Mengyuan Zhao1, Wenyu Dai2, Hongzhe Wang2, Chaoran Xue2, Jie Feng2, Yiruo He2, Peiqi Wang2, Sijia Li2, Ding Bai2, Rui Shu3. 1. Institute of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing, PR China. 2. Department of Orthodontics, West China School of Stomatology, State Key Laboratory of Oral Diseases, Sichuan University, ChengDu, Sichuan, PR China. 3. Department of Orthodontics, West China School of Stomatology, State Key Laboratory of Oral Diseases, Sichuan University, ChengDu, Sichuan, PR China. Electronic address: shuruispecial@163.com.
Abstract
OBJECTIVES: This study evaluated the role of human periodontal ligament fibroblasts (hPDLFs)-derived exosomes in periodontitis progression and discovered whether hPDLFs influence bone remodeling activity via exosome secretion. MATERIALS AND METHODS: Exosomes were isolated and quantified from Porphyromonas gingivalis lipopolysaccharide (LPS)-treated primary hPDLFs and evaluated by western blotting, dynamic light scattering, and transmission electron microscopy. GW4869 was used to block exosome secretion in conditioned medium (CM). hPDLFs-derived CM, CM containing GW4869 (CM + GW4869) and exosomes were used to stimulate MG-63 cell lines. The expression levels of proinflammatory mediators, osteogenic genes, and osteoclastogenesis-related genes were measured by quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, western blotting, and alkaline phosphatase staining. RESULTS: Exosome-enriched protein and total exosomal protein levels were higher in the LPS-treated group than in the vehicle controls. hPDLFs-derived exosomes were incorporated into MG-63 osteoblasts and slightly upregulated the expression of Interleukin-6 and tumor necrosis factor-alpha. CM and exosomes inhibited alkaline phosphatase, Collagen-I, Runt-related transcription factor 2, and Osteoprotegerin expression as well as ALP activity, and blocking exosome secretion by GW4869 eliminated the inhibitory effects. CONCLUSION: These results indicate that LPS-pretreated hPDLFs induce inflammation and inhibit osteogenic activity of osteoblasts through secreting exosomes. This study provides a potential mechanism by which localized periodontal inflammation may influence bone remodeling by release exosomes.
OBJECTIVES: This study evaluated the role of human periodontal ligament fibroblasts (hPDLFs)-derived exosomes in periodontitis progression and discovered whether hPDLFs influence bone remodeling activity via exosome secretion. MATERIALS AND METHODS: Exosomes were isolated and quantified from Porphyromonas gingivalislipopolysaccharide (LPS)-treated primary hPDLFs and evaluated by western blotting, dynamic light scattering, and transmission electron microscopy. GW4869 was used to block exosome secretion in conditioned medium (CM). hPDLFs-derived CM, CM containing GW4869 (CM + GW4869) and exosomes were used to stimulate MG-63 cell lines. The expression levels of proinflammatory mediators, osteogenic genes, and osteoclastogenesis-related genes were measured by quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, western blotting, and alkaline phosphatase staining. RESULTS: Exosome-enriched protein and total exosomal protein levels were higher in the LPS-treated group than in the vehicle controls. hPDLFs-derived exosomes were incorporated into MG-63 osteoblasts and slightly upregulated the expression of Interleukin-6 and tumor necrosis factor-alpha. CM and exosomes inhibited alkaline phosphatase, Collagen-I, Runt-related transcription factor 2, and Osteoprotegerin expression as well as ALP activity, and blocking exosome secretion by GW4869 eliminated the inhibitory effects. CONCLUSION: These results indicate that LPS-pretreated hPDLFs induce inflammation and inhibit osteogenic activity of osteoblasts through secreting exosomes. This study provides a potential mechanism by which localized periodontal inflammation may influence bone remodeling by release exosomes.
Authors: Zhenhong Ni; Siru Zhou; Song Li; Liang Kuang; Hangang Chen; Xiaoqing Luo; Junjie Ouyang; Mei He; Xiaolan Du; Lin Chen Journal: Bone Res Date: 2020-06-19 Impact factor: 13.567
Authors: Zhenhong Ni; Siru Zhou; Song Li; Liang Kuang; Hangang Chen; Xiaoqing Luo; Junjie Ouyang; Mei He; Xiaolan Du; Lin Chen Journal: Bone Res Date: 2020-06-19 Impact factor: 13.567