Zdrehus Razvan-Septimiu, Mocan Teodora, Sabau Lavinia Ioana, Matea Cristian Tudor, Tăbăran Flaviu, Pop Teodora, Delcea Cristian, Mosteanu Ofelia, Mocan Lucian
Nanomedicine Department, Regional Institute of Gastroenterology and Hepatology, "Iuliu Hatieganu" University of Medicine and Pharmacy, 400347 Cluj-Napoca, Romania.
Physiology Department, "Iuliu Hatieganu" University of Medicine and Pharmacy, 400012 Cluj-Napoca, Romania.
Vaccines (Basel). 2025 Jun 21;13(7):668. doi: 10.3390/vaccines13070668.
Gold nanoparticles (AuNPs) offer promising potential as nanocarriers in vaccine development due to their biocompatibility, tunable surface properties and capacity to enhance antigen presentation. This study aimed to evaluate the in vitro cytocompatibility, cellular uptake and antigen processing of carcinoembryonic antigen (CEA)-functionalized AuNPs as a nanovaccine candidate. AuNPs were synthesized by citrate reduction and subsequently functionalized with CEA through physical adsorption. Nanoparticle size, morphology, and surface charge were characterized using UV-Vis spectroscopy, dynamic light scattering (DLS), and transmission electron microscopy (TEM). Cytocompatibility was assessed via MTT assay on RAW 264.7 murine macrophages. Cellular uptake and antigen processing were evaluated using hyperspectral dark-field microscopy and fluorescence microscopy with proteasomal pathway markers. The synthesized AuNPs displayed a uniform spherical morphology with a mean hydrodynamic diameter of ~50 nm and a stable zeta potential. CEA conjugation slightly altered the surface charge and spectral profile. MTT assays confirmed good cytocompatibility across tested concentrations. Hyperspectral and confocal microscopy revealed the efficient uptake of CEA-AuNPs by RAW 264.7 cells and colocalization with lysosomal compartments, suggesting successful antigen processing. The in vitro data support the safety and biological interaction of CEA-functionalized AuNPs with macrophages. These findings highlight their potential as a nanovaccine delivery platform and warrant further in vivo evaluation to assess immunogenicity and protective efficacy.
金纳米颗粒（AuNPs）因其生物相容性、可调节的表面性质以及增强抗原呈递的能力，在疫苗开发中作为纳米载体具有广阔的应用前景。本研究旨在评估癌胚抗原（CEA）功能化的AuNPs作为纳米疫苗候选物的体外细胞相容性、细胞摄取和抗原加工情况。通过柠檬酸盐还原法合成AuNPs，随后通过物理吸附用CEA对其进行功能化。使用紫外可见光谱、动态光散射（DLS）和透射电子显微镜（TEM）对纳米颗粒的大小、形态和表面电荷进行表征。通过MTT法对RAW 264.7小鼠巨噬细胞进行细胞相容性评估。使用高光谱暗场显微镜和带有蛋白酶体途径标记物的荧光显微镜评估细胞摄取和抗原加工情况。合成的AuNPs呈现出均匀的球形形态，平均流体动力学直径约为50 nm，且具有稳定的zeta电位。CEA偶联略微改变了表面电荷和光谱特征。MTT分析证实了在测试浓度范围内具有良好的细胞相容性。高光谱和共聚焦显微镜显示RAW 264.7细胞有效摄取了CEA-AuNPs，并与溶酶体区室共定位，表明抗原加工成功。体外数据支持CEA功能化的AuNPs与巨噬细胞的安全性和生物相互作用。这些发现突出了它们作为纳米疫苗递送平台的潜力，值得进一步进行体内评估以评估免疫原性和保护效果。
