Effect of Porphyromonas gingivalis lipopolysaccharide on innate immune responses in bone-marrow derived macrophages and osteoclasts
10.3760/cma.j.issn.1002-0098.2020.01.007
- VernacularTitle: 牙龈卟啉单胞菌脂多糖对骨髓来源的巨噬细胞和破骨细胞先天免疫反应的影响
- Author:
Xiaojing KE
1
;
Houxuan LI
1
;
Fuhua YAN
1
;
Lang LEI
2
Author Information
1. Department of Periodontology, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing 210008, China
2. Department of Orthodontics, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing 210008, China
- Publication Type:Journal Article
- Keywords:
Macrophages;
Osteoclasts;
Immune response
- From:
Chinese Journal of Stomatology
2020;55(1):32-37
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To analyze the immune responses of bone-marrow derived macrophages and osteoclasts to lipopolysaccharide from Porphyromonas gingivalis (Pg-LPS) so as to provide reference for host immunomodulatory therapy of periodontitis.
Methods:Bone marrow mononuclear cells from C57BL/6 mice were obtained and induced into bone marrow macrophages (BMM) and osteoclasts (OC) by conditioned medium. The BMMs and OCs were divided into blank control group and Pg-LPS treated experimental group. Pg-LPS (10 μg/ml) was used to stimulate BMM and OC. The mRNA and protein expression of TLR-2 and TLR-4 and secretion of inflammatory cytokines were detected, respectively. Furthermore, the effect of Pg-LPS on the differentiation of BMM into OC was evaluated.
Results:Mouse bone marrow mononuclear cells were successfully induced into BMM and mature OC. In the presence of Pg-LPS, the gene level of TLR-2 in BMM was significantly up-regulated by (41.41±13.07) folds (P<0.01), while the level of TLR-4 mRNA was not significantly changed. TLR-2 mRNA and TLR-4 mRNA levels in OC increased by (2.24±0.23) times (P<0.05) and (4.83±1.07) times (P<0.01), respectively. Flow cytometry results showed that TLR-2 protein expression in BMM was robustly enhanced (P<0.01), as mean fluorescence intensities (MFI) were (39.85±5.27) in blank group and (221.57±13.13) in experimental group, respectively. The MFI of TLR-2 in OC also increased as (83.31±2.69) in blank group and (108.65±6.32) in experimental group, respectively (P<0.01). However, no significant TLR-4 expressions were observed in both BMM and OC (P>0.05). Moreover, the productions of tumor necrosis factor-α and interleukin-6 in response to LPS in BMM and OC remarkably increased (P<0.01). Lower cytokine expression was observed in OC than that in BMM (P<0.01). In addition, Pg-LPS obviously inhibited the number and size of OC formation, with a reduction in area percentage of approximately 47%.
Conclusions:Pg-LPS generated a robust immune inflammatory response to BMM, while OC had a relatively weaker immune response to the Pg-LPS. Pg-LPS might inhibite the differentiation of BMM into OC.
