMA fibers were present in all monolayer cultures and became more pronounced with TGF beta 2. This study demonstrates that the physical environment determined by the substrate used co-determines the response of cells to TGF beta. The presence of mechanical stress, determined with alpha SMA-staining, is probably responsible for the increase in collagen cross-linking upon addition of TGF beta.