A Novel Chitosan-Gelatin Scaffold and Cell Spray Therapy for Treating Limbal Stem Cell Deficiency

Limbal stem cell deficiency (LSCD) is a severe ocular condition that impairs vision and causes chronic pain. Current therapeutic approaches, such as stem cell transplantation and biomaterial scaffolds, are under continuous refinement. This study evaluated a novel treatment strategy combining a biocompatible and biodegradable chitosan-gelatin (CS-Gel) electrospun scaffold with a topical limbal stem cell (LSC) spray for treating alkaline-induced LSCD in a rabbit model. The scaffold was fabricated via electrospinning, stabilized with water vapor, and characterized using SEM, ATR-FTIR, contact angle, weight loss, and swelling analyses. Limbal stem cells were isolated from cadaveric corneas, cultured, and prepared as a spray suspension (10⁴ cells/mL). Nine rabbits with induced alkaline burns were divided into three groups: an untreated control, a group treated with the CS-Gel scaffold alone, and a group receiving the CS-Gel scaffold followed by the LSC spray one week later. Clinical and histopathological evaluations assessed epithelialization, stromal integrity, neovascularization, and inflammation. The results demonstrated that the combined CS-Gel + cell spray treatment significantly enhanced corneal regeneration, reduced inflammation, and improved healing outcomes compared to the other groups. This study confirms the potential of the CS-Gel + cell spray as a promising platform for LSCD therapy, warranting further investigation. Clinical Registration: This study is an animal-based research and does not involve human clinical trials.