Construction and characterization of a tissue‐engineered oral mucosa equivalent based on a chitosan‐fish scale collagen composite

Abstract

This study was designed to (1) assess the in vitro biocompatibility of a chitosan–collagen composite scaffold (C3) constructed by blending commercial chitosan and tilapia scale collagen with oral mucosa keratinocytes, (2) histologically and immunohistochemically characterize an ex vivo‐produced oral mucosa equivalent constructed using the C3 (EVPOME‐C), and (3) compare EVPOME‐C with oral mucosa constructs utilizing AlloDerm® (EVPOME‐A), BioMend® Extend™ (EVPOME‐B), and native oral mucosa. C3 scaffold had a well‐developed fibril network and a sufficiently small porosity to prevent keratinocytes from growing inside the scaffold after cell‐seeding. The EVPOME oral mucosa constructs were fabricated in a chemically defined culture system. After culture at an air‐liquid interface, EVPOME‐C and EVPOME‐B had multilayered epithelium with keratinization, while EVPOME‐A had a more organized stratified epithelium. Ki‐67 and p63 immunolabeled cells in the basal layer of all EVPOMEs suggested a regenerative ability. Compared with native oral mucosa, the keratin 15 and 10/13 expression patterns in all EVPOMEs showed a less‐organized differentiation pattern. In contrast to the β1‐integrin and laminin distribution in EVPOME‐A and native oral mucosa, the subcellular deposition in EVPOME‐C and EVPOME‐B indicated that complete basement membrane formation failed. These findings demonstrated that C3 has a potential application for epithelial tissue engineering and provides a new potential therapeutic device for oral mucosa regenerative medicine. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2012.