Aspirin Enhances Osteogenic Potential of Periodontal Ligament Stem Cells (PDLSCs) and Modulates the Expression Profile of Growth Factor–Associated Genes in PDLSCs

2016 ◽  
Vol 87 (7) ◽  
pp. 837-847 ◽  
Author(s):  
Fazliny Abd Rahman ◽  
Johari Mohd Ali ◽  
Mariam Abdullah ◽  
Noor Hayaty Abu Kasim ◽  
Sabri Musa
Keyword(s):  
Stem Cells ◽  
Growth Factor ◽  
Expression Profile ◽  
Periodontal Ligament ◽  
Osteogenic Potential ◽  
Periodontal Ligament Stem Cells

scholarly journals 3D Clumps/Extracellular Matrix Complexes of Periodontal Ligament Stem Cells Ameliorate the Attenuating Effects of LPS on Proliferation and Osteogenic Potential

2021 ◽  
Vol 11 (6) ◽  
pp. 528
Author(s):  
Spoorthi Ravi Banavar ◽  
Swati Yeshwant Rawal ◽  
Shaju Jacob Pulikkotil ◽  
Umer Daood ◽  
Ian C. Paterson ◽  
...  
Keyword(s):  
Stem Cells ◽  
Periodontal Ligament ◽  
3D Culture ◽  
Osteogenic Potential ◽  
Culture Methods ◽  
Periodontal Ligament Stem Cells ◽  
Raman Analysis ◽  
Atp Production ◽  
Tnf Α ◽  
Inflammatory Milieu

Background: The effects of lipopolysaccharide (LPS) on cell proliferation and osteogenic potential (OP) of MSCs have been frequently studied. Objective: to compare the effects of LPS on periodontal-ligament-derived mesenchymal stem cells (PDLSCs) in monolayer and 3D culture. Methods: The PDLSCs were colorimetrically assessed for proliferation and osteogenic potential (OP) after LPS treatment. The 3D cells were manually prepared by scratching and allowing them to clump up. The clumps (C-MSCs) were treated with LPS and assessed for Adenosine triphosphate (ATP) and OP. Raman spectroscopy was used to analyze calcium salts, DNA, and proline/hydroxyproline. Multiplexed ELISA was performed to assess LPS induced local inflammation. Results: The proliferation of PDLSCs decreased with LPS. On Day 28, LPS-treated cells showed a reduction in their OP. C-MSCs with LPS did not show a decrease in ATP production. Principal bands identified in Raman analysis were the P–O bond at 960 cm−1 of the mineral component, 785 cm−1, and 855 cm−1 showing qualitative changes in OP, proliferation, and proline/hydroxyproline content, respectively. ELISA confirmed increased levels of IL-6 and IL-8 but with the absence of TNF-α and IL-1β secretion. Conclusions: These observations demonstrate that C-MSCs are more resistant to the effects of LPS than cells in monolayer cell culture. Though LPS stimulation of C-MSCs creates an early pro-inflammatory milieu by secreting IL-6 and IL-8, PDLSCs possess inactivated TNF promoter and an ineffective caspase-1 activating process.

scholarly journals TRIM16 Positively Regulates Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Modulating the Ubiquitination and Degradation of RUNX2

2020 ◽  
Author(s):  
Yi Zhao ◽  
Qiaoli Zhai ◽  
Hong Liu ◽  
Xun Xi ◽  
Shuai Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Alkaline Phosphatase ◽  
Bone Regeneration ◽  
Osteogenic Differentiation ◽  
Periodontal Ligament ◽  
Osteogenic Potential ◽  
Common Disease ◽  
Periodontal Ligament Stem Cells ◽  
Alizarin Red ◽  
Periodontal Tissues

Abstract BackgroundPeriodontal disease is a common disease that compromises the integrity of tooth-supporting tissues. Bone regeneration is the ultimate goal of periodontal therapies, in which osteogenic differentiation of human periodontal ligament stem cells plays a critical role. The tripartite motif (TRIM)16 is downregulated in periodontal tissues of patients with periodontitis and involved in osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMSCs).However, the role of TRIM16 in the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) is largely unknown.MethodshPDLSCs were isolated and identified by immunophenotype assays using flow cytometry. Overexpression plasmids and specific short-hairpin RNAs (shRNAs) were constructed to manipulate the expression of target molecules. Alkaline phosphatase (ALP) staining, alizarin red staining (ARS) and enzyme‐linked immunosorbent assays (ELISA) were used to evaluate osteogenic potential capacity. Reverse transcription quantitative PCR (RT-qPCR) and Western blot analysis were performed to determine the expression of osteogenic-related markers and activation of relevant signaling pathways. Co-immunoprecipitation assays were performed to confirm the interactions between proteins and the ubiquitination of RUNX2. A LC-MS/MS analysis was performed to explore the different expression proteins in present of TRIM16.ResultsTRIM16 significantly promoted alkaline phosphatase activity and mineralized nodule formation, and positively regulated the osteogenic differentiation of hPDLSCs by enhancing protein expression of RUNX2, COL1A1 and OCN. Mechanistically, TRIM16 serves as a pivotal factor that stabilizes RUNX2 protein levels by decreasing CHIP-mediated K48-linked ubiquitination degradation of the RUNX2 protein. Besides, TRIM16 significantly increased expression of COL1A1 via activation of p38MAPK/RUNX2.ConclusionThis study identified a novel mechanism of TRIM16 in regulating stability of the RUNX2 protein, which may promote the osteogenic differentiation of hPDLSCs. TRIM16 may be a potential target of stem cell based-bone regeneration for periodontal therapies.

scholarly journals Transforming Growth Factor-β3/Chitosan Sponge (TGF-β3/CS) Facilitates Osteogenic Differentiation of Human Periodontal Ligament Stem Cells

2019 ◽  
Vol 20 (20) ◽  
pp. 4982 ◽  
Author(s):  
Yangfan Li ◽  
Zhifen Qiao ◽  
Fenglin Yu ◽  
Huiting Hu ◽  
Yadong Huang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factor ◽  
Periodontal Disease ◽  
Osteogenic Differentiation ◽  
Periodontal Ligament ◽  
Transforming Growth Factor ◽  
Calcium Content ◽  
Hard Tissue ◽  
Periodontal Ligament Stem Cells ◽  
Tissue Defects

Periodontal disease is the main reason for tooth loss in adults. Tissue engineering and regenerative medicine are advanced technologies used to manage soft and hard tissue defects caused by periodontal disease. We developed a transforming growth factor-β3/chitosan sponge (TGF-β3/CS) to repair periodontal soft and hard tissue defects. We investigated the proliferation and osteogenic differentiation behaviors of primary human periodontal ligament stem cells (hPDLSCs) to determine the bioactivity and potential application of TGF-β3 in periodontal disease. We employed calcein-AM/propidium iodide (PI) double labeling or cell membranes (CM)-Dil labeling coupled with fluorescence microscopy to trace the survival and function of cells after implantation in vitro and in vivo. The mineralization of osteogenically differentiated hPDLSCs was confirmed by measuring alkaline phosphatase (ALP) activity and calcium content. The levels of COL I, ALP, TGF-βRI, TGF-βRII, and Pp38/t-p38 were assessed by western blotting to explore the mechanism of bone repair prompted by TGF-β3. When hPDLSCs were implanted with various concentrations of TGF-β3/CS (62.5–500 ng/mL), ALP activity was the highest in the TGF-β3 (250 ng/mL) group after 7 d (p < 0.05 vs. control). The calcium content in each group was increased significantly after 21 and 28 d (p < 0.001 vs. control). The optimal result was achieved by the TGF-β3 (500 ng/mL) group. These results showed that TGF-β3/CS promotes osteogenic differentiation of hPDLSCs, which may involve the p38 mitogen-activated protein kinase (MAPK) signaling pathway. TGF-β3/CS has the potential for application in the repair of incomplete alveolar bone defects.

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 Epithelial Cell Rests of Malassez Provide a Favorable Microenvironment for Ameliorating the Impaired Osteogenic Potential of Human Periodontal Ligament Stem Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Yanjiao Li ◽  
Anqi Liu ◽  
Liqiang Zhang ◽  
Zhiwei Wang ◽  
Nana Hui ◽  
...  
Keyword(s):  
Stem Cells ◽  
Epithelial Cell ◽  
Periodontal Ligament ◽  
Wnt Pathway ◽  
Periodontal Regeneration ◽  
Osteogenic Potential ◽  
Periodontal Ligament Stem Cells ◽  
Root Sheath ◽  
Healthy Donors ◽  
Stromal Stem Cells

Human periodontal ligament stromal/stem cells (PDLSCs) are ideal candidates for periodontal regeneration and are of significant importance in clinical practice. However, PDLSCs derived from diseased microenvironments exert impaired behavior, which leads to the failure of periodontal regeneration. The epithelial cell rests of Malassez (ERM), which are involved in periodontal homeostasis, are residual cells from Hertwig's epithelial root sheath (HERS). However, the function of ERM remains largely unknown. Therefore, the aim of this study was to evaluate the effect of ERM on the osteogenic potential of PDLSCs from an impaired microenvironment. PDLSCs from healthy donors (H-PDLSCs), periodontitis donors (P-PDLSCs) and human ERM were harvested. Osteogenic evaluation showed a lower osteogenic potential of P-PDLSCs compared to that of H-PDLSCs. Then, we co-cultured ERM with P-PDLSCs, and the data showed that ERM promoted the expression of osteogenic genes and proteins in P-PDLSCs. In addition, we collected the PDLSCs from aged donors (A-PDLSCs) and analyzed the osteogenesis capacity of the A-PDLSCs and A-PDLSCs + ERM groups, which displayed similar results to P-PDLSCs. Finally, we evaluated the Wnt pathway, which is associated with osteogenic differentiation of stromal/stem cells, in A-PDLSCs + ERM and P-PDLSCs + ERM groups, which indicated that suppression of the Wnt pathway may result in an increase in the osteogenic properties of A-PDLSCs + ERM and P-PDLSCs + ERM groups. Taken together, the above findings shed new light on the function of ERM and provide a novel therapeutic for optimizing PDLSCs-based periodontal regeneration.

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