Electro-spun Membranes as Scaffolds for Human Corneal Endothelial Cells

• Published in: Current eye research Vol. 43; no. 1; pp. 1 - 11
• Main Authors: Kruse, Magnus, Walter, Peter, Bauer, Benedict, Rütten, Stephan, Schaefer, Karola, Plange, Niklas, Gries, Thomas, Jockenhoevel, Stefan, Fuest, Matthias
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 02-01-2018
• Subjects: Aged, 80 and over; Biocompatible Materials; Cells, Cultured; Cornea; Corneal Endothelial Cell Loss - pathology; Corneal Endothelial Cell Loss - surgery; Corneal Transplantation - methods; electro-spinning; endothelial cells; Endothelium, Corneal - ultrastructure; Female; Humans; Materials Testing; Microscopy, Electron, Scanning; Polymethyl Methacrylate; scaffold; tissue engineering; Tissue Engineering - methods; Tissue Scaffolds; electro-spinning; Cornea; endothelial cells; tissue engineering; scaffold
• ISSN: 0271-3683; 1460-2202
• DOI: 10.1080/02713683.2017.1377258

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 µm) showed significantly higher diameters than PCL (2.29 ± 0.11 µm; p = 0.003) and PLGA (1.84 ± 0.21 µm; p < 0.001), while fibers from PCL also showed larger diameters than those from PLGA (p = 0.002). PMMA scaffolds (26.77 ± 17.48 µm) had significantly larger interstitial spaces than those from PCL (13.30 ± 5.47 µm; p = 0.04) and PLGA (10.42 ± 6.15 µm; 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.