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™ 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.

中文翻译：

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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。

与免疫细胞培养的 ACP 纳米粒子 (ACP NPs) 相关的潜在风险及其对成骨的间接影响尚未深入研究。该项目旨在评估 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/巨噬细胞调节环境对成骨能力的影响骨髓间充质干细胞。采用CCK-8和Annexin V凋亡检测试剂盒检测细胞增殖率和凋亡率。通过ROS测试试剂盒和蛋白质印迹（WB）评估ROS和自噬表达。通过扫描电镜、细胞骨架染色、RT-PCR 和 ELISA 确定巨噬细胞极化和细胞因子表达。TMT™ 定量蛋白质分析用于评估蛋白质表达。通过ALP染色、茜素红溶液染色和RT-PCR检测BMSC成骨分化。ACP NPs 对巨噬细胞是安全的，但在高浓度时会促进自噬并诱导 ROS 产生。ACP NPs改变巨噬细胞的形态并诱导极化为M1型，从而促进炎症细胞因子的表达。ACP NPs/巨噬细胞调节的环境削弱了 BMSCs 的成骨能力。ACP NPs 将巨噬细胞极化为 M1 表型并改变细胞因子的分泌模式。ACP NPs/巨噬细胞调节的环境削弱了 BMSCs 的成骨能力。ACP NPs 可能引起无菌性炎症并减弱成骨作用。ACP NPs 对巨噬细胞是安全的，但在高浓度时会促进自噬并诱导 ROS 产生。ACP NPs改变巨噬细胞的形态并诱导极化为M1型，从而促进炎症细胞因子的表达。ACP NPs/巨噬细胞调节的环境削弱了 BMSCs 的成骨能力。ACP NPs 将巨噬细胞极化为 M1 表型并改变细胞因子的分泌模式。ACP NPs/巨噬细胞调节的环境削弱了 BMSCs 的成骨能力。ACP NPs 可能引起无菌性炎症并减弱成骨作用。ACP NPs 对巨噬细胞是安全的，但在高浓度时会促进自噬并诱导 ROS 产生。ACP NPs改变巨噬细胞的形态并诱导极化为M1型，从而促进炎症细胞因子的表达。ACP NPs/巨噬细胞调节的环境削弱了 BMSCs 的成骨能力。ACP NPs 将巨噬细胞极化为 M1 表型并改变细胞因子的分泌模式。ACP NPs/巨噬细胞调节的环境削弱了 BMSCs 的成骨能力。ACP NPs 可能引起无菌性炎症并减弱成骨作用。ACP NPs 将巨噬细胞极化为 M1 表型并改变细胞因子的分泌模式。ACP NPs/巨噬细胞调节的环境削弱了 BMSCs 的成骨能力。ACP NPs 可能引起无菌性炎症并减弱成骨作用。ACP NPs 将巨噬细胞极化为 M1 表型并改变细胞因子的分泌模式。ACP NPs/巨噬细胞调节的环境削弱了 BMSCs 的成骨能力。ACP NPs 可能引起无菌性炎症并减弱成骨作用。