为了评估其作为治疗乳腺癌候选药物的潜力，利用超临界液体技术开发了甲氨蝶呤（MTX）的新型超顺磁性氧化铁纳米颗粒（SPIONs），并采用Box-Behnken设计进行优化。聚（乳酸-缩乙二醇酸）-聚乙二醇400包被的MTX-SPIONs具有500 nm的聚集尺寸和46.8 ± 3.9%的封装效率。傅里叶变换红外光谱分析揭示了主要带的移位，因为分子间氢键，而差示扫描量热法分析揭示了MTX熔化放热峰的缺失，表明完全封装了氧化铁纳米颗粒。Zeta电位结果显示为4.98 mV，而体外释放研究显示在12小时后有35.1 ± 2.78% 的可观释放。使用流式细胞术评估了对照组、MTX和MTX-SPIONs的细胞凋亡，其中MTX-SPIONs的细胞凋亡显著高于对照组和MTX。此外，MTX-SPIONs抑制了细胞分裂和内容物组织，并显着增加了处于G1和G2期的细胞比例，相对于对照组。与单独应用MTX相比，MTX-SPIONs对MCF-7细胞系表现出了持久的抗癌效果，表明SPION传递的化疗药物可能增加细胞毒性。药物稳定性良好，封装药物流失量低，证明基于超临界液体技术的方法是生成药物-聚合物磁性复合纳米颗粒用于癌症治疗的一种有前途的途径。©2023年作者授权，由美国化学学会出版。

Novel superparamagnetic iron oxide nanoparticles (SPIONs) of Methotrexate (MTX) were developed using supercritical liquid technology and optimized with a Box-Behnken design in order to assess its potential as a candidate for the treatment of breast cancer. MTX-SPIONs coated with poly(lactic-co-glycolic acid)-polyethylene glycol 400 had an aggregate size of 500 nm and an encapsulation efficiency of 46.8 ± 3.9%. The Fourier-transformed infrared spectroscopy analysis revealed a shift in the main bands due to intermolecular hydrogen bonds, whereas the differential scanning calorimetry analysis revealed the absence of the MTX melting endotherm, indicating complete encapsulation with oxide nanoparticles. The zeta potential results indicated a value of 4.98 mV, whereas the in vitro release study revealed an initial burst release followed by a considerable release of 35.1 ± 2.78% after 12 h. Using flow cytometry, control, MTX, and MTX-SPIONs were evaluated for apoptosis, with MTX-SPIONs exhibiting greater apoptosis than the control group and MTX. In addition, MTX-SPIONs inhibited cell division and content organization while substantially increasing the proportion of cells in the G1 and G2 phases relative to the control group. MTX-SPIONs exhibited prolonged anticancer effects against MCF-7 cell lines compared to MTX alone, indicating that SPION-delivered chemotherapeutics may increase cytotoxicity. The medication was stable with low encapsulated drug loss, suggesting that the supercritical liquid technology-based method is a promising way for generating drug-polymer magnetic composite nanoparticles for cancer treatment.© 2023 The Authors. Published by American Chemical Society.