Induction of predominant tenogenic phenotype in human dermal fibroblasts via synergistic effect of TGF-β and elongated cell shape
Induction of predominant tenogenic phenotype in human dermal fibroblasts via synergistic effect of TGF-β and elongated cell shape
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通过 TGF-β 和细长细胞形状的协同作用诱导人真皮成纤维细胞的主要肌腱表型。
DOI：
10.1152/ajpcell.00300.2015
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发表时间：
2016-03-01
影响因子：
5.5
通讯作者：
Liu, Wei
中科院分区：
文献类型：
--
作者：
Wang, Wenbo;Li, Jie;Liu, Wei
微图形拓扑学因其在调节干细胞分化中的作用而被广泛研究，但在成熟细胞类型之间的表型转换方面仍未被探索。本研究探讨了人工延长培养细胞在人真皮成纤维细胞(HDFS)中诱导伸展表型的可能性。我们的结果表明，平行的微槽地形可以将铺展的HDF转化为细长的形状，并诱导出优势的伸展表型，因为生物标志物如硬化轴、腱调节蛋白、I、III、VI和核心蛋白的表达显著增强。可促进转化生长因子-β-1的表达，但不能促进α-平滑肌肌动蛋白的表达。延长的HDFS不能诱导其他表型，如成骨、成软骨、成神经性和肌成性谱系。相反，在细长的人软骨细胞中不能诱导出伸展表型，尽管软骨形成表型被抑制。外源性转化生长因子-β-1在低剂量(2 ng/ml)时可促进延长的hDFS的伸展表型，但在高剂量(10 ng/ml)时可促进hDFS的肌成纤维细胞转分化，而与细胞形态无关。拉长的形状还导致RhoA活性降低，Rho相关蛋白激酶(ROCK)活性增加。用Y27632拮抗转化生长因子-β或抑制ROCK活性，或用细胞松弛素D解聚肌动蛋白都能显著抑制张力表型的诱导，尤其是在加长的hdf中。总之，培养的真皮成纤维细胞的延长可能通过转化生长因子-β和细胞骨架信号的协同作用而诱导出主要的张力表型。
Micropattern topography is widely investigated for its role in mediating stem cell differentiation, but remains unexplored for phenotype switch between mature cell types. This study investigated the potential of inducing tenogenic phenotype in human dermal fibroblasts (hDFs) by artificial elongation of cultured cells. Our results showed that a parallel microgrooved topography could convert spread hDFs into an elongated shape and induce a predominant tenogenic phenotype as the expression of biomarkers was significantly enhanced, such as scleraxis, tenomodulin, collagens I, III, VI, and decorin. It also enhanced the expression of transforming growth factor (TGF)-β1, but not α-smooth muscle actin. Elongated hDFs failed to induce other phenotypes, such as adiopogenic, chondrogenic, neurogenic, and myogenic lineages. By contrast, no tenogenic phenotype could be induced in elongated human chondrocytes, although chondrogenic phenotype was inhibited. Exogenous TGF-β1 could enhance the tenogenic phenotype in elongated hDFs at low dose (2 ng/ml), but promoted myofibroblast transdifferentiation of hDFs at high dose (10 ng/ml), regardless of cell shape. Elongated shape also resulted in decreased RhoA activity and increased Rho-associated protein kinase (ROCK) activity. Antagonizing TGF-β or inhibiting ROCK activity with Y27632 or depolymerizing actin with cytochalasin D could all significantly inhibit tenogenic phenotype induction, particularly in elongated hDFs. In conclusion, elongation of cultured dermal fibroblasts can induce a predominant tenogenic phenotype likely via synergistic effect of TGF-β and cytoskeletal signaling.