June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Epidermal growth factor-loaded collagen gels to enhance corneal wound healing: Effect of matrix crosslinking chemistry
Author Affiliations & Notes
  • Youngyoon Amy Seo
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
  • Gabriella Maria Fernandes Cunha Rogers
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
  • David Myung
    Ophthalmology, Stanford University School of Medicine, Stanford, California, United States
    Chemical Engineering, Stanford University, Stanford, California, United States
  • Footnotes
    Commercial Relationships   Youngyoon Amy Seo None; Gabriella Rogers None; David Myung None
  • Footnotes
    Support  National Institutes of Health (National Eye Institute K08EY028176 and a Departmental P30- EY026877 core grant); a core grant and Career Development Award from Research to Prevent Blindness (RPB); the Matilda Ziegler Foundation; VA Rehabilitation Research and Development Small Projects in Rehabilitation Effectiveness (SPiRE) program (I21 RX003179)
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3230 – A0265. doi:
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    • Get Citation

      Youngyoon Amy Seo, Gabriella Maria Fernandes Cunha Rogers, David Myung; Epidermal growth factor-loaded collagen gels to enhance corneal wound healing: Effect of matrix crosslinking chemistry. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3230 – A0265.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Millions of people suffer from corneal diseases, but less than 2% of patients have access to corneal transplantation due to the shortage of cadaveric donor corneal tissue. Moreover, rapid and complete epithelial wound healing is critical to prevent infection and scarring that can lead to blindness. Biomaterial matrices have been investigated as a way to fill corneal defects and support epithelialization. Loading growth factors into defect-filling hydrogels is a promising approach toward enhancing corneal epithelialization. Here, we incorporate epidermal growth factor (EGF) into in situ-forming collagen gels to study the effect of their controlled release from within defect-stabilizing matrices on epithelial cell adhesion and proliferation.

Methods : EGF was added to a neutralized collagen solution and crosslinked either via N-hydroxysuccinimide chemistry using 8-arm PEG or via UV light using riboflavin as a photosensitizer. We compared the gels’ degradation as well as the proliferation, migration, and phenotype of corneal epithelial cells (CECs) seeded on these gels and the epithelial wound healing response in ex vivo organ culture model. Cell viability and cytotoxicity were measured using colorimetric assays. We also executed an in vitro scratch assay which mimics cell migration during wound healing in vivo. The presence of biomarkers that promote epithelial healing were compared across three groups: cells without treatment, collagen gels alone, and EGF encapsulated within PEG- or UV-crosslinked collagen hydrogels.

Results : Encapsulated EGF accelerated in vitro corneal epithelial cell proliferation and migration. Gels loaded with EGF also exhibited a lower degradation rate and accelerated corneal wound healing in our organ culture studies. Immunohistochemical analysis exhibited expression of 4-hydroxynonenal within the epithelium and alpha-smooth muscle actin within the surrounding stroma for 8-arm PEG collagen and UV riboflavin gels with EGF. Immunohistochemical studies demonstrated that both types of EGF-loaded collagen gels fostered tight junction formation and maintained normal epithelial phenotype.

Conclusions : Our study suggests that collagen gels with encapsulated EGF can fill corneal defects and enhance corneal epithelial regeneration. Further work to investigate the effects of EGF-loaded collagen gels in vivo are merited.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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