A novel cytocompatible thermoresponsive co‐polymer for corneal tissue engineering

Purpose Using thermoresponsive polymers as a substrate to expand healthy sheets of transplantable cells for the immediate cover of wounds is an attractive novel treatment for ocular surface reconstruction. However, the cytocompatibility of thermoresponsive polymer and the effect on cell expansion must be assessed. Changes in the expression of secretory proteins in response to different microenvironments can be used to determine a more precise suitability of polymers in cell culture at molecular level. We assess the specific cytocompatibility of a novel thermoresponsive co‐polymer with the future goal of its use in corneal reconstruction. Methods Protein was extracted from conditioned medium of SV‐40 immortalised corneal cell line (CCL) cultured on both thermoresponsive polymer and tissue culture plastic. Proteins were separated by two‐dimensional gel electrophoresis and the resulting spot map profiles were differentially compared. Differences in the protein profiles were characterised by mass spectrometry and western blot analysis. Results Differential analysis of whole cell extract, and conditioned medium from CCL on plastic identified over 50 potentially secreted proteins. Mass spec characterisation confirmed these proteins as secretory, but also suggested healthy cell expansion was occurring. Comparison of CCL on plastic and thermoresponsive polymer revealed few differences suggesting the polymer did not effect cell expansion. Conclusion Thermoresponsive polymer is cytocompatible for CEC cell growth. This novel co‐polymer maybe a potential substrate for the future ex‐vivo engineering of transplantable cell sheets, for improved corneal reconstruction