Garcia Vinícius Barreto, de Carvalho Thaís Gomes, da Silva Gasparotto Luiz Henrique, da Silva Heloiza Fernanda Oliveira, de Araújo Aurigena Antunes, Guerra Gerlane Coelho Bernardo, Schomann Timo, Cruz Luis J, Chan Alan B, de Araújo Júnior Raimundo Fernandes
Department of Morphology, Federal University of Rio Grande do Norte, Natal, RN, 59072-970, Brazil.
Post-Graduation Programme in Health Science, Federal University of Rio Grande do Norte, Natal, RN, 59072-970, Brazil.
Nanoscale Res Lett. 2019 May 17;14(1):166. doi: 10.1186/s11671-019-2986-y.
For many cancers, early detection is the key to improve survival and reduce the morbidity, which is associated with radical resections due to late diagnosis. Here, we describe the efficiency of primary antibody-conjugated gold nanoparticles (AuNPs) to specifically target chronic inflammatory processes, specially M2 macrophages, in tissue sections of ulcerative colitis (UC) and steatohepatitis in rats which may lead to colorectal cancer and liver carcinoma, respectively. In this study, we demonstrate that AuNPs synthesized by a simple, inexpensive, and environmentally compatible method can be easily conjugated with the antibodies anti-COX-2, anti-MIF, and Alexa Fluor® 488 (ALEXA) to perform immunofluorescence staining in inflamed tissues. Moreover, we showed that primary antibody-conjugated gold nanoparticles (AuNPs) can be used to target M2 macrophages by flow cytometry. We designed three immunofluorescence staining protocols of tissue section with AuNPs for 30 min and overnight incubation, as well as one flow cytometry protocol of M2 macrophage labeling with AuNPs for 30 min. Immunofluorescence and flow cytometry results suggest that conjugation was achieved by direct adsorption of antibodies on the AuNPs surface. When compared to the standard ALEXA protocol in immunofluorescence (IF) and flow cytometry (FC), our 30-min incubation protocol using AuNPs instead of ALEXA decreased from approximately 23 h to 5 h for IF and from 4 h to 1 h for FC, proving to be less laborious, which makes the method eligible for inflammation-induced cancer diagnostic.
对于许多癌症来说,早期检测是提高生存率和降低发病率的关键,而晚期诊断导致的根治性切除会带来发病率。在此,我们描述了初级抗体偶联金纳米颗粒(AuNPs)在溃疡性结肠炎(UC)和大鼠脂肪性肝炎组织切片中特异性靶向慢性炎症过程,特别是M2巨噬细胞的效率,这两种炎症可能分别导致结直肠癌和肝癌。在本研究中,我们证明通过一种简单、廉价且环境兼容的方法合成的AuNPs可以很容易地与抗COX - 2、抗MIF和Alexa Fluor® 488(ALEXA)抗体偶联,以在炎症组织中进行免疫荧光染色。此外,我们表明初级抗体偶联金纳米颗粒(AuNPs)可用于通过流式细胞术靶向M2巨噬细胞。我们设计了三种用AuNPs对组织切片进行30分钟和过夜孵育的免疫荧光染色方案,以及一种用AuNPs对M2巨噬细胞进行30分钟标记的流式细胞术方案。免疫荧光和流式细胞术结果表明,抗体通过直接吸附在AuNPs表面实现偶联。与免疫荧光(IF)和流式细胞术(FC)中的标准ALEXA方案相比,我们使用AuNPs代替ALEXA的30分钟孵育方案使IF从约23小时减少到5小时,FC从4小时减少到1小时,证明工作量更小,这使得该方法适用于炎症诱导癌症的诊断。