Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, Iran.
Department of Oral & Maxillofacial Surgery, VU University Medical Center, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands.
Pharm Res. 2017 Dec;34(12):2891-2900. doi: 10.1007/s11095-017-2272-6. Epub 2017 Nov 6.
To employ Doxorubicin-loaded liposomes, modified with YSA-peptide to target EphA2, to reduce adverse effects against primary bone cells and maximize toxicity against Saos-2 osteosarcoma cells.
PEGylated liposomes were prepared by thin film method using Dipalmitoylphosphatidylcholine (DPPC), cholesterol and distearylphosphatidylethanolamine-polyethyleneglycol conjugate (DSPE-mPEG) in 67.9:29.1:3 M ratios, and loaded with DOX (L-DOX) by pH-gradient method. Targeted liposomes (YSA-L-DOX), were prepared by conjugating YSA-peptide to DSPE-mPEG. Liposomes were physicochemically characterized and tested in cellular toxicity assays.
YSA conjugation efficiency was >98%. Size and polydispersity index of both L-DOX and YSA-L-DOX were around 88 nm and 0.188, respectively. Both had similar zeta potential, and 85% DOX loading efficiencies. DOX release kinetics followed the Korsmeyer-Peppa model, and showed comparable release for both formulations from 1-8 h, and a plateau of 29% after 48 h. Both formulations could be stably stored for ≥6 months at 4°C in the dark. Toxicity assays showed a significant 1.91-fold higher cytotoxicity compared to free DOX in the Saos-2 cells, and 2-fold lesser toxicity in primary bone cells compared to the Saos-2 cells. Cellular uptake studies showed higher and more nuclear uptake in YSA-L-DOX compared to L-DOX treated cells.
YSA-L-DOX vesicles might be effective for targeted treatment of osteosarcoma.
使用载多柔比星脂质体，用 YSA-肽进行修饰以针对 EphA2，减少对原代骨细胞的不良反应，并最大限度地提高对 Saos-2 骨肉瘤细胞的毒性。
采用薄膜法制备 PEG 化脂质体，用二棕榈酰磷脂酰胆碱（DPPC）、胆固醇和二硬脂酰基磷脂酰乙醇胺-聚乙二醇缀合物（DSPE-mPEG）以 67.9:29.1:3 M 的比例制备，并通过 pH 梯度法装载 DOX（L-DOX）。通过将 YSA-肽缀合到 DSPE-mPEG 上来制备靶向脂质体（YSA-L-DOX）。对脂质体进行物理化学特性表征，并在细胞毒性测定中进行测试。
YSA 偶联效率>98%。L-DOX 和 YSA-L-DOX 的粒径和多分散指数均约为 88nm 和 0.188。两者具有相似的 Zeta 电位和 85%的 DOX 载药效率。DOX 释放动力学符合 Korsmeyer-Peppa 模型，两种制剂在 1-8 小时内具有相似的释放率，48 小时后达到 29%的平台。两种制剂在 4°C 暗处稳定储存≥6 个月。毒性测定显示，与游离 DOX 相比，Saos-2 细胞的细胞毒性显著提高了 1.91 倍，而与 Saos-2 细胞相比，原代骨细胞的毒性降低了 2 倍。细胞摄取研究表明，与 L-DOX 处理的细胞相比，YSA-L-DOX 处理的细胞具有更高和更多的核摄取。
YSA-L-DOX 囊泡可能是骨肉瘤靶向治疗的有效方法。
