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首页> 外文期刊>Dermatologic surgery >Time- and dose-dependent effects of chronic wound fluid on human adult dermal fibroblasts.
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Time- and dose-dependent effects of chronic wound fluid on human adult dermal fibroblasts.

机译:慢性伤口液对人成年真皮成纤维细胞的时间和剂量依赖性。

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摘要

BACKGROUND: Wound healing is a biologic process that is altered in patients affected by chronic venous ulcers. The wound microenvironment is reflected in the chronic wound fluid (CWF), an exudate containing serum components and tissue-derived proteins. OBJECTIVES: We investigated the effects of increasing doses of CWF collected from patients suffering from chronic venous ulcers on human adult dermal fibroblasts cultured in vitro and the relationship among CWF effects and treatment length. METHODS: Fibroblasts were treated with 60, 240, and 720 microg/mL CWF for 3 and 7 days. We evaluated cell proliferation and viability by MTT and Trypan blue assay, cell morphology by light microscopy, F-actin microfilaments organization by tetramethylrhodamine B isothiocyanate-conjugated phalloidin, alpha-smooth muscle actin expression by immunofluorescence, and senescence-associated beta-galactosidase activity. RESULTS: CWF induced an increase in cell proliferation in the first 3 days of treatment. In contrast, at 7 days, a strong decrease in cell viability was observed. These changes were related to a cytoskeletal F-actin reorganization and not to fibroblast-myofibroblast differentiation nor to changes in cellular senescence. CONCLUSIONS: This study shows a dose-dependent and biphasic effect of CWF on dermal fibroblasts, suggesting that a continuous exposure to chronic wounds microenvironment may induce late cellular dysfunctions possibly involved in the delayed wound healing.
机译:背景:伤口愈合是受慢性静脉溃疡影响的患者的生物学过程。伤口微环境反映在慢性伤口液(CWF)中,这是一种含有血清成分和组织衍生蛋白的渗出液。目的:我们研究了从慢性静脉溃疡患者中收集的增加剂量的CWF对体外培养的成人成年真皮成纤维细胞的影响,以及CWF效应与治疗时间之间的关系。方法:成纤维细胞分别用60、240和720 microg / mL CWF处理3天和7天。我们通过MTT和台盼蓝试验评估细胞增殖和活力,通过光学显微镜评估细胞形态,通过四甲基若丹明B异硫氰酸酯结合的鬼笔环肽F-肌动蛋白微丝组织,通过免疫荧光法检测α-平滑肌肌动蛋白表达,以及与衰老相关的β-半乳糖苷酶活性。结果:CWF在治疗的前三天诱导细胞增殖增加。相反,在第7天,观察到细胞活力大大降低。这些变化与细胞骨架F-肌动蛋白的重组有关,与成纤维细胞-成肌纤维细胞的分化无关,也与细胞衰老的变化无关。结论:这项研究表明CWF对皮肤成纤维细胞具有剂量依赖性和两相作用,这表明持续接触慢性伤口微环境可能诱发晚期细胞功能障碍,可能与伤口愈合延迟有关。

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