Collagen Vitrigel Scaffold Production Method for Corneal Engineering Purposes

Abstract

Purpose : The main challenge in corneal scaffold production is to obtain optical transparency and mechanical properties through a feasible and reproducible method. In this study, we analyze the characteristics of a collagen vitrigel membranes (CVM) manufactured with a low-cost process for corneal endothelium engineering.

Methods : CVM of two different concentrations were synthesized (1x and 3x) in desiccator inside an oven (Matryoshka system) with a regulated relative humidity by means of K2CO3. Transmittance was analyzed with UV–VIS spectra (380–700 nm). Tensile strength and deformation were assessed using a biaxial test system. The ultrastructure of 37-day samples were observed and measured with an EVO MA Scanning Electron Microscope. Cytotoxicity and viability were tested using Cell Titer Blue on NIH3T3 cells. Each analysis was made by triplicates.

Results : Transparency of 1x and 3x CVM were 92.6% and 89.6% respectively, which is consistent with human corneal transparency (86%-94%). Tensile strength showed a maximum rupture load of ~0.8mm for both concentrations (400-fold increase than in reported literature). SEM showed a laminar, organized structure of collagen fibers. Cell proliferation increased 12% in both collagen concentrations.

Conclusions : Matryoshka system is a feasible method to produce CVM with enhanced transparency, tensile strength and cell proliferation. Making this a useful tool for corneal engineering.

This is a 2020 ARVO Annual Meeting abstract.