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Synthesis of Gold Functionalised Nanoparticles with the Eranthis hyemalis Lectin and Preliminary Toxicological Studies on Caenorhabditis elegans

机译:hy虫凝集素合成金功能化纳米粒子及秀丽隐杆线虫的初步毒理学研究

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摘要

The lectin found in the tubers of the Winter Aconite (Eranthis hyemalis) plant (EHL) is a Type II Ribosome Inactivating Protein (RIP). Type II RIPs have shown anti-cancer properties and have great potential as therapeutic agents. Similarly, colloidal gold nanoparticles are successfully used in biomedical applications as they can be functionalised with ligands with high affinity and specificity for target cells to create therapeutic and imaging agents. Here we present the synthesis and characterization of gold nanoparticles conjugated with EHL and the results of a set of initial assays to establish whether the biological effect of EHL is altered by the conjugation. Gold nanoparticles functionalised with EHL (AuNPs@EHL) were successfully synthesised by bioconjugation with citrate gold nanoparticles (AuNPs@Citrate). The conjugates were analysed by UV-Vis spectroscopy, Dynamic Light Scattering (DLS), Zeta Potential analysis, and Transmission Electron Microscopy (TEM). Results indicate that an optimal functionalisation was achieved with the addition of 100 µL of EHL (concentration 1090 ± 40 µg/mL) over 5 mL of AuNPs (concentration [Au0] = 0.8 mM). Biological assays on the effect of AuNPs@EHL were undertaken on Caenorhabditis elegans, a free-living nematode commonly used for toxicological studies, that has previously been shown to be strongly affected by EHL. Citrate gold nanoparticles did not have any obvious effect on the nematodes. For first larval stage (L1) nematodes, AuNPs@EHL showed a lower biological effect than EHL. For L4 stage, pre-adult nematodes, both EHL alone and AuNPs@EHL delayed the onset of reproduction and reduced fecundity. These assays indicate that EHL can be conjugated to gold nanoparticles and retain elements of biocidal activity.
机译:在冬季乌头(Eranthis hyemalis)植物(EHL)的块茎中发现的凝集素是II型核糖体失活蛋白(RIP)。 II型RIP已显示出抗癌特性,具有作为治疗剂的巨大潜力。类似地,胶体金纳米颗粒可以成功地用于生物医学应用,因为它们可以通过对靶细胞具有高亲和力和特异性的配体进行功能化,从而产生治疗剂和显像剂。在这里,我们介绍了与EHL结合的金纳米颗粒的合成和表征,以及一系列初步测定的结果,以确定EHL的生物学作用是否因结合而改变。通过与柠檬酸金纳米颗粒(AuNPs @ Citrate)生物共轭,成功合成了具有EHL功能的金纳米颗粒(AuNPs @ EHL)。通过UV-Vis光谱,动态光散射(DLS),Zeta电位分析和透射电子显微镜(TEM)分析缀合物。结果表明,在5 mL AuNPs(浓度[Au 0 ] = 0.8 mM)上添加100 µL EHL(浓度1090±40 µg / mL)可获得最佳功能。在秀丽隐杆线虫上进行了AuNPs @ EHL效果的生物学测定,秀丽隐杆线虫是一种通常用于毒理学研究的自由生活的线虫,以前已证明它会受到EHL的强烈影响。柠檬酸金纳米颗粒对线虫没有明显影响。对于第一个幼虫阶段(L1)线虫,AuNPs @ EHL的生物学效应低于EHL。对于L4期,成虫前的线虫,单独的EHL和AuNPs @ EHL都延迟了生殖的开始并降低了繁殖力。这些测定表明EHL可以与金纳米颗粒结合并保留杀生物活性的元素。

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