Corneal reconstruction in chemically damaged cornea using temperature sensitive surface assisted mesenchymal stem cell transplantation in rabbits

Abstract Background: The study aims to assess the potential of corneal reconstruction using mesenchymal stem cells (MSCs) transplantation using temperature sensitive membranes as carriers and compare with fibrin glue method in chemically damaged rabbit cornea. Methods: MSCs were cultured from rabbit’s bone marrow and transplanted over rabbit’s cornea, using either temperature sensitive membranes or fibrin glue method. Endogenous levels of MSCs were assessed to decide the optimal time point for transplanting cells. MSC transplanted corneas were harvested 15, 30 and 60 days post transplantation. Corneal repair markers were evaluated using histopathology, immunohistochemistry (IHC) and real time PCR. The quality of cornea reconstructed was evaluated using corneal opacity scoring and IHC. Results: Endogenous levels of MSCs were significantly higher 48 hours post corneal injury. Transplantation of MSCs using temperature sensitive method resulted in uniform and homogenous spread of MSCs over the damaged surface. Corneal transparency improved 7 days onward in rabbit chemical injured cornea after MSC transplantation. Immunohistochemistry (IHC) of damaged cornea showed complete absence of corneal epithelium as seen by CK3 expression, while re-epithelialization was observed after MSC transplantation. Histopathology showed complete re-epithelialization 15 days of transplantation. Evaluation of corneal quality using epithelial and stromal markers showing restoration of vimentin, α-smooth muscle actin and collagen levels in MSC transplanted cornea compared to damaged corneas. CK3 expression showed differentiation of MSC into corneal epithelial cells. MSC transplanted corneal clarity seemed a little better with temperature sensitive method compared to fibrin glue method. No toxic effects of MSC transplantation were observed on other organs or on resident corneal cells. Conclusions: Temperature sensitive surface MSC transplantation method results in uniform and homogenous delivery of cells on corneal surface and a transparent clear cornea. Epithelial and stromal markers showing restoration of vimentin, α-smooth muscle actin and collagen levels in MSC transplanted cornea compared to chemically damaged corneas. Our data also show the differentiation of MSC into corneal epithelial cells on corneal surface. Overall temperature surface MSC transplantation is an easy and safe alternative method to fibrin glue for exploring corneal reconstruction.