Therapeutic efficacy of BSA formulated hydrogels in corneal wound healing and epithelial cell regeneration: an ex vivo study

Corneal injury requires both epithelial regeneration and stromal repair, and formulated biomaterials established to repair damaged corneas can be used in regenerative medicine. The challenge is to ensure that biomaterials can be incorporated into the host tissue and delivered intracellularly without causing rapid material deprivation, thus maintaining corneal transparency. Bovine serum albumin-formulated hydrogels (BHG) were prepared by dissolving with riboflavin, retinoic acid, and 2.5% glutaraldehyde solutions. Periphery-centered wounds from camel corneas (8mm diameter and 250 µm depth) were mounted on a dome-shaped agarose gel in six-well plates containing BHG-supplemented serum-free Medium 199. The plates were then incubated at 37 °C for 24, 48, and 72 h. A complete set of corneoscleral rings was procured and processed for histopathological, electron microscopy, and immunohistochemistry assays. Histological and electron microscopy results showed that all epithelial layers and anterior stroma developed faster in the BHG-treated wounds than in the untreated wounded corneas. Compared to untreated wounded corneas, BHG-treated corneas accumulated higher levels of fibronectin and ki-67 and lower levels of alpha-smooth muscle actin inductions. BHG-treated corneal wounds healed faster than untreated wounded corneas. Overall, BHG enhances epithelial regeneration and strengthens the stromal architecture by upregulating ECM and growth factors. Hence, BHG is a promising therapeutic hydrogel for wounded corneas, and further studies on corneal stromal wound healing and epithelial cell reimbursement in an in vivo model are required.