Yuchen Zhang1, Shiying Li1, Shujing Yuan1, Huifeng Zhang1, Jingying Liu2. 1. Department of Stomatology, Tianjin Union Medical Center, Tianjin 300121, China. 2. Department of Stomatology, Tianjin Union Medical Center, Tianjin 300121, China. Electronic address: a3635121@163.com.
Abstract
OBJECTIVES: To investigate the role of microRNA-23a (miR-23a) in the osteogenesis of periodontal mesenchymal stem cells (PDLSCs) in periodontitis. MATERIALS AND METHODS: Gingival crevicular fluid samples were collected from 21 control subjects and 29 patients with chronic periodontitis. MiR-23a was determined by quantitative real-time PCR. PDLSCs were transfected with miR-23a overexpressing lentiviruses. Subsequently, PDLSCs were induced with osteogenic differentiation media. Osteogenic differentiation of PDLSCs was assessed by alkaline phosphatase activity assay, Alizarin red staining, and qRT-PCT detection of osteogenic gene expression. Western blot was performed to detect the protein levels of the SMAD family member 1/5/9 (Smad1/5/9) and their phosphorylation level. TargetScan was used to predict the target gene of miR-23a. Cotransfections of bone morphogenetic protein receptor type 1B (BMPR1B) and miR-23a applied to explore the relationship between BMPR1B and miR-23a. RESULTS: MiR-23a was significantly increased in PDLSCs and gingival crevicular fluid of periodontitis patients. Patients with gingival crevicular fluid miR-23a levels above a threshold showed more clinical indicators of periodontitis. After periodontal therapy, miR-23a levels significantly decreased. Overexpression miR-23a inhibited osteogenesis of PDLSCs, which was evidenced by reduced Alizarin Red S and osteogenic gene expressions. In addition, miR-23a inhibited the phosphorylation of Smad1/5/9. TargetScan predicted that BMPR1B is a target gene of miR-23a. Overexpression of BMPR1B abolished the effects caused by overexpression of miR-23a. CONCLUSION: Our study provides novel evidence that miR-23a acts as a negative regulator of osteogenesis in periodontitis patients'PDLSCs and that miR-23a may serve as a biomarker and potential target of periodontitis.
OBJECTIVES: To investigate the role of microRNA-23a (miR-23a) in the osteogenesis of periodontal mesenchymal stem cells (PDLSCs) in periodontitis. MATERIALS AND METHODS:Gingival crevicular fluid samples were collected from 21 control subjects and 29 patients with chronic periodontitis. MiR-23a was determined by quantitative real-time PCR. PDLSCs were transfected with miR-23a overexpressing lentiviruses. Subsequently, PDLSCs were induced with osteogenic differentiation media. Osteogenic differentiation of PDLSCs was assessed by alkaline phosphatase activity assay, Alizarin red staining, and qRT-PCT detection of osteogenic gene expression. Western blot was performed to detect the protein levels of the SMAD family member 1/5/9 (Smad1/5/9) and their phosphorylation level. TargetScan was used to predict the target gene of miR-23a. Cotransfections of bone morphogenetic protein receptor type 1B (BMPR1B) and miR-23a applied to explore the relationship between BMPR1B and miR-23a. RESULTS:MiR-23a was significantly increased in PDLSCs and gingival crevicular fluid of periodontitispatients. Patients with gingival crevicular fluid miR-23a levels above a threshold showed more clinical indicators of periodontitis. After periodontal therapy, miR-23a levels significantly decreased. Overexpression miR-23a inhibited osteogenesis of PDLSCs, which was evidenced by reduced Alizarin Red S and osteogenic gene expressions. In addition, miR-23a inhibited the phosphorylation of Smad1/5/9. TargetScan predicted that BMPR1B is a target gene of miR-23a. Overexpression of BMPR1B abolished the effects caused by overexpression of miR-23a. CONCLUSION: Our study provides novel evidence that miR-23a acts as a negative regulator of osteogenesis in periodontitispatients'PDLSCs and that miR-23a may serve as a biomarker and potential target of periodontitis.
Authors: Chiara Mazziotta; Carmen Lanzillotti; Maria Rosa Iaquinta; Francesca Taraballi; Elena Torreggiani; John Charles Rotondo; Lucia Otòn-Gonzalez; Elisa Mazzoni; Francesca Frontini; Ilaria Bononi; Monica De Mattei; Mauro Tognon; Fernanda Martini Journal: Int J Mol Sci Date: 2021-02-27 Impact factor: 6.208