Chinese Journal of Tissue Engineering Research ›› 2022, Vol. 26 ›› Issue (24): 3840-3845.doi: 10.12307/2022.563

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Effects of red deer horn powder extract on the biological behavior of bone marrow derived macrophages

Shan Shuai1, Dong Yi1, Liu Jialin1, Han Xiangzhen1, He Huiyu1, 2   

  1. 1Department of Prosthodontics and Implant, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 2Xinjiang Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Received:2021-09-23 Accepted:2021-11-03 Online:2022-08-28 Published:2022-01-24
  • Contact: He Huiyu, Chief physician, Professor, Doctoral supervisor, Master’s supervisor, Department of Prosthodontics and Implant, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; Xinjiang Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • About author:Shan Shuai, Master candidate, Department of Prosthodontics and Implant, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    the Science and Technology Support Project of Xinjiang Uygur Autonomous Region, No. 2018E02060 (to HHY); Postgraduate Research and Innovation Project of Xinjiang Uygur Autonomous Region, No. XJ2021G196 (to LJL)

Abstract: BACKGROUND: Foreign body reactions will lead to the failure of material implantation. In recent years, research on immunity has gradually become a hot spot. It is of great significance to select scaffold materials with good immune properties for the development of bone defect repair.  
OBJECTIVE: To investigate the effects of calcinated antler cancellous bone and untreated antler bone on the biological behavior of bone marrow-derived M1 macrophages.
METHODS:  Calcinated antler cancellous bone was prepared by physical and chemical methods. The physical and chemical properties of calcinated antler cancellous bone and untreated antler bone were detected by X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy. The extracts of calcinated antler cancellous bone and untreated antler bone were prepared and the contents of Ca, P and Mg in the extracts were determined. Bone marrow-derived macrophages were cultured in vitro and polarized into M1 macrophages under lipopolysaccharide and interferon γ stimulation. These cells were then co-cultured with different extracts in calcinated antler cancellous bone and untreated antler bone groups, while co-cultured with an IMDM medium in control group, followed by cell counting kit-8 detection of cell survival, flow cytometry detection of M1 markers, and qPCR detection of macrophage-related genes.  
RESULTS AND CONCLUSION: (1) The contents of Ca, P and Mg were higher in the calcinated antler cancellous bone group than the untreated antler bone group. (2) The main functional groups in these two groups included OH-, PO43-, and CO32 -. (3) There was no significant difference in cell survival rate among the three groups for 1, 3, and 7 days of co-culture. (4) The expression rate of CD16/32 was lower in the calcinated antler cancellous bone group than the control group after 1-day co-culture. (5) The expression levels of pro-inflammatory cytokines, tumor necrosis factor-α and interleukin-6 were lower in the calcinated antler cancellous bone group than the control group after 1 and 3 days of co-culture, while there was no significant difference between the untreated antler bone group and the control group. The expression of CD206, a main marker of M2 macrophages, was significantly higher in the calcinated antler cancellous bone group than the control group at 3 days of co-culture, while there was no significant difference between the untreated antler bone group and the control group at 1 and 3 days of co-culture. (6) To conclude, calcinated antler cancellous bone powder extract can reduce the expression of pro-inflammatory cytokines in M1 macrophages and promote their polarization to M2 macrophages.

Key words: antler, calcinated bone, extract, bone marrow-derived macrophages, foreign body reaction, macrophage polarization

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