摘要
趋化因子12[chemokine(C-X-C motif)ligand 12,CXCL12]作为一种化学引诱剂,具有促进干细胞增殖的作用,在干细胞治疗中可用于治疗心肌梗死等疾病。但由于CXCL12蛋白分子结构复杂,半衰期较短,注射给药后作用时间短暂。本试验在保持CXCL12生物活性的前提下,制备其长效微球。首先,将CXCL12溶于含葡聚糖和聚乙二醇(PEG)的混合溶液中,通过冷冻相分离法制备蛋白-葡聚糖颗粒,从而维持蛋白的分子构象。然后,分别通过水包油包固(s/o/w)法和水包油包水(w/o/w)法,以具有生物可降解性的聚乳酸-羟基乙酸共聚物(PLGA)为载体制备载CXCL12-葡聚糖颗粒或CXCL12的微球(F1和F2微球)。结果显示,s/o/w法和w/o/w法制备的微球粒径为(63.17±21.54)和(28.98±10.29)μm,包封率分别为(46.07±1.75)%和(30.86±1.17)%。F1微球中的药物可持续释放超过60 d,且较符合零级释放模型,突释低(3%);而F2微球的突释率高(9%),持续释放时间仅为25 d。通过大鼠骨髓间充质干细胞的增殖试验测定了体外释放过程中蛋白的生物活性,结果F1微球中蛋白的生物活性基本维持在50%以上。
Chemokine(C-X-C motif)ligand 12(CXCL12)is a chemical attractant that can induce mobilization of many cell types,which can be used in stem cell therapy to treat myocardial infarction(MI).Due to the complex molecular structure and short half-life of CXCL12,the effects are short-lived after injection.This study was aimed to prepare CXCL12-loaded long-acting microspheres,which could prolong the persistence of CXCL12 at the site of injection,while maintaining its bioactivity.Firstly,CXCL12 was dissolved in a solution of dextran and polyethylene glycol(PEG)to prepare dextran microparticles loaded with CXCL12 by the method of lyophilized induced phase separation,which could efficiently maintain the molecular conformation of the protein.Two kinds of poly(lactic-coglycolic acid)(PLGA)microspheres,namely F1 and F2 microspheres respectively loaded with CXCL12-dextran microparticles and CXCL12,were prepared by solid-in-oil-in-water(s/o/w)and water-in-oil-in-water(w/o/w)methods,and their physical characterizations were also investigated.The results showed that the average particle size and encapsulation efficiency of F1 microspheres were(63.17±21.54)μm and(46.07±1.75)%,while for F2 microspheres were(28.98±10.29)μm and(30.86±1.17)%.The F1 microspheres had a nearly zero-order release profile with a minimal first-day burst(3%)and released CXCL12 over a 60-day timeframe.While,the first-day burst of F2 microspheres was higher(9%)and CXCL12 was entirely released within 25 d.The rat bone marrow mesenchymal stem cells(BMSCs)proliferation tests were used to determine the bioactivity of the protein during the in vitro release.The bioactivity of CXCL12 released from F1 microspheres was basically over 50%.
作者
杜其然
王猛
吴飞
金拓
DU Qiran;WANG Meng;WU Fei;JIN Tuo(School of Pharmacy,Shanghai Jiao Tong University,Shanghai 200240)
出处
《中国医药工业杂志》
CAS
CSCD
北大核心
2020年第8期1002-1009,共8页
Chinese Journal of Pharmaceuticals
