Oroxylin A Shows Protective Effects on Biological Activity of Lipopolysaccharide Treated Human Periodontal Ligament Stem Cells

2019 ◽  
Vol 9 (10) ◽  
pp. 1362-1368
Author(s):  
Ting Wang ◽  
Xinqiang Liu ◽  
Chunmiao Jiang ◽  
Dapeng Ren ◽  
Yuli Gao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Biological Activity ◽  
Periodontal Ligament ◽  
Time Dependent ◽  
Dependent Manner ◽  
Protective Effects ◽  
Oroxylin A ◽  
Periodontal Ligament Stem Cells ◽  
Dose Dependent

The abnormal proliferation and apoptosis of human periodontal ligament stem cells (hPDLSCs) serves a crucial role in the development of periodontitis. Oroxylin A has shown protective effects in a variety of inflammatory diseases. The present study was aimed to investigate the effects of oroxylin A on lipopolysaccharide (LPS) treated hPDLSCs. In the present study, cells were exposed to different concentrations (10, 20, 40 uM) of oroxylin A for 24 h or 48 h, co-treated with LPS. The cell proliferation capacity was assessed using cell counting kit-8 (CCK-8), and the cell apoptosis was evaluated by flow cytometry. The Ki67 expression was measured using immunofluorescence and NO production was detected by enzyme linked immunosorbent assay (ELISA) respectively. Western blot analyses were used to investigate the level of cell proliferation related proteins (PCNA, CDK2 and p21) as well as NF-κB, I-κBα and downstream molecules iNOS, IL-6 and TNF-α. The results demonstrated that oroxylin A increased cell survival of LPS treated hPDLSCs in a dose-dependent and time-dependent manner. In addition, oroxylin A treatment inhibited cell apoptosis in hPDLSCs. Furthermore, the levels of NO, NF-κB, iNOS, IL-6 and TNF-α were significantly reduced. And the expression of Ki67, I-κBα, PCNA and CDK2 were significantly increased. Taken together, these findings indicate that oroxylin A promote proliferation and suppress apoptosis in a dose-dependent and time-dependent manner. Oroxylin A may affects LPS induced biological activity via inhibiting NF-κB activation and proinflammatory cytokines expression in hPDLSCs.

scholarly journals Impact of Magnetic Stimulation on Periodontal Ligament Stem Cells

2021 ◽  
Vol 23 (1) ◽  
pp. 188
Author(s):  
Valentina Peluso ◽  
Laura Rinaldi ◽  
Teresa Russo ◽  
Olimpia Oliviero ◽  
Anna Di Vito ◽  
...  
Keyword(s):  
Magnetic Field ◽  
Stem Cells ◽  
Cell Proliferation ◽  
Osteogenic Differentiation ◽  
Periodontal Ligament ◽  
Magnetic Stimulation ◽  
Laser Scanning Microscopy ◽  
Time Dependent ◽  
Cellular Respiration ◽  
Periodontal Ligament Stem Cells

The aim of this study was to evaluate the effect of a time-dependent magnetic field on the biological performance of periodontal ligament stem cells (PDLSCs). A Western blot analysis and Alamar Blue assay were performed to investigate the proliferative capacity of magnetically stimulated PDLSCs (PDLSCs MAG) through the study of the MAPK cascade (p-ERK1/2). The observation of ALP levels allowed the evaluation of the effect of the magnetic field on osteogenic differentiation. Metabolomics data, such as oxygen consumption rate (OCR), extracellular acidification rate (ECAR) and ATP production provided an overview of the PDLSCs MAG metabolic state. Moreover, the mitochondrial state was investigated through confocal laser scanning microscopy. Results showed a good viability for PDLSCs MAG. Magnetic stimulation can activate the ERK phosphorylation more than the FGF factor alone by promoting a better cell proliferation. Osteogenic differentiation was more effectively induced by magnetic stimulation. The metabolic panel indicated significant changes in the mitochondrial cellular respiration of PDLSCs MAG. The results suggested that periodontal ligament stem cells (PDLSCs) can respond to biophysical stimuli such as a time-dependent magnetic field, which is able to induce changes in cell proliferation and differentiation. Moreover, the magnetic stimulation also produced an effect on the cell metabolic profile. Therefore, the current study demonstrated that a time-dependent magnetic stimulation may improve the regenerative properties of PDLSCs.

scholarly journals Pharmacodynamics of siRNARANKL and Oestrogen Loaded MSNs-CADY on Human Periodontal Ligament Stem Cells with Porphyromonas gingivalis infection

2020 ◽  
Author(s):  
Liang Song ◽  
Xiaojun Shi ◽  
Fengling Hu ◽  
Huijuan Chen ◽  
Bin Xu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Drug Release ◽  
Mesoporous Silica ◽  
Porphyromonas Gingivalis ◽  
Periodontal Ligament ◽  
Mesoporous Silica Nanoparticles ◽  
Cell Penetrating Peptide ◽  
Periodontal Ligament Stem Cells ◽  
Cell Penetrating

Abstract Background: Periodontitis irreversibly invades and destroys periodontal supporting tissues, loses the ability of periodontal regeneration and restoration, and eventually leads to tooth loosening and loss. periodontal ligament stem cells (PDLSCs) hold great promises for periodontal tissue regeneration which was the potential target of periodontitis treatment, siRNARANKL and oestrogen can help PDLSCs maintain normal function, however, it was very difficult for siRNARANKL and oestrogen to get into PDLSCs. Here, Cell penetrating peptide CADY was modified on the surface of siRNARANKL and oestrogen loaded mesoporous silica nanoparticles (MSNs) to carry them into Porphyromonas gingivalis infected PDLSCs, Then further affect the proliferation of PDLSCs. Methods: 120-150 nm Mesoporous silica nanoparticles (MSNs) was prepared, and the biocompatibility, loading capacity and drug release properity were tested; MSNs was modified by penetrating peptide CADY and the prepared MSNs/CADY was loaded with siRNARANKL and oestrogen; In vitro drug release of siRNARANKL/MSNs-CADY and oestrogen/MSNs-CADY was tested by using semi-permeable dialysis bag diffusion; Cellular uptake and internalization of FITC-Labeled MSNs and FITC-Labeled MSNs-CADY was observed by use of Laser confocal microscopy; Finally, the effect of siRNARANKL and oestrogen loaded MSNs-CADY on cell proliferation of Porphyromonas gingivalis infected human periodontal ligament stem cells was tested by MTT assay. Results: according to the results, MSNs-CADY with a concentration of 6.25-200 ug/mL have no toxic to PDLSCs; 24.6 mg oestrogen and 0.5 mM siRNARANKL can be loaded into 1mg of MSNs-CADY; and drug loaded MSNs-CADY nanodrug carriers can release siRNARANKL and oestrogen stably for at least 48 h; After modification with cell penetrating peptide CADY, more MSNs-CADY can be taken by PDLSCs. siRNARANKL/oestrogen/MSNs-CADY can increase the proliferation of PDLSCs significantly. Conclusion: siRNARANKL/oestrogen/MSNs-CADY constructed can significantly improve the cell proliferation of P-gingivalis infected PDLSCs, this nano drug carrier has the potential to be used in PDLSCs -based periodontitis treatment, this work provided a useful theoretical basis and therapeutic ideas for the treatment of periodontitis.

scholarly journals CoCl2 induces apoptosis via a ROS-dependent pathway and Drp1-mediated mitochondria fission in periodontal ligament stem cells

2018 ◽  
Vol 315 (3) ◽  
pp. C389-C397 ◽  
Author(s):  
Yuting He ◽  
Xueqi Gan ◽  
Ling Zhang ◽  
Beilei Liu ◽  
Zhuoli Zhu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Periodontal Ligament ◽  
Mitochondrial Membrane ◽  
Mitochondrial Fission ◽  
Orthodontic Appliances ◽  
Dependent Manner ◽  
Periodontal Ligament Stem Cells ◽  
Atp Level

Oxygen deficiency is associated with various oral diseases, including chronic periodontitis, age-related alveolar bone loss, and mechanical stress-linked cell injury from orthodontic appliances. Nevertheless, our understanding of the impact of hypoxia on periodontal tissues and its biochemical mechanism is still rudimentary. The purpose of this research was to elucidate the effects of hypoxia on the apoptosis of human periodontal ligament stem cells (PDLSCs) in vitro and the underlying mechanism. Herein, we showed that cobalt chloride (CoCl2) triggered cell dysfunction in human PDLSCs in a concentration-dependent manner and resulted in cell apoptosis and oxidative stress overproduction and accumulation in PDLSCs. In addition, CoCl2 promoted mitochondrial fission in PDLSCs. Importantly, CoCl2 increased the expression of dynamin-related protein 1 (Drp1), the major regulator in mitochondrial fission, in PDLSCs. Mitochondrial division inhibitor-1, pharmacological inhibition of Drp1, not only inhibited mitochondrial fission but also protected against CoCl2-induced PDLSC dysfunction, as shown by increased mitochondrial membrane potential, increased ATP level, reduced reactive oxygen species (ROS) level, and decreased apoptosis. Furthermore, N-acety-l-cysteine, a pharmacological inhibitor of ROS, also abolished CoCl2-induced expression of Drp1 and protected against CoCl2-induced PDLSC dysfunction, as shown by restored mitochondrial membrane potential, ATP level, inhibited mitochondrial fission, and decreased apoptosis. Collectively, our data provide new insights into the role of the ROS-Drp1-dependent mitochondrial pathway in CoCl2-induced apoptosis in PDLSCs, indicating that ROS and Drp1 are promising therapeutic targets for the treatment of CoCl2-induced PDLSC dysfunction.

scholarly journals Comparison of Biocompatibility of Calcium Silicate-Based Sealers and Epoxy Resin-Based Sealer on Human Periodontal Ligament Stem Cells

Materials ◽  
2020 ◽  
Vol 13 (22) ◽  
pp. 5242
Author(s):  
Hanseul Oh ◽  
Egan Kim ◽  
Sukjoon Lee ◽  
Soyeon Park ◽  
Dongzi Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Cell Viability ◽  
Epoxy Resin ◽  
Calcium Silicate ◽  
Periodontal Ligament ◽  
Expression Level ◽  
Osteogenic Potential ◽  
Periodontal Ligament Stem Cells ◽  
Ah Plus

The aim of this study was to evaluate the biocompatibility of calcium silicate-based sealers (CeraSeal and EndoSeal TCS) and epoxy resin-based sealer (AH-Plus) in terms of cell viability, inflammatory response, expression of mesenchymal phenotype, osteogenic potential, cell attachment, and morphology, of human periodontal ligament stem cells (hPDLSCs). hPDLSCs were acquired from the premolars (n = 4) of four subjects, whose ages extended from 16 to 24 years of age. Flow cytometry analysis showed stemness of hPDLSCs was maintained in all materials. In cell viability test, AH-Plus showed the lowest cell viability, and CeraSeal showed significantly higher cell viability than others. In ELISA test, AH-Plus showed higher expression of IL-6 and IL-8 than calcium silicate-based sealers. In an osteogenic potential test, AH-Plus showed a lower expression level than other material; however, EndoSeal TCS showed a better expression level than others. All experiments were repeated at least three times per cell line. Scanning electronic microscopy studies showed low degree of cell proliferation on AH-Plus, and high degree of cell proliferation on calcium silicate-based sealers. In this study, calcium silicate-based sealers appear to be more biocompatible and less cytotoxic than epoxy-resin based sealers.

scholarly journals Protease-Activated Receptor Type 1 Activation Enhances Osteogenic Activity in Human Periodontal Ligament Stem Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Emanuel Silva Rovai ◽  
Lucas Macedo Batitucci Ambrósio ◽  
Bruno Nunes de França ◽  
Letícia Rodrigues de Oliveira ◽  
Letícia Miquelitto Gasparoni ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Protein Expression ◽  
Periodontal Ligament ◽  
Calcium Concentration ◽  
Periodontal Ligament Stem Cells ◽  
Osteogenic Activity ◽  
Gene And Protein Expression ◽  
New Strategy ◽  
Calcium Deposits

Protease-activated receptor 1 (PAR1) has been associated to tissue repair and bone healing. The aim of the present study was to evaluate the effect of PAR1 activation on the osteogenic activity of human periodontal ligament stem cells (PDLSCs). PDLSCs were cultured in the presence of PAR1-selective agonist peptide (100 nM), thrombin (0.1 U/mL), or PAR1 antagonist peptide (100 nM). Calcium deposits, calcium concentration (supernatant), alkaline phosphatase activity (ALP), cell proliferation, and gene (qPCR) and protein expression (ELISA assay) of osteogenic factors were assessed at 2, 7, and 14 days. PAR1 activation led to increased calcium deposits (p<0.05), calcium concentration (p<0.05), ALP activity (p<0.05), and cell proliferation (p<0.05). Further, PAR1 activation may increase gene and protein expression of Runx2 (p<0.05) and OPG (p<0.05). In conclusion, PAR1 activation increases osteogenic activity of PDLSCs, providing a possible new strategy for periodontal regenerative therapies.

scholarly journals Stemness Characteristics of Periodontal Ligament Stem Cells from Donors and Multiple Sclerosis Patients: A Comparative Study

2017 ◽  
Vol 2017 ◽  
pp. 1-14 ◽  
Author(s):  
Francesca Diomede ◽  
Thangavelu Soundara Rajan ◽  
Marco D’Aurora ◽  
Placido Bramanti ◽  
Ilaria Merciaro ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Stem Cells ◽  
Cell Proliferation ◽  
Stem Cell ◽  
Cell Therapy ◽  
Stem Cell Therapy ◽  
Periodontal Ligament ◽  
Stem Cell Marker ◽  
Periodontal Ligament Stem Cells ◽  
Multiple Sclerosis Patients

Multiple sclerosis (MS) is the most prevalent and progressive autoimmune disease that affects the central nervous system, and currently, no drug is available for the treatment. Stem cell therapy has received substantial attention in MS treatment. Recently, we demonstrated the immunosuppressive effects of mesenchymal stem cells derived from neural crest-originated human periodontal ligament tissue (hPDLSCs) in an in vivo model of MS. In the present study, we comparatively investigated the stemness properties of hPDLSCs derived from healthy donors and relapsing-remitting MS patients. Stem cell marker expression, cell proliferation, and differentiation capacity were studied. We found that both donor- and MS patient-derived hPDLSCs at early passage 2 showed similar expression of surface antigen markers and cell proliferation rate. Significant level of osteogenic, adipogenic, chondrogenic, and neurogenic differentiation capacities was observed in both donor- and MS patient-derived hPDLSCs. Interestingly, these cells maintained the stemness properties even at late passage 15. Senescence markers p16 and p21 expression was considerably enhanced in passage 15. Our results propose that hPDLSCs may serve as simple and potential autologous stem cell niche, which may help in personalized stem cell therapy for MS patients.

scholarly journals Galuteolin suppresses proliferation and inflammation in TNF-α-induced RA-FLS cells by activating HMOX1 to regulate IKKβ/NF-κB pathway

2020 ◽  
Vol 15 (1) ◽  
Author(s):  
Yin Guan ◽  
Xiaoqian Zhao ◽  
Weiwei Liu ◽  
Yue Wang
Keyword(s):  
Cell Proliferation ◽  
Caspase 3 ◽  
Inflammatory Factors ◽  
Tunel Staining ◽  
Dependent Manner ◽  
Protective Effects ◽  
Tnf Α ◽  
Apoptosis And Inflammation ◽  
Dose Dependent ◽  
Anti Inflammation

Abstract Objective Galuteolin (Galu) is a substance extracted and purified from honeysuckle. The purpose of this study was to explore the effects of Galu on the TNF-α-induced RA-FLS cells (synoviocytes) and reveal its potential molecular mechanism from the perspectives of anti-apoptosis and anti-inflammation. Methods After TNF-α stimulation, cell proliferation of RA-FLS was assessed by CCK-8 assay. TUNEL staining was used to detect the apoptosis. Western blot was used to detect the expressions of Iκκβ, p-p65, p65, p-IκB, IκB, Cleaved-caspase3, Caspase-3, Bcl-2, and Bax. HO-1 were determined by RT-PCR. The contents of pro-inflammatory cytokines IL-1β, IL-6, IL-8, and MMP-1 were determined by ELISA. Results Galu significantly suppressed cell proliferation in a dose-dependent manner. Additionally, Galu obviously promotes cell apoptosis rate of RA-FLS cells and elevated the expression levels of HO-1, caspase-3, and Bax, while reducing the expression level of Bcl-2. Furthermore, Galu apparently inhibited the levels of Iκκβ, p-p65, and p-IκB. Moreover, Galu also significantly reduced the levels of pro-inflammatory factors IL-1β, IL-6, IL-8, and MMP-1 in RA-FLS cells. Conclusion Galuteolin exerts protective effects against TNF-α-induced RA-FLS cells by inhibiting apoptosis and inflammation, which can guide the clinical use of rheumatoid arthritis.

Mechanism for depression of heart sarcolemmal Ca2+ pump by oxygen free radicals

1989 ◽  
Vol 257 (3) ◽  
pp. H804-H811 ◽  
Author(s):  
M. Kaneko ◽  
V. Elimban ◽  
N. S. Dhalla
Keyword(s):  
Free Radicals ◽  
Oxygen Free Radicals ◽  
Sulfhydryl Group ◽  
Sulfhydryl Groups ◽  
Time Dependent ◽  
Dependent Manner ◽  
Protective Effects ◽  
Inhibitory Effects ◽  
Sulfhydryl Reagents ◽  
Dose Dependent

To understand the involvement of changes in sulfhydryl groups in causing depression of the sarcolemmal Ca2+-pump activities, this study was undertaken to examine the effects of oxygen free radicals on rat heart sarcolemmal sulfhydryl groups, Ca2+-stimulated adenosinetriphosphatase (ATPase), and ATP-dependent Ca2+ accumulation. In addition, the effects of sulfhydryl reagents such as dithiothreitol, cysteine, and N-ethylmaleimide on Ca2+-pump activities were investigated. The inhibition of sarcolemmal Ca2+-pump activities by O2-. (xanthine + xanthine oxidase) and H2O2 was decreased by the addition of dithiothreitol or cysteine in a dose-dependent manner. N-ethylmaleimide also showed inhibitory effects on Ca2+-pump activities both in a dose- and time-dependent manner; dithiothreitol and cysteine prevented changes in Ca2+-pump activities because of N-ethylmaleimide. Heart sarcolemmal sulfhydryl groups were depressed by O2-., H2O2, and .OH (H2O2 + Fe2+) both in a dose- and time-dependent manner. Superoxide dismutase, catalase, and D-mannitol showed protective effects on the sulfhydryl group depression by O2-., H2O2, and .OH, respectively. A significant correlation between changes in sarcolemmal Ca2+-stimulated ATPase activity and sarcolemmal sulfhydryl groups was seen. These results indicate that oxygen free radicals may depress the heart sarcolemmal Ca2+-pump activities by modifying the sulfhydryl groups in the sarcolemmal membrane.

scholarly journals ET-1 Promotes Differentiation of Periodontal Ligament Stem Cells into Osteoblasts through ETR, MAPK, and Wnt/β-Catenin Signaling Pathways under Inflammatory Microenvironment

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Li Liang ◽  
Wei Zhou ◽  
Nan Yang ◽  
Jifeng Yu ◽  
Hongchen Liu
Keyword(s):  
Stem Cells ◽  
Western Blot ◽  
Signaling Pathways ◽  
Periodontal Ligament ◽  
Cytokine Expression ◽  
Osteogenic Potential ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Periodontal Ligament Stem Cells ◽  
Inflammatory Microenvironment

Periodontitis is a kind of chronic inflammatory disease that affects the tooth-supporting tissues. ET-1 is related to periodontitis and involved in the regulation of cytokines, but the mechanisms remain unclear. The aim of this study is to investigate how ET-1 affects proinflammatory cytokine expression and differentiation in human periodontal ligament stem cells (PDLSCs). PDLSCs were isolated from the periodontal ligament tissues of periodontitis patients and then treated with ET-1 (1, 10, or 100 nM) for 12 h, 24 h, or 72 h. The osteogenic potential of PDLSCs was tested using ALP staining. TNF-α, IL-1β, and IL-6 levels were evaluated by ELISA and western blot. Runx2, OCN, and COL1 mRNA and western levels were detected by RT-PCR and western blot, respectively. To examine the signaling pathways and molecular mechanisms involved in ET-1-mediated cytokine expression and osteogenic differentiation, ETR pathway, MAPKs pathway, Wnt/β-catenin pathway, and Wnt/Ca2+pathway were detected by RT-PCR and western blot, respectively. ET-1 promoted differentiation of PDLSCs into osteoblasts by increasing secretion of TNF-α, IL-1β, and IL-6 in a dose- and time-dependent manner. ET-1 also increased expression of Runx2, OCN, and COL1. ET-1 promotes differentiation of PDLSCs into osteoblasts through ETR, MAPK, and Wnt/β-catenin signaling pathways under inflammatory microenvironment.

Sign in / Sign up

Export Citation Format

Share Document