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Amorphous Calcium Phosphate NPs Mediate the Macrophage Response and Modulate BMSC Osteogenesis

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Abstract

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.

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Funding

This work was supported financially by the National Natural Science Fund (81600904 and 81771102), the High Technology Research and Development Program of China (863 program, 2015AA033502) and the incubation programme of Guangzhou Medical University (No. B185004177).

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Author notes

  1. Hongchen Liu and Longquan Shao contributed equally to this work.

Authors and Affiliations

  1. Institute of Stomatology & Oral Maxilla Facial Key Laboratory, Chinese PLA General Hospital, 28 Fuxing Road, Beijing, 100853, China

    Liangjiao Chen, Pengyan Qiao & Hongchen Liu

  2. Department of Orthodontics, Affilicated Stomatology Hospital of Guangzhou Medical University, Guangzhou Key Laboratory of Basic and Applied Research of Oral Regeneartive Medicine, Guangzhou, 510140, China

    Liangjiao Chen

  3. Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China

    Longquan Shao

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  1. Liangjiao Chen
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Contributions

LJC, HCL and LQS designed the research. LJC was responsible for characterizing ACP NPs and detecting macrophage polarization and cytokine expression. ROS and autophagy expression were evaluated by LJC and PYQ. TMT™ quantitative protein analysis and BMSC osteogenic differentiation were detected by LJC and PYQ. LJC generated the figures and drafted the manuscript, which was critically revised by HCL and LQS. All authors read, corrected and approved the manuscript.

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Correspondence to Hongchen Liu or Longquan Shao.

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Chen, L., Qiao, P., Liu, H. et al. Amorphous Calcium Phosphate NPs Mediate the Macrophage Response and Modulate BMSC Osteogenesis. Inflammation 44, 278–296 (2021). https://doi.org/10.1007/s10753-020-01331-9

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