3D打印技术制备聚磷酸钙/淫羊藿苷骨支架诱导骨髓间充质干细胞成骨分化治疗骨缺损  

A 3D-printed calcium polyphosphate/icariin bone scaffold induces osteogenic differentiation of bone marrow mesenchymal stem cells for treating bone defects

作　　者：石永新 逄增金 羊明智[3] 彭立军[3] 李富琴 宋晖 罗福薇[4] Shi Yongxin;Pang Zengjin;Yang Mingzhi;Peng Lijun;Li Fuqin;Song Hui;Luo Fuwei(Shenzhen Hospital,University of Chinese Academy of Sciences,Shenzhen 518106,Guangdong Province,China;Songgang People’s Hospital,Shenzhen 518105,Guangdong Province,China;the First Affiliated Hospital of University of South China,Hengyang 421001,Hunan Province,China;Shenzhen Maternity & Child Healthcare Hospital,Shenzhen 518017,Guangdong Province,China)

机构地区：[1]中国科学院大学深圳医院(光明),广东省深圳市518106 [2]深圳市宝安区松岗人民医院,广东省深圳市518105 [3]南华大学附属第一医院,湖南省衡阳市421001 [4]深圳市妇幼保健院,广东省深圳市518017

出　　处：《中国组织工程研究》2019年第21期3309-3315,共7页Chinese Journal of Tissue Engineering Research

基　　金：2018广东省中医药局建设专项资金立项资助科研课题(20181239);项目负责人:石永新;深圳市宝安区科创局医疗卫生基础研究项目(2017JD071);项目负责人:逄增金~~

摘　　要：背景:随着组织工程学的快速发展,寻找最佳的支架材料、诱导细胞成骨分化的因子以及分化增殖能力强、来源充足的干细胞,成为骨科领域研究的热点。目的:探讨聚磷酸钙/淫羊藿苷骨支架诱导骨髓间充质干细胞成骨分化治疗骨缺损的效果。方法:采用3D打印技术制备聚磷酸钙/淫羊藿苷复合骨支架,分离和培养兔骨髓间充质干细胞。36只新西兰大白兔根据随机数字法分为对照组和实验组,每组18只,均建立股骨髁骨缺损模型并植入聚磷酸钙/淫羊藿苷骨支架复合体,然后实验组大白兔经股静脉注入骨髓间充质干细胞1.5 m L(1×10~9L^(-1)),对照组大白兔经股静脉注入等量生理盐水。术后4,8,12周每组取6只大白兔,分离培养骨髓间充质干细胞,Transwell小室测定骨髓间充质干细胞迁移能力,q RT-PCR法测定骨髓间充质干细胞Ⅰ型胶原蛋白、CD44 mRNA水平;然后取兔股骨髁标本,X射线片观察复合骨支架以及周围骨痂形成情况,Van-Gieson染色观察骨组织学变化,免疫组化染色法测定骨痂神经生长因子水平。结果与结论:①实验组兔骨髓间充质干细胞迁移数均高于对照组(P <0.05);实验组兔骨髓间充质干细胞Ⅰ型胶原蛋白、CD44 mRNA水平均高于对照组(P <0.05);②X射线检查和组织学观察结果显示实验组骨痂形成速度和效果优于对照组;③实验组骨痂神经生长因子水平均高于对照组(P <0.05);④结果表明3D打印技术制备的聚磷酸钙/淫羊藿苷骨支架可诱导骨髓间充质干细胞向成骨细胞分化,在骨缺损治疗中具有较好效果。BACKGROUND:With the rapid development of tissue engineering,seeking for optimal scaffold materials,osteogenic factors and abundant stem cells with strong differentiation and proliferation potential has been an issue of concern in the field of orthopedics.OBJECTIVE:To explore the effect of calcium polyphosphate/icariin bone scaffold to induce osteogenic differentiation of bone marrow mesenchymal stem cells for the treatment of bone defects.METHODS:The CPP/ICA composite bone scaffold was prepared using 3D printing technology.The rabbit bone marrow mesenchymal stem cells were isolated and cultured.Bone defect models were made in 36 New Zealand white rabbits,and equally randomized into control group and experimental group,followed by implantation of calcium polyphosphate/icariin composite bone scaffold.Afterwards,in the experimental group,1.5 mL of bone marrow mesenchymal stem cells(1×10^9/L)was injected through the femoral vein into the rabbits,while the rabbits in the control group were given the same amount of normal saline via the femoral vein.At 4,8 and 12 weeks postoperatively,six rabbits from each group were taken to isolate and culture bone marrow mesenchymal stem cells.Transwell chamber assay was used to measure the migration ability of bone marrow mesenchymal stem cells.Fluorescence quantification-reverse transcriptase polymerase chain reaction was used to determine the type I collagen and CD44 mRNA levels in the cells.Rabbit femoral condyle specimens were taken,and X-ray films were used to observe the formation of epiphyses around the composite scaffold.Van-Gieson staining was used to observe the histological changes of bone tissues.Immunohistochemical staining was used to determine the level of epiphyseal nerve growth factors.RESULTS AND CONCLUSION:The number of migrated bone marrow mesenchymal stem cells was significantly higher in the experimental group than the control group(P<0.05).The type I collagen and CD44 mRNA levels in the cells were also higher in the experimental group than the control group(

关 键 词：股骨髁 骨缺损 聚磷酸钙 淫羊藿苷 复合骨支架 3D打印技术 骨髓间充质干细胞 成骨分化 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]