Agren M S, Steenfos H H, Dabelsteen S, Hansen J B, Dabelsteen E
Coloplast Research, Humlebaek, Department of Dermatology, Bispebjerg Hospital, Copenhagen, Denmark.
J Invest Dermatol. 1999 Apr;112(4):463-9. doi: 10.1046/j.1523-1747.1999.00549.x.
Several pathophysiologic mechanisms have been proposed to explain slow-healing leg ulcers, but little is known about the growth behavior of cells in these wounds. Platelet-derived growth factor-BB applied topically to chronic wounds has shown beneficial effects, although the effects have been less pronounced than would have been expected based on studies on acute wounds. The objective of this study was to compare fibroblasts in culture obtained from chronic wounds (non-healing chronic venous leg ulcers), acute wounds and normal dermis regarding growth, mitogenic response to platelet-derived growth factor-BB and levels ofplatelet-derived growth factor alpha-receptor and beta-receptor. Fibroblasts were obtained by an explant technique and expanded in vitro using fibroblast growth medium supplemented with 10% fetal bovine serum and used for the assays at their third passage. Growth of chronic wound fibroblasts (n = 8) was significantly (p < 0.05) decreased compared with those from acute wounds (n = 10) and normal dermis (n = 5). Fibroblasts from ulcers older than 3 y grew significantly (p < 0.01) slower than those from ulcers that had been present for less than 3 y. Morphology and size of fibroblasts from the oldest chronic wounds deviated substantially from those of acute wounds and normal dermis, and resembled in vitro aged or senescent fibroblasts. Mitogenic response of chronic wound fibroblasts to human recombinant platelet-derived growth factor-BB was also reduced with ulcer age. No significant differences were found in the amount of either platelet-derived growth factor alpha-receptor or beta-receptor among the three groups. The features decreased growth related to ulcer age, altered morphology, and reduced response to platelet-derived growth factor, indicating that fibroblasts in some chronic wounds have approached or even reached the end of their lifespan (phase III). This might provide one explanation for the non-healing state and therapy resistance to topical platelet-derived growth factor-BB of some venous leg ulcers.
已经提出了几种病理生理机制来解释腿部溃疡愈合缓慢的现象，但对于这些伤口中细胞的生长行为却知之甚少。局部应用于慢性伤口的血小板衍生生长因子 -BB 已显示出有益效果，尽管基于对急性伤口的研究，其效果不如预期明显。本研究的目的是比较从慢性伤口（不愈合的慢性静脉性腿部溃疡）、急性伤口和正常真皮中获取的培养成纤维细胞在生长、对血小板衍生生长因子 -BB 的促有丝分裂反应以及血小板衍生生长因子α受体和β受体水平方面的差异。通过外植技术获取成纤维细胞，并在补充有 10%胎牛血清的成纤维细胞生长培养基中进行体外扩增，并在第三代时用于检测。与急性伤口（n = 10）和正常真皮（n = 5）来源的成纤维细胞相比，慢性伤口成纤维细胞（n = 8）的生长显著降低（p < 0.05）。病程超过 3 年的溃疡来源的成纤维细胞生长速度明显（p < 0.01）慢于病程小于 3 年的溃疡来源的成纤维细胞。最陈旧慢性伤口来源的成纤维细胞的形态和大小与急性伤口和正常真皮来源的成纤维细胞有很大差异，类似于体外老化或衰老的成纤维细胞。随着溃疡年龄的增加，慢性伤口成纤维细胞对人重组血小板衍生生长因子 -BB 的促有丝分裂反应也降低。三组之间血小板衍生生长因子α受体或β受体的数量没有显著差异。与溃疡年龄相关的生长减少、形态改变以及对血小板衍生生长因子反应降低等特征表明，一些慢性伤口中的成纤维细胞已接近甚至达到其寿命末期（III 期）。这可能为一些静脉性腿部溃疡的不愈合状态和对局部血小板衍生生长因子 -BB 的治疗抵抗提供一种解释。
