基于骨组织工程技术比较骨碎补总黄酮两种给药方式修复大鼠大段骨缺损模型的效果  

Comparison of the effects between two routes of total flavones of Rhizoma Drynariae administration on large segmental bone defects in rats based on bone tissue engineering technique

作　　者：申震 郭英 姜自伟[2] 张严 李紫阁 陈泽华 叶翔凌 陈国茜 Shen Zhen;Guo Ying;Jiang Ziwei;Zhang Yan;Li Zige;Chen Zehua;Ye Xiangling;Chen Guoqian(Department of Orthopedics,Third Affiliated Hospital of Yunnan University of Chinese Medicine(Kunming Municipal Hospital of Traditional Chinese Medicine),Kunming 650011,Yunnan Province,China;Department of Orthopedics,First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong Province,China;Department of Orthopedics,Guangdong Second Traditional Chinese Medicine Hospital,Guangzhou 510095,Guangdong Province,China)

机构地区：[1]云南中医药大学第三附属医院(昆明市中医医院)骨伤科,云南省昆明市650011 [2]广州中医药大学第一附属医院骨伤科,广东省广州市510405 [3]广东省第二中医院骨伤科,广东省广州市510095

出　　处：《中国组织工程研究》2022年第27期4346-4352,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81974575),项目负责人:姜自伟。

摘　　要：背景:复合材料支架治疗大段骨缺损已被证实有效,但骨修复缓慢、成骨质量不佳是其主要问题。骨碎补总黄酮可促进成骨、加速骨愈合,但骨碎补总黄酮给药途径依然局限于灌胃形式,局部载药直接作用于骨缺损部位方式的研究仍相对不足。目的:构建骨碎补总黄酮缓释微球/β-磷酸三钙复合支架,观察骨碎补总黄酮局部给药与灌胃两种方式修复大鼠胫骨大段骨缺损的差异。方法:利用3D打印技术构建多孔β-磷酸三钙支架;采用超声乳化溶剂透析法制备骨碎补总黄酮缓释微球;采用冷冻干燥法制备骨碎补总黄酮缓释微球/β-磷酸三钙支架。将40只SD大鼠随机分为4组,应用环形外固定支架构建大鼠胫骨3 mm骨缺损模型,空白组骨缺损处不植入任何材料,空白支架组植入单纯的β-磷酸三钙支架,灌胃支架组植入β-磷酸三钙支架并联合骨碎补总黄酮灌胃处理,载药支架组植入骨碎补总黄酮缓释微球/β-磷酸三钙复合支架,术后8周进行缺损部位影像学检测、组织学染色与免疫组化染色。结果与结论:①X射线片与Micro-CT检测显示,空白组缺损区几乎无骨痂生成;空白支架组可见生成的骨痂连接截骨端,截骨线依稀可见;灌胃支架组截骨线基本消失,可见较多新骨连接截骨两端;载药支架组可见大量骨痂生成,截骨线完全消失,皮质重建良好,髓腔再通;②苏木精-伊红、Masson和番红固绿染色显示,空白组缺损区域内可见少量血管及大量结缔组织填充;空白支架组可见较多骨基质形成,但成熟度不高;灌胃支架组可见大量骨基质形成且成熟度较高;载药支架组截骨缺损间隙内生成大量骨样组织,软骨组织成熟度高,且髓腔再通趋势明显;③免疫组化染色显示,相比于空白支架组和空白组,灌胃支架组与载药支架组的转化生长因子β与骨形态发生蛋白2表达更高(P<0.05),且载药支架组高于灌胃支架组(P<0BACKGROUND:Composite scaffold has been proven to be effective in treating large segmental bone defects.However,slow bone repair and poor osteogenesis remain to be the main problem.Total flavones of Rhizoma Drynariae can promote osteogenesis and accelerate bone healing,but the route of administration is still confined to intragastric administration and the research about local drug delivery into bone defects is still relatively insufficient.OBJECTIVE:To observe the difference between the local and intragastric administration of total flavones of Rhizoma Drynariae for large segmental bone defects by constructing the scaffold composed of total flavones of Rhizoma Drynariae controlled release microsphere-β-tricalcium phosphate.METHODS:Three-dimensional printing technology was utilized to construct porousβ-tricalcium phosphate scaffold.The ultrasonic emulsification solvent dialysis method was used to prepare total flavones of Rhizoma Drynariae sustained-release microspheres.The total flavones of Rhizoma Drynariae sustainedrelease microsphere/β-tricalcium phosphate scaffold was prepared by freeze-drying method.A total of forty SD rats were randomly divided into four groups,and a 3 mm long bone defect model of the tibia was constructed using the circular external fixation.No material was implanted in the bone defect of the blank group,and a simpleβ-tricalcium phosphate scaffold was placed in the blank scaffold group.Aβ-tricalcium phosphate scaffold was implanted in the gavage group that was combined with intragastric administration of total flavones of Rhizoma Drynariae,whereas the scaffold composed of total flavones of Rhizoma Drynariae controlled release microsphere-β-tricalcium phosphate was implanted in the drug loaded group.At 8 weeks after operation,imaging test,histological staining,and immunohistochemical staining were conducted.RESULTS AND CONCLUSION:(1)X-ray films and Micro-CT examination showed that there was almost no callus formation in the defect area of the blank group.The callus generated in the bl

关 键 词：骨碎补总黄酮 Β-磷酸三钙 复合支架 给药方式 大段骨缺损 骨修复 组织工程 

分 类 号：R456[医药卫生—治疗学] R318[医药卫生—临床医学]