Abstract
There is a significant clinical need to improve current therapeutic approaches to treat ocular surface injuries and disease, which affect hundreds of millions of people annually worldwide. The work presented here demonstrates that the presence of Silk-Derived Protein (SDP) on the healing rabbit corneal surface, administered in an eye drop formulation, corresponds with an enhanced epithelial wound healing profile. Rabbit corneas were denuded of their epithelial surface, and then treated for 72-hours with either PBS or PBS containing 5 or 20 mg/mL SDP in solution four times per day. Post-injury treatment with SDP formulations was found to accelerate the acute healing phase of the injured rabbit corneal epithelium. In addition, the use of SDP corresponded with an enhanced tissue healing profile through the formation of a multi-layered epithelial surface with increased tight junction formation. Additional biological effects were also revealed that included increased epithelial proliferation, and increased focal adhesion formation with a corresponding reduction in the presence of MMP-9 enzyme. These in vivo findings demonstrate for the first time that the presence of SDP on the injured ocular surface may aid to improve various steps of rabbit corneal wound healing, and provides evidence that SDP may have applicability as an ingredient in therapeutic ophthalmic formulations.
Highlights
Over 50 million people worldwide suffer from blindness due to corneal disease, and nearly 2.5 million new eye injuries occur each year in the United States, representing the greatest ocular environmental health problem today [1]
Formulation of Silk-Derived Protein (SDP) eye drops and fluorescein staining of rabbit corneal wounds To determine the longitudinal stability of aqueous SDP, a hermetically sealed SDP solution was stored at 4 ̊C for up to 6 months and evaluated by visual inspection and for molecular weight distribution by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)
Solution samples were drawn from each time point, Silk-Derived Protein enhances rabbit corneal epithelial wound healing and SDS-PAGE analysis was performed to determine the molecular weight distribution (MWD) and material stability over time (Fig 1A)
Summary
Over 50 million people worldwide suffer from blindness due to corneal disease, and nearly 2.5 million new eye injuries occur each year in the United States, representing the greatest ocular environmental health problem today [1]. It is estimated that annually, another 337 million people suffer from the debilitating symptoms of dry eye disease worldwide, representing another significant threat to the ocular surface [2]. Silk-Derived Protein enhances rabbit corneal epithelial wound healing specific role of these authors are articulated in the ‘author contributions’ section. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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