Ouyang Cuiling, Deng Minxin, Tan Xiaowei, Liu Ziyi, Huang Tuo, Yu Siyu, Ge Zan, Zhang Yafang, Ding Yujun, Chen Hezhang, Chu Hui, Chen Jian
School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan, Hunan 411201, People's Republic of China.
Biomed Mater. 2024 May 30;19(4). doi: 10.1088/1748-605X/ad4e86.
The traditional chemotherapeutic agents' disadvantages such as high toxicity, untargeting and poor water solubility lead to disappointing chemotherapy effects, which restricts its clinical application. In this work, novel size-appropriate and glutathione (GSH)-responsive nano-hydrogels were successfully prepared via the active ester method between chitosan (containing -NH) and cross-linker (containing NHS). Especially, the cross-linker was elaborately designed to possess a disulfide linkage (SS) as well as two terminal NHS groups, namely NHS-SS-NHS. These functionalities endowed chitosan-based cross-linked scaffolds with capabilities for drug loading and delivery, as well as a GSH-responsive mechanism for drug release. The prepared nano-hydrogels demonstrated excellent performance applicable morphology, excellent drug loading efficiency (∼22.5%), suitable size (∼100 nm) and long-term stability. The prepared nano-hydrogels released over 80% doxorubicin (DOX) after incubation in 10 mM GSH while a minimal DOX release less than 25% was tested in normal physiological buffer (pH = 7.4). The unloaded nano-hydrogels did not show any apparent cytotoxicity to A 549 cells. In contrast, DOX-loaded nano-hydrogels exhibited marked anti-tumor activity against A 549 cells, especially in high GSH environment. Finally, through fluorescent imaging and flow cytometry analysis, fluorescein isothiocyanate-labeled nano-hydrogels show obvious specific binding to the GSH high-expressing A549 cells and nonspecific binding to the GSH low-expressing A549 cells. Therefore, with this cross-linking approach, our present finding suggests that cross-linked chitosan nano-hydrogel drug carrier improves the anti-tumor effect of the A 549 cells and may serve as a potential injectable delivery carrier.
传统化疗药物存在高毒性、非靶向性和水溶性差等缺点，导致化疗效果不尽人意，限制了其临床应用。在本研究中，通过壳聚糖（含-NH）与交联剂（含NHS）之间的活性酯法成功制备了尺寸合适且对谷胱甘肽（GSH）有响应的新型纳米水凝胶。特别地，交联剂经过精心设计，具有二硫键（SS）以及两个末端NHS基团，即NHS-SS-NHS。这些功能赋予了基于壳聚糖的交联支架药物负载和递送能力，以及药物释放的GSH响应机制。所制备的纳米水凝胶表现出优异的性能，包括适用的形态、出色的载药效率（约22.5%）、合适的尺寸（约100nm）和长期稳定性。在10mM GSH中孵育后，所制备的纳米水凝胶释放了超过80%的阿霉素（DOX），而在正常生理缓冲液（pH = 7.4）中测试时，DOX的释放量极小，不到25%。未负载药物的纳米水凝胶对A549细胞未表现出明显的细胞毒性。相比之下，负载DOX的纳米水凝胶对A549细胞表现出显著的抗肿瘤活性，尤其是在高GSH环境中。最后，通过荧光成像和流式细胞术分析，异硫氰酸荧光素标记的纳米水凝胶对高表达GSH的A549细胞表现出明显的特异性结合，对低表达GSH的A549细胞表现出非特异性结合。因此，通过这种交联方法，我们目前的研究结果表明，交联壳聚糖纳米水凝胶药物载体提高了对A549细胞的抗肿瘤效果，可能作为一种潜在的可注射递送载体。
