The Construction of 3D Printed Oral Mucosa Equivalent: Using a Skin 3D Printing Concept

Abstract

Abstract There are multiple ways to construct oral mucosal tissue via various tissue-engineering techniques. However, although 3D bioprinting has been used in the construction of skin equivalents, to date, there have been no attempts to use 3D bioprinting for oral mucosa equivalent production. Therefore, the aim of this project was to tissue engineer oral mucosa tissue using 3D bioprinting. In this project an Allevi 2 3D printer was used to bioprint a gelatin hydrogel scaffold populated with fibroblast primary cells at different densities. The printed fibroblast/GelMA was then seeded with 1 x 105 cells/ml of keratocytes, cultured submerged for 3 days then incubated at an air-to-liquid interface for a further 14 days. After the incubation period, the tissue engineered oral mucosa was validated against a native oral mucosal tissue via histological and immunohistochemical analysis as well as cell viability assessment. Increased fibroblast density was associated with increased connective tissue thickness; in addition to improved keratocytes stratification. In comparison between native and tissue engineered oral mucosa, the tissue engineered oral mucosa showed a high similarity to the native oral mucosa in terms of immunohistochemical and histological analysis.