Germain L, Jean A, Auger F A, Garrel D R
Laboratoires des grands brûles/LOEX, Hôpital du Saint-Sacrement, Sainte-Foy, Québec, Canada.
J Surg Res. 1994 Aug;57(2):268-73. doi: 10.1006/jsre.1994.1143.
Contractile and phenotypic properties of human fibroblasts from healing wounds were compared to those of dermal fibroblasts using in vitro models. Wound fibroblasts were recovered from implants, made of a polyvinyl alcohol sponge threaded into a perforated silicone tube, 12 days after their subcutaneous implantation in human volunteers. Dermal fibroblasts were isolated from the skin of healthy subjects. Two morphologically different fibroblast populations were observed in cells cultured from implants. In order to characterize these fibroblast populations, intracellular alpha-actin expression was studied by immunofluorescence labeling of cells cultured in monolayer. This protein was detected in less than 1% of the dermal fibroblasts. By contrast, 30 to 40% of wound fibroblasts were labeled and contained fiber networks of alpha-actin. These results confirm the presence of myofibroblasts in human wound healing tissues. The contractile property of fibroblasts and myofibroblasts was evaluated using a three-dimensional cell culture model (fibroblast populated collagen gels). Cells were incorporated in a collagen matrix and cultured for 14 days. The surface area of collagen gels was measured every day. Our results show that wound fibroblasts strongly contract collagen gels during the first 24 hr (surface area at 24 hr = 20-55% of initial surface area) in comparison to dermal fibroblasts (surface area at 24 hr = 70-75% of initial surface area). This superior level of contraction was observed until the fifth day of culture.(ABSTRACT TRUNCATED AT 250 WORDS)
利用体外模型,将愈合伤口处人类成纤维细胞的收缩特性和表型特性与真皮成纤维细胞进行了比较。伤口成纤维细胞是在聚乙烯醇海绵穿入多孔硅胶管制成的植入物皮下植入人类志愿者12天后,从植入物中获取的。真皮成纤维细胞则从健康受试者的皮肤中分离得到。在从植入物培养的细胞中观察到两种形态不同的成纤维细胞群体。为了表征这些成纤维细胞群体,通过对单层培养细胞进行免疫荧光标记来研究细胞内α - 肌动蛋白的表达。在不到1%的真皮成纤维细胞中检测到了这种蛋白。相比之下,30%至40%的伤口成纤维细胞被标记,并含有α - 肌动蛋白纤维网络。这些结果证实了人类伤口愈合组织中存在肌成纤维细胞。使用三维细胞培养模型(成纤维细胞填充胶原凝胶)评估成纤维细胞和肌成纤维细胞的收缩特性。将细胞掺入胶原基质中并培养14天。每天测量胶原凝胶的表面积。我们的结果表明,与真皮成纤维细胞(24小时时的表面积 = 初始表面积的70 - 75%)相比,伤口成纤维细胞在最初24小时内强烈收缩胶原凝胶(24小时时的表面积 = 初始表面积的20 - 55%)。这种较高水平的收缩在培养的第五天之前都能观察到。(摘要截断于250字)