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Exposure of the murine RAW 264.7 macrophage cell line to dicalcium silicate coating: assessment of cytotoxicity and pro-inflammatory effects

  • Biomaterials Synthesis and Characterization
  • Original Research
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Abstract

Inflammatory effects are significant elements of the immune response to biomaterials. Previously, we reported inflammatory effects in response to dicalcium silicate (Ca2SiO4, C2S) particles. However, the immunological effects of C2S coatings have not been studied. C2S often used as coatings materials in orthopedic and dentistry applications. It may have different effect from C2S particles. Further, it remains unclear whether C2S coating is equally biocompatible as 45S5 coating. The aim of this study was to test the cytotoxicity and pro-inflammatory effects of C2S coating on RAW 264.7 macrophages. C2S and 45S5 coatings were characterized using scanning electron microscopy (SEM), atomic force microscopy (AFM), energy dispersive analysis (EDS) and X-ray diffraction (XRD). inductively coupled plasma optical emission spectroscopy (ICP-OES) was used to detect ionic concentrations after soaking coated discs in medium. The cytotoxicity of C2S and 45S5 coatings against RAW 264.7 macrophages was measured using the LDH Cytotoxicity Assay Kit, Cell Counting Kit-8 (CCK-8) assays and flow cytometry for apoptosis assays. The gene and protein expression of TNF-α, IL-6 and IL-1β were detected using RT-q PCR and ELISA, respectively. The tested coating materials are not cytotoxic to macrophages. The C2S-coated surface stimulated macrophages to express pro-inflammatory mediators, such as TNF-α, IL-6 and IL-1β, and C2S coating caused less IL-6 but greater IL-1β production than the 45S5 coating. C2S coating have no cytotoxicity when directly cultured with macrophages. C2S and 45S5 coatings both have the potential to induce pro-inflammatory effects, and the biocompatibility of C2S is similar to that of 45S5.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31070857 and 50973045) and the Project on the Integration of Industry, Education and Research of Guangdong Province, China (2012B091000147). This work was also supported by Medical Research Foundation of Guangdong Province (A2015132) and Scientific research projects of Guangzhou Medical University (2015C43).

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    1. Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou, 510140, People’s Republic of China

      Liangjiao Chen

    2. Department of Stomatology, Nanfang Hospital, Southern Medical University, 1838 North Guangzhou Road, Guangzhou, 510515, People’s Republic of China

      Yanli Zhang, Jia Liu, Limin Wei, Bin Song & Longquan Shao

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    1. Liangjiao Chen
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    Correspondence to Longquan Shao.

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    Liangjiao Chen and Yanli Zhang have contributed equally to this work and should be considered co-first authors.

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    Chen, L., Zhang, Y., Liu, J. et al. Exposure of the murine RAW 264.7 macrophage cell line to dicalcium silicate coating: assessment of cytotoxicity and pro-inflammatory effects. J Mater Sci: Mater Med 27, 59 (2016). https://doi.org/10.1007/s10856-016-5668-7

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