Tissue engineering of the human corneal endothelium using thermo-responsive carriers based on poly(vinyl methyl ether)

Abstract

Purpose: Generation of transplantable human corneal endothelial cell sheets using thermo-responsive cell culture carriers with tunable biochemical and mechanical properties.

Methods: The human corneal endothelial cell line HCEC-12 was cultured on electron beam cross-linked thin films of poly(vinyl methyl ether) blended with the alternating copolymer of vinyl methyl ether and maleic acid [Acta Biomaterialia 9 (2013) 5031], and complemented with an additional poly (N-isopropylacrylamide) (PNiPAAm) component. This thermo-responsive coating combines a wide parameter range for adjusting thickness, stiffness and swelling behavior as well as reactive sites for biomolecular functionalization with the outstanding interaction profile of PNiPAAm. Cell morphology, vitality and functionality before and after transfer are carefully characterized by light microscopy, immunofluorescence microscopy, vitality assays and transepithelial resistance measurements in a simplified model system. The potential application in ophthalmology is demonstrated by transplantation onto de-endothelialized porcine corneas in vitro.

Results: The PVME-based carrier supported the in vitro cultivation of HCEC and formation of morphological prerequisites for tissue function. Moreover, it allowed for the detachment and transfer of HCEC sheets onto the target tissue.

Conclusions: The novel cell culture carrier is successfully applied to the generation of transplantable sheets of human corneal endothelial cells. The developed method to engineer corneal endothelial tissue may significantly improve therapeutic options.

Keywords: 481 cornea: endothelium • 741 transplantation • 480 cornea: basic science