Polyhydroxyalkanoate (PHA) and Wharton Jelly-mesenchymal Stem Cells (MSCs) for Corneal Regenerative Therapeutic Strategy.

Introduction: Corneal regenerative medicine is a contemporary approach involving the use of biomaterials, biological factors and stem cells to reduce dependence on native tissue supply for transplantation. This study aims to investigate cellularized polyhydroxyalkanoate (PHA) scaffold with human telomerase-immortalized cornea epithelial cells (HTCEC) in combination with Wharton Jelly-Mesenchymal stem cells (MSC) or its conditioned media (CM) for ocular surface regeneration. Materials and methods: The PHA employed in this investigation, P(3HB-co-4HB-co-5HV-co-3HHx), was biosynthesized from Cupriavidus necator. The water contact angle as a measure of scaffold wettability was determined, and in vitro PHA biodegradation by lipase enzyme was conducted. The effects of MSC on the focal adhesion proteins were determined by immunofluorescence and immune regulatory proteins were studied. Common corneal genes' expression was evaluated using qPCR. Results: There was a significant loss of PHA dry weight due to lipase biodegradation. MSC-conditioned media (CM) significantly improved HTCEC viability (121%) as compared to control (100%), p < 0.005. The effect of HTCEC/MSC co-culture on focal adhesion protein expression was significantly higher as compared to HTCEC single culture (p < 0.05). MSC co-culture with HTCEC showed increased secretion of IL-1β and TGF-β1 more than single HTCEC culture in a pro-inflammatory stimulated HTCEC model. Gene expressions for common corneal markers ITGB1, ABCG2, ABCB5, CK3, CK12, CX43, and ΔNP63 were upregulated in the presence of CM p < 0.05. Conclusion: Cellularized PHA scaffold used in combination with MSC is a novel regenerative medicine approach which has a huge potential for anterior ocular surface diseases. [ABSTRACT FROM AUTHOR]