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Mark Rosenblatt, Brian Lawrence, Aihong Liu, Brigette Cole, Waleed Abdel-Naby; Silk fibroin stimulates corneal epithelial cellular migration and growth during wound healing. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):727.
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© ARVO (1962-2015); The Authors (2016-present)
To characterize the ability of soluble silk fibroin to accelerate wound healing by promoting epithelial cell migration and growth.
We utilized an in vitro wound scratch model system to mimic the cell behavior of the human eye surface during a corneal abrasion. A human corneal limbal epithelial (HCLE) cell line was grown as a confluent monolayer, and the rate of scratch closure monitored with pretreatment or concurrent treatment with silk fibroin using live cell time-lapse imaging of cells. The migration rate of individual cells during wound closure was also evaluated to assess the effects of silk fibroin treatment on cell migration. Isolated cells were also analyzed for cell spreading, as well as the distribution of focal adhesions using immunocytochemistty for vinculin. Cellular proliferation of cells treated with silk fibroin was evaluated using MTT assay and FACS cell cycle analysis, to determine if the accelerated wound healing effect of silk can be partially attributed to acceleration in epithelial cell proliferation.
Analysis of our results demonstrates a dose-dependent improvement in wound healing by corneal epithelial cells treated with silk fibroin relative to control cells. The effect was independent of whether cells were pretreated or treated concurrently with silk fibroin. Treatment with silk fibroin significantly increased cell spreading and redistributed focal adhesions to the cell periphery. Silk fibroin stimulated cell proliferation by over 50%, relative to control cells. Cell cycle analysis also revealed that silk fibroin caused an increase in the amount of cells in S phase of the cell cycle.
Our data showed that cell migration and proliferation was increased when cells were grown in the presence of silk fibroin solution, compared to cells grown in the absence of silk. Stimulation of these biological processes contributed to an enhanced wound healing rate, as evaluated by our in vitro wound scratch model. These findings add to our understanding of corneal epithelial cell biology as well as provide working evidence for the potential use and application of silk fibroin based materials for the treatment of ocular surface wounds.
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