Oral Mucosal Reconstruction Using Autologous Cultured Oral Epithelial Sheets

Objectives: Human amniotic membrane (AM), a thin intrauterine placental membrane obtained from multigravida women, has high biocompatibility, anti-inflammatory, anti-scarring and anti-infection properties. Using AM, we developed a novel material for cultivating mucosal epithelial cell sheets. This study investigated autologous transplantation of oral mucosal epithelial cells onto AM and examined the usability of this material for oral mucosal reconstruction.

Methods: The protocol for this experiment was approved by the Kyoto Prefectural University of Medicine ethics committee. Specimens of AM were obtained from women undergoing Caesarean section. Four patients undergoing oral surgery were studied. All four patients gave informed consent to this study. Using oral mucosal biopsy specimens obtained from these patients, oral epithelial cells were cultivated for 2-3 weeks on an AM carrier with 3T3 fibroblast co-culture, then air-lifted. The resultant oral epithelial sheet on AM was transplanted onto the mucosal defects.

Results: After 2-3 weeks in culture, the cultured oral epithelial cells were well-differentiated and showed stratification into 5-7 layers on AM. Immunohistochemistry demonstrated that the cultured oral epithelial cells expressed highly specific mucosal epithelial cell markers (keratins 4/13) and basement membrane proteins (integrina6, laminin 10). The oral mucosal defect was reconstructed with an oral epithelial sheet cultured on AM. Clinically, the oral epithelial sheets had sufficient strength to be handled. After the surgical procedure, the reconstructed sites did not show infection, bleeding, rejection or sheet detachment, and achieved a new oral mucous membrane.

Conclusions: The oral epithelial sheets cultured on AM maintained the properties of mucous membrane and expressed basement membrane proteins. Autologous transplantation was performed using oral epithelial sheet cultured on AM and the transplanted tissue showed adherence to the oral mucosal defect. These findings showed that this novel oral epithelial sheet material is a useful biomaterial for oral mucosal reconstruction.