PURPOSE:Peptide growth factors are known accelerators of corneal wound healing, probably mediated through increased proliferation of the cells; however, information about their effect on keratocyte motility is lacking. The influence of peptide growth factors on keratocyte migratory activity was investigated, using the following growth factors: platelet derived growth factor (PDGF-BB), epidermal growth factor (EGF), acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF), insulin-like growth factor-I (IGF-I) and transforming growth factor-beta-1 (TGF-beta 1).

METHODS:Keratocytes were seeded on gels of type 1 collagen, growth factor added, and the cells left to migrate for 72 hours. Subsequently, the number of keratocytes at the different levels in the collagen gel was evaluated by optically sectioning the gel at 20 microns, intervals, with an inverted phase contrast microscope.

RESULTS:PDGF, EGF and bFGF at 10 ng/ml, all increased the number of keratocytes at the different levels of the gel as compared to a non-stimulated control (p < 0.05 or p < 0.01, students t-test). TGF-beta proved to be a strong inhibitor of keratocyte migration, decreasing the number of keratocytes observed at every level in the gel (p < 0.05 and p < 0.01, students t-test), whereas no effect of IGF-I and aFGF was found. During the 72 hours of migration, no contraction of the collagen gels was observed. Autoradiography of histological sections of the gels showed that during the 72-hour period only TGF-beta and 10% fetal bovine serum induced an increase in keratocyte proliferation.

CONCLUSION:PDGF, EGF and bFGF increase keratocyte migration, independent of proliferation in a collagen gel invasion assay and might promote corneal wound healing, not only by increasing cell proliferation, but also through increased motility.

译文

目的:肽生长因子是角膜伤口愈合的已知促进剂,可能是通过细胞增殖的增加来介导的。然而,缺乏有关它们对角膜细胞运动性影响的信息。使用以下生长因子研究了肽生长因子对角膜细胞迁移活性的影响:血小板衍生生长因子(PDGF-BB),表皮生长因子(EGF),酸性成纤维细胞生长因子(aFGF),碱性成纤维细胞生长因子(bFGF) ),胰岛素样生长因子I(IGF-I)和转化生长因子β-1(TGF-beta 1)。

方法:将角质形成细胞接种在1型胶原蛋白,添加了生长因子,细胞迁移了72小时。随后,通过倒置相差显微镜以20微米的间隔对凝胶进行光学切片,评估胶原蛋白凝胶中不同水平的角质形成细胞的数量。

结果:PDGF与未刺激的对照组相比,10 ng / ml的EGF和bFGF均增加了凝胶水平不同时的角膜细胞数量(p <0.05或p <0.01,学生t检验)。 TGF-β被证明是一种强烈的角膜细胞迁移抑制剂,可减少凝胶中各个水平上观察到的角膜细胞数量(p <0.05和p <0.01,学生t检验),而IGF-I和aFGF则没有作用成立。在迁移的72小时内,未观察到胶原凝胶的收缩。凝胶组织学切片的放射自显影显示,在72小时内,只有TGF-β和10%的胎牛血清诱导了角膜细胞增殖的增加。

结论:PDGF,EGF bFGF和bFGF可增加角膜细胞迁移,而不受胶原凝胶入侵试验中的增殖的影响,并且不仅可以通过增加细胞增殖,而且可以通过增加运动性来促进角膜伤口愈合。

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