壳多糖-胶原凝胶对口腔黏膜成纤维细胞生物学行为的影响

Influence of chitosan-collagen gels on the biological behavior of oral mucosa fibroblasts

目的:观察壳多糖-胶原凝胶对口腔黏膜成纤维细胞生物学行为的影响。方法:实验于2003-10/2004-04在中国医科大学第二医院卫生部小儿先天畸形实验室和病毒室完成。①大鼠口腔黏膜成纤维细胞原代培养。②壳多糖-胶原凝胶制作:参照文献方法Plate1的配方在冰浴条件下,先后加入一定量的I型鼠尾胶原,两倍浓缩DMEM、胎牛血清、壳多糖,充分搅拌混匀,用0.1mol/LNaOH,调pH值至7.3左右,1h后凝固。③浸提液的制作:按体积分数为0.1的胎牛血清DMEM液,与壳多糖-胶原凝胶表面积之比为10mL/cm2加入培养液,于4℃下静置24h。④用噻唑蓝比色分析法,在光镜下观察、检测大鼠口腔黏膜成纤维细胞在壳多糖-胶原浸提液中和体积分数为0.1的胎牛血清DMEM液中生长、形态、增殖情况。结果:大鼠口腔黏膜成纤维细胞在壳多糖-胶原浸提液中能较长时间保持增殖活力,与含血清基础培养基中生长良好的大鼠口腔黏膜成纤维细胞,在细胞形态、吸光度值方面相似(P>0.05)。结论:壳多糖-胶原凝胶对口腔黏膜成纤维细胞的生长繁殖无抑制作用。

AIM： To observe the effect of chitosan-collagen gels on the biological behavior of oral mucosa fibroblasts （OMFs）. METHODS： The experiment was performed at the Children Congenital Deformity Laboratory and Virus Room of Department of Health, Second Hospital of China Medical University from October 2003 to April 2004. （1） OMFs of rats were cultured primarily. （2）Preparation of chitosan-collagen gels： Based on the Platel prescription in reference, 1 type collagen was added early or late, twice concentrated DMEM, fetal calf serum, chitosan, mixed evenly under iced bath condition. With 0.1 mol/L NaOH, pH value was adjusted to about 7.3, and conagulated 1 hour later. （3）Preparation of leaching liquor： According to 0.1 volume fraction DMEM liquor of fetal calf serum was moved into culture solution based on 10 mL/cm^2 of chitosan-collagen gel at 4℃ for 24 hours. （4）Under light microscope growth, formation and proliferation of OMFs in chitosan-collagen gel leaching liquor and 0.1 volume fraction fetal calf serum with MTT microassay. RESULTS： OMFs of rats in chitosan-collagen gel leaching liquor could keep proliferation activity as well as those in serum containing medium, no significant difference in morphology and absorbance （A） （P 〉 0.05）. CONCLUSION： Chitosan-collagen gels have no inhibitive effect on the proliferation of OMFs.