移植BMMSCs-蚕丝蛋白生物支架对急性大鼠脊髓半切损伤修复的影响  

Effects of Combinates Transplantation of Bone Marrow Mesenchymal Stem Cells with Silk Fibroin Porous Scaffolds on Repair of Acute Hemi-transection Spinal Cord Injury in Rats

作　　者：屈涛[1] 王世勇[1] 张军华[1] 文益民[1] 

机构地区：[1]兰州军区兰州总医院脊柱外科,甘肃兰州730050

出　　处：《现代生物医学进展》2013年第20期3818-3822,3839,共6页Progress in Modern Biomedicine

摘　　要：目的：以蚕丝蛋白支架（silk fibroin porous scaffolds SFPS）接种骨髓基质干细胞（bone marrow mesenchymal stem cells,BMMSCs）移植入SD大鼠脊髓半切损伤模型内,观察BMMSCs-SFPS复合生物支架对损伤脊髓的修复作用。方法：密度梯度离心法提取、贴壁法培养BMMSCs,取第三代对数生长期细胞,采用注射法制备BMMSCs-SFPS复合支架,复合14天进行生物相容性检测。40只SD大鼠复制脊髓半切损伤模型后随机分配为四组（n=10）：A组BMMSCs-SFPS联合移植、B组单独移植BMMSCs、C组单独移植SFPS、D组为空白对照组,移植后分别于1、2、3、4周进行运动功能评分和术后4周进行HE染色观察、免疫荧光检测。结果：BMMSCs-SFPS复合支架体外培养14天后,扫描电镜可见BMMSCs附于SFPS支架内表面生长,细胞贴附良好并互有接触。移植入脊髓半切损伤模型后4周进行HE染色,结果显示A组脊髓空洞较其余三组小,免疫荧光检测结果示A组NF200、Nestin阳性表达高于B、C、D组,A组GFAP表达则明显低于其余三组。A组术后2~4周Basso-Beattie-Bresnahan（BBB）评分均高于同期B、C、D组,比较差异有统计学意义（P〈0.01）,D组评分明显低于同期其他3组,差异有统计学意义（P〈0.05）。结论：BMMSCs-SPFS具有良好生物相容性,复合支架保证BMMSCs的存活数量、能抑制胶质瘢痕。BMMSCS-SFPS复合生物支架能发挥协同作用,促进脊髓半切损伤的大鼠运动功能恢复。Objective： To investigate the effects of transplantation of BMMSCs-SFPS on repairing acute hemi- transection SCI in rats. Methods： BMMSCs were harvested by density gradient centrifugation and adherent-cultured. The third generation BMMSCs in logarithmic growth phase were injected into SFPS to construct the BMMSCs-SFPS scaffolds, which biocompatibility was observed in vitro at 2 weeks ,40 SD adult rats were subjected hemi-transect at T11 level to build SCI model then divided randomly into 4 groups （each group, n=10）, BMMSCs-SFPS, BMMSCs and SFPS that were implanted in group A, B, and C, respectively. Group D was blank control group. At 1, 2, 3 and 4 weeks after surgery, the functional recovery of the hind limbs was evaluated by the Basso-Beattie- Bresnahan （BBB） locomotors rating score, And HE staining and immun-fluorescence assay were adopted at one month after surgery. Results： Abundance BMMSC has been confirmed adhering to the internal surface of scaffolds by SEM after co-cultivated 14 days, cells attached well to each other. After the surgery ofhemi- transection 1 month, HE staining showed that the cavity of spinal marrow in group A was smaller than that in group B, C and D. Immunofluorescence staining showed that more neurofilament 200 （NF-200） positive fibers and Nestin positive cells were detected in group A, but GFAP positive cells significantly less than that in other three groups. The BBB scores at 2, 3 and 4 weeks after surgery in group A was Top Rating than those in other 3 groups, and group D was the lowest at each test time, and the differences between groups were statistically significant （P 〈 0.01）. Conclusions： SFPS which can be biocompatible with BMMSCs in vitro, BMMSCs-SFPS scaffolds can ensure the number of survivalling BMMSCs and inhibit glial scar after transplanted into spinal cord, Synergistic effect of co-transplant BMMSCs-SFPS scaffolds can promote SCI rat motor function recover.

关 键 词：骨髓基质干细胞 蚕丝蛋白支架 联合移植 急性脊髓损伤 

分 类 号：Q953[生物学—动物学] R651.2[医药卫生—外科学]