载美洲大蠊提取物蛛丝蛋白膜的制备、表征及体外释药性和细胞毒性考察  

Preparation,characterization,in vitro drug release property and cytotoxicity of Periplaneta americana extract-loaded spider fibroin membrane

作　　者：曾惠娜 卿晨 薛楠楠 杨自忠 巫秀美 李和伟 赵昱 李启艳 ZENG Huina;QING Chen;XUE Nannan;YANG Zizhong;WU Xiumei;LI Hewei;ZHAO Yu;LI Qiyan(Yunnan Provincial Key Laboratory of Entomological Biopharmaceutical R&D,Dali University,Yunnan Dali 671000,China;National-Local Joint Engineering Research Center of Entomoceutics,Dali University,Yunnan Dali 671000,China;Collaborative Innovation Center for the Development and Utilization of Medicinal Insect and Spider Resources in Southwest China,Dali University,Yunnan Dali 671000,China;School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Dept.of Periodontology,the First People’s Hospital of Yunnan Province,Kunming 650032,China)

机构地区：[1]大理大学云南省昆虫生物医药研发重点实验室,云南大理671000 [2]大理大学药用特种昆虫开发国家地方联合工程研究中心,云南大理671000 [3]大理大学中国西南药用昆虫及蛛形类资源开发利用协同创新中心,云南大理671000 [4]南京中医药大学药学院,南京210023 [5]云南省第一人民医院牙周科,昆明650032

出　　处：《中国药房》2023年第2期168-172,211,共6页China Pharmacy

基　　金：云南省科学技术院项目-生物资源数字化开发应用(No.202002AA100007);云南省“万人计划”名医专项(No.YNWR-MY-2018-017);云南省医学领军人才项目(No.L-2017019)。

摘　　要：目的制备一种载美洲大蠊提取物的蛛丝蛋白膜,并对其进行表征及体外释药性和细胞毒性考察。方法以天然敬钊缨毛蛛蛛丝为原料,美洲大蠊提取物为模型药物,采用溶剂浇铸法制备载药蛛丝蛋白膜(后文称“载药膜”),同法制备不含美洲大蠊提取物的载药材料基质蛛丝蛋白膜(后文称“空白膜”)。经静态水接触角、傅里叶红外色谱、X-射线衍射以及扫描电镜等技术对膜剂进行表征,从不同角度分析膜剂结构;通过体外模拟药物在人工唾液中的释放特性评价药物的缓释性能;以MTT法验证载药膜的体外细胞毒性。结果制备得到的载药膜的静态水接触角均小于90°,且小于空白膜;载药膜在1500~1700 cm^(-1)存在美洲大蠊提取物多肽的特征吸收峰;同时X-射线衍射和扫描电镜也证实美洲大蠊提取物已成功负载到膜剂中。该膜剂可在人工唾液中释药时长逾200 min。MTT实验结果显示,空白膜和载药膜作用后的细胞增殖率分别为84.6%、79.4%(均大于70%),无显著的潜在细胞毒性。结论以天然蛛丝所制备的载药膜在人工唾液中具有一定的缓释效果,有望作为一种具优异生物相容性的药物缓释载体应用于临床。OBJECTIVE To prepare spider fibroin membrane loaded with Periplaneta americana extract,and investigate its characterization,in vitro drug release property and cytotoxicity.METHODS Using natural spider silk collected from Chilobrachys guangxiensis as raw material,P.americana extract as model drug,the drug-loaded spider fibroin membrane(hereinafter referred to as drug-loaded membrane)was prepared by solvent casting method.The material matrix spider fibroin membrane without P.americana extract(hereinafter referred to as blank membrane)was prepared with same method.The membrane structure was characterized by static water contact angle,Fourier infrared chromatography,X-ray diffraction and scanning electron microscopy from different angles;drug release characteristics in artificial saliva were simulated in vitro to evaluate the drug sustained-release performance.MTT assay was adopted to validate the cytotoxicity of drug-loaded membrane.RESULTS The drug-loaded membrane was prepared,and the static water contact angle was less than 90°,which was less than that of blank membrane.The drug-loaded membrane showed the characteristic absorption peak to polypeptide of P.americana extract at 1500-1700 cm^(-1).X-ray diffraction and scanning electron microscopy also proved that the drug was successfully loaded into the pellicle.The release time of the pellicle in artificial saliva was more than 200 min.The MTT test results showed that the cell proliferation rates of blank membrane and drug-loaded membrane were 84.6% and 79.4%(both greater than 70%),respectively,without significant potential cytotoxicity.CONCLUSIONS Drug-loaded membrane prepared with natural spider silk has a certain sustained-release effect in artificial saliva,which can be further developed as a drug sustained-release carrier with excellent biological characteristics and biocompatibility.

关 键 词：美洲大蠊 口腔膜剂 天然蛛丝 敬钊缨毛蛛 蛛丝蛋白膜 药物缓释 含量测定 细胞毒性 

分 类 号：R943[医药卫生—药剂学]