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硫酸钡微颗粒被三种不同的细胞类型摄取:HeLa、THP-1 和 hMSC。

Barium sulphate microparticles are taken up by three different cell types: HeLa, THP-1, and hMSC.

机构信息

Inorganic Chemistry and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, Universitätsstr. 5-7, 45117 Essen, Germany.

Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147 Essen, Germany.

出版信息

Acta Biomater. 2023 Jul 1;164:577-587. doi: 10.1016/j.actbio.2023.03.043. Epub 2023 Apr 4.

Abstract

Cytotoxicity and cellular uptake of spherical barium sulphate microparticles (diameter 1 µm) were studied with three different cell lines, i.e. THP-1 cells (monocytes; model for a phagocytosing cell line), HeLa cells (epithelial cells; model for a non-phagocytosing cell line), and human mesenchymal stem cells (hMSCs; model for non-phagocytosing primary cells). Barium sulphate is a chemically and biologically inert solid which allows to distinguish two different processes, e.g. the particle uptake and potential adverse biological reactions. Barium sulphate microparticles were surface-coated by carboxymethylcellulose (CMC) which gave the particles a negative charge. Fluorescence was added by conjugating 6-aminofluorescein to CMC. The cytotoxicity of these microparticles was studied by the MTT test and a live/dead assay. The uptake was visualized by confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). The particle uptake mechanism was quantified by flow cytometry with different endocytosis inhibitors in THP-1 and HeLa cells. The microparticles were easily taken up by all cell types, mostly by phagocytosis and micropinocytosis, within a few hours. STATEMENT OF SIGNIFICANCE: The interaction of particles and cells is of primary importance in nanomedicine, drug delivery, and nanotoxicology. It is commonly assumed that cells take up only nanoparticles unless they are able to phagocytosis. Here, we demonstrate with chemically and biologically inert microparticles of barium sulphate that even non-phagocytosing cells like HeLa and hMSCs take up microparticles to a considerable degree. This has considerable implication in biomaterials science, e.g. in case of abrasive debris and particulate degradation products from implants like endoprostheses.

摘要

细胞毒性和细胞摄取 1 µm 球形硫酸钡微颗粒(直径)用三种不同的细胞系进行了研究,即 THP-1 细胞(单核细胞;吞噬细胞系模型)、HeLa 细胞(上皮细胞;非吞噬细胞系模型)和人骨髓间充质干细胞(hMSCs;非吞噬原代细胞模型)。硫酸钡是一种化学和生物学惰性的固体,可区分两种不同的过程,例如颗粒摄取和潜在的不良生物反应。硫酸钡微颗粒表面涂覆有羧甲基纤维素(CMC),使其带负电荷。通过将 6-氨基荧光素缀合到 CMC 上来添加荧光。通过 MTT 试验和活/死试验研究这些微颗粒的细胞毒性。通过共聚焦激光扫描显微镜(CLSM)和扫描电子显微镜(SEM)观察摄取情况。通过在 THP-1 和 HeLa 细胞中使用不同的内吞作用抑制剂,通过流式细胞术定量测量颗粒摄取机制。这些微颗粒很容易被所有细胞类型摄取,主要是通过吞噬作用和微吞噬作用,在数小时内。重要性陈述:颗粒与细胞的相互作用在纳米医学、药物输送和纳米毒理学中至关重要。通常假设细胞仅摄取纳米颗粒,除非它们能够吞噬。在这里,我们用化学和生物学惰性的硫酸钡微颗粒证明,即使是非吞噬细胞如 HeLa 和 hMSCs 也会在相当程度上摄取微颗粒。这在生物材料科学中具有重要意义,例如在植入物(如假体)的磨料碎屑和颗粒降解产物的情况下。

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