柠檬酸铑功能化磁性纳米颗粒上蛋白电晕的形成及其与人巨噬细胞的相互作用

Weskly F Lago, N. V. Carvalho, N. Chaves, M. A. Radicchi, M. H. Sousa, W. Fontes, M. Sousa, S. Báo
{"title":"柠檬酸铑功能化磁性纳米颗粒上蛋白电晕的形成及其与人巨噬细胞的相互作用","authors":"Weskly F Lago, N. V. Carvalho, N. Chaves, M. A. Radicchi, M. H. Sousa, W. Fontes, M. Sousa, S. Báo","doi":"10.35248/2157-7439.19.10.534","DOIUrl":null,"url":null,"abstract":"Nanoscience and Nanotechnology enable innovations for Medicine and for Biomedical Science. Among the current nanoparticles, maghemite associated with rhodium citrate (Magh-RhCit) has been shown to be promising, because it reduces side effects of drugs while maintaining cytotoxicity for tumor cells. However, NPs in contact with biological fluids are immediately coated by proteins (protein corona) that are unique to each nanomaterial. In this study, the adsorption of the most abundant binding proteins was studied in vitro using a three-step analysis: (1) characterization of the magnetic fluid (Magh-RhCit) before and after incubation with serum; (2) identification and physical and biochemical analysis of protein corona; and (3) the cellular internalization of these nanoparticles in human macrophages. Magh-RhCit was initially obtained in the magnetite phase (Fe3O4) via alkaline coprecipitation of Fe2+ and Fe3+ ions and subsequently oxidized to maghemite by the bubbling of oxygen gas in the suspension. Later, rhodium citrate was associated with nanoparticles. Dynamic Light Scattering data were used for characterization of the hydrodynamic diameter and zeta potential. The morphological characterization and measurement of the particles were obtained from Scanning and Transmission Electron Microscopy, and the X-ray Diffraction technique. The identification of the proteins was performed by Liquid Chromatograph coupled to Mass Spectrometry. Human blood serum altered the characteristics of the nanoparticle, making it less polydisperse, larger and with less negative zeta potential, indicating the formation of the protein corona. Forty-nine proteins (which mostly promote opsonization, phagocytosis and endocytosis in cells of the immune system) were identified and characterized: albumin, IgGs, apoliproteins, serpins, complement (C5), kinases, haptoglobin, glycoproteins and transferrin. Nanoparticle characterization and mass spectrometric data of the digested protein corona suggest improved biocompatibility. Moreover, results regarding nanoparticles’ interaction with macrophages suggest that the corona may have profoun implications for in vitro and in vivo extrapolations and will require some consideration in the future.","PeriodicalId":16532,"journal":{"name":"Journal of Nanomedicine & Nanotechnology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Formation of Protein Corona on Rhodium Citrate-Functionalized Magnetic Nanoparticles and Their Interaction with Human Macrophages\",\"authors\":\"Weskly F Lago, N. V. Carvalho, N. Chaves, M. A. Radicchi, M. H. Sousa, W. Fontes, M. Sousa, S. Báo\",\"doi\":\"10.35248/2157-7439.19.10.534\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Nanoscience and Nanotechnology enable innovations for Medicine and for Biomedical Science. Among the current nanoparticles, maghemite associated with rhodium citrate (Magh-RhCit) has been shown to be promising, because it reduces side effects of drugs while maintaining cytotoxicity for tumor cells. However, NPs in contact with biological fluids are immediately coated by proteins (protein corona) that are unique to each nanomaterial. In this study, the adsorption of the most abundant binding proteins was studied in vitro using a three-step analysis: (1) characterization of the magnetic fluid (Magh-RhCit) before and after incubation with serum; (2) identification and physical and biochemical analysis of protein corona; and (3) the cellular internalization of these nanoparticles in human macrophages. Magh-RhCit was initially obtained in the magnetite phase (Fe3O4) via alkaline coprecipitation of Fe2+ and Fe3+ ions and subsequently oxidized to maghemite by the bubbling of oxygen gas in the suspension. Later, rhodium citrate was associated with nanoparticles. Dynamic Light Scattering data were used for characterization of the hydrodynamic diameter and zeta potential. The morphological characterization and measurement of the particles were obtained from Scanning and Transmission Electron Microscopy, and the X-ray Diffraction technique. The identification of the proteins was performed by Liquid Chromatograph coupled to Mass Spectrometry. Human blood serum altered the characteristics of the nanoparticle, making it less polydisperse, larger and with less negative zeta potential, indicating the formation of the protein corona. Forty-nine proteins (which mostly promote opsonization, phagocytosis and endocytosis in cells of the immune system) were identified and characterized: albumin, IgGs, apoliproteins, serpins, complement (C5), kinases, haptoglobin, glycoproteins and transferrin. Nanoparticle characterization and mass spectrometric data of the digested protein corona suggest improved biocompatibility. Moreover, results regarding nanoparticles’ interaction with macrophages suggest that the corona may have profoun implications for in vitro and in vivo extrapolations and will require some consideration in the future.\",\"PeriodicalId\":16532,\"journal\":{\"name\":\"Journal of Nanomedicine & Nanotechnology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2019-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Nanomedicine & Nanotechnology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.35248/2157-7439.19.10.534\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Nanomedicine & Nanotechnology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.35248/2157-7439.19.10.534","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

摘要

纳米科学和纳米技术使医学和生物医学科学的创新成为可能。在目前的纳米颗粒中,与柠檬酸铑相关的磁铁矿(Magh-RhCit)已被证明是有前途的,因为它可以减少药物的副作用,同时保持对肿瘤细胞的细胞毒性。然而,与生物流体接触的NPs立即被每种纳米材料特有的蛋白质(蛋白质冠)包裹。本研究采用三步分析法研究了最丰富的结合蛋白在体外的吸附作用:(1)对血清培养前后的磁流体(Magh-RhCit)进行表征;(2)蛋白冠鉴定及理化分析;(3)这些纳米颗粒在人巨噬细胞中的细胞内化。在磁铁矿相(Fe3O4)中,通过Fe2+和Fe3+离子的碱性共沉淀法获得磁铁矿- rhcit,然后通过悬浮液中氧气的鼓泡氧化为磁铁矿。后来,柠檬酸铑与纳米粒子联系在一起。动态光散射数据用于表征流体动力直径和zeta势。通过扫描电镜、透射电镜和x射线衍射技术对颗粒进行了形态表征和测量。采用液相色谱-质谱联用技术对蛋白质进行鉴定。人血清改变了纳米颗粒的特性,使其分散性降低,体积增大,负zeta电位减小,表明蛋白质冠的形成。鉴定并鉴定了49种蛋白(主要促进免疫系统细胞的调节、吞噬和内吞作用):白蛋白、igg、载脂蛋白、蛇形蛋白、补体(C5)、激酶、接触珠蛋白、糖蛋白和转铁蛋白。消化蛋白冠的纳米粒子表征和质谱数据表明生物相容性得到改善。此外,关于纳米颗粒与巨噬细胞相互作用的结果表明,冠状颗粒可能对体外和体内外推有深远的影响,未来需要一些考虑。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Formation of Protein Corona on Rhodium Citrate-Functionalized Magnetic Nanoparticles and Their Interaction with Human Macrophages
Nanoscience and Nanotechnology enable innovations for Medicine and for Biomedical Science. Among the current nanoparticles, maghemite associated with rhodium citrate (Magh-RhCit) has been shown to be promising, because it reduces side effects of drugs while maintaining cytotoxicity for tumor cells. However, NPs in contact with biological fluids are immediately coated by proteins (protein corona) that are unique to each nanomaterial. In this study, the adsorption of the most abundant binding proteins was studied in vitro using a three-step analysis: (1) characterization of the magnetic fluid (Magh-RhCit) before and after incubation with serum; (2) identification and physical and biochemical analysis of protein corona; and (3) the cellular internalization of these nanoparticles in human macrophages. Magh-RhCit was initially obtained in the magnetite phase (Fe3O4) via alkaline coprecipitation of Fe2+ and Fe3+ ions and subsequently oxidized to maghemite by the bubbling of oxygen gas in the suspension. Later, rhodium citrate was associated with nanoparticles. Dynamic Light Scattering data were used for characterization of the hydrodynamic diameter and zeta potential. The morphological characterization and measurement of the particles were obtained from Scanning and Transmission Electron Microscopy, and the X-ray Diffraction technique. The identification of the proteins was performed by Liquid Chromatograph coupled to Mass Spectrometry. Human blood serum altered the characteristics of the nanoparticle, making it less polydisperse, larger and with less negative zeta potential, indicating the formation of the protein corona. Forty-nine proteins (which mostly promote opsonization, phagocytosis and endocytosis in cells of the immune system) were identified and characterized: albumin, IgGs, apoliproteins, serpins, complement (C5), kinases, haptoglobin, glycoproteins and transferrin. Nanoparticle characterization and mass spectrometric data of the digested protein corona suggest improved biocompatibility. Moreover, results regarding nanoparticles’ interaction with macrophages suggest that the corona may have profoun implications for in vitro and in vivo extrapolations and will require some consideration in the future.
求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信