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 : 肽生长因子是已知的角膜伤口愈合的促进剂,可能是通过细胞增殖增加介导的; 但是,缺乏有关它们对角质细胞运动的影响的信息。用以下生长因子: 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-β1 (tgf-β1)。
方法 : 将角质细胞接种在1型胶原蛋白的凝胶上,添加生长因子,细胞迁移72小时。随后,通过用倒置相差显微镜以20微米的间隔光学切片来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,EGF和bFGF为10 ng/ml,与未刺激的对照相比,在凝胶的不同水平下,所有这些都增加了角质细胞的数量 (p <0.05或p <0.01,学生t检验)。TGF-β 被证明是角质细胞迁移的强抑制剂,减少了在凝胶中每个水平观察到的角质细胞的数量 (p <0.05和p <0.01,学生t检验),而没有发现igf-i和aFGF的作用。在迁移的72小时内,未观察到胶原蛋白凝胶的收缩。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% 胎牛血清诱导角质细胞增殖增加。
结论 :PDGF,EGF和bFGF增加角质细胞迁移,在胶原蛋白凝胶侵袭试验中独立于增殖,并且可能不仅通过增加细胞增殖,而且通过增加运动性来促进角膜伤口愈合。

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