负载木通皂苷D的纳米羟基磷灰石/壳聚糖支架修复骨缺损  

Nano hydroxyapatite/chitosan scaffold loaded with Akebia saponin D in bone defect repair

作　　者：贠霄[1] 丁童[1] 杨卫强[1] 郭新军[1] Yun Xiao;Ding Tong;Yang Weiqiang;Guo Xinjun(Department of Orthopedics,Xinxiang Central Hospital,Xinxiang 453000,Henan Province,China)

机构地区：[1]新乡市中心医院骨科,河南省新乡市453000

出　　处：《中国组织工程研究》2022年第27期4293-4299,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:木通皂苷D具有促进成骨细胞的增殖与分化、提高成骨细胞活性与数量、促进基质钙化与骨痂生长等诸多作用,主要被用于治疗骨质疏松与促进骨折愈合,将其应用于骨缺损修复的研究较少见。目的:以纳米羟基磷灰石/壳聚糖支架为载体,将包裹木通皂苷D的缓释微球负载于其中,观察其骨缺损修复作用。方法:采用W/O/W方法制作包裹木通皂苷D的缓释微球,采用冷冻干燥方法制备负载包裹木通皂苷D缓释微球的纳米羟基磷灰石/壳聚糖支架(以下简称缓释支架)与单纯的纳米羟基磷灰石/壳聚糖支架(以下简称空白支架),检测缓释微球与缓释支架的体外释药能力。将小鼠来源前成骨细胞MC3T3-E1分别接种于两种支架上,以单独培养的细胞为对照,分析细胞的黏附、增殖与分化情况。在24只成年新西兰大白兔双侧桡骨中段制造1.5 cm的骨缺损,分别植入空白支架与缓释支架,术后4,12周时进行大体观察、Micro-CT扫描影像学检查及组织学观察。结果与结论:①包裹木通皂苷D的缓释微球与缓释支架均具有缓释作用,其中缓释支架的药物释放速率更加平稳、持久;②CCK-8实验显示,缓释支架上的细胞增殖速率快于空白支架、对照组(P<0.05);扫描电镜下可见,小鼠来源前成骨细胞MC3T3-E1覆盖在两组支架表面,其中缓释支架上的细胞数量要多于空白支架;③培养7,14 d时,缓释支架组的碱性磷酸酶活性、Runx2 mRNA表达高于空白支架组(P<0.05);培养第21天时,缓释支架组的骨桥蛋白、骨钙素蛋白表达量高于空白支架组(P<0.05);④影像学检查与组织学观察结果显示,术后4周时,缓释支架组材料周围可见大量的新生骨,其新生骨量明显多于空白支架组;术后12周时,缓释支架内部也可见大量的新生骨长入,空白支架内部仅见少量的新生骨长入,并且缓释支架组的材料残余明显少于空白支架组(P<0.05);⑤结果表明,�BACKGROUND:Akebia saponin D can promote the proliferation and differentiation of osteoblasts,increase the activity and quantity of osteoblasts,and promote matrix calcification and callus growth.Akebia saponin D is mainly used to treat osteoporosis and promote fracture healing,and its application in bone defect repair is rare.OBJECTIVE:To observe bone defects repaired by loading the sustained-release microspheres containing Akebia saponin D onto the nano hydroxyapatite/chitosan scaffold.METHODS:The sustained-release microspheres containing Akebia saponin D were prepared by W/O/W method.The nano hydroxyapatite/chitosan scaffold of sustained-release microspheres containing Akebia saponin D(hereinafter referred to as the sustained-release scaffold)and the simple nano hydroxyapatite/chitosan scaffold(hereinafter referred to as blank scaffold)were prepared by freeze drying method to evaluate the in vitro drug release ability of sustainedrelease microspheres and the scaffold.Mouse-derived preosteoblast MC3T3-E1 was seeded on two kinds of scaffolds to analyze cell adhesion,proliferation,and differentiation.Individually cultured cells were used as controls.A 1.5-cm bone defect was made in the middle of bilateral radius of 24 adult New Zealand white rabbits.Blank scaffold and sustained-release scaffold were implanted,separately.Gross observation,Micro-CT imaging examination,and histological observation were performed at 4 and 12 weeks after operation.RESULTS AND CONCLUSION:(1)Both the sustained-release microspheres and sustained-release scaffolds containing Akebia saponin D had sustained-release effects.The drug release rate of sustained-release scaffold was more stable and lasting.(2)CCK-8 assay demonstrated that the cell proliferation rate on the sustained-release scaffold was significantly faster than that on the blank scaffold and control groups(P<0.05).Under the scanning electron microscope,mouse-derived preosteoblast MC3T3-E1 was covered on the scaffolds of the two groups,and the number of cells on the sustained-rele

关 键 词：木通皂苷D 纳米羟基磷灰石 壳聚糖 支架 骨生物材料 骨缺损修复 骨组织工程 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]