Proliferation and collagen production of human patellar tendon fibroblasts in response to cyclic uniaxial stretching in serum-free conditions
Proliferation and collagen production of human patellar tendon fibroblasts in response to cyclic uniaxial stretching in serum-free conditions
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DOI：
10.1016/j.jbiomech.2004.01.005
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发表时间：
2004-10-01
影响因子：
2.4
通讯作者：
Wang, JHC
中科院分区：
文献类型：
--
作者：
Yang, GG;Crawford, RC;Wang, JHC
我们研究了无血清条件下循环机械拉伸对人髌腱成纤维细胞增殖、胶原 mRNA 表达和蛋白质产生的影响。还研究了转化生长因子-β1 (TGF-β1) 在周期性拉伸的肌腱成纤维细胞产生胶原蛋白中的作用。肌腱成纤维细胞在微槽硅胶培养皿中生长，其中细胞高度拉长并与微槽对齐。对硅胶皿进行恒定频率和持续时间（0.5 Hz，4 小时）但不同拉伸幅度（无拉伸、4% 和 8%）的循环单轴拉伸。拉伸一段时间后，将细胞在拉伸条件培养基中静置20小时以允许细胞增殖。在单独的实验中，细胞被拉伸 4 小时，然后再休息 4 小时。培养基、总细胞 RNA 和蛋白质样品用于分析胶原蛋白和 TGF-β1 基因表达和产生。研究发现，与未拉伸的成纤维细胞相比，拉伸 4% 和 8% 时，成纤维细胞增殖略有增加，而拉伸 8% 时，成纤维细胞增殖显着。研究还发现，I 型胶原蛋白和 TGF-β1 的基因表达和蛋白质产量以拉伸幅度依赖性方式增加。而且，尽管蛋白质的基因表达水平略有增加，但 111 型胶原蛋白的水平没有改变。此外，外源添加抗TGF-β1抗体消除了循环单轴拉伸条件下I型胶原蛋白产生的增加。结果表明，机械拉伸可以在无血清的情况下调节人肌腱成纤维细胞的增殖，并增加 1 型胶原蛋白的细胞生成，这至少部分是由 TGF-β1 介导的。 (C) 2004 Elsevier Ltd. 保留所有权利。
We studied the effect of cyclic mechanical stretching on the proliferation and collagen mRNA expression and protein production of human patellar tendon fibroblasts under serum-free conditions. The role of transforming growth factor-beta1 (TGF-beta1) in collagen production by cyclically stretched tendon fibroblasts was also investigated. The tendon fibroblasts were grown in microgrooved silicone dishes, where the cells were highly elongated and aligned with the microgrooves. Cyclic uniaxial stretching with constant frequency and duration (0.5 Hz, 4 h) but varying magnitude of stretch (no stretch, 4%, and 8%) was applied to the silicone dishes. Following the period of stretching, the cells were rested for 20 h in stretching-conditioned medium to allow for cell proliferation. In separate experiments, the cells were stretched for 4 h and then rested for another 4 h. Samples of the medium, total cellular RNA and protein were used for analysis of collagen and TGF-beta1 gene expression and production. It was found that there was a slight increase in fibroblast proliferation at 4% and 8% stretch, compared to that of non-stretched fibroblasts, where at 8% stretch the increase was significant. It was also found that the gene expression and protein production of collagen type I and TGF-beta1 increased in a stretching-magnitude-dependent manner. And, levels of collagen type 111 were not changed, despite gene expression levels of the protein being slightly increased. Furthermore, the exogenous addition of anti-TGF-beta1 antibody eliminated the increase in collagen type I production under cyclic uniaxial stretching conditions. The results suggest that mechanical stretching can modulate proliferation of human tendon fibroblasts in the absence of serum and increase the cellular production of collagen type 1, which is at least in part mediated by TGF-beta1. (C) 2004 Elsevier Ltd. All rights reserved.