鹿瓜多肽及骨髓基质干细胞经股动脉注入治疗骨折不愈合的实验研究  

Experimental study on the treating nonunion of bone by femoral artery injection of cervus and cucumis polypeptide combined with transfected green fluorescent bone marrow stromal cells

作　　者：蔡俊[1] 郑光磊 沈晓强[1] 顾晓民[1] CAI Jun;ZHENG Guanglei;SHEN Xiaoqiang(Department of Orthopaedics,the Affiliated Hospital of Hangzhou Normal University,Hangzhou 310015,China)

机构地区：[1]杭州师范大学附属医院骨科

出　　处：《浙江医学》2019年第22期2384-2388,I0005,共6页Zhejiang Medical Journal

基　　金：杭州市卫生科技计划A类项目(2016A16)

摘　　要：目的观察经股动脉注入鹿瓜多肽和骨髓基质干细胞(BMSC)治疗骨折不愈合的疗效。方法将21只新西兰兔分为对照组(A组)、穿刺组(B组)、经股动脉组(C组)3组,每组7只,锯断单侧股骨造成横行骨折,再将骨折端切除0.5cm,骨折远近端剥除约0.5cm骨膜,固定后形成骨折不愈合模型。术后每周A组经股动脉导管注入0.9%氯化钠注射液10ml,B组经皮穿刺直接在骨折端注入鹿瓜多肽注射液(0.5ml/kg)及转染的荧光BMSC(1×10^6个细胞,0.5ml)混合液,混合液稀释至10ml,C组经股动脉导管注入等量鹿瓜多肽注射液(0.5ml/kg)及转染的荧光BMSC(1×10^6个细胞,0.5ml)混合液,均共7次。术后第1、4、7周行骨痂X线检查及Nordsletten评分评定疗效,并抽取静脉血行碱性磷酸酶(ALP)、骨钙素(OCN)水平检测。第7周处死动物后取骨折端的BMSC,用MATLAB图像分析软件处理各组荧光细胞图像,并计算B、C两组荧光细胞所占面积(△S)/图像总面积(S)比值。结果术后第1、4、7周,C组骨痂生成量均高于A组和B组,骨痂Nordsletten评分除第1周C组和B组差异无统计学意义外(P>0.05),其余C组均高于A组和B组,差异均有统计学意义(均P<0.05);C组ALP、OCN水平均较A、B组明显升高,差异均有统计学意义(均P<0.05);B、C组在荧光显微镜下均可见明显呈活性状态的BMSC,且C组的△S/S比值高于B组,差异有统计学意义(P<0.05)。结论鹿瓜多肽和BMSC经股动脉注入操作简单,疗效安全,能有效促进骨折愈合。Objective To study the effects of cervus and cucumis polypeptide combined with bone marrow stromal cells by femoral artery injection for nonunion of bone.Methods Twenty-one New Zealand rabbits were randomly divided into three groups,were modelled 0.5 cm bone defects in the unilateral femur.Both end of the osteotomy were stripped of 0.5 cm periosteum,and the bone defects were fixed with kirschner wires.Once a week,the group A was injected 0.9%sodium chloride 10 ml by femoral artery;the group B was injected the same amount of cervus and cucumis polypeptide(0.5 ml/kg)combined with transfected green fluorescent BMSC(1×10^6 cells,0.5 ml)by percutaneous puncture;the group C was injected the same amount of cervus and cucumis polypeptide(0.5 ml/kg)combined with transfected green fluorescent BMSC(1×10^6 cells,0.5 ml)by femoral artery.The change of X-ray was evaluated,the changes of the ALP and OCN activity were examined at 1,4 and 7 weeks postoperatively.All rabbits were executed after 7 weeks postoperatively,and were intercepted 1.5 cm bone segments around fracture.Cells were derived from bone marrow.Transfected green fluorescent cells were detected and analysed by fluorescent microscopy.Results At 1,4 and 7 weeks postoperatively,the mean X-ray Nordsletten scores in group C was higher than in group A and B(P<0.05),and the expression of ALP and OCN in group C also increased compared with group A and B(P<0.05).Transfected green fluorescent cells grew well,and more transfected green fluorescent cells aggregation were detected and analysed by fluorescent microscopy in group C(P<0.05).Conclusion Injection of BMSC combined with cervus and cucumis polypeptide by femoral artery is safe treatment pathway,and can promote effectively regeneration of bone defect.

关 键 词：鹿瓜多肽 骨髓基质干细胞 骨折 不愈合 

分 类 号：R68[医药卫生—骨科学]