Amorphous Calcium Phosphate NPs Mediate the Macrophage Response and Modulate BMSC Osteogenesis
Amorphous Calcium Phosphate NPs Mediate the Macrophage Response and Modulate BMSC Osteogenesis
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无定形磷酸钙纳米颗粒介导巨噬细胞反应并调节 BMSC 成骨
DOI：
10.1007/s10753-020-01331-9
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发表时间：
2020-09-16
期刊：
影响因子：
5.1
通讯作者：
Shao, Longquan
中科院分区：
文献类型：
--
作者：
Chen, Liangjiao;Qiao, Pengyan;Shao, Longquan
与免疫细胞培养的ACP纳米颗粒（ACP NPs）相关的潜在风险及其对骨生成的间接影响尚未深入研究。本项目旨在评价ACP NPs在巨噬细胞中的安全性、巨噬细胞对ACP NPs的反应（巨噬细胞极化、巨噬细胞的细胞因子分泌模式和细胞内稳态）以及ACP NPs/巨噬细胞调节的环境对BMSCs成骨能力的影响。CCK-8和Annexin V凋亡检测试剂盒检测细胞增殖率和凋亡。用ROS检测试剂盒和Western blot（WB）检测ROS和自噬表达。扫描电镜、细胞骨架染色、RT-PCR和ELISA检测巨噬细胞极化和细胞因子表达。TMT（TM）定量蛋白分析用于评估蛋白表达。ALP染色、茜素红染色、RT-PCR检测BMSC成骨分化。ACP NP对巨噬细胞是安全的，但在高浓度下促进自噬并诱导ROS产生。ACP NPs改变了巨噬细胞的形态，诱导其向M1型极化，从而促进炎性细胞因子的表达。ACP NPs/巨噬细胞调节的环境削弱了BMSCs的成骨能力。ACP NPs使巨噬细胞转化为M1表型，并改变细胞因子分泌模式。ACP NPs/巨噬细胞调节的环境削弱了BMSCs的成骨能力。ACP NP可引起无菌性炎症并减弱骨生成。
The potential risk associated with ACP nanoparticles (ACP NPs) cultured with immune cells and their indirect effects on osteogenesis have not been studied deeply. This project aims to evaluate the safety of ACP NPs in macrophages, the responses of macrophages (macrophage polarization, the cytokine secretion pattern of macrophages and intracellular homeostasis) to ACP NPs and the effect of ACP NPs/macrophage-modulated environments on the osteogenic ability of BMSCs. The cell proliferation rate and apoptosis were detected by CCK-8 and Annexin V Apoptosis Detection kits. ROS and autophagy expression were evaluated by ROS test kits and Western blot (WB). Macrophage polarization and cytokine expression were determined by SEM, cytoskeletal staining, RT-PCR and ELISA. TMT (TM) quantitative protein analysis was used to evaluate protein expression. BMSC osteogenic differentiation was detected by ALP staining, Alizarin Red solution staining and RT-PCR. ACP NPs were safe to macrophages but promoted autophagy and induced ROS production at high concentrations. ACP NPs changed morphology of macrophages and induced polarization into M1 type, thus promoting the expression of inflammatory cytokines. ACP NPs/macrophage-modulated environments weakened the osteogenic ability of BMSCs. ACP NPs polarize macrophages into the M1 phenotype and change the cytokine secretion pattern. ACP NPs/macrophage-modulated environments weaken the osteogenic ability of BMSCs. ACP NPs may cause aseptic inflammation and attenuate osteogenesis.