人脐带间充质干细胞来源外泌体治疗SD大鼠骨质疏松性股骨骨折  

Exosomes derived from human umbilical cord mesenchymal stem cells in treatment of osteoporotic femoral fractures in SD rats

作　　者：卜宪敏[1] 梁迪 张斌[3] 徐英杰 丁豪 吴彬[4] 田荣华 Bu Xianmin;Liang Di;Zhang Bin;Xu Yingjie;Ding Hao;Wu Bin;Tian Ronghua(Department of Pathology,Jining No.1 People’s Hospital,Jining 272067,Shandong Province,China;School of Clinical Medicine,Jining Medical University,Jining 272067,Shandong Province,China;Department of Laboratory,Affiliated Hospital of Jining Medical University,Jining 272067,Shandong Province,China;Department of Orthopedic Trauma,Affiliated Hospital of Jining Medical University,Jining 272067,Shandong Province,China)

机构地区：[1]济宁市第一人民医院病理科,山东省济宁市272067 [2]济宁医学院临床医学院,山东省济宁市272067 [3]济宁医学院附属医院检验科,山东省济宁市272067 [4]济宁医学院附属医院创伤骨科,山东省济宁市272067

出　　处：《中国组织工程研究》2025年第31期6634-6641,共8页Chinese Journal of Tissue Engineering Research

基　　金：济宁医学院高层次科研项目培育计划(JYGC2021FKJ016),项目负责人:吴彬;济宁市重点研发计划基金(2021YXNS029),项目负责人:吴彬;济宁市重点研发计划基金(2022YXNS129),项目负责人:卜宪敏;济宁医学院附属医院主诊医师团队专项临床研究计划(ZZTD-MS-2023-04),项目负责人:吴彬。

摘　　要：背景:人脐带间充质干细胞来源外泌体已广泛用于骨修复和重建研究,能显著促进成骨和血管生成。目的:探究人脐带间充质干细胞来源外泌体治疗骨质疏松性骨折的机制。方法:采用组织块培养法提取人脐带间充质干细胞及超速离心法提取人脐带间充质干细胞来源外泌体并进行鉴定。30只12周龄雌性SD大鼠随机分为假手术组(6只)和去势组(24只)。去势组采用经典的去势法构建骨质疏松模型,术后12周随机选取去势组6只大鼠与假手术组6只大鼠行Micro-CT及苏木精-伊红染色进行模型验证。验证骨质疏松模型构建成功后,将去势组剩余18只大鼠随机分为3组,均构建骨质疏松性股骨骨折模型,骨折端分别注射PBS(PBS组)、1.5×10^(11)particles/mL(低浓度外泌体组)、3×10^(11)particles/mL(高浓度外泌体组)外泌体。骨折术后4周行影像学及组织学观察评估骨折愈合和骨组织血管生成情况。结果与结论:①大体标本观察:与PBS组相比,外泌体组骨折愈合速度明显增快,骨折端骨痂生成量显著增加;②X射线片评分:外泌体组显著高于PBS组,差异有显著性意义(P<0.05);③Micro-CT三维成像:与PBS组相比,外泌体组骨折愈合加快,骨痂生成量显著增加;外泌体组骨体积分数显著高于PBS组,差异有显著性意义(P<0.01);④苏木精-伊红染色和Masson染色:与PBS组相比,外泌体组有更多的骨小梁生成,有更多的新生骨组织生长;⑤免疫组织化学染色:与PBS组相比,外泌体组骨钙素与CD31的表达更加显著;高浓度外泌体组较低浓度外泌体组新生血管密度更大,骨痂生成量更多,骨折愈合增快,呈现浓度依赖性。结果表明,人脐带间充质干细胞来源外泌体可通过促进血管生成和成骨,最终促进骨质疏松性骨折修复,其机制可能与CD31和骨钙素表达增加有关。BACKGROUND:Exosomes derived from human umbilical cord mesenchymal stem cells are widely used for bone repair and reconstruction,significantly enhancing osteogenesis and promoting angiogenesis.OBJECTIVE:To explore the mechanisms of exosomes derived from human umbilical cord mesenchymal stem cells in the treatment of osteoporotic fractures.METHODS:Human umbilical cord mesenchymal stem cells were extracted using tissue block culture method.Exosomes derived from human umbilical cord mesenchymal stem cells were extracted using ultracentrifugation method for identification.Thirty 12-week-old female SD rats were randomly divided into sham-operated group(n=6)and ovariectomized group(n=24).Osteoporosis model was established by castration in the ovariectomized group.12 weeks after modeling,6 rats in the ovariectomized group and 6 rats in the sham-operated group were randomly selected for Micro-CT and hematoxylin-eosin staining to verify the models.After verification,the remaining 18 rats in the ovariectomized group were randomly assigned to three groups to establish osteoporotic fracture models.The fracture end was separately injected with PBS(PBS group),exosomes at 1.5×10^(11) particles/mL(low-concentration exosome group),and 3×10^(11) particles/mL(high-concentration exosome group).Four weeks after operation,fracture healing and bone angiogenesis were evaluated by imaging and histological observations.RESULTS AND CONCLUSION:(1)In the gross specimens,compared with the PBS group,the exosome group had faster fracture healing and more callus formation.(2)The X-ray score of fracture healing in the exosome group was significantly higher than that in the PBS group,and the difference was significant(P<0.05).(3)Micro-CT three-dimensional imaging:Compared with the PBS group,the fracture healing in the exosome group was accelerated and the callus formation was significantly increased;the bone volume fraction in the exosome group was significantly higher than that in the PBS group,and the difference was significant(P<0.01).(4)Hemat

关 键 词：骨质疏松性骨折 人脐带间充质干细胞 外泌体 骨折愈合 骨钙素 CD31 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R683.42