Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Department of Plastic Surgery, National Taiwan University Hospital, Taiwan.
Biomaterials. 2016 Jun;90:12-26. doi: 10.1016/j.biomaterials.2016.03.005. Epub 2016 Mar 3.
Ovarian cancers are the leading cause for mortality among gynecologic malignancies with five-year survival rate less than 30%. The purpose of this study is to develop a redox and pH-sensitive self-assembling hyaluronic acid nanoparticle with active targeting peptide for anticancer drug delivery. Anti-cancer drug is grafted onto hyaluronic acid (HA) via cis-aconityl linkage and disulfide bond to possess pH sensitivity and redox property, respectively. This conjugate is amphiphilic and can self-assemble into nanoparticle (NP) in aqueous solution. The results show that the nanoconjugate is successfully developed and the grafting ratio of cystamine (cys) is 17.8% with drug loading amount about 6.2% calculated by (1)H NMR spectra. The particle size is approximately 229.0 nm using dynamic light scatting measurement, and the morphology of nanoparticles is observed as spherical shape by transmission electron microscope. The pH and redox sensitivities are evaluated by changing either pH value or concentration of dithiothreitol in the medium. It is proved that the drug carrier is capable of achieving sustained controlled release of anti-cancer drug to 95% within 150 h. The intracellular uptake is observed by fluorescent microscope and the images show that conjugating luteinizing hormone-releasing hormone (LHRH) peptide can enhance specific uptake of nanoparticles by OVCAR-3 cancer cells; thus, resulting in inhibitory cell growth to less than 20% in 72 h in vitro. Orthotopic ovarian tumor model is also established to evaluate the therapeutic and diagnostic efficacy using non-invasive in vivo imaging system. The representative results demonstrate that LHRH-conjugated NPs possess a preferable tumor imaging capability and an excellent antitumor ability to almost 30% of original size in 20 days.
卵巢癌是妇科恶性肿瘤死亡的主要原因，五年生存率低于 30%。本研究旨在开发一种具有活性靶向肽的氧化还原和 pH 敏感的自组装透明质酸纳米粒子用于抗癌药物输送。抗癌药物通过顺式丙烯酰基键和二硫键接枝到透明质酸（HA）上，分别具有 pH 敏感性和氧化还原性质。该缀合物具有两亲性，可以在水溶液中自组装成纳米颗粒（NP）。结果表明，成功开发了纳米缀合物，半胱胺（cys）的接枝率为 17.8%，通过（1）H NMR 谱计算药物载药量约为 6.2%。动态光散射测量的粒径约为 229.0nm，透射电子显微镜观察到纳米颗粒的形态为球形。通过改变介质中的 pH 值或二硫苏糖醇浓度来评估 pH 和氧化还原敏感性。证明药物载体能够在 150 h 内实现抗癌药物的持续控释，达到 95%。通过荧光显微镜观察细胞内摄取，图像显示连接黄体生成素释放激素（LHRH）肽可以增强 LHRH 肽对 OVCAR-3 癌细胞的特异性摄取；从而导致细胞生长抑制率在 72 h 内低于 20%。还建立了卵巢肿瘤原位模型，使用非侵入性体内成像系统评估治疗和诊断效果。代表性结果表明，LHRH 缀合的 NPs 具有更好的肿瘤成像能力和极好的抗肿瘤能力，在 20 天内几乎减少到原始大小的 30%。
