摘要
背景:前期研究证实多尺度羟基磷灰石/壳聚糖微管成骨优良,而血管再生在成骨过程中较为关键,探索多尺度微管结构在其成血管中的作用及相关机制意义重大。目的:探索多尺度羟基磷灰石/壳聚糖微管结构对兔骨缺损修复及成血管的影响。方法:构建多尺度羟基磷灰石/壳聚糖微管结构与羟基磷灰石/壳聚糖单体结构,将两种材料分别与骨髓间充质干细胞共培养,21 d后电镜下观察材料对细胞的吸附能力及细胞状态。在48只成年新西兰大白兔桡骨部位制作骨缺损模型,实验组(n=24)植入多尺度羟基磷灰石/壳聚糖微管结构材料,对照组(n=24)植入羟基磷灰石/壳聚糖单体结构材料,术后4,8,12周分别进行放射性核素骨显像、X射线片检查、组织形态学、CD31免疫组化染色及血管内皮生长因子蛋白检测。动物实验得到江西省人民医院伦理委员会批准。结果与结论:(1)电镜下可见,多尺度羟基磷灰石/壳聚糖微管结构材料孔隙相通,具有良好的空间结构,其表面吸附的骨髓间充质干细胞多于羟基磷灰石/壳聚糖单体结构材料。(2)放射性核素骨显像显示,实验组术后各时间点的血池成像均高于对照组(P<0.05)。(3)X射线片显示,实验组各时间点的骨痂形成多于对照组。(4)苏木精-伊红染色显示,实验组术后12周时骨缺损植骨区域内可见大量的新生血管及成骨,对照组术后12周时仅见骨折断端少许再生血管及成骨细胞附着;CD31免疫组化染色显示,实验组各时间点的微血管密度高于对照组(P<0.05)。(5)Western Blot检测显示,实验组各时间点的血管内皮生长因子蛋白表达均高于对照组(P<0.05)。(6)结果表明,多尺度羟基磷灰石/壳聚糖微管结构材料较其单体结构具有更加良好的成骨与成血管能力。
BACKGROUND:Early studies have confirmed that multi-scale hydroxyapatite/chitosan microtubules are excellent in osteogenesis.Angiogenesis is critical in the process of osteogenesis.It is significant to explore the role of tube structure with multi-scale microtubules in its vascularization and related mechanisms.OBJECTIVE:To explore the effect of multi-scale hydroxyapatite/chitosan microtubule structure on rabbit bone defect repair and vascularization.METHODS:Multi-scale hydroxyapatite/chitosan microtubule structure and hydroxyapatite/chitosan monomer structure were constructed and co-cultured with bone marrow mesenchymal stem cells for 21 days.The adsorption ability of the material to cells and the cell state were observed under the electron microscope.Bone defect models were made on the radius of 48 adult New Zealand white rabbits.Rabbits in the experimental group(n=24)were implanted with multi-scale hydroxyapatite/chitosan microtubule structure material.Rabbits in the control group(n=24)were implanted with hydroxyapatite/chitosan monomer structure material.Emission computed tomography,X-ray,histomorphological examination,and immunohistochemistry for CD31 and vascular endothelial growth factor protein detection were performed at 4,8,and 12 weeks after surgery.The animal experiment was approved by the Ethics Committee of Jiangxi Provincial People’s Hospital.RESULTS AND CONCLUSION:(1)Under the electron microscope,the multi-scale hydroxyapatite/chitosan microtubule structure materials were interlinked and had a good spatial structure.The number of bone marrow mesenchymal stem cells on the multi-scale hydroxyapatite/chitosan microtubule structure material was more than that on the hydroxyapatite/chitosan monomer structure material.(2)Emission computed tomography exhibited that the blood pool image was higher in the experimental group than that in the control group at various time points after surgery(P<0.05).(3)X-ray films showed that the formation of multi-scale microtubules was more obvious in the experimental group than that in the control group.(4)Hematoxylin-eosin staining demonstrated that at 12 weeks after surgery,the appearance of osteoblasts and angiogenesis was obvious in the experimental group;only a few regenerated blood vessels and osteoblast attachment were found at the fractured end in the control group.Immunohistochemistry for CD31 exhibited that the microvessel density of the experimental group was higher than that of the control group at each time point(P<0.05).(5)Western blot assay results displayed that the expression of vascular endothelial growth factor protein in the experimental group was higher than that in the control group at each time point(P<0.05).(6)The results showed that the multi-scale hydroxyapatite/chitosan microtubule structure had better osteogenesis and vascularization ability than its monomer structure.
作者
龙智生
熊龙
龚飞鹏
李经堂
曾建华
邓颖
兰敏
孔维豪
陈钢
Long Zhisheng;Xiong Long;Gong Feipeng;Li Jingtang;Zeng Jianhua;Deng Ying;Lan Min;Kong Weihao;Chen Gang(Department of Orthopedics,Jiangxi Provincial People’s Hospital Affiliated to Nanchang University,Nanchang 330006,Jiangxi Province,China;Department of Orthopedics,Second Affiliated Hospital of Nanchang University,Nanchang 330006,Jiangxi Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2022年第34期5436-5441,共6页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金项目(32060222),项目负责人:龙智生
国家自然科学基金项目(81560355),项目负责人:曾建华。
关键词
成骨
成血管
多尺度微管结构
组织工程骨
羟基磷灰石
壳聚糖
人工骨
osteogenesis
angiogenesis
multi-scale microtubule structure
tissue-engineered bone
hydroxyapatite
chitosan
artificial bone
