Lokesh B Siva, Ajmeera Suresh, Choudhary Rajat, Moharana Sanjaya Kumar, Purohit C S, Konkimalla V Badireenath
School of Biological Sciences, National Institute of Science Education and Research, HBNI, Jatni, Odisha 752050, India.
Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai 400094, India.
J Mater Chem B. 2025 Jan 22;13(4):1437-1458. doi: 10.1039/d4tb01236d.
Tunable redox-sensitive polymeric-lipid hybrid nanocarriers (RS-PLHNCs) were fabricated using homogenization and nanoprecipitation methods. These nanocarriers were composed of novel redox-cholesterol with disulfide linkages and synthesized by conjugating cholesterol with dithiodipropionic acid esterification. Berberine (BBR) was loaded into the fabricated nanocarriers to investigate the selective uptake of BBR by cancer cells as well as its release and enhanced cytotoxicity. The optimized BBR nanocarriers BBR NP-17 and -18 exhibited a spherical shape and uniform distribution, with a particle size of 124.7 ± 1.2 nm and 185.2 ± 1.6 nm and a zeta potential of -5.9 ± 2.5 mV and -20.3 ± 1.1 mV, respectively. These NCs released >80% BBR in a simulated intracellular tumor microenvironment (TME), while only 30%-45% was released under normal physiological conditions. The accelerated drug release in the TME was due to disulfide bond cleavage and ester bond hydrolysis in the presence of GSH and acidic pH, whereas under normal conditions, the NCs remained stable/undissociated. Cellular uptake studies confirmed enhanced BBR uptake in GSH-rich cancer cells (H1975) compared with normal cells (BEAS-2B and HEK293A). Following uptake, compared with the free form of the drug, the optimized nanocarriers displayed significant selective cytotoxicity and apoptosis in cancer cells by notably downregulating anti-oxidant (NFE2L2, HO-1, NQO1, and TXRND1) and anti-apoptotic (MCL-1) genes while upregulating pro-apoptotic genes (PUMA and NOXA). This resulted in increased oxidative stress, thereby inducing selective apoptosis in the GSH-rich lung cancer cells. These results suggest that the synthesized novel NCs hold great potential for specifically delivering drugs to cancer cells (with a reduced environment) while sparing normal cells, thus ensuring safe and efficient cancer therapy.
采用匀化和纳米沉淀法制备了可调节的氧化还原敏感型聚合物-脂质杂化纳米载体（RS-PLHNCs）。这些纳米载体由具有二硫键的新型氧化还原胆固醇组成，通过胆固醇与二硫代二丙酸的酯化反应合成。将小檗碱（BBR）负载到制备的纳米载体中，以研究癌细胞对BBR的选择性摄取及其释放和增强的细胞毒性。优化后的BBR纳米载体BBR NP-17和-18呈球形且分布均匀，粒径分别为124.7±1.2 nm和185.2±1.6 nm，ζ电位分别为-5.9±2.5 mV和-20.3±1.1 mV。这些纳米载体在模拟的细胞内肿瘤微环境（TME）中释放了>80%的BBR，而在正常生理条件下仅释放30%-45%。TME中药物的加速释放是由于在谷胱甘肽（GSH）和酸性pH存在下二硫键断裂和酯键水解，而在正常条件下，纳米载体保持稳定/未解离。细胞摄取研究证实，与正常细胞（BEAS-2B和HEK293A）相比，富含GSH的癌细胞（H1975）对BBR的摄取增强。摄取后，与游离药物形式相比，优化后的纳米载体通过显著下调抗氧化（NFE2L2、HO-1、NQO1和TXRND1）和抗凋亡（MCL-1）基因，同时上调促凋亡基因（PUMA和NOXA），在癌细胞中表现出显著的选择性细胞毒性和凋亡。这导致氧化应激增加，从而在富含GSH的肺癌细胞中诱导选择性凋亡。这些结果表明，合成的新型纳米载体在特异性地将药物递送至癌细胞（处于还原环境）同时保护正常细胞方面具有巨大潜力，从而确保安全有效的癌症治疗。
