Schäffer M R, Tantry U, Ahrendt G M, Wasserkrug H L, Barbul A
Department of Surgery, Eberhard-Karls-Universität, Tübingen, Germany.
Int J Biochem Cell Biol. 1997 Jan;29(1):231-9. doi: 10.1016/s1357-2725(96)00136-7.
Fibroblast proliferation and fibroblast-mediated matrix contraction are critical to wound healing. Different cytokines have been shown to modulate fibroblast functions but little is known about the physiological role of these soluble factors during wound repair. In these experiments we characterized a fibroblast stimulating factor in wound fluid. Wound fluid was obtained from subcutaneously implanted polyvinyl alcohol sponges harvested 10 days post-wounding (pool of 100 Lewis rats). Normal dermal fibroblasts were obtained from Lewis rats by an explant technique, while wound fibroblasts were isolated from sponges harvested 10 days post-wounding. Proliferation in response to 0.5% and 10% fetal bovine serum was assessed by [3H]-thymidine incorporation. A fibroblast-populated collagen lattice was used for assaying contractile properties. Wound fibroblasts demonstrated markedly diminished proliferative and enhanced contractile properties compared to normal dermal fibroblasts. 10% wound fluid (v/v) stimulated proliferation of normal dermal fibroblasts (119%, p < 0.001) and wound fibroblasts (103%, p < 0.001). Wound fluid also stimulated collagen gel contraction by normal dermal fibroblasts (24% at 24 hr and 16% at 72 hr, p < 0.01), but not by wound fibroblasts. Separation by Sephadex G-100 gel filtration identified the active factor in wound fluid to have a molecular weight of about 100 kDa. Characterization of the soluble factor showed it to be a protein (ammonium sulfate precipitation), sensitive to trypsin digestion, heat resistant (56 degrees C, 30 min), and neuraminidase resistant. The isoelectric point appeared to be 7.0, as determined by ion exchange chromatography. Mitogenic proliferation of thymic lymphocytes was not affected by the active factor, suggesting cell target specificity. These data demonstrate that the wound environment contains high molecular weight protein(s) that promote fibroblast functions, essential for the healing process.
成纤维细胞增殖和成纤维细胞介导的基质收缩对伤口愈合至关重要。不同的细胞因子已被证明可调节成纤维细胞功能，但对于这些可溶性因子在伤口修复过程中的生理作用知之甚少。在这些实验中，我们对伤口渗出液中的一种成纤维细胞刺激因子进行了特性分析。伤口渗出液取自受伤10天后皮下植入的聚乙烯醇海绵（100只Lewis大鼠的样本池）。正常真皮成纤维细胞通过外植技术从Lewis大鼠获取，而伤口成纤维细胞则从受伤10天后采集的海绵中分离得到。通过[3H] - 胸腺嘧啶核苷掺入法评估对0.5%和10%胎牛血清的增殖反应。使用成纤维细胞填充的胶原晶格来测定收缩特性。与正常真皮成纤维细胞相比，伤口成纤维细胞的增殖能力明显减弱，收缩特性增强。10%（体积/体积）的伤口渗出液刺激正常真皮成纤维细胞增殖（119%，p < 0.001）和伤口成纤维细胞增殖（103%，p < 0.001）。伤口渗出液还刺激正常真皮成纤维细胞的胶原凝胶收缩（24小时时为24%，72小时时为16%，p < 0.01），但对伤口成纤维细胞无此作用。通过Sephadex G - 100凝胶过滤分离鉴定出伤口渗出液中的活性因子分子量约为100 kDa。对该可溶性因子的特性分析表明它是一种蛋白质（硫酸铵沉淀），对胰蛋白酶消化敏感，耐热（56℃，30分钟），且对神经氨酸酶有抗性。通过离子交换色谱法测定其等电点似乎为7.0。活性因子对胸腺淋巴细胞的有丝分裂增殖无影响，表明具有细胞靶点特异性。这些数据表明伤口环境中含有促进成纤维细胞功能的高分子量蛋白质，这对愈合过程至关重要。
