碱性成纤维细胞生长因子缓释微球对许旺细胞在小肠粘膜下层细胞粘附活性的影响  

Study on basic fibroblast growth factor controlled release microspheres combined with schwann cells and small intestinal submucosa

作　　者：肖睿[1] 张开伟[2] 李同相[1] 项舟[3] 段宏[3] 舒运兵[1] 

机构地区：[1]宜宾市第一人民医院骨二科,四川宜宾644000 [2]贵阳中医学院第一附属医院 [3]四川大学华西医院

出　　处：《西部医学》2012年第8期1441-1445,共5页Medical Journal of West China

基　　金：四川省宜宾市重点科技计划项目(编号:200803010)

摘　　要：目的观察碱性成纤维细胞生长因子(bFGF)缓释微球对许旺细胞(SC)在小肠粘膜下层(SIS)上的粘附影响。方法运用双酶两步消化法对SC进行体外分离、培养、纯化,将bFGF-PLGA缓释微球(bFGF-PLGA-MS)与SC及SIS进行体外复合培养,并以游离bFGF组及单纯培养液组进行对比,观察bFGF-PLGA缓释微球对SC在SIS上粘附影响。结果运用双酶两步消化的体外原代培养方法获取的雪旺细胞数量较多,细胞纯度可达90%以上。细胞的体外增殖活性良好,细胞增殖周期约为6～7天,培养后2～7天为对数生长期,第7天后进入生长平台期;bFGF缓释微球能持续地促进雪旺细胞在SIS上的粘附、增殖及迁移;细胞增殖周期缩短,细胞能较稳定地保持着良好的活性;提高了细胞增殖指数,并使SIS上的SC持续保持较高的增殖活性;细胞数量与bFGF含量的相关系数为0.988(P=0.02),表明两者间有明显正相关关系。结论 bFGF-PLGA微球的药物缓释促进了SC在SIS上持续而稳定的粘附,为以SC复合SIS构建人工神经提供了有利条件。Objective To observe the effects of basic fibroblast growth factor（bFGF） micropheres on adhesion and proliferation and migration of schwann cells（SC） on scaffold of small intestinal submucosa（SIS）,and the activities of SC proliferation was observed.Methods Schwann cells was isolated,cultured and purified in vitro using two-step enzymatic digestion method.The effects of bFGF-PLGA-MS on adhesion and proliferation and migration of schwann cell（SC） on the scaffolds of SIS in vitro were observed,and the free bFGF group and simple medium group was used as control.Results A larger number of active Schwann cells with the cell purity above 90% were obtained using two-step enzyme digestion method a of primary cells culture in vitro.The good activity of cells in vitro were confirmed.The cell cycle was about 6-7 days and the logarithmic growth phase is 2-7 days after culutre,The bFGF-MS was observed continuously promoting the adhesion and proliferation and migration of Schwann cells on SIS.The cell cycle of SC be shortened by bFGF-MS with a more stable and good activity of cells,according to flow cytometry.The bFGF microspheres increased cell proliferation index,and continuously maintained a higher proliferative activity of SC on SIS.It showed a clear positive correlation between the content of bFGF released from microspheres and cells counts,with the correlation coefficient of 0.988（P=0.02）.Conclusion The drug delivery of bFGF-PLGA-MS promotes Schwann cells sustained and stable adhesion and proliferation on the scaffold of small intestinal submucosa,matched with the drug release rate.Schwann cells are provided to be constructed artificial nerve combined with SIS.

关 键 词：碱性成纤维细胞生长因子 缓释微球 bFGF-PLGA缓释微球 许旺细胞 小肠粘膜下层 

分 类 号：R656.7[医药卫生—外科学]