Redox-sensitive and hyaluronic acid functionalized liposomes for cytoplasmic drug delivery to osteosarcoma in animal models
Redox-sensitive and hyaluronic acid functionalized liposomes for cytoplasmic drug delivery to osteosarcoma in animal models
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DOI：
10.1016/j.jconrel.2017.06.027
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发表时间：
2017-09-10
影响因子：
10.8
通讯作者：
Wu, Zimei
中科院分区：
文献类型：
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作者：
Chi, Yingying;Yin, Xuelei;Wu, Zimei
本研究旨在开发氧化还原敏感和靶向cd44的脂质体来改善骨肉瘤的化疗。用一种新型的可分离聚乙二醇（PEG2000）通过生物可还原二硫连接剂（胆- ss - mpeg）与胆固醇偶联制备了阳离子脂质体，并对其进行了稳定。透明质酸（HA，分子量20-40 kDa）是CD44的配体，非共价包被在阳离子脂质体上。多柔比星（DOX）作为模型药物在脂质体中被主动加载。研究了HA和choll - ss - mpeg对细胞内药物传递效率和抗肿瘤效果的影响。用傅里叶变换红外光谱和核磁共振（1H NMR）对其结构进行了确证。脂质体choll - ss - mpeg /HA-L的平均直径为165 nm， zeta电位为-28.9 mV，在还原或酸性（pH 5-6）条件下不稳定。DOX的体外释放在生理条件下得到很好的控制，但与非氧化还原敏感脂质体（胆- mpeg / HA-L和胆- mpeg - l）相比，在10 mM谷胱甘肽（GSH）存在下，其释放量达到60%。MTT细胞活力测定显示，与非还原敏感或非ha包被脂质体相比，载药量为15.0% （w/w）的双功能choll - ss - mpeg /HA-L对MG63骨肉瘤细胞具有显著更高的细胞毒性（p < 0.01），这与共聚焦显微镜和流式细胞术的细胞摄取和细胞内运输研究一致。此外，ha包被的gsh响应脂质体优先内化到MG63而不是人肝细胞LO2。在大鼠中，用choll - ss - mpeg或choll - mpeg稳定的脂质体，无论是否有HA，都使DOX的半衰期延长了10倍。在MG63异种移植小鼠模型中，与其他脂质体相比，胆- ss - mpeg /HA-L显示出最有效的肿瘤抑制作用，肝脏摄取最少。与无dox治疗的动物相比，所有用脂质体制剂治疗的动物都存活了下来。总之，易于制备的choll - ss - mpeg /HA-L被证明是一种优秀的cd44介导的细胞内递送系统，具有长循环和gsh触发的胞质药物释放能力。需要进一步的转化和多学科研究，使其对癌症患者真正的临床益处。
This study aimed to develop redox-sensitive and CD44-targeted liposomes to improve chemotherapy of osteosarcoma. Cationic liposomes were prepared and stabilized with a novel detachable polyethylene glycol (PEG2000) conjugated with cholesterol through a bio-reducible disulfide linker (Chol-SS-mPEG). Hyaluronic acid (HA, MW 20-40 kDa), a ligand to CD44, was non-covalently coated on the cationic liposomes. Doxorubicin (DOX) was actively loaded in the liposomes as a model drug. The roles of HA and Chol-SS-mPEG on intracellular drug delivery efficiency, and antitumor efficacy were studied. The structure of Chol-SS-mPEG was confirmed with Fourier-transform infrared and nuclear magnetic resonance (1H NMR). The liposomes, Chol-SS-mPEG/HA-L had a mean diameter of 165 nm, zeta potential -28.9 mV, and destabilized in reducing or acidic (pH 5-6) conditions. In vitro release of DOX was well-controlled at physiological conditions, but a burst release of 60% was observed in the presence of 10 mM glutathione (GSH), in contrast to non-redox sensitive liposomes (Chol-mPEG/ HA-L and Chol-mPEG-L). MTT cell viability assay showed that the dual-functional Chol-SS-mPEG/HA-L with a drug loading of 15.0% (w/w) had significantly higher cytotoxicity to MG63 osteosarcoma cells compared with non-reduction sensitive or non-HA coated liposomes (p < 0.01), consistent with the cellular uptake and intracellular trafficking studies using confocal microscopy and flow cytometry. Furthermore, the HA-coated GSHresponsive liposomes preferentially internalized to MG63 over human liver cells LO2. In rats, liposomes stabilized with either Chol-SS-mPEG or Chol-mPEG, with or without HA, increased the half-life of DOX by> 10-fold. In a MG63 xenograft mouse model, Chol-SS-mPEG/HA-L showed the most effective tumor suppression with minimal uptake by the liver compared with other liposomes. All animals treated with liposomal formulations survived, in contrast to those free-DOX treated. In conclusion, the easily prepared Chol-SS-mPEG/HA-L was demonstrated as an excellent CD44-mediated intracellular delivery system capable of long-circulation and GSHtriggered cytoplasmic drug release. Further translational and multidisciplinary research is required to make it real clinical benefits to cancer patients.