microRNA-146a reverses the inhibitory effects of <italic>Porphyromonas gingivalis</italic> lipopolysaccharide on osteogenesis of human periodontal ligament cells

Dongwang Zhu; Dong Xue; Wen Lai; Wanning XU; Shaoyun Jiang

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microRNA-146a reverses the inhibitory effects of Porphyromonas gingivalis lipopolysaccharide on osteogenesis of human periodontal ligament cells

VernacularTitle: 微RNA-146a反转牙龈卟啉单胞菌脂多糖对人牙周膜细胞成骨的抑制作用
Author: Dongwang ZHU ¹ ; Dong XUE ² ; Wen LAI ³ ; Wanning XU ⁴ ; Shaoyun JIANG ⁵
Author Information

1. Department of Oral and Maxillofacial Surgery, Institute of Stomatology, School and Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China
2. Department of Periodontics, Institute of Stomatology, School and Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China(Present address: Department of Dentistry, Xuanwu Hospital, Capital Medical University, Beijing 100053, China)
3. Department of Periodontics, Institute of Stomatology, School and Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China
4. Department of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University & State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, Chengdu 610041, China
5. Department of Periodontics, Institute of Stomatology, School and Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China(Present address: Center of Stomatology, Shenzhen Hospital, Peking University, Shenzhen Guangdong 518036, China)
Publication Type:Journal Article
Keywords: Periodontitis; MicroRNAs; Osteogenic differentiation; Human periodontal ligament cells
From: Chinese Journal of Stomatology 2018;53(11):753-759
CountryChina
Language:Chinese
Abstract: Objective:To investigate the effects and mechanisms of microRNA-146a (miR-146a) on osteogenic differentiation of human periodontal ligament cells (hPDLC) stimulated by lipopolysaccharide (LPS) of Porphyromonas gingivalis (Pg).
Methods:hPDLC were cultured in vitro and induced to the phase of osteogenic differentiation. These cells were divided into five groups: non-osteogenic differentiation cells, osteogenic differentiation cells, osteogenic differentiation cells treated with Pg LPS, osteogenic differentiation cells treated with Pg LPS and miR-146a mimic, osteogenic differentiation cells treated with Pg LPS and miR-146a negative control. Osteogenic markers and mineralization were detected via quantitative real-time PCR (qPCR) and alizarin red staining, respectively. Meanwhile, non-radioactive transcription factor assay was applied to explore the nuclear activity of nuclear factor kappa B (NF-κB) p65 in nuclear extracts of hPDLC.
Results:Compared with cells of osteogenic differentiation in non-LPS-stimulated groups, Pg LPS could decrease the markers of osteogenic differentiation of hPDLC such as collagen Ⅰ (Col-Ⅰ), alkaline phosphatase (ALP), Runt-related transcription factor-2 (RUNX2) and osteocalcin (OCN) (P<0.05), inhibit mineralization, and stimulate NF-κB p65 nuclear activity expression (non-LPS stimulated group: 1.023±0.217, LPS stimulated group: 6.252±0.613, P=0.008). However, compared with cells in Pg LPS/miR-146a negative control group, miR-146a increased Col-Ⅰ (P=0.007) and OCN (P=0.049) mRNA expression, rather than ALP (P=0.167) and RUNX2 (P=0.580) at day 3; miR-146a also upregulated mRNA levels of Col-Ⅰ, ALP, RUNX2 and OCN (P<0.05) at day 7 and day 14, and enhance mineralization. Meanwhile, miR-146a mimic could decrease the nuclear activity of NF-κB p65 induced by Pg LPS in hPDLC (miR-146a: 2.427±0.354, negative control: 5.863±0.482, P=0.019).
Conclusions:miR-146a could reverse the inhibitory effects of Pg LPS on osteogenic differentiation of hPDLC through enhancing the expression of osteogenic markers and decreasing inflammatory pathway in hPDLC.