Effect of fibroblasts on epidermal regeneration
Effect of fibroblasts on epidermal regeneration
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DOI：
10.1046/j.1365-2133.2002.04871.x
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发表时间：
2002-08-01
影响因子：
10.3
通讯作者：
Ponec, M
中科院分区：
文献类型：
--
作者：
El-Ghalbzouri, A;Gibbs, S;Ponec, M
关于真皮成纤维细胞和表皮角质形成细胞之间特异性相互作用的信息很少。使用由角质形成细胞和成纤维细胞的器官型共培养组成的工程皮肤等效物为此类研究提供了一种有吸引力的方法。目的探讨成纤维细胞在重建表皮形成和维持中的作用。方法将人角质形成细胞接种于成纤维细胞增多的胶原基质上，在气液界面培养2周。结果在无成纤维细胞的情况下，形成层状表皮，仅有3 ~ 4层活细胞。在成纤维细胞存在的情况下，角质形成细胞增殖受到刺激，表皮形态得到改善。表皮形态的发生也明显改善了表皮的形成，在皮肤等量物培养基中生长的器官型角质形成细胞单一培养物或器官型角质形成细胞-成纤维细胞共培养物中生长的表皮。这些观察结果清楚地表明，从成纤维细胞释放的可溶性因子具有促增殖活性。在所有实验条件下，角蛋白10在所有基底上细胞层的表达表明角化细胞分化的开始。随着胶原基质中成纤维细胞数量的增加，与角化细胞活化相关的标志物，如角蛋白6、16和17以及角化包膜前体SKALP的表达减少，天合蛋白的定位向颗粒层转移。当成纤维细胞在胶原基质中预培养1周而不是过夜时，这种成纤维细胞介导的效果更加明显。基底膜蛋白胶原VII和层粘连蛋白5存在于上皮-基质边界。整合素alpha6beta4和E-cadherin的表达与天然皮肤相当，不受成纤维细胞的显著调节。在所有实验条件下，整合素亚基alpha2、alpha3和beta1的表达均上调，表明角质形成细胞被激活。结论胶原基质中成纤维细胞的数量及其功能状态是正常表皮形态发生的关键因素。
Background There is little information on specific interactions between dermal fibroblasts and epidermal keratinocytes. The use of engineered skin equivalents consisting of organotypic cocultures of keratinocytes and fibroblasts offers an attractive approach for such studies.Objectives To examine the role fibroblasts play in generation and maintenance of reconstructed epidermis.Methods Human keratinocytes were seeded on collagen matrices populated with increasing numbers of fibroblasts and cultured for 2 weeks at the air-liquid interface.Results In the absence of fibroblasts, stratified epidermis with only three or four viable cell layers was formed. In the presence of fibroblasts, keratinocyte proliferation was stimulated and epidermal morphology was improved. Epidermal morphogenesis was also markedly improved in epidermis generated in organotypic keratinocyte monocultures grown in medium derived from dermal equivalents or from organotypic keratinocyte-fibroblast cocultures. These observations clearly indicate the proliferation-stimulating activity of soluble factors released from fibroblasts. Under all experimental conditions, onset of keratinocyte differentiation was shown by the expression of keratin 10 in all suprabasal cell layers. With increasing numbers of fibroblasts incorporated into the collagen matrix, the expression of markers associated with keratinocyte activation, e.g. keratins 6, 16 and 17 and the cornified envelope precursor SKALP decreased, and involucrin localization shifted toward the granulosum layer. This fibroblast-mediated effect was even more pronounced when the fibroblasts were precultured in the collagen matrices for 1 week instead of overnight. The basement membrane proteins collagen VII and laminin 5 were present at the epithelial-matrix border. The expression of integrin alpha6beta4 and of E-cadherin was comparable with that seen in native skin and was not significantly modulated by fibroblasts. Under all experimental conditions the expression of integrin subunits alpha2, alpha3 and beta1 was upregulated, indicating keratinocyte activation.Conclusions Our results illustrate that numbers of fibroblasts in the collagen matrix and their functional state is a critical factor for establishment of normal epidermal morphogenesis.