THE WOUND ENVIRONMENT AS A REGULATOR OF FIBROBLAST PHENOTYPE
THE WOUND ENVIRONMENT AS A REGULATOR OF FIBROBLAST PHENOTYPE
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DOI：
10.1016/0022-4804(91)90022-e
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发表时间：
1991-05-01
影响因子：
2.2
通讯作者：
BARBUL, A
中科院分区：
文献类型：
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作者：
REGAN, MC;KIRK, SJ;BARBUL, A
成纤维细胞是伤口成功愈合的基础。我们假设各种成纤维细胞功能（增殖、胶原合成和重塑）的诱导和调节是由伤口环境决定的。我们检查了伤口液（WF）作为伤口环境的反映，对正常真皮（NF）和伤口收获的成纤维细胞（WHF）表型表达的影响。 WF 和 WHF 是从植入 10 天伤口的聚乙烯醇海绵中获得的。 NF 和 WHF 用于一到三代之间。使用血清刺激在微培养系统中测定增殖功能（n=12）。添加 20% WF 后，NF 和 WHF 对血清的增殖反应均显着降低（NF 为 17,261 ± 1231 cpm 对比 2704 ± 1215 cpm，P < 0.05；WHF 为 15,391 ± 3735 cpm 对比 1701 ± 816 cpm，血清和 WF 中 P< 0.05，分别）。两种成纤维细胞类型的总蛋白质合成（通过[3H]脯氨酸掺入测量）相同；然而，相对胶原合成（胶原酶可消化部分）明显不同（NF 为 2.2 ± 0.9%，WHF 为 11.4 ± 2%，P< 0.05）。添加 WF 显着增强 NF 胶原合成至 9.4 ± 2%，但对 WHF 没有影响。使用成纤维细胞填充的胶原蛋白晶格测定机械和重塑功能。在血清中，WHF 比 NF 收缩晶格更快（24 小时时为 499 ± 14 mm2 与 770 ± 30 mm2，P < 0.05；72 小时时为 301 ± 18 mm2 与 540 ± 21 mm2，P < 0.05）。 WF 的添加进一步增加了 WHF 的晶格收缩，并诱导 NF 的收缩活性增强。因此，与 NF 相比，WHF 表现出增殖减少、胶原合成和重塑增强。通过添加 10 天大的 WF，可以将 NF 的表型改变为类似于 WHF。因此，伤口环境改变成纤维细胞表型，通过最大化胶原蛋白合成和重塑来促进伤口修复，同时下调增殖。
Fibroblasts are fundamental to successful wound healing. We hypothesized that the induction and regulation of various fibroblast functions (proliferation, collagen synthesis, and remodeling) are determined by the wound environment. We examined the effect of wound fluid (WF), as a reflection of the wound environment, on the phenotypic expression of normal dermal (NF) and wound-harvested fibroblasts (WHF). WF and WHF were obtained from implanted polyvinyl alcohol sponges in 10-day-old wounds. NF and WHF were used between one and three passages. Proliferative function was assayed in a microculture system using serum stimulation (n= 12). The proliferative response of both NF and WHF to serum was significantly reduced by the addition of 20% WF (17,261 ± 1231 cpm vs 2704 ± 1215 cpm for NF,P< 0.05; and 15,391 ± 3735 cpm vs 1701 ± 816 cpm for WHF,P< 0.05 in serum and WF, respectively). Total protein synthesis (measured by [3H]proline incorporation) was equal in both fibroblast types; however, the relative collagen synthesis (collagenase-digestible fraction) was markedly different (2.2 ± 0.9% for NF vs 11.4 ± 2% for WHF,P< 0.05). Addition of WF markedly enhanced NF collagen synthesis to 9.4 ± 2%, but had no effect on WHF. Mechanical and remodeling functions were assayed using fibroblast-populated collagen lattices. In serum, WHF contracted the lattices faster than NF (499 ± 14 mm2vs 770 ± 30 mm2at 24 hr,P< 0.05, and 301 ± 18 mm2vs 540 ± 21 mm2at 72 hr,P< 0.05). Addition of WF further increased lattice contraction by WHF and induced enhanced contractile activity in NF. Thus, WHF demonstrate reduced proliferation and enhanced collagen synthesis and remodeling in comparison to NF. The phenotype of NF can be altered to resemble WHF by the addition of 10-day-old WF. Thus, the wound environment alters fibroblast phenotype to facilitate wound repair by maximizing collagen synthesis and remodeling, while down-regulating proliferation.