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可生物降解镁颗粒与细胞相互作用的延时评估

Time-Lapse Evaluation of Interactions Between Biodegradable Mg Particles and Cells.

作者信息

Alvarez Florencia, Lozano Puerto Rosa M, Pérez-Maceda Blanca, Grillo Claudia A, Fernández Lorenzo de Mele Mónica

机构信息

1Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas (INIFTA),1900 La Plata,Argentina.

2Cell-Biomaterial Recognition Group,Cellular and Molecular Biology Department,Centro de Investigaciones Biológicas (CIB-CSIC),28040 Madrid,Spain.

出版信息

Microsc Microanal. 2016 Feb;22(1):1-12. doi: 10.1017/S1431927615015597. Epub 2016 Jan 26.

Abstract

Mg-based implants have promising applications as biodegradable materials in medicine for orthopedic, dental, and cardiovascular therapies. During wear and degradation microdebris are released. Time-lapse multidimensional microscopy (MM) is proposed here as a suitable tool to follow, in fixed intervals over 24-h periods, the interaction between cells and particles. Results of MM show interactions of macrophages (J774) with the magnesium particles (MgPa) that led to modifications of cell size and morphology, a decrease in duplication rate, and cell damage. Corrosion products were progressively formed on the surface of the particles and turbulence was generated due to hydrogen development. Changes were more significant after treating MgPa with potassium fluoride. In order to complement MM observations, membrane damage as detected by a lactase dehydrogenase (LDH) assay and mitochondrial activity as detected by a WST-1 assay with macrophages and osteoblasts (MC3T3-E1) were compared. A more significant concentration-dependent effect was detected for macrophages exposed to MgPa than for osteoblasts. Accordingly, complementary data showed that viability and cell cycle seem to be more altered in macrophages. In addition, protein profiles and expression of proteins associated with the adhesion process changed in the presence of MgPa. These studies revealed that time-lapse MM is a helpful tool for monitoring changes of biodegradable materials and the biological surrounding in real time and in situ. This information is useful in studies related to biodegradable biomaterials.

摘要

镁基植入物作为可生物降解材料在医学的骨科、牙科和心血管治疗中具有广阔的应用前景。在磨损和降解过程中会释放出微碎片。本文提出使用延时多维显微镜(MM)作为一种合适的工具,在24小时内以固定间隔跟踪细胞与颗粒之间的相互作用。MM结果显示巨噬细胞(J774)与镁颗粒(MgPa)之间的相互作用导致细胞大小和形态发生改变、复制率降低以及细胞损伤。颗粒表面逐渐形成腐蚀产物,并且由于氢气产生而产生湍流。用氟化钾处理MgPa后变化更为显著。为了补充MM观察结果,比较了用乳糖脱氢酶(LDH)测定法检测的膜损伤以及用WST-1测定法检测的巨噬细胞和成骨细胞(MC-3T3-E1)的线粒体活性。与成骨细胞相比,暴露于MgPa的巨噬细胞检测到更显著的浓度依赖性效应。因此,补充数据表明巨噬细胞的活力和细胞周期似乎改变更大。此外,在存在MgPa的情况下,与粘附过程相关的蛋白质谱和蛋白质表达发生了变化。这些研究表明,延时MM是实时原位监测可生物降解材料及其生物环境变化的有用工具。这些信息在与可生物降解生物材料相关的研究中很有用。

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