摘要
背景:研究表明,骨髓干细胞可在损伤脊髓中存活、向损伤部位迁移并向神经元和星形胶质细胞分化,促进损伤脊髓功能的恢复,是治疗脊髓损伤的一条有效途径。自体骨髓干细胞动员移植与手术移植治疗脊髓损伤均具有更广阔的临床应用前景,但两者的疗效与治疗机制是否存在区别还不清楚。目的:比较自体骨髓干细胞动员移植与手术移植治疗脊髓损伤的效果,并以定性定量化指标评价。设计、时间及地点:随机对照动物实验,于2007-06/2008-04在河南省人民医院完成。材料:10周龄SD大鼠90只,雌雄各半,体质量(240±10)g,用于制备脊髓损伤模型。方法:动物造模前注射5-溴2-脱氧尿嘧啶核苷50mg/(kg?d)×3d后抽取自体骨髓,体外分离自体骨髓干细胞;NYUImpactor制作脊髓损伤模型。90只模型大鼠按随机数字表法分为3组,每组30只。动员移植组:应用重组粒细胞刺激因子皮下注射,20mg/(kg?d)×7d;手术移植组为损伤局部移植0.3mL(1×1010L-1)骨髓间充质干细胞;对照组:脊髓损伤后给予相同体积(0.3mL)的生理盐水。各组均从术前3d开始,连续10d腹腔注射5-溴2-脱氧尿嘧啶核苷50mg/(kg?d)。主要观察指标:采用Basso-Beattie-Bresnahan(BBB)评分检测伤后3d,1,2,4,8周大鼠后肢的运动功能;伤后1,2,4,8周通过体感诱发电位和运动诱发电位检测脊髓上、下行神经传导通路,判断脊髓损伤和恢复程度;病理和免疫组织化学观察脊髓损伤组织细胞结构变化及5-溴2-脱氧尿嘧啶核苷、胶质纤维酸性蛋白和神经元特异性烯醇化酶分布表达。结果:①脊髓损伤后1,2,4,8周动员移植组和手术移植组BBB评分均较对照组升高(P<0.01);动员移植组和手术移植组相比,差异均无显著性意义(P>0.05)。②脊髓损伤1,2,4,8周后,与对照组相比,动员移植组和手术移植组体感诱发电位和运动诱发电位潜伏期均降低(P<0.05～0.01),波幅均增高(P<0.05～0.01);动员移植组和手术移植组各时间点相比,差异均无显著性意义(P>0.05)。③组织病理学显示动员移植组和手术移植组较对照组有更少的空洞、坏死及胶质纤维酸性蛋白瘢痕组织,较多的5-溴2-脱氧尿嘧啶核苷阳性细胞和特异性烯醇化酶阳性细胞。结论:自体骨髓干细胞动员移植和手术移植两种方法均能明显减轻脊髓损伤的程度,促进损伤后脊髓功能的恢复,两者对比,前者更为方便、无创,实用性强。
BACKGROUND： The transplanted bone marrow stem cells （BMSCs） survive, migrate to the injury site and differentiate into neurons and astrocytes. Transplantation of BMSCs following spinal cord injury （SCI） may improve the recovery of spinal function and may be an effect way for treating spinal cord injury. The treatment of spinal cord injury by autologous BMSCs mobilization and surgical transplantation has a wide prospect of clinic application. However, it is still unclear whether outcomes and mechanisms of them are different.
OBJECTIVE： To compare the efficacy of treating SCI in rats by mobilization and surgical transplantation of autologous BMSCs, and assess the two methods by qualitative indexes.
DESIGN, TIME AND SETTING： A randomized controlled animal experiment was conducted at the Henan Province People＇s Hospital from June 2007 to April 2008.
MATERIALS： A total of 90 healthy Sprague Dawley rats aged 10 weeks old, half male and half female, weighing （240±10） g, were subjected to make spinal cord injury models. MATHODS： After 3 days injection with bromodeoxyuridine 50 mg/kg per day, BMSCs were isolated from bone marrow of healthy rats. All the rats were underwent spinal cord injury by NYU impactor. All 90 rats were randomly divided into 3 groups, with 30 for each group. Rats in the mobilization group were subcutaneously injected with granulocyte-colony stimulating factor for 7 days, 20 mg/kg per day. In surgical transplantation group, a total of 0.3 mL （1 × 10^10/L） BMSCs were transplanted into injured area of spinal cord. Rats in control group were given the same volume of saline （0.3 mL） into injured area of spinal cord. All the rats were injected with 50 mg/kg bromodeoxyuridine daily at day 3 before surgery for 10 days in each group.
MAIN OUTCOME MEASURES： The Basso-Beattie-Bresnahan （BBB） locomotor score was used to evaluate functional recovery in rat hindlima at 3 days, 1, 2, 4 and 8 weeks after SCI. Motion evoked potential （MEP） and somatosensory evoked potential （SEP） tests were performed to detect the neural pathway so as to evaluate recovery of injured spinal cord. The cell structural changes and the expression and distribution of Brdu, glial fibrillary acidic protein （GFAP）, and neuron specific enolase （NSE） were observed by pathological and immunohistochemical methods. RESULTS： ①BBB score was higher in mobilization group and surgical transplantation group than the control group at 1, 2, 4, and 8 weeks after injury （P 〈 0.05）, but there was no significant difference between the mobilization group and surgical transplantation group （P 〉 0.05）.②The latency period in SEP＆MEP 1, 2, 4, and 8 weeks was decreased and wave amplitude was increased in the mobilization group and surgical transplantation group compared with the control group （P 〈 0.05-0.01）, but there was no significant difference between the mobilization group and surgical transplantation group （P 〉 0.05）. ③ Histopathology showed that there were less empty, necrosis and GFAP-positive glial scar tissue, more Brdu-positive cells and NSE-positive cells in the mobilization group and surgical transplantation group than in the control group.
CONCLUSION： Autologous BMSCs mobilization transplantation and surgery transplantation could significantly reduce the degree of SCI, and promote the recovery of the spinal cord function after injury for treating SCI. Compared between the two, mobilization of autologous BMSCs are more convenient, non-invasive, more likely to seize the opportunity to treatment.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第45期8911-8916,共6页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
河南省科技攻关重点基金(0424410054)
河南省杰出人才计划(084200410011)~~
