Electro-spun Membranes as Scaffolds for Human Corneal Endothelial Cells.

Background: Corneal endothelial dysfunction remains the most frequent indication for corneal transplantation, limited by donor material shortage, poor long-term graft survival, or allogeneic graft rejection. Therefore, tissue-engineered endothelial grafts (TEEG) represent a promising alternative to human donor tissue. In this study, we generated electro-spun scaffolds and tested these for their suitability for human corneal endothelial cell (hCEC) cultivation. Methods: The polymers poly(methyl-methacrylate) (PMMA), poly(lactic-co-glycolic acid) (PLGA), and polycaprolactone (PCL) were spun with equal parameters. HCEC-12 was cultured on the scaffolds for 3 to 7 days. Scaffolds were evaluated by light microscopy, porometry, light transmission, scanning electron microscopy (SEM), live/dead staining and cell viability assay. Results: Electro-spun fibers from PMMA (2.99 +/- 0.24 mu m) showed significantly higher diameters than PCL (2.29 +/- 0.11 mu m; p = 0.003) and PLGA (1.84 +/- 0.21 mu m; p < 0.001), while fibers from PCL also showed larger diameters than those from PLGA (p = 0.002). PMMA scaffolds (26.77 +/- 17.48 mu m) had significantly larger interstitial spaces than those from PCL (13.30 +/- 5.47 mu m; p = 0.04) and PLGA (10.42 +/- 6.15 mu m; p = 0.002), while PCL and PLGA did not differ significantly (p = 0.26). SEM analysis revealed that only PLGA fibers preserved a normal HCEC-12 morphology. PLGA and PCL did not differ in cell number, death, or viability after 7 days of HCEC-12 cultivation. PMMA showed significantly higher cytotoxicity (p < 0.001; PLGA: 1626.2 +/- 183.8 RLU; PMMA: 841.9 +/- 92.7 RLU; PCL: 1580.2 +/- 171.02 RLU). Conclusions: The biodegradable PLGA and PCL electro-spun scaffolds resulted in equal biocompatibility, while PMMA showed cytotoxicity. Only PLGA preserved hCEC morphology and consequently seems to be a promising candidate for TEEG construction.