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 Porphyromonas gingivalis lipopolysaccharide stimulation in human periodontal ligament stem cells: role of epigenetic modifications to the inflammation

2017 ◽  
Vol 61 (3) ◽  
Author(s):  
Francesca Diomede ◽  
Soundara Rajan Thangavelu ◽  
Ilaria Merciaro ◽  
Monica D'Orazio ◽  
Placido Bramanti ◽  
...  
Keyword(s):  
Stem Cells ◽  
Porphyromonas Gingivalis ◽  
Inflammatory Disease ◽  
Periodontal Ligament ◽  
Model System ◽  
Epigenetic Mechanism ◽  
Periodontal Ligament Stem Cells ◽  
Porphyromonas Gingivalis Lipopolysaccharide

<p>Periodontitis is a chronic oral inflammatory disease produced by bacteria. Gingival retraction and bone and connective tissues resorption are the hallmarks of this disease. Chronic periodontitis may contribute to the risk of onset or progression of neuroinflammatory pathological conditions, such as Alzheimer’s disease. The main goal of the present study was to investigate if the role of epigenetic modulations is involved in periodontitis using human periodontal ligament stem cells (hPDLSCs) as an <em>in vitro</em> model system. hPDLSCs were treated with lipopolysaccharide of <em>Porphyromonas gingivalis</em> and the expression of proteins associated with DNA methylation and histone acetylation, such as DNMT1 and p300, respectively, and inflammatory transcription factor NF-kB, were examined. Immunofluorescence, Western blot and next generation sequencing results demonstrated that <em>P. gingivalis </em>lipopolysaccharide significantly reduced DNA methylase DNMT1, while it markedly upregulated the level of histone acetyltransferase p300 and NF-kB in hPDLSCs. Our results showed that <em>P. gingivalis </em>lipopolysaccharide markedly regulate the genes involved in epigenetic mechanism, which may result in inflammation induction. We propose that <em>P. gingivalis </em>lipopolysaccharide-treated hPDLSCs could be a potential in vitro model system to study epigenetics modulations associated with periodontitis, which might be helpful to identify novel biomarkers linked to this oral inflammatory disease.</p>

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.

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.

Melatonin Promotes Osteogenic Differentiation in Lipopolysaccharide-Stimulated Human Periodontal Ligament Stem Cells Through Bone Morphogenic Proteins-2-Related Signaling

2019 ◽  
Vol 9 (5) ◽  
pp. 679-686
Author(s):  
Na Yu ◽  
Jinghui Zhang ◽  
Lijuan Han ◽  
Cunjirigala Na ◽  
Xiaoguang Yuan
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Wnt Signaling ◽  
Osteogenic Differentiation ◽  
Periodontal Ligament ◽  
Biological Activities ◽  
Phase Cell ◽  
Periodontal Ligament Stem Cells ◽  
Alizarin Red ◽  
Protein Expressions

Periodontitis is one of the most widespread infectious diseases that troubled the majority of adults. Human periodontal ligament stem cells (hPDLSCs) have been reported as a promising therapy for the treatment of periodontitis. Melatonin, an indoleamine hormone from pineal gland, has various biological activities such as anti-inflammation, anti-cancer and so on. However, whether it is functional in periodontitis is still unclear. The aim of this study was to investigate the effect of melatonin in periodontitis and elucidate the molecular mechanism. Lipopolysaccharide (LPS) was used to stimulate hPDLSCs, and viability of hPDLSCs that was treated with melatonin (0, 1, 10, 50 and 100 μmol/L) for 24 h or 48 h was determined by MTT assay. Flow cytometry analysis was carried out to detect the influence of melatonin on cell proliferation. Osteogenic differentiation ability of melatonin was determined by Alkaline phosphatase (ALP) assay kit and Alizarin Red Staining. Lastly, western blot was used for the determination of protein expressions related to proliferation, differentiation and ERK/Wnt signaling activity. The results showed that LPS significantly inhibited cell viability, which was reversed by melatonin, especially at 10 μM for 48 h and at 50 μM for 24 h. Melatonin (10 μM, 48 h) and melatonin (50 μM, 24 h) notably induced G0/G1 phase cell arrest, increased the expression of CDK2, cyclin E and decreased the expression of p27 in LPS-stimulated hPDLSCs. Besides, melatonin significantly promoted cell differentiation through increasing ALP activity, mineralization and protein expressions of Oct4, Sox-2, Runx2 and bone morphogenic protein-2 (BMP-2). Additionally, BMP-2 related ERK and Wnt signaling was activated with the treatment of melatonin in LPS-stimulated hPDLSCs. Collectively, melatonin could improve cell proliferation and osteogenic differentiation in LPS-stimulated hPDLSCs, partly through regulating BMP2-related ERK/Wnt pathway.

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