Tissue engineered human oral mucosa has the potential to fill tissue deficits caused by facial trauma or malignant lesion surgery. It can also help elucidate the biology of oral mucosa and serve as an alternative to in vivo testing of oral care products. The aim of this thesis was to construct a tissue engineered full-thickness human oral mucosa closely mimicking the native tissue. To this end, the feasibility of the concept was tested by co-culturing fibroblasts and epithelial cells isolated from normal human oral mucosa biopsies in a collagen-glycosaminoglycan-chitosan scaffold, developed in our laboratory to construct a skin equivalent. An oral mucosal equivalent closely mimicking the native one was obtained and characterized by histology, immunohistochemistry and transmission electron microscopy. Using the same model, the influence of mesenchymal cells on oral epithelial development was investigated by culturing epithelial cells on lamina propria, corneal stroma and dermal equivalents. They were found to significantly influence the thickness and the ultrastructure of the epithelium. Finally, in order to improve the adhesiveness of conventional scaffolds, an elastin-like recombinamer (ELR) containing the cell adhesion tripeptide, RGD, was used in the production of novel bilayer scaffolds employing lyophilization and electrospinning. These scaffolds were characterized by mercury porosimetry, scanning electron microscopy and mechanical testing. In vitro tests revealed positive contribution of ELR on the proliferation of both fibroblasts and epithelial cells. It was thus possible to construct a viable oral mucosa equivalent using the principles of tissue engineering.

L’ingénierie de la muqueuse orale humaine (MOH) a pour but le comblement des pertes de substances suite à un traumatisme facial ou à la chirurgie des lésions malignes. Elle a aussi des applications en recherche pour élucider les mécanismes biologiques de la MO et en pharmacotoxicologie comme alternative à l’expérimentation animale. L'objectif de cette thèse était de reconstruire une MOH proche du tissu normal. À cette fin, la faisabilité du concept a d'abord été testée par co-culture de fibroblastes de la lamina propria et de cellules épithéliales de MOH dans le substrat de collagène-chitosan glycosaminoglycane, développé pour la production de peaux reconstruites. La caractérisation de la MOH reconstruite par histologie, immunohistochimie et microscopie électronique à transmission a montré la présence d’une LP équivalente avec un épithélium pluristratifié et non kératinisé très proche du tissu d’origine. Grâce à ce modèle, nous avons ensuite démontré que l’origine des fibroblastes (MO, cornée, peau) influence significativement l’épaisseur et l’ultrastructure de l'épithélium obtenu par culture de cellules épithéliales orales. Enfin, afin d'améliorer les propriétés adhésives du substrat à base collagène, nous avons ajouté au collagène, une élastine-like recombinante (ELR) contenant le tri-peptide d’adhésion cellulaire, RGD, et produit un nouveau substrat bicouche, poreux par lyophilisation et recouvert d’une couche fibreuse par électrofilage. Ces substrats ont été caractérisés par porosimétrie au mercure, microscopie électronique à balayage et essais mécaniques. Nous avons démontré l’effet stimulant de ELR sur la prolifération des fibroblastes et des cellules épithéliales / Tissue engineered human oral mucosa has the potential to fill tissue deficits caused by facial trauma or malignant lesion surgery. It can also help elucidate the biology of oral mucosa and serve as an alternative to in vivo testing of oral care products. The aim of this thesis was to construct a tissue engineered full-thickness human oral mucosa closely mimicking the native tissue. To this end, the feasibility of the concept was tested by co-culturing fibroblasts and epithelial cells isolated from normal human oral mucosa biopsies in a collagen-glycosaminoglycan-chitosan scaffold, developed in our laboratory to construct a skin equivalent. An oral mucosal equivalent closely mimicking the native one was obtained and characterized by histology, immunohistochemistry and transmission electron microscopy. Using the same model, the influence of mesenchymal cells on oral epithelial development was investigated by culturing epithelial cells on lamina propria, corneal stroma and dermal equivalents. They were found to significantly influence the thickness and the ultrastructure of the epithelium. Finally, in order to improve the adhesiveness of conventional scaffolds, an elastin-like recombinamer (ELR) containing the cell adhesion tripeptide, RGD, was used in the production of novel bilayer scaffolds employing lyophilization and electrospinning. These scaffolds were characterized by mercury porosimetry, scanning electron microscopy and mechanical testing. In vitro tests revealed positive contribution of ELR on the proliferation of both fibroblasts and epithelial cells. It was thus possible to construct a viable oral mucosa equivalent using the principles of tissue engineering