The effect of interferon alpha 2b on the expression of cytoskeletal proteins in an in vitro model of wound contraction.

B Nedelec, Y J Shen, A Ghahary, P G Scott, E E Tredget
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Abstract

Wound contraction is an essential component of wound healing. However, the development of scar contractures in tissues and organs disrupts normal organ integrity and produces functional deformities. Although interferons alpha and gamma inhibit extracellular matrix protein production by fibroblasts, their effects on cytoskeletal protein mediated-wound contraction are as yet unclear. The fibroblast-populated collagen lattice is an in vitro assay that simulates wound contraction. When matched pairs of human hypertrophic scar and normal dermal fibroblast cultures established from patients recovering from a thermal injury were used, interferon-alpha 2b exposure before lattice formation was found to significantly inhibit contraction in a treatment time-dependent manner (p < 0.05). Fibroblasts generated contractile forces that were triphasic and serum sensitive (p < 0.01). Comparison of hypertrophic scar and normal dermal fibroblasts revealed no significant differences in ability to induce lattice contraction. Northern blot analysis of mRNAs for the intracellular contractile proteins revealed that interferon-alpha 2b significantly down-regulated mRNA levels of the actin isoforms beta and gamma (50% to 60%) but had no significant effect on alpha-tubulin, vimentin, and alpha-actinin. Fibroblast-populated collagen lattices were stained with rhodamine-labeled phalloidin to reveal filamentous actin proteins. Marked morphologic alterations of the stress fibers were associated with reductions in lattice contraction after interferon-alpha 2b treatment. Thus interferon-alpha 2b's inhibition of wound contraction in vitro is associated with reductions in mRNA for beta and gamma actin and distinct morphologic alterations in fibroblast stress fiber morphology.

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干扰素α 2b对体外创面收缩模型细胞骨架蛋白表达的影响。
伤口收缩是伤口愈合的重要组成部分。然而,组织和器官中瘢痕挛缩的发展破坏了正常器官的完整性并产生功能畸形。尽管干扰素α和γ抑制成纤维细胞产生细胞外基质蛋白,但它们对细胞骨架蛋白介导的伤口收缩的影响尚不清楚。成纤维细胞填充的胶原晶格是一种模拟伤口收缩的体外实验。当使用从热损伤恢复的患者中建立的人增生性瘢痕和正常真皮成纤维细胞培养物配对时,发现在晶格形成之前暴露于干扰素- α 2b以治疗时间依赖的方式显著抑制收缩(p < 0.05)。成纤维细胞产生的收缩力具有三期性和血清敏感性(p < 0.01)。增生性瘢痕与正常真皮成纤维细胞的比较显示其诱导晶格收缩的能力无显著差异。细胞内收缩蛋白mRNA的Northern blot分析显示,干扰素- α 2b显著下调了肌动蛋白异构体β和γ的mRNA水平(50%至60%),但对α -微管蛋白、波形蛋白和α -肌动蛋白没有显著影响。用罗丹明标记的phalloidin染色成纤维细胞填充的胶原晶格,显示丝状肌动蛋白。在干扰素2b处理后,应力纤维的显著形态改变与晶格收缩的减少有关。因此,干扰素- α 2b对体外伤口收缩的抑制与β和γ肌动蛋白mRNA的减少以及成纤维细胞应激纤维形态的明显形态学改变有关。
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