Department of Pharmaceutical Engineering & Technology, Indian Institute of Technology, Banaras Hindu University, Varanasi, 221005, Uttar Pradesh, India.
AAPS PharmSciTech. 2023 Aug 8;24(6):166. doi: 10.1208/s12249-023-02624-6.
Exosomes are biological nanovesicles that are intrinsically loaded with thousands of biomacromolecules and are principally responsible for cell-to-cell communication. Inspired by the natural payload, they have been extensively investigated as drug delivery vehicles; however, the drug distribution, whether into or onto exosomes, is still debatable. In the present work, we have tried to investigate it systemically by selecting 5-fluorouracil (5-FU) (hydrophilic) and paclitaxel (PAC) (hydrophobic), drugs with very different physicochemical characteristics, for the loading to the exosomes. Exosomes were obtained from bovine milk, and the drugs were loaded using three different methods: incubation, sonication, and triton x-100. The particle size was found to be approximately 100 nm in all the cases; however, the highest drug loading was found in the sonication method. Fluorescence spectrophotometer, EDX analysis, EDX mapping, XPS, and XRD analysis indicated the possible presence of more drugs over the surface in the case of the incubation method. Drugs loaded by the sonication method had more controlled release than simple incubation and triton x-100. The method of drug loading had an insignificant effect on the cytotoxicity while in line with our previous observation, the combination (PAC and 5-FU) exhibited synergism as evidenced by ROS assay, colony formation assay, and mitochondrial membrane potential assay.
外泌体是天然存在的生物纳米囊泡，内部负载着数千种生物大分子，主要负责细胞间的通讯。受其天然载药的启发，外泌体被广泛研究作为药物传递载体；然而，药物的分布（进入或负载到外泌体中）仍然存在争议。在本工作中，我们尝试通过选择具有非常不同理化特性的药物 5-氟尿嘧啶（5-FU）（亲水性）和紫杉醇（PAC）（疏水性）来系统地研究这一问题，将它们负载到外泌体中。外泌体从牛乳中获得，使用三种不同的方法进行药物负载：孵育、超声和 Triton X-100。在所有情况下，粒径均约为 100nm；然而，超声法的药物负载量最高。荧光分光光度计、EDX 分析、EDX 映射、XPS 和 XRD 分析表明，在孵育法中，表面可能存在更多的药物。与简单的孵育和 Triton X-100 相比，超声法负载的药物具有更可控的释放。药物负载方法对细胞毒性的影响不大，但与我们之前的观察一致，联合（PAC 和 5-FU）表现出协同作用，这可以通过 ROS 测定、集落形成测定和线粒体膜电位测定来证明。
