Microdeformation of Three-Dimensional Cultured Fibroblasts Induces Gene Expression and Morphological Changes
Microdeformation of Three-Dimensional Cultured Fibroblasts Induces Gene Expression and Morphological Changes
复制标题
三维培养成纤维细胞的微变形诱导基因表达和形态变化
DOI：
10.1097/sap.0b013e3181ea1e9b
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发表时间：
2011-03-01
影响因子：
1.5
通讯作者：
Orgill, Dennis P.
中科院分区：
文献类型：
--
作者：
Lu, Feng;Ogawa, Rei;Orgill, Dennis P.
背景：真空辅助闭合诱导创面微变形，加速复杂创面愈合；然而，缺乏对细胞机械转导生物学的透彻理解。我们假设成纤维细胞的形状和功能可以在体外真空辅助闭合装置中改变。方法：将三维纤维蛋白基质与培养的小鼠成纤维细胞和中间的聚氨酯泡沫暴露于125 mm Hg的吸液中，并与不吸液的类似孔进行比较。我们使用荧光显微镜测量成纤维细胞的增殖和形态，并使用实时逆转录酶聚合酶链反应在24、48和72小时测量基因表达变化。结果：抽吸孔诱导成纤维细胞显著增殖，形态变化明显，表现为更大、更圆、明显的树突样分支和过程延伸。1型胶原α - 1 （COL1A1）、成纤维细胞生长因子2 （FGF2、bFGF）、转化生长因子β 1 （TGF&bgr;1）在抽吸48h后均表达上调。72小时后，平滑肌肌动蛋白α 2 （Acta2, &agr;-SMA）上调。结论：聚氨酯泡沫和吸力联合产生的微变形与成纤维细胞增殖增加和成纤维细胞基因表达上调有关。
Background:Vacuum-assisted closure induces microdeformations of the wound surface and accelerates healing of complex wounds; however, a thorough understanding of the biology of cellular mechanotransduction is lacking. We hypothesized that fibroblast shape and function can be altered in an in vitro vacuum-assisted closure device. Methods:A 3-dimensional fibrin matrix with cultured murine fibroblasts and an intervening polyurethane foam was exposed to 125 mm Hg suction and compared with similar wells without suction. We measured fibroblast proliferation and morphology using fluorescence microscopy and gene expression change using real-time reverse-transcriptase polymerase chain reaction at 24, 48, and 72 hours. Results:Wells exposed to suction induced significant proliferation of fibroblasts and morphologic changes visible by larger, rounder, and notable dendrite-like branching and process extensions. Type 1 collagen alpha 1 (COL1A1), fibroblast growth factor 2 (FGF2, bFGF), and transforming growth factor beta 1 (TGF&bgr;1) were all up-regulated after 48 hours of exposure to suction. Smooth muscle actin alpha 2 (Acta2, &agr;-SMA) was up-regulated after 72 hours. Conclusions:Microdeformations produced by the combination of polyurethane foam and suction are associated with increased fibroblast proliferation and up-regulation of gene expressions in fibroblasts.