黄连素-柚皮苷双重载药复合微球的制备及其抗菌-成骨性能评估研究  

Preparation of berberine-naringin dual drug-loaded composite microspheres and evaluation of their antibacterial-osteogenic properties

作　　者：熊伟 袁灵梅[2] 王梁霞 钱国文 梁超轶 潘斌 郭灵 魏文强 邱勋祥 邓文芳 曾志奎 XIONG Wei;YUAN Lingmei;WANG Liangxia;QIAN Guowen;LIANG Chaoyi;PAN Bin;GUO Ling;WEI Wenqiang;QIU Xunxiang;DENG Wenfang;ZENG Zhikui(Graduate School,Jiangxi University of Traditional Chinese Medicine,Nanchang Jiangxi,330004,P.R.China;Department of Ophthalmology,Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine,Nanchang Jiangxi,330006,P.R.China;Department of Traumatic Orthopedics,Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine,Nanchang Jiangxi,330006,P.R.China;Institute of International Innovation,Jiangxi University of Science and Technology,Nanchang Jiangxi,330013,P.R.China;National Engineering Research Center of Traditional Chinese Medicine Solid Preparation Manufacturing Technology of Jiangxi University of Traditional Chinese Medicine,Nanchang Jiangxi,330004,P.R.China)

机构地区：[1]江西中医药大学研究生院,南昌330004 [2]江西中医药大学附属医院眼科,南昌330006 [3]江西中医药大学附属医院创伤骨科,南昌330006 [4]江西理工大学国际创新学院,南昌330013 [5]江西中医药大学中药固体制剂制造技术国家工程研究中心,南昌330004

出　　处：《中国修复重建外科杂志》2023年第12期1505-1513,共9页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(81960880);江西中医药大学科技创新团队项目(CXTD22009);江西省临床医学中心资助项目(20212BCG74004)。

摘　　要：目的研制一种可同时释放抗菌药物黄连素(berberine,BBR)与促成骨药物柚皮苷(naringin,NG)的载药复合微球,以期用于治疗感染性骨缺损。方法将NG负载于介孔生物玻璃微球(mesoporous bioactive glasses,MBG)获得载药微球(NG-MBG),随后使用聚多巴胺(polydopamine,PDA)对微球进行包裹,以及使用BBR对包裹的PDA涂层进行修饰,获得双重载药复合微球(NG-MBG@PDA-BBR)。取上述制备复合微球,利用扫描电镜、X射线衍射、比表面积及孔容分析仪、傅里叶红外光谱仪进行理化性能表征;测量NG、BBR载药率以及释放量;与金黄色葡萄球菌、大肠杆菌共培养12 h后对菌落计数并计算抑菌率,观察其抗菌性能;与大鼠BMSCs共培养,24、72 h后通过活/死细胞荧光染色与细胞计数试剂盒8法评估生物相容性,7、14 d后分别行ALP染色与茜素红染色评估其成骨性能。结果成功构建NG-MBG@PDA-BBR和3种对照微球(MBG、MBG@PDA及NG-MBG@PDA)。扫描电镜观察示,NG-MBG@PDA-BBR呈粗糙片层结构,而MBG微球表面光滑,MBG@PDA和NG-MBG@PDA呈包裹团聚结构;比表面积分析显示MBG存在介孔结构,具备载药潜力;小角度X线衍射示NG成功负载于MBG;而X射线衍射图谱对比示各组微球均呈现非晶态;傅里叶红外光谱示NG-MBG@PDA-BBR中存在NG和BBR特征峰。且NG-MBG@PDA-BBR具有良好药物缓释能力,NG和BBR均呈现早期突释与后期缓释表现。NG-MBG@PDA-BBR能抑制金黄色葡萄球菌及大肠杆菌生长,抗菌能力高于MBG、MBG@PDA及NG-MBG@PDA(P<0.05);4种微球生物相容性在72 h时差异有统计学意义(P<0.05)。ALP和茜素红染色示NG-MBG@PDA-BBR的ALP阳性面积和钙结节数量高于MBG、NGMBG(P<0.05),与NG-MBG@PDA差异无统计学意义(P>0.05)。结论NG-MBG@PDA-BBR对NG与BBR均具有缓释作用,提示其具有理想成骨与抗菌双重性能。Objective To develop a drug-loaded composite microsphere that can simultaneously release the berberine(BBR)and naringin(NG)to repair infectious bone defects.Methods The NG was loaded on mesoporous microspheres(MBG)to obtain the drug-loaded microspheres(NG-MBG).Then the dual drug-loaded compound microspheres(NG-MBG@PDA-BBR)were obtained by wrapping NG-MBG with polydopamine(PDA)and modifying the coated PDA with BBR.The composite microspheres were characterized by scanning electron microscopy,X-ray diffraction,specific surface area and pore volume analyzer,and Fourier transform infrared spectroscopy;the drug loading rate and release of NG and BBR were measured;the colony number was counted and the bacterial inhibition rate was calculated after co-culture with Staphylococcus aureus and Escherichia coli for 12 hours to observe the antibacterial effect;the biocompatibility was evaluated by live/dead cell fluorescence staining and cell counting kit 8 assay after co-culture with rat’s BMSCs for 24 and 72 hours,respectively,and the osteogenic property was evaluated by alkaline phosphatase(ALP)staining and alizarin red staining after 7 and 14 days,respectively.Results NG-MBG@PDA-BBR and three control microspheres(MBG,MBG@PDA,and NG-MBG@PDA)were successfully constructed.Scanning electron microscopy showed that NG-MBG@PDA-BBR had a rough lamellar structure,while MBG had a smooth surface,and MBG@PDA and NG-MBG@PDA had a wrapped agglomeration structure.Specific surface area analysis showed that MBG had a mesoporous structure and had drug-loading potential.Low angle X-ray diffraction showed that NG was successfully loaded on MBG.The X-ray diffraction pattern contrast showed that all groups of microspheres were amorphous.Fourier transform infrared spectroscopy showed that NG and BBR peaks existed in [e-mail]-MBG@PDA-BBR had good sustained drug release ability,and NG and BBR had early burst release and late sustained release.NGMBG@PDA-BBR could inhibit the growth of Staphylococcus aureus and Escherichia coli,and the an

关 键 词：介孔生物玻璃微球 聚多巴胺 黄连素 抗菌性能 成骨性能 

分 类 号：R318.08[医药卫生—生物医学工程]