Amorphous Calcium Phosphate NPs Mediate the Macrophage Response and Modulate BMSC Osteogenesis
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ORIGINAL ARTICLE
Amorphous Calcium Phosphate NPs Mediate the Macrophage Response and Modulate BMSC Osteogenesis Liangjiao Chen,1,2 Pengyan Qiao,1 Hongchen Liu ,1,4 and Longquan Shao3,4 Received 7 April 2020; accepted 24 August 2020
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 Abstract—
Hongchen Liu and Longquan Shao contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10753-020-01331-9) contains supplementary material, which is available to authorized users. 1
Institute of Stomatology & Oral Maxilla Facial Key Laboratory, Chinese PLA General Hospital, 28 Fuxing Road, Beijing, 100853, China 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 3 Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China 4 To whom correspondence should be addressed to Hongchen Liu at Institute of Stomatology & Oral Maxilla Facial Key Laboratory, Chinese PLA General Hospital, 28 Fuxing Road, Beijing, 100853, China. E-mail: [email protected]; and ; [email protected]
0360-3997/20/0000-0001/0 # 2020 Springer Science+Business Media, LLC, part of Springer Nature
Chen, Qiao, Liu, and Shao pattern. ACP NPs/macrophage-modulated environments weaken the osteogenic ability of BMSCs. ACP NPs may cause aseptic inflammation and attenuate osteogenesis. KEY WORDS: ACP nanoparticles; macrophage polarization; cytokine; autophagy; osteogenic differentiation.
INTRODUCTION Calcium phosphate (CaP) is often used as a bone substitute or coating material. It has been proven safe and biocompatible [1]. Howeve
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