Andresen J L, Ledet T, Hager H, Josephsen K, Ehlers N
Department of Ophthalmology, Aarhus University Hospital, Denmark.
Exp Eye Res. 2000 Jul;71(1):33-43. doi: 10.1006/exer.2000.0850.
Motivated by the alterations seen in the corneal matrix composition after photorefractive keratectomy and the migration of corneal keratocytes seen following this procedure, the locomotor response of corneal stromal fibroblasts to various extracellular matrix proteins was determined. In addition, the involvement of integrin mediated attachment to the matrix proteins was investigated. Quantitative invasion assays were performed using collagen gels, supplemented with either fibronectin, tenascin, collagen type V, collagen type VI, chondroitin sulfate or keratan sulfate. The ultrastructure of the gels was visualized by scanning electron microscopy and related to the migration results. The extent of alpha(1)beta(1), alpha(2)beta(1), alpha(3)beta(1)and alpha(5)beta(1)integrin mediated attachment to the matrix proteins was evaluated using blocking antibodies. Fibronectin increased corneal fibroblast migration significantly, and served as an excellent substrate for cellular attachment, mediated by the alpha(5)beta(1)integrin. Addition of tenascin to the fibronectin-containing gels disrupted these effects, while attachment to this matrix also involved the integrins alpha(2)beta(1)and alpha(3)beta(1). Chondroitin sulfate and collagen types V and VI primarily altered the structure of the collagen matrix, resulting in an inhibition of migration by the collagens and an increase by chondroitin sulfate. They all served as poor substrates for attachment. Thus, the migratory activity of corneal fibroblasts in vitro is influenced by the composition of the surrounding extracellular matrix, either by integrin mediated cell-matrix interactions or through matrix-matrix interactions. This study provides evidence that the provisional matrix deposited in a corneal stromal wound may facilitate the entry of migrating corneal fibroblasts.
鉴于在准分子激光原位角膜磨镶术后观察到的角膜基质成分改变以及该手术后角膜基质细胞的迁移情况，我们测定了角膜基质成纤维细胞对各种细胞外基质蛋白的运动反应。此外，还研究了整合素介导的与基质蛋白附着的参与情况。使用补充了纤连蛋白、腱生蛋白、Ⅴ型胶原、Ⅵ型胶原、硫酸软骨素或硫酸角质素的胶原凝胶进行定量侵袭试验。通过扫描电子显微镜观察凝胶的超微结构，并将其与迁移结果相关联。使用阻断抗体评估α(1)β(1)、α(2)β(1)、α(3)β(1)和α(5)β(1)整合素介导的与基质蛋白附着的程度。纤连蛋白显著增加了角膜成纤维细胞的迁移，并作为由α(5)β(1)整合素介导的细胞附着的优良底物。向含纤连蛋白的凝胶中添加腱生蛋白会破坏这些作用，而与该基质的附着还涉及整合素α(2)β(1)和α(3)β(1)。硫酸软骨素以及Ⅴ型和Ⅵ型胶原主要改变了胶原基质的结构，导致胶原抑制迁移而硫酸软骨素促进迁移。它们均作为附着的不良底物。因此，体外角膜成纤维细胞的迁移活性受周围细胞外基质组成的影响，这要么通过整合素介导的细胞 - 基质相互作用，要么通过基质 - 基质相互作用。本研究提供了证据表明，沉积在角膜基质伤口中的临时基质可能促进迁移的角膜成纤维细胞的进入。
