在当前的研究中，我们报道了甲氨蝶呤缀合的氧化锌纳米粒子（MTX-ZnONPs）的合成及其对肺癌细胞的高效作用。 MTX 与 ZnONP 的结合通过紫外可见光谱、动态光散射 (DLS)、傅里叶变换红外 (FTIR) 光谱和透射电子显微镜 (TEM) 进行验证。这种药物纳米缀合物还表现出高载药效率。进一步在体外测试了MTX-ZnONPs对A549细胞的治疗效果，MTT和LDH释放实验结果表明，除游离MTX外，MTX-ZnONPs对A549细胞也能有效发挥细胞毒作用；然而，与游离 MTX 相比，MTX-ZnONPs 的有效性在极低剂量下显着增强。此外，与 MTX-ZnONPs 和 MTX 相比，单独的 ZnONPs 在更高浓度下显着抑制 A549 细胞的细胞活力。此外，所有处理组中 A549 细胞的细胞形态学特征为细胞收缩和从表面脱离。类似地，所有处理组中的 A549 细胞都显示出碎片和浓缩的细胞核，表明细胞凋亡的开始。 A549 细胞中的线粒体膜电位 (ψm) 在所有治疗组中均显示逐渐丧失。定性和定量分析的结果显示 A549 细胞中活性氧 (ROS) 水平增加。 Caspase活性测定结果显示，MTX-ZnONPs和游离MTX引起A549细胞中caspase-9、-8和-3的显着激活；然而，与游离 MTX 相比，MTX-ZnONP 在极低剂量下的作用更为深远。因此，我们的结果显示了 MTX-ZnONP 的高效能，表明 ZnONP 作为纳米载体可以有效地在细胞内递送药物。版权所有 © 2023 Mishra、Ali Ahmad、Al-Keridis、Saeed、Alshammari、Alabdallah、Tiwari、Ahmad、Verma、Fatima 和安萨里。

In the current study, we report the synthesis of methotrexate-conjugated zinc oxide nanoparticles (MTX-ZnONPs) and their high efficacy against lung cancer cells. Conjugation of MTX with ZnONPs was authenticated by UV-vis spectroscopy, dynamic light scattering (DLS), Fourier-transform infrared (FTIR) spectroscopy, and transmission electron microscopy (TEM). This drug-nanoconjugate also showed high drug-loading efficiency. The therapeutic efficacy of MTX-ZnONPs was further tested in vitro against A549 cells, and the results of MTT and LDH release assays showed that MTX-ZnONPs, in addition to free MTX, were efficient in exerting cytotoxic effect on A549 cells; however, the effectiveness of MTX-ZnONPs was found to be considerably enhanced at very low doses compared to that of free MTX. Moreover, ZnONPs alone significantly inhibited the cell viability of A549 cells at a much higher concentration compared to MTX-ZnONPs and MTX. Furthermore, the cytomorphology of A549 cells was characterized by cellular shrinkage and detachment from the surface in all the treatment groups. Similarly, A549 cells, in all the treatment groups, showed fragmented and condensed nuclei, indicating the initiation of apoptosis. Mitochondrial membrane potential (ψm) in A549 cells showed a gradual loss in all the treatment groups. Results of the qualitative and quantitative analyses depicted increased reactive oxygen species (ROS) levels in A549 cells. The results of the caspase activity assay showed that MTX-ZnONPs andfree MTX caused significant activation of caspase-9, -8, and -3 in A549 cells; however, the effect of MTX-ZnONPs was more profound at very low doses compared to that of free MTX. Thus, our results showed high efficacy of MTX-ZnONPs, suggesting efficient intracellular delivery of the drug by ZnONPs as nanocarriers.Copyright © 2023 Mishra, Ali Ahmad, Al-Keridis, Saeed, Alshammari, Alabdallah, Tiwari, Ahmad, Verma, Fatima and Ansari.