Rickard K A, Taylor J, Rennard S I, Spurzem J R
Department of Internal Medicine, University of Nebraska Medical Center, Omaha 68198-2465.
Am J Respir Cell Mol Biol. 1993 Jan;8(1):63-8. doi: 10.1165/ajrcmb/8.1.63.
Migration of epithelial cells is an important feature of wound healing. Components of extracellular matrix stimulate migration of other cells, and we hypothesized that basement membrane components, laminin and type IV collagen, stimulate migration of bovine bronchial epithelial cells (BBEC). BBEC cultured for 3 days were used in migration assays using the blindwell chamber technique. BBEC migrated in a concentration-dependent manner to laminin, type IV collagen, and fibronectin. "Checkerboard" analysis demonstrated that the migration was directional for each of the components studied. We also evaluated the stimulatory activity of fibronectin fragments. A fragment containing the RGDS peptide was stimulatory of migration while other fragments were much less so, suggesting a role for RGDS-sensitive, integrin-mediated mechanisms. In order to evaluate whether bound or soluble matrix components were required to direct cell migration, we performed haptotaxis assays with precoated filters in the migration chambers. Fibronectin-precoated filters demonstrated significant stimulation of migration, suggesting that some of the migration in our original assays with fibronectin present in the lower chambers could be termed haptotactic. laminin- and type IV collagen-precoated filters were less active. When insulin was used as a chemoattractant, the fibronectin-precoated filters were more facilitatory of migration than the other matrix coatings. In summary, BBEC can chemotactically migrate to extracellular matrix components. Thus, the composition of the provisional matrix that forms at sites of epithelial injury may play an important role in the repair processes that occur after injury to the bronchial epithelium.
上皮细胞迁移是伤口愈合的一个重要特征。细胞外基质成分可刺激其他细胞迁移,我们推测基底膜成分、层粘连蛋白和IV型胶原可刺激牛支气管上皮细胞(BBEC)迁移。使用盲孔室技术对培养3天的BBEC进行迁移试验。BBEC以浓度依赖的方式向层粘连蛋白、IV型胶原和纤连蛋白迁移。“棋盘”分析表明,所研究的每种成分的迁移都是定向的。我们还评估了纤连蛋白片段的刺激活性。含有RGDS肽的片段可刺激迁移,而其他片段的刺激作用则小得多,这表明RGDS敏感的整合素介导机制发挥了作用。为了评估引导细胞迁移需要结合的还是可溶性的基质成分,我们在迁移室中用预包被的滤膜进行了趋触性试验。预包被纤连蛋白的滤膜对迁移有显著刺激作用,这表明在我们最初用下室中存在纤连蛋白的试验中,一些迁移可称为趋触性迁移。预包被层粘连蛋白和IV型胶原的滤膜活性较低。当使用胰岛素作为趋化剂时,预包被纤连蛋白的滤膜比其他基质包被更有利于迁移。总之,BBEC可通过趋化作用向细胞外基质成分迁移。因此,上皮损伤部位形成的临时基质的组成可能在支气管上皮损伤后的修复过程中起重要作用。