关键词: 组织工程, 口腔黏膜, 上皮细胞, 成纤维细胞

Abstract:

objective　To engineer and investigate three-dimensional tissue engineered oral mucosal composite sheets that closely resemble native mucosa. Methods　The tissues sized 1.0 cm×0.5 cm were collected from retromolar trigone while the impacted tooth being extracted in 10 patients. The tissue was explanted with epithelial layer and lamina propria together. The epithelial cells and fibroblasts were dissociated by trypsin digestion method after tissue cells proliferated．The cells were identified with immunofluorescence technique and Hematoxylin-eosin staining．First，1 × 106/mL fibroblasts were seeded onto the acellular dermal matrix（ADM）scaffold. The ADM scaffold containing the fibroblasts was kept in a submerged condition for 7 days with medium. Then the proliferated epithelial cells were co-cultured onto the ADM scaffold containing the proliferated fibroblasts at a density of 1 × 106/mL for an additional 7 days. The construct was raised to the air–liquid interface to induce stratification of keratinocytes by lowering the medium level for 7 days. Overall morphology was assessed by hematoxylin-eosin stain. At the same time，epidermal components were analysed by immunohistochemistry. Results　The newly developed TEM construct had a well-developed，multilayered and stratified squamous epithelium，consisting of a basal，intermediate and superficial layer and resembles native oral mucosa. But the structure of the lamina propria is not clear. Conclusion　Three-dimensional tissue engineered oral mucosal composite sheets which histologically resemble human oral mucosa can be established in vitro.

Key words: tissue engineering, oral mucosa, keratinocyte, fibroblast