Department of Pharmacy, Birla Institute of Technology and Science (BITS PILANI), Pilani Campus, Pilani, Rajasthan, 333031, India.
Sci Rep. 2024 Sep 17;14(1):21669. doi: 10.1038/s41598-024-72226-0.
A novel core-shell nanocarrier system has been designed for co-delivery of a small anticancer drug, docetaxel (DTX) and tumor suppressor (TS) miR-34a named as Exo(PAN). The core is formed by pH dependent polyamine salt aggregates (PSA) containing both the payloads and the shell is formed by RAW 264.7 cell derived exosomal fragments. Herein, phosphate driven polyallylamine hydrochloride (PAH, MW:17,500 Da) PSA was formed in presence of miR-34a and DTX to form PAN. The formulation exhibited pH dependent DTX release with only 33.55 ± 2.12% DTX release at pH 7.2 and 75.21 ± 1.8% DTX release till 144 h at pH 5.5. At 1.21 molar ratio of phosphate to the amine (known as R value), efficient complexation of miR-34a (3.6 μM) in the PAN particles was obtained. PAN demonstrated particle size (163.86 ± 12.89 nm) and zeta-potential value of 17.53 ± 5.10 mV which upon exosomal fragment layering changed to - 7.23 ± 2.75 mV which is similar to the zeta-potential of the exosomal fragments, i.e., - 8.40 ± 1.79 mV. The final formulation Exo(PAN), loaded with 40 ng/mL DTX and 50 nM miR-34a exhibited 48.20 ± 4.59% cytotoxicity in triple negative breast cancer (TNBC) cells, 4T1. Co-localization of CM-DiI (red fluorescence) stained exosomal fragments and FAM-siRNA (green fluorescence) in the cytoplasm of 4T1 cells after 6 h of Exo(PAN) treatment confirmed the efficiency of the designed system to co-deliver two actives. Exo(PAN) also reduced BCL-2 expression (target gene for miR-34a) by 8.98 folds in comparison to free DTX confirming promising co-delivery and apoptosis inducing effect of Exo(PAN) in 4T1.
一种新型的核壳纳米载体系统已被设计用于共递送一种小分子抗癌药物多西紫杉醇（DTX）和肿瘤抑制因子（TS）miR-34a，名为 Exo(PAN)。该核由 pH 依赖性聚胺盐聚集体（PSA）形成，其中包含两种有效载荷，壳由 RAW 264.7 细胞衍生的外泌体片段形成。在此，在 miR-34a 和 DTX 的存在下形成磷酸盐驱动的聚烯丙基盐酸盐（PAH，MW：17500 Da）PSA，以形成 PAN。该制剂表现出 pH 依赖性 DTX 释放，在 pH 7.2 时仅释放 33.55±2.12%的 DTX，在 pH 5.5 时释放 75.21±1.8%的 DTX 直到 144 小时。在磷酸盐与胺的摩尔比为 1.21（称为 R 值）时，可获得有效复合物化 miR-34a（3.6 μM）在 PAN 颗粒中。PAN 表现出粒径（163.86±12.89 nm）和zeta 电位值为 17.53±5.10 mV，在外泌体片段层化后变为 -7.23±2.75 mV，与外泌体片段的 zeta 电位值相似，即-8.40±1.79 mV。最终的制剂 Exo(PAN)，负载 40 ng/mL 的 DTX 和 50 nM 的 miR-34a，在三阴性乳腺癌（TNBC）细胞 4T1 中表现出 48.20±4.59%的细胞毒性。在 Exo(PAN)处理 6 小时后，CM-DiI（红色荧光）染色的外泌体片段和 FAM-siRNA（绿色荧光）在 4T1 细胞的细胞质中的共定位证实了设计的系统共递两种活性物质的效率。与游离 DTX 相比，Exo(PAN)还将 BCL-2 表达（miR-34a 的靶基因）降低了 8.98 倍，证实了 Exo(PAN)在 4T1 中具有有前途的共递和诱导凋亡的效果。
