Development of composite cultured oral mucosa utilizing collagen sponge matrix and contracted collagen gel: a preliminary study for clinical applications.

Abstract

A new type of cultured mucosa was developed as a mucosal substitute. This composite cultured oral mucosa (CCOM) was composed of (1) a lamina propria in which fibroblasts were embedded in contacted collagen gel and honeycomb structured collagen sponge and (2) stratified epithelial cell layers on the surface of the cultured lamina propria. CCOM had a well-stratified and differentiated epithelial cell layer, and its involucrin and laminin expression resembled that of normal oral mucosa. Desmosomes were recognizable with transmission electron microscopic examination. In the lamina propria, contracted collagen gel had pooled away from the sponge wall, leaving a sparse structure inside the collagen sponge. Transplantation of CCOM to nude mice was performed by creating full-thickness wound and then applying CCOM (n = 12). Murine skin allograft (n = 4) and no-graft conditions (n = 5) served as controls. The mice were sacrificed for histological evaluation and assessed for wound contraction 28 days after transplantation. The epithelium of the CCOM-treated group had five to 10 cell layers, and the dermis contained many fibroblasts and a large amount of collagen bundles. The wound contraction of the CCOM-treated group was statistically less than that of the no-graft group. These results indicate that CCOM has barrier functions against various stresses and can induce a fibrovascular ingrowth from the surrounding wound bed, and that CCOM could be applied in a clinical setting.