Application of chitosan hydrogel and amniotic membrane carriers - impact on structure of epithelial cells cultures

Abstract

Purpose: Comparison of cultured epithelium structure on different culture carries. Comparison amniotic membrane (AM) with new chitosan hydrogel scaffolds (CH). A hydrogel scaffold in the form of a membrane was obtained from chitosan - uniquely biocompatible, biologically inert, stable in the natural environmental and anti-bacterial polysaccharide derived from chitin. A biodegradable and safe for human chitosan film was cross-linked with genipin - naturally occurring and nontoxic agent

Methods: 26 patients were donors of epithelial cells; 13 from the corneal limbus, 13 from oral mucosa. Cells from biopsy were seeded on AM and CH. Cultures were carried in supplemented DMEM in presence of 3T3 fibroblasts coculture. After 14 days of culture stratified epithelium was analized under light microscope, moreover specimens were fixed to perform immunohistochemical recognition of epithelial cytokeratins and to perform histologic examination.

Results: AM revealed regular layers of both epithelia with mean number of layers: 6.7 + 2.1 of limbal and 9.3 + 4.2 of oral origin. CH with number of layers 4.3 + 0.9 of limbal and 5.0 + 1.1 of oral epithelium

Conclusions: AM is more effective carrier to receive regularly. Nevertheless polymeric membranes based on biopolymers seems to be a promising material for further tests directed towards the development of implantable corneal epithelium tissue

Keywords: 482 cornea: epithelium