壳多糖-胶原凝胶构建组织工程口腔黏膜固有层

背景：有研究探讨了以胶原/壳聚糖聚合物为支架材料构建组织工程人工皮肤的可行性。 目的：拟探讨以壳多糖-胶原凝胶支架材料与口腔黏膜成纤维细胞在体外构建人工固有层的可行性。 方法：口腔黏膜成纤维细胞取自成年鼠颊部、牙龈或腭部。以2×DMEM培养液、胎牛血清、胶原溶液、壳多糖等或2×DMEM培养液、胎牛血清、胶原溶液等按一定比例配置成凝胶溶液，将大鼠口腔黏膜成纤维细胞与上述凝胶溶液迅速混合制成口腔黏膜人工固有层，动态观察细胞生长状况和凝胶收缩情况。 结果与结论：胶原-口腔黏膜成纤维细胞复合物、壳多糖-胶原-口腔黏膜成纤维细胞复合物体外培养1 d，口腔黏膜成纤维细胞在支架材料中生长良好，细胞呈长梭形，排列无一定方向。随时间推移，口腔黏膜成纤维细胞逐渐增多，凝胶颜色变浅，厚度变薄，分别于2，3 d后发生收缩。结果证实，口腔黏膜成纤维细胞可在壳多糖-胶原凝胶形成的三维空间结构生长并分泌基质，形成类似黏膜固有层的致密结缔组织。壳多糖-胶原凝胶是构建组织工程化口腔黏膜固有层的合适材料。 关键词：口腔黏膜成纤维细胞；胶原；凝胶；壳多糖；组织工程 doi:10.3969/j.issn.1673-8225.2010.25.016

Chitosan and collagen gel constructed tissue-engineered oral mucous propria

BACKGROUND: Studies have explored the feasibility of tissue engineered artificial human skin using chitosan-collagen as scaffold material. OBJECTIVE: To investigate the feasibility of in vitro construction of artificial lamina propria using artificial scaffold of chitosan-collagen and oral mucous fibroblasts (OMF). METHODS: The OMF was extracted from adult rat buccal division, gingival or pars palatalis and cultured in 2XDMEM solution, serum of embryo cattle, collagen solution and chitosan, or 2XDMEM solution, serum of embryo cattle and collagen solution were mixed together into gel solution pro rata. OMFs were added into above gel solution to form oral mucous of artificial lamina propria. OMFs were observed; chitosan-collagen gel substrate and collagen gel substrate were also investigated. RESULTS AND CONCLUSION: After 1 day of collagen-OMF complex (FPCL) and chitosa-collagen-OMF complex (FPCCL) culture, OMF adhered to artificial scaffold and grew in rendered olive-like and unordered. OMFs proliferated with time, and the gel became pale and thin, constrict at second and third days respectively. OMFs may grow and secrete substrate in three-dimensional space formed by chitosan-collagen gel, which forms compact connective tissue similar to oral mucous propria. Chitosan-collagen gel is a suitable material for constructing oral mucous propria by tissue engineering.