中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2009年
45期
8911-8916
,共6页
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50 mg/(kg·d)×3 d后抽取自体骨髓,体外分离自体骨髓干细胞;NYU Imlpactor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,20 mg/(kg·d)×7 d:手术移植组为损伤局部移植0.3 mL(1×10~(10)L~(-1))骨髓间充质干细胞;对照组:脊髓损伤后给予相同体积(0.3 mL)的生理盐水.各组均从术前3 d开始,连续10 d腹腔注射5-溴2-脱氧尿嘧啶核苷50 mg/(kg·d).主要观察指标:采用Basso-Beattie-Bresnahan(BBB)评分检测伤后3 d,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.
揹景:研究錶明,骨髓榦細胞可在損傷脊髓中存活、嚮損傷部位遷移併嚮神經元和星形膠質細胞分化,促進損傷脊髓功能的恢複,是治療脊髓損傷的一條有效途徑.自體骨髓榦細胞動員移植與手術移植治療脊髓損傷均具有更廣闊的臨床應用前景,但兩者的療效與治療機製是否存在區彆還不清楚.目的:比較自體骨髓榦細胞動員移植與手術移植治療脊髓損傷的效果,併以定性定量化指標評價.設計、時間及地點:隨機對照動物實驗,于2007-06/2008-04在河南省人民醫院完成.材料:10週齡SD大鼠90隻,雌雄各半,體質量(240±10)g,用于製備脊髓損傷模型.方法:動物造模前註射5-溴2-脫氧尿嘧啶覈苷50 mg/(kg·d)×3 d後抽取自體骨髓,體外分離自體骨髓榦細胞;NYU Imlpactor製作脊髓損傷模型.90隻模型大鼠按隨機數字錶法分為3組,每組30隻.動員移植組:應用重組粒細胞刺激因子皮下註射,20 mg/(kg·d)×7 d:手術移植組為損傷跼部移植0.3 mL(1×10~(10)L~(-1))骨髓間充質榦細胞;對照組:脊髓損傷後給予相同體積(0.3 mL)的生理鹽水.各組均從術前3 d開始,連續10 d腹腔註射5-溴2-脫氧尿嘧啶覈苷50 mg/(kg·d).主要觀察指標:採用Basso-Beattie-Bresnahan(BBB)評分檢測傷後3 d,1,2,4,8週大鼠後肢的運動功能;傷後1,2,4.8週通過體感誘髮電位和運動誘髮電位檢測脊髓上、下行神經傳導通路,判斷脊髓損傷和恢複程度;病理和免疫組織化學觀察脊髓損傷組織細胞結構變化及5-溴2-脫氧尿嘧啶覈苷、膠質纖維痠性蛋白和神經元特異性烯醇化酶分佈錶達.結果:①脊髓損傷後1,2,4,8週動員移植組和手術移植組BBB評分均較對照組升高(P＜0.01);動員移植組和手術移植組相比,差異均無顯著性意義(P＞0.05).②脊髓損傷1,2,4,8週後,與對照組相比,動員移植組和手術移植組體感誘髮電位和運動誘髮電位潛伏期均降低(P＜0.05～0.01),波幅均增高(P＜0.05～0.01);動員移植組和手術移植組各時間點相比,差異均無顯著性意義(P＞0.05).③組織病理學顯示動員移植組和手術移植組較對照組有更少的空洞、壞死及膠質纖維痠性蛋白瘢痕組織,較多的5-溴2-脫氧尿嘧啶覈苷暘性細胞和特異性烯醇化酶暘性細胞.結論:自體骨髓榦細胞動員移植和手術移植兩種方法均能明顯減輕脊髓損傷的程度,促進損傷後脊髓功能的恢複,兩者對比,前者更為方便、無創,實用性彊.
배경:연구표명,골수간세포가재손상척수중존활、향손상부위천이병향신경원화성형효질세포분화,촉진손상척수공능적회복,시치료척수손상적일조유효도경.자체골수간세포동원이식여수술이식치료척수손상균구유경엄활적림상응용전경,단량자적료효여치료궤제시부존재구별환불청초.목적:비교자체골수간세포동원이식여수술이식치료척수손상적효과,병이정성정양화지표평개.설계、시간급지점:수궤대조동물실험,우2007-06/2008-04재하남성인민의원완성.재료:10주령SD대서90지,자웅각반,체질량(240±10)g,용우제비척수손상모형.방법:동물조모전주사5-추2-탈양뇨밀정핵감50 mg/(kg·d)×3 d후추취자체골수,체외분리자체골수간세포;NYU Imlpactor제작척수손상모형.90지모형대서안수궤수자표법분위3조,매조30지.동원이식조:응용중조립세포자격인자피하주사,20 mg/(kg·d)×7 d:수술이식조위손상국부이식0.3 mL(1×10~(10)L~(-1))골수간충질간세포;대조조:척수손상후급여상동체적(0.3 mL)적생리염수.각조균종술전3 d개시,련속10 d복강주사5-추2-탈양뇨밀정핵감50 mg/(kg·d).주요관찰지표:채용Basso-Beattie-Bresnahan(BBB)평분검측상후3 d,1,2,4,8주대서후지적운동공능;상후1,2,4.8주통과체감유발전위화운동유발전위검측척수상、하행신경전도통로,판단척수손상화회복정도;병리화면역조직화학관찰척수손상조직세포결구변화급5-추2-탈양뇨밀정핵감、효질섬유산성단백화신경원특이성희순화매분포표체.결과:①척수손상후1,2,4,8주동원이식조화수술이식조BBB평분균교대조조승고(P＜0.01);동원이식조화수술이식조상비,차이균무현저성의의(P＞0.05).②척수손상1,2,4,8주후,여대조조상비,동원이식조화수술이식조체감유발전위화운동유발전위잠복기균강저(P＜0.05～0.01),파폭균증고(P＜0.05～0.01);동원이식조화수술이식조각시간점상비,차이균무현저성의의(P＞0.05).③조직병이학현시동원이식조화수술이식조교대조조유경소적공동、배사급효질섬유산성단백반흔조직,교다적5-추2-탈양뇨밀정핵감양성세포화특이성희순화매양성세포.결론:자체골수간세포동원이식화수술이식량충방법균능명현감경척수손상적정도,촉진손상후척수공능적회복,량자대비,전자경위방편、무창,실용성강.
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 wees 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 grenulocyte-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-Brasnahan (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 defect 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.and 8 weeks after injury (P ＜ 0.05),but there was no significant difference between the mobilization group and surgical was increased in the mobilization group and surgical transplantation group compared with the control group (P ＜ 0.05-0.01),but 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.
