组织工程化口腔粘膜的构建技术初步研究

目的　探索组织工程化口腔粘膜的构建方法。方法　用 Dispase中性蛋白酶分离提取未长毛的 SD乳鼠硬腭口腔粘膜细胞 ,用角化细胞培养液对口腔粘膜细胞进行培养 ,以自制藻酸钠薄膜作为支架材料构建组织工程化的口腔粘膜。结果　 Dispase中性蛋白酶可较好地分离提纯口腔粘膜细胞 ,用 Dispase消化酶分离获取上皮后 ,再用胰蛋白酶消化上皮为单细胞的最佳时间为 10 m in,过低的细胞密度不易形成克隆 ,鼠口腔粘膜细胞培养的最适密度为 1.5× 10 5/ cm2 。角化细胞培养液能对鼠口腔粘膜细胞进行培养 ,且没有成纤维细胞污染 ;口腔粘膜细胞在藻酸钠薄膜上生长良好。结论　利用角化细胞培养液作为培养基 ,藻酸钠薄膜作为支架材料可成功构建组织工程化口腔粘膜

Preliminary Research for Constructing Tissue-engineered Oral Mucosa

OBJECTIVE: To search for a method of constructing tissue-engineered oral mucosa. METHODS: Hard palate mucoperiosteum were excised and extracted from raw SD milk rat. Tissue-engineered oral mucosa was made with the cultured oral keratinocytes that had been digested by Dispase and cultured in the serum-free keratinocytes medium, the supportive membrane being made from sodium alginate. RESULTS: Rat oral mucosal epithelial cells could be obtained with Dispase digestion. It was found that the best time for oral mucosa membrane to be digested to independent cells by Dispase again is ten minutes after the mucosa membrane has been obtained by Dispase; the best density of rat oral mucosa cells cultured is 1.5 x 10(5)/cm2, and the mucosa cells will be difficult to form the colon if the density of cells is too low. The keratinocytes can be cultured in serum-free keratinocytes medium without fibroblast contamination; the mucosa cells grow well on the sodium alginate membrane. CONCLUSION: Tissue-engineered oral mucosa can be constructed with the cultured oral keratinocytes in the serum-free keratinocytes medium and the self-made sodium alginate membrane.