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Research Article: Comparison of the regenerative potential of different functionalized gelatin-based hydrogels as fillers of rabbit corneal wounds

Date Published: 2025-09-24

Abstract:
Persistent epithelial defects (PEDs) and chronic corneal ulcers are lesions resistant to treatment for over 2 weeks, risking inadequate healing, reduced sensitivity, and corneal lysis or perforation. This study evaluates the regenerative potential of functionalized gelatin-based hydrogels for treating rabbit corneal wounds as a non-surgical alternative. Thirty female New Zealand white rabbits underwent anterior stromal keratectomy and were assigned to five groups: control (0.2% HA artificial tears) and four hydrogel treatment groups. Hydrogels included non-functionalized gelatin-RFP (H) and functionalized versions with infliximab (H-Ab), autologous serum (H-AS), and human amniotic membrane extracts (H-HAMe). Crosslinking was performed in situ with blue light. Corneas were evaluated at 7 and 21?days for re-epithelialization, fibrosis, and inflammation using histology, qPCR and immunohistochemistry, focusing on markers of proliferation (Ki67), differentiation (CK3), stemness ( PAX6, p63, CD44 ), adhesion (integrin ?4), and fibrosis ( ? -SMA). All treatments supported re-epithelialization by day 7 and restored barrier function (ZO-1), with H-AS achieving the fastest closure. Expression of the adhesion marker integrin ?4 improved over time across all groups. Hydrogel formulations promoted limbal activation ( PAX6, CD44 ), with H-AS and H-HAMe showing elevated p63 expression at day 7. All hydrogels reduced fibrosis ( ? -SMA), though extracellular matrix organization varied. H-Ab and H-HAMe reduced inflammation ( IL-1? ), while H-AS showed minimal irritability. Functionalized gelatin-RFP hydrogels promote re-epithelialization, reduce fibrosis and inflammation, and restore ocular integrity, offering a promising solution for corneal wound repair.

Introduction:
The human cornea is a transparent, avascular tissue that accounts for approximately two-thirds of the eye’s total refractive power. Despite its critical function, it is particularly vulnerable to environmental insults such as trauma, infections, and chemical injuries ( 1 ). Damage to the corneal epithelium can result in persistent epithelial defects (PEDs) and corneal ulcers, which ulcers represent significant therapeutic challenges in ophthalmological practice, frequently resulting in severe complications such as…

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