Ipriflavone promotes proliferation and osteogenic differentiation of periodontal ligament cells by activating GPR30/PI3K/AKT signaling pathway

Abstract

ObjectivesThis study was performed to investigate the effects and mechanism of ipriflavone (IP) on the proliferation and osteoblastic differentiation of periodontal ligament cells in vitro and periodontal tissue remodeling following orthodontic tooth movement (OTM) in vivo.Materials and methodsHuman periodontal ligament cells (hPDLCs) were cultured in vitro and cell counting kit-8, alkaline phosphatase (ALP) activity assay, plate clone formation assay, and alizarin red staining were used to test proliferation and osteogenic differentiation of hPDLCs. What is more, the expression of ALP, Runx2, and GPR30 was examined by real-time polymerase chain reaction and Western blot. To find out if PI3K/AKT signaling pathway was involved in the process, AKT and p-AKT were examined by Western blot. LY294002 (PI3K signaling pathway inhibitor) and small interfering RNA targeting GPR30 mRNA (siGPR30) were used to verify the function of GPR30-mediated PI3K/AKT pathway in this process. Twenty-four male Wistar rats were randomized into 2 groups, the control group with force application and the IP group with force application plus IP. Morphological changes in the periodontal tissue between roots of teeth were investigated using hematoxylin and eosin (HE) staining and bone morphogenetic protein-2 was detected to assess bone remodeling by immunohistochemical staining.ResultsIn vitro, 10−7 M IP was selected significantly promoting proliferation, ALP activity, colony forming efficiency, and mineral deposition (P<0.05) on hPDLCs. Gene expressions of ALP, Runx2, GPR30, and p-AKT were all upregulated than the control group (P<0.05). According to the mechanism, promotion of ALP and Runx2 interdicted by LY294002 and siGPR30 reduced the activation of PI3K/AKT signaling pathway. In addition, HE staining and immunohistochemical staining results showed that the IP group had more new bone formation in the periodontal tissue compared to the control group in vivo.ConclusionIP can promote the expression of ALP and Runx2 which was probably related to the GPR30-mediated PI3K/AKT signaling pathway. Moreover, IP coordination seemed to have the potential to prevent relapsing following OTM.

G Protein-Coupled Estrogen Receptor 1 Inhibits Angiotensin II-Induced Cardiomyocyte Hypertrophy via the Regulation of PI3K-Akt-mTOR Signalling and Autophagy

“…In our study, we found that G1 treatment activated the Akt-mediated signalling pathway, which is believed to be associated with cardiomyocyte growth, hypertrophy and survival. This finding regarding the regulation of Akt signaling pathway by GPER1 is consistent with other report in human periodontal ligament cells 38, 39. mTOR is an important protein molecule downstream from Akt-mediated signalling.…”

DNA methylation of noncoding RNAs: new insights into osteogenesis and common bone diseases

“…The P13K-Akt signaling pathway can control many differentiation processes including chondrocyte proliferation, osteogenic differentiation, adipogenic differentiation, and cell apoptosis [47][48][49]. Various studies have proposed the interacted regulation role between PTEN and P13K-Akt signaling in many diseases [50,51].…”

Periodontal Destruction and Regeneration in Experimental Models: Combined Research Approaches

“…Сonsidering the evidence from animal models, implantation of periodontal ligament stem cells is emerging as a potential periodontal regenerative procedure [8]. The advantages and feasibility of periodontal tissue regeneration using platelet-rich fibrin combined with rat periodontal ligament stem cells, and the effects and mechanism of ipriflavone on the proliferation and osteoblastic differentiation of periodontal ligament cells in vitro and periodontal tissue remodeling following orthodontic tooth movement in vivo, were investigated by Duan X. et al (2018) and Han Y. et al (2018) [9,10]. Increasing evidence indicates, that bone remodeling is under the control of factors related to neuronal regulation, but the underlying mechanisms remain largely unknown [11].…”

“…The results obtained were the subject of histomorphomorphological/-metrical [10, 13, 14, 17, 19, 20, 21-25, 27-29, 30], immunohisto (-cyto)chemical [15,16,10,20,7,28,21,29,6,30,5,19] and histopathological [2,4,5,9,31,22] analysis, micro-computed tomography [9,15,16,18,19,21,24,28]. The laboratory methods involved were real-time quantitative reverse-transcription polymerase chain reaction [11,15,16,21] enzyme-linked immunosorbent assay [16,17], scanning [3], and transmission electron microscopy [32], polarizing light microscopy [29], confocal microscopy and spectrophotometry [15], radiographic and biomechanical analysis [18], descriptive histology and computer-assisted image analysis [12].…”

“…The mild decalcification method allowed Alves A. et al (2019) in situ maintenance of the periodontal soft and hard tissue integrity, and histomorphometrical data showed that qualitatively, cellular and extracellular matrix morphologies were well preserved compared to non-decalcified periodontal soft tissue biopsies [12]. The effects of fluoxetine, a selective serotonin reuptake inhibitor, on the formation of the periodontal ligament collagen fibers in rats, and ipriflavone on new bone formation in the periodontal tissue in vivo, were assessed by Regueira L. S. et al (2017) and Han Y. et al [10,29]. The effects of denervation on neuropeptide expression and bone remodeling-related factors in the rats, both of which govern the periodontal alveolar bone regeneration processes, were assessed by Yu X. et al (2015) [11].…”