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[成年大鼠骨髓基质细胞移植联合软骨素酶ABC微量注射修复亚急性脊髓挤压伤]

[Repair of subacute spinal cord crush injury by bone marrow stromal cell transplantation and chondroitinase ABC microinjection in adult rats].

作者信息

Zhang Chun, Yao Cong, He Xi-jing, Li Hao-peng

机构信息

Department of Orthopedics, Second Hospital of Xi'an Jiaotong University College of Medicine, Xi'an 710004, China..

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2010 Sep;30(9):2030-5.

Abstract

OBJECTIVE

To investigate the possibility of repairing spinal cord injury by bone marrow stromal cell (MSC) transplantation and microinjection of chondroitinase ABC (ChABC) in adult rats.

METHODS

MSCs isolated from the femoral and tibial bones of new-born Wistar rats were cultured and the cell density was adjusted to 1×10(5)µl before transplantation. SD rats with T8 spinal cord crush injury were divided into 4 groups, namely group A (control) and groups B, C and D with injections of the MSCs, ChABC and MSCs+ChABC at the injury site, respectively. At 0 h, 1 day, 2 days, 3 days, 1 week, and 2 weeks after the injury, the BBB score system was used to evaluate the motion function. At 14 days after the injury, the maximal transverse diameter and actual area of necrosis were evaluated on HE stained sections. GFAP-CS56, GFAP-GAP43 and GFAP-NF160 immunofluorescence double labeling staining were carried out to evaluate the regeneration of the nerve fibers.

RESULTS

At the 14th day after the injury, BBB scores showed significant differences between group A and groups B, C and D (P<0.05), between the groups B and D and between groups C and D, butnot between groups B and C. HE staining showed cavity formation at the injury site in each group, with significant differences between group A and groups B, C and D and also between the latter 3 groups. Immunofluorescence staining revealed more intense expression of GFAP in group A than in the other groups with apparent cavity formation at the lesion site, which was only moderate in groups B and C. The expression of GAP-43 was more intense in group D than in groups B and C. The expression of NF-160 was more intense in group D than in the other 3 groups.

CONCLUSION

The strategy of MSC transplantation combined with ChABC can be effective for repairing spinal cord injury in adult rats.

摘要

目的

探讨成年大鼠骨髓基质细胞(MSC)移植及微量注射软骨素酶ABC(ChABC)修复脊髓损伤的可能性。

方法

从新生Wistar大鼠股骨和胫骨分离出骨髓基质细胞进行培养,移植前将细胞密度调整为1×10(5)µl。将T8脊髓挫伤损伤的SD大鼠分为4组,即A组(对照组)和B、C、D组,分别在损伤部位注射骨髓基质细胞、ChABC以及骨髓基质细胞+ChABC。在损伤后0小时、1天、2天、3天、1周和2周,采用BBB评分系统评估运动功能。损伤后14天,在苏木精-伊红(HE)染色切片上评估坏死的最大横径和实际面积。进行胶质纤维酸性蛋白(GFAP)-CS56、GFAP-生长相关蛋白43(GAP43)和GFAP-神经丝蛋白160(NF160)免疫荧光双标记染色,以评估神经纤维的再生情况。

结果

损伤后第14天,BBB评分显示A组与B、C、D组之间存在显著差异(P<0.05),B组与D组之间以及C组与D组之间也存在显著差异,但B组与C组之间无显著差异。HE染色显示每组损伤部位均形成空洞,A组与B、C、D组之间以及后3组之间存在显著差异。免疫荧光染色显示,A组GFAP表达比其他组更强烈,病变部位形成明显空洞,而B组和C组仅为中度空洞。D组GAP-43的表达比B组和C组更强烈。D组NF-160的表达比其他3组更强烈。

结论

骨髓基质细胞移植联合ChABC策略可有效修复成年大鼠脊髓损伤。

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