透明质酸温敏水凝胶介导的重组人甲状旁腺激素(1-34)可控释放与细胞生物学研究  

Controlled release and cytobiology of recombinant human parathyroid hormone 1-34 mediated by hyaluronic acid thermo-sensitive hydrogels

作　　者：宣少燕[1] 朱月 张林[1] 唐志华[1] 徐锋 陈滕 汪祖华 XUAN Shao-yan;ZHU Yue;ZHANG Lin;TANG Zhi-hua;XU Feng;CHEN Teng;WANG Zu-hua(Shaoxing People's Hospital,Shaoxing 312000,China;Nano-drug Technology Research Center,College of Pharmaceutical Sciences,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;Guizhou Engineering Technology Research Center for Processing and Preparation of Traditional Chinese Medicine and Ethnic Medicine,Guiyang 550025,China)

机构地区：[1]绍兴市人民医院,浙江绍兴312000 [2]贵州中医药大学药学院,纳米药物技术研究中心,贵州贵阳550025 [3]贵州中药炮制与制剂工程技术研究中心,贵州贵阳550025

出　　处：《药学学报》2022年第3期809-817,共9页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81760643);贵州省基础研究计划资助项目(黔科合基础[2019] 1039号);浙江省医药卫生科技计划资助项目(2018 KY833);贵州省药物新剂型新工艺科技创新人才团队(黔科合平台人才[2017] 5655)。

摘　　要：促成骨剂重组人甲状旁腺激素(1-34)[PTH (1-34)]的半衰期短和生物利用率低。本研究制备了一种可生物降解的透明质酸-聚N-异丙基丙烯酰胺温敏水凝胶(AHA-g-PNIPAAm),并探究其作为药物载体对PTH (1-34)释放和细胞行为的影响。采用核磁共振氢谱和红外光谱对AHA-g-PNIPAAm的结构进行确证;利用物理溶胀法制备载药温敏水凝胶,并进行稳定性考察;采用扫描电镜观察水凝胶的形貌;采用紫外分光光度法考察水凝胶的最低临界转变温度和体外药物释放行为;四甲基偶氮唑盐微量酶反应比色法(MTT法)研究载药温敏水凝胶对小鼠单核巨噬细胞RAW264.7和小鼠颅骨前成骨细胞MC3T3-E1细胞的毒性与增殖作用;抗酒石酸酸性磷酸酶检测法考察载药水凝胶对RAW264.7分化的影响。结果显示,制备的载PTH (1-34)温敏水凝胶呈现规整的三维蜂窝状结构,具有良好的稳定性、温敏性和药物缓控释性能,可有效促进MC3T3-E1细胞的增殖,抑制RAW264.7向破骨细胞的分化。The bone formation promoter recombinant human parathyroid hormone 1-34 [PTH(1-34)] has a short half-life and low bioavailability. In this study, we prepared a biodegradable and temperature-sensitive hyaluronic acid-poly-N-isopropyl acrylamide(AHA-g-PNIPAAm), and further investigated its effects of PTH(1-34) release and cell behavior as drug carrier. The structure of AHA-g-PNIPAAM was confirmed by hydrogen nuclear magnetic resonance spectroscopy and infrared spectroscopy. Next, PTH(1-34) loaded thermo-sensitive hydrogels were prepared by physical swelling method and their stability was investigated. The morphology of hydrogel was observed by scanning electron microscope. The minimum critical transition temperature and drug release behavior of hydrogels were investigated by ultraviolet spectrophotometry. The tetrazolium-based colorimetric assay(MTT assay) was used to investigate the toxicity and proliferation effects of PTH(1-34)-loaded thermo-sensitive hydrogel on mouse mononuclear macrophage RAW264.7 and mouse precranial osteoblasts MC3T3-E1. The effect of PTH(1-34)-loaded thermo-sensitive hydrogel on the differentiation of RAW264.7 was investigated by the tartrateresistant acid phosphatase assay. The results showed that the PTH(1-34)-loaded thermo-sensitive hydrogel prepared in this study displayed regular three-dimensional honeycomb structure, and had good stability, thermo-sensitivity and sustained and controlled release properties, which could promote the proliferation of MC3T3-E1 cells more effectively and inhibit the differentiation of RAW264.7 into osteoclasts.

关 键 词：透明质酸 温敏水凝胶 药物缓控释 破骨细胞 重组人甲状旁腺激素(1-34) 

分 类 号：R943[医药卫生—药剂学]