甲氨蝶呤（MTX）是一种常用于癌症治疗的叶酸拮抗剂，其特点是水溶性差、皮肤渗透性低。这些问题可以通过使用药物输送系统来缓解，例如以其多功能性和独特特性而闻名的功能化金纳米颗粒 (AuNP)。本研究旨在开发用 MTX 功能化的多壳金纳米粒子，以改善 MTX 的抗肿瘤、抗氧化和生物相容性特性。合成了稳定的膦包被的 AuNP，并用定制的聚乙二醇 (PEG) 和短支化聚乙烯亚胺 (PEI) 部分进行功能化，然后进行 MTX 共价结合。通过紫外可见光谱和傅里叶变换红外光谱 (FTIR)、动态光散射 (DLS)、扫描透射电子显微镜 (STEM) 和 X 射线光电子能谱 (XPS) 进行的物理化学表征证实了每个步骤的合成。使用DPPH自由基清除测定、铁离子降低抗氧化能力(FRAP)和铜降低抗氧化能力(CUPRAC)测定来测定功能化AuNPs的抗氧化活性。使用 MTT 和 LDH 测定对 HaCaT 人角质形成细胞和 CAL27 鳞状细胞癌评估生物相容性和细胞毒性。与单个成分相比，MTX 功能化的 AuNPs 表现出增强的抗氧化活性和对肿瘤细胞的显着细胞毒性作用，凸显了它们改善癌症治疗的潜力。

Methotrexate (MTX) is a folic acid antagonist routinely used in cancer treatment, characterized by poor water solubility and low skin permeability. These issues could be mitigated by using drug delivery systems, such as functionalized gold nanoparticles (AuNPs), known for their versatility and unique properties. This study aimed to develop multi-shell AuNPs functionalized with MTX for the improvement of MTX antitumoral, antioxidant, and biocompatibility features. Stable phosphine-coated AuNPs were synthesized and functionalized with tailored polyethylene glycol (PEG) and short-branched polyethyleneimine (PEI) moieties, followed by MTX covalent binding. Physicochemical characterization by UV-vis and Fourier-transform infrared spectroscopy (FTIR) spectroscopy, dynamic light scattering (DLS), scanning transmission electron microscopy (STEM), and X-ray photoelectron spectroscopy (XPS) confirmed the synthesis at each step. The antioxidant activity of functionalized AuNPs was determined using DPPH radical scavenging assay, ferric ions' reducing antioxidant power (FRAP), and cupric reducing antioxidant capacity (CUPRAC) assays. Biocompatibility and cytotoxicity were assessed using MTT and LDH assays on HaCaT human keratinocytes and CAL27 squamous cell carcinoma. MTX functionalized AuNPs demonstrated enhanced antioxidant activity and a pronounced cytotoxic effect on the tumoral cells compared to their individual components, highlighting their potential for improving cancer therapy.