Weiss E, Yamaguchi Y, Falabella A, Crane S, Tokuda Y, Falanga V
Department of Dermatology and Cutaneous Surgery, University of Miami School of Medicine, Florida, 33136, USA.
J Cell Physiol. 1998 Jan;174(1):58-65. doi: 10.1002/(SICI)1097-4652(199801)174:1<58::AID-JCP7>3.0.CO;2-F.
Wound repair is characterized by the presence of a fibrin-rich matrix, but the effect of fibrin on re-epithelialization remains unclear. In this study, we determined the effects of different fibrin matrices on cultured human neonatal keratinocytes. Using purified fibrinogen and fibrin gels generated by the enzymatic action of thrombin, batroxobin (it leads to retention of fibrinopeptide B), or Agkistrodon contortrix thrombin-like enzyme (ACTE; it leads to retention of fibrinopeptide A), we determined the effect of each of these matrices on keratinocyte morphology, attachment, spreading, and replication as compared to tissue culture plastic. Morphologically, keratinocytes seeded on fibrin surfaces were more rounded and formed three-dimensional structures. Specific cell attachment, as measured at either 37 degrees C or 4 degrees C, was not altered on the different fibrin substrates (P > .05) but was increased on fibrinogen and factor XIII cross-linked fibrin (P < .01). However, keratinocytes seeded on fibrin, regardless of the presence or absence of fibrinopeptides A or B, showed a marked decrease (up to 71%) in cell numbers by days 5 (P = .0357) and 10 (P = .0114). Keratinocyte spreading was decreased by 78.8% (P = .0006), 80.3% (P = .0001), and 89.2% (P = .0001) on thrombin-, batroxobin-, and ACTE-generated fibrin, respectively, but not on fibrinogen-coated dishes. However, either the addition of fibronectin or cross-linking of fibrin with factor XIII allowed full keratinocyte spreading to occur (P = .0002 and P = .0013, respectively). We conclude that fibrin inhibits keratinocyte spreading in the absence of other matrix or plasma proteins or cross-linking by factor XIII.
伤口修复的特征是存在富含纤维蛋白的基质，但纤维蛋白对再上皮化的影响仍不清楚。在本研究中，我们确定了不同纤维蛋白基质对培养的人新生儿角质形成细胞的影响。使用纯化的纤维蛋白原以及由凝血酶、巴曲酶（它导致纤维蛋白肽B保留）或五步蛇凝血酶样酶（ACTE；它导致纤维蛋白肽A保留）的酶促作用产生的纤维蛋白凝胶，我们确定了与组织培养塑料相比，这些基质中的每一种对角质形成细胞形态、附着、铺展和增殖的影响。在形态学上，接种在纤维蛋白表面的角质形成细胞更圆，并形成三维结构。在37℃或4℃下测量的特异性细胞附着在不同的纤维蛋白底物上没有改变（P>.05），但在纤维蛋白原和因子XIII交联的纤维蛋白上增加（P<.01）。然而，接种在纤维蛋白上的角质形成细胞，无论有无纤维蛋白肽A或B，在第5天（P=.0357）和第10天（P=.0114）细胞数量均显著减少（高达71%）。在凝血酶、巴曲酶和ACTE产生的纤维蛋白上，角质形成细胞铺展分别减少了78.8%（P=.0006）、80.3%（P=.0001）和89.2%（P=.0001），但在纤维蛋白原包被的培养皿上没有减少。然而，添加纤连蛋白或用因子XIII交联纤维蛋白均可使角质形成细胞完全铺展（分别为P=.0002和P=.0013）。我们得出结论，在没有其他基质或血浆蛋白或因子XIII交联的情况下，纤维蛋白会抑制角质形成细胞的铺展。
