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Cornelia W Peterson, Heather L Chandler; INSULIN FACILITATES IN VITRO CORNEAL WOUND HEALING IN THE DIABETIC ENVIRONMENT. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1392.
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© ARVO (1962-2015); The Authors (2016-present)
There are more than 30 million people living with diabetes in the US alone, and it is projected that this number will increase to more than 500 million people globally by 2030. Half of these patients will develop corneal changes, termed diabetic keratopathy, during the course of their disease, predisposing them to pain and delayed wound healing. Due to the consistently increasing prevalence and estimated costs to treat diabetic keratopathy, there has been a growing effort to identify novel therapies to minimize discomfort and sight-threatening scarring. Topical insulin has been shown to reduce wound area and improve corneal sensitivity in diabetic rats. In the current study, we aimed to evaluate the effects of exogenous insulin on wound healing in human corneal epithelial cells (hCEC) in vitro.
Confluent hCEC were treated with glucose-supplemented (0, 0.92, or 6mM) media for 24 hours (h), then incubated with 0 or 2 units (u) of human recombinant insulin, and a standard scratch test was performed. Wounds were imaged at 12h intervals from 24-48h, and software was used to determine the wound area. To evaluate protein synthesis, hCEC were again pre-treated with glucose, incubated with 0 or 2u of insulin, and then labeled with a fluorescent marker of nascent proteins with or without cycloheximide, an inhibitor of translation. A fluorescence assay was performed on treated hCEC.
Insulin-treated hCEC showed a significant reduction in the mean wound area when compared to the vehicle at all time points in the 0mM glucose conditions. At the termination of the scratch test, 31.0 +/- 10.3% mean wound area remained in the vehicle-treated hCEC and 6.0 +/- 3.9% in the insulin-treated hCEC (T test, p ≤ 0.05). Insulin-treated hCEC showed a significant increase in protein synthesis when compared to vehicle control across all glucose concentrations (T test; see Table 1).
These results suggest that insulin facilitates in vitro wound healing of hCEC at specific glucose concentrations by stimulating protein synthesis. Following further characterization of the mechanism of action, insulin may be developed into a new therapy for diabetic keratopathy.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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