scholarly journals Differential Expression of Osteo-Modulatory Molecules in Periodontal Ligament Stem Cells in Response to Modified Titanium Surfaces

2014 ◽  
Vol 2014 ◽  
pp. 1-12 ◽  
Author(s):  
So Yeon Kim ◽  
Ji-Yeon Yoo ◽  
Joo-Young Ohe ◽  
Jung-Woo Lee ◽  
Ji-Hoi Moon ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Wnt Signaling ◽  
Osteogenic Differentiation ◽  
Mrna Expression ◽  
Periodontal Ligament ◽  
Signaling Molecules ◽  
Periodontal Ligament Stem Cells ◽  
Osteogenic Activity ◽  
Wnt Pathways

This study assessed differential gene expression of signaling molecules involved in osteogenic differentiation of periodontal ligament stem cells (PDLSCs) subjected to different titanium (Ti) surface types. PDLSCs were cultured on tissue culture polystyrene (TCPS), and four types of Ti discs (PT, SLA, hydrophilic PT (pmodPT), and hydrophilic SLA (modSLA)) with no osteoinductive factor and then osteogenic activity, including alkaline phosphatase (ALP) activity, mRNA expression of runt-related gene 2, osterix, FOSB, FRA1, and protein levels of osteopontin and collagen type IA, were examined. The highest osteogenic activity appeared in PDLSCs cultured on SLA, compared with the TCPS and other Ti surfaces. The role of surface properties in affecting signaling molecules to modulate PDLSC behavior was determined by examining the regulation of Wnt pathways. mRNA expression of the canonical Wnt signaling molecules, Wnt3a andβ-catenin, was higher on SLA and modSLA than on smooth surfaces, but gene expression of the calcium-dependent Wnt signaling molecules Wnt5a, calmodulin, and NFATc1 was increased significantly on PT and pmodPT. Moreover, integrinα2/β1, sonic hedgehog, and Notch signaling molecules were affected differently by each surface modification. In conclusion, surface roughness and hydrophilicity can affect differential Wnt pathways and signaling molecules, targeting the osteogenic differentiation of PDLSCs.

scholarly journals Influence of photobiomodulation and vitamin D on osteoblastic differentiation of human periodontal ligament stem cells and bone-like tissue formation through enzymatic activity and gene expression

2020 ◽  
Vol 11 (1) ◽  
pp. 172-181
Author(s):  
Latifa Mohamed Abdelgawad ◽  
Asmaa Mohamed Abdelaziz ◽  
Dina Sabry ◽  
Marwa Abdelgwad
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Vitamin D ◽  
Cell Growth ◽  
Osteogenic Differentiation ◽  
Growth Curve ◽  
Mtt Assay ◽  
Periodontal Ligament ◽  
Mrna Levels ◽  
Periodontal Ligament Stem Cells

Abstract(1)BackgroundHuman periodontal ligament stem cells (HPDLSCs) are a unique population of mesenchymal stem cells (MSCs). Recently, the positive effects of photobiomodulation on the regulation of MSCs proliferation and osteogenic differentiation have gained significant attention. This study aimed to assess the effects of photobiomodulation and vitamin D (as an anabolic factor) on HPDLSCs for bone regeneration.(2)MethodsHPDLSCs were collected, isolated, and characterized and then divided into six groups: groups I and II, control and (10−7 Mol) vitamin D, respectively; group III, irradiation at 1 J/cm2 of 808-nm diode laser; group IV, irradiation at 1 J/cm2 and culture with vitamin D; group V, irradiation at 2 J/cm2, and group VI, irradiation at 2 J/cm2 and culture with vitamin D. Cell viability assay was measured through MTT assay and cell growth curve. Alkaline phosphatase (ALP) enzyme activity and mRNA levels of RUNX2, collagen 1 (Col-1), ALP, and osteonectin were also assessed.(3)ResultsPhotobiomodulation at 1 and 2 J/cm2 combined with vitamin D significantly promoted HPDLSC proliferation (in MTT assay and cell growth curve results) and osteogenic differentiation (through the gene expression of RUNX2, Col-1, ALP, and osteonectin levels (p < 0.05).(4)ConclusionLaser irradiation at 2 J/cm2 combined with vitamin D3 enhanced osteoblast differentiation and proliferation of cultured HPDLSCs and thus could further substitute bone grafting.

scholarly journals Comparison of 2- and 3-Dimensional Cultured Periodontal Ligament Stem Cells; a Pilot Study

2021 ◽  
Vol 11 (3) ◽  
pp. 1083
Author(s):  
Yun Yeong Jeong ◽  
Mi Sun Kim ◽  
Ko Eun Lee ◽  
Ok Hyung Nam ◽  
Ji-Hyun Jang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Rna Sequencing ◽  
Periodontal Ligament ◽  
Cultured Cells ◽  
Control Group ◽  
Periodontal Ligament Stem Cells ◽  
3 Dimensional ◽  
2D And 3D

This study compared the characteristics of periodontal ligament stem cells (PDLSCs) cultured using 3-dimensional (3D) versus conventional 2-dimensional (2D) methods. PDLSCs were cultured in either a 3D culture with a non-adhesive culture plate (Stemfit 3D®) or a conventional 2D culture using a 6-well plate. Morphology, viability, proliferation ability, and osteogenic differentiation were analyzed to characterize the differences induced in identical PDLSCs by 3D and 2D culture environments. In addition, gene expression was analyzed using RNA sequencing to further characterize the functional differences. The diameter and the viability of the 3D-cultured PDLSCs decreased over time, but the shape of the spheroid was maintained for 20 days. Although osteogenic differentiation occurred in both the 2D- and 3D-cultured PDLSCs, compared to the control group it was 20.8 and 1.6 higher in the 3D- and 2D-cultured cells, respectively. RNA sequencing revealed that PDLSCs cultured using 2D and 3D methods have different gene expression profiles. The viability of the 3D-cultured cells was decreased, but they showed superior osteogenic differentiation compared to 2D-cultured cells. Within the limitations of this study, the results demonstrate that the structure and function of PDLSCs are influenced by the cell culture method.

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.

scholarly journals Gene expression profiling on effect of aspirin on osteogenic differentiation of periodontal ligament stem cells

BDJ Open ◽  
2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Fazliny Abd Rahman
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Periodontal Ligament ◽  
Expression Profiles ◽  
Periodontal Ligament Stem Cells ◽  
Extracellular Matrix Components ◽  
Regenerative Dentistry

AbstractPeriodontal ligament (PDL) contains a unique population of mesenchymal stem cells (MSCs), also known as PDL stem cells (PDLSCs). The regenerative properties of PDLSCs hold great potential for its use in stem cells based therapy, particularly for periodontal or bone regeneration. The present study investigated the global gene expression profile in PDLSCs during osteogenic differentiation. MSCs from PDL were isolated from normal permanent human teeth (n = 3). Microarray analysis was used to study the effects of ASA (200, 500, and 1000 μM) on the gene expression profiles in PDLSCs during osteogenic differentiation. Microarray study revealed that ASA was able to modulate PDLSCs gene expression profile. At 200 µM, 315 genes were dysregulated genes (DE), involving 151 upregulated and 164 downregulated genes. At 500 µM, 794 genes were DE, involving of 364 upregulated and 430 downregulated genes. At 1000 µM, the number of DE genes increased to 2035, of which 735 were upregulated and 1300 were downregulated. Bioinformatics analyses of the gene expression data revealed that the majority of DE genes (for 500 and 1000 µM ASA treatment) are involved in osteogenic differentiation. The gene network analysis was carried out using Ingenuity Pathway Analysis (IPA) software, and this revealed that the number of gene groups involved in cell adhesion and extracellular matrix components were increased. This study indicated that ASA could enhance PDLSCs functions and provide evidence for the potential use of ASA with PDLSCs for regenerative dentistry applications, particularly in the areas of periodontal health and regeneration. Periodontal ligament stem cells (PDLSCs) Aspirin (ASA) Microarray Osteogenic

scholarly journals In Vitro Characterization of Periodontal Ligament Stem Cells Derived from Supernumerary Teeth in Three-Dimensional Culture Method

2021 ◽  
Vol 11 (13) ◽  
pp. 6040
Author(s):  
Yun Yeong Jeong ◽  
Mi Sun Kim ◽  
Ko Eun Lee ◽  
Ok Hyung Nam ◽  
Ji-Hyun Jang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Periodontal Ligament ◽  
Three Dimensional ◽  
3D Culture ◽  
Culture Method ◽  
Supernumerary Teeth ◽  
Periodontal Ligament Stem Cells

Objective: The aim of this study was to compare the characteristics of periodontal ligament stem cells derived from supernumerary teeth (sPDLSCs), cultured using a three-dimensional (3D) method and a conventional two-dimensional (2D) method. Methods: The morphology, viability, and osteogenic differentiation of the cells were analyzed. In addition, gene expression was analyzed by RNA sequencing, to characterize the functional differences. Results: The diameter of the 3D-cultured sPDLSCs decreased over time, but the spheroid shape was maintained for 7 days. The osteogenic differentiation was similar in the 2D and 3D. The gene expression related to the extracellular matrix (7.3%), angiogenesis (5.6%), cell proliferation (4.6%), inflammatory response (3.7%), and cell migration (3.5%) differed (p < 0.05). Conclusions: Within the limitations of this study, sPDLSCs varied in formation and function, depending on the culture method. In future, it is necessary to study tissue engineering using the advantages of 3D culture and the fewer ethical problems of supernumerary teeth.

scholarly journals EphrinB2 regulates osteogenic differentiation of periodontal ligament stem cells and alveolar bone defect regeneration in beagles

2019 ◽  
Vol 10 ◽  
pp. 204173141989436 ◽  
Author(s):  
Penglai Wang ◽  
Wen Wang ◽  
Tengyu Geng ◽  
Yi Liu ◽  
Shaoyue Zhu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Bone Regeneration ◽  
Osteogenic Differentiation ◽  
Periodontal Ligament ◽  
Alveolar Bone ◽  
Bone Homeostasis ◽  
Periodontal Ligament Stem Cells ◽  
Osteogenic Gene Expression ◽  
Osteogenic Gene

EphrinB2, a membrane protein regulating bone homeostasis, has been demonstrated to induce osteogenic gene expression in periodontal ligament fibroblasts. The aim of this study was to explore the effects of ephrinB2 on osteogenic differentiation of periodontal ligament stem cells and on alveolar bone regeneration in vivo. We assessed the osteogenic gene expression and osteogenic differentiation potential of ephrinB2-modified human and canine periodontal ligament stem cells, in which ephrinB2 expression was upregulated via lentiviral vector transduction. EphrinB2-modified canine periodontal ligament stem cells combined with PuraMatrix were delivered to critical-sized alveolar bone defects in beagles to evaluate bone regeneration. Results showed that ephrinB2 overexpression enhanced osteogenic gene transcription and mineral deposition in both human and canine periodontal ligament stem cells. Animal experiments confirmed that ephrinB2-modified canine periodontal ligament stem cells + PuraMatrix resulted in greater trabecular bone volume per tissue volume and trabecular thickness compared with other groups. Our study demonstrated that ephrinB2 promoted osteogenic differentiation of periodontal ligament stem cells and alveolar bone repair in beagles, highlighting its therapeutic potential for the treatment of alveolar bone damage.

scholarly journals Effects of Naringin on Proliferation and Osteogenic Differentiation of Human Periodontal Ligament Stem Cells In Vitro and In Vivo

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Lihua Yin ◽  
Wenxiao Cheng ◽  
Zishun Qin ◽  
Hongdou Yu ◽  
Zhanhai Yu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Periodontal Ligament ◽  
Trabecular Structure ◽  
Control Group ◽  
Periodontal Ligament Stem Cells ◽  
Expression Levels ◽  
Proliferation And Differentiation

This study is to explore the osteogenesis potential of the human periodontal ligament stem cells (hPDLSCs) induced by naringin in vitro and in vitro. The results confirmed that 1 μM naringin performs the best effect and a collection of bone-related genes (RUNX2,COL1A2, OPN, and OCN) had significantly higher expression levels compared to the control group. Furthermore, a typical trabecular structure was observed in vivo, surrounded by a large amount of osteoblasts. These results demonstrated that naringin, at a concentration of 1 μM, can efficiently promote the proliferation and differentiation of hPDLSCs both in vitro and in vivo.

scholarly journals Azithromycin Promotes the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells after Stimulation with TNF-α

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Tingting Meng ◽  
Ying Zhou ◽  
Jingkun Li ◽  
Meilin Hu ◽  
Xiaomeng Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Alkaline Phosphatase ◽  
Osteogenic Differentiation ◽  
Alkaline Phosphatase Activity ◽  
Periodontal Ligament ◽  
Caspase 3 ◽  
Caspase 8 ◽  
Periodontal Ligament Stem Cells ◽  
Alizarin Red ◽  
Induced Apoptosis

Background and Objective. This study investigated the effects and underlying mechanisms of azithromycin (AZM) treatment on the osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) after their stimulation with TNF-α in vitro. Methods. PDLSCs were isolated from periodontal ligaments from extracted teeth, and MTS assay was used to evaluate whether AZM and TNF-α had toxic effects on PDLSCs viability and proliferation. After stimulating PDLSCs with TNF-α and AZM, we analyzed alkaline phosphatase staining, alkaline phosphatase activity, and alizarin red staining to detect osteogenic differentiation. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis was performed to detect the mRNA expression of osteogenic-related genes, including RUNX2, OCN, and BSP. Western blotting was used to measure the NF-κB signaling pathway proteins p65, phosphorylated p65, IκB-α, phosphorylated IκB-α, and β-catenin as well as the apoptosis-related proteins caspase-8 and caspase-3. Annexin V assay was used to detect PDLSCs apoptosis. Results. TNF-α stimulation of PDLSCs decreased alkaline phosphatase and alizarin red staining, alkaline phosphatase activity, and mRNA expression of RUNX2, OCN, and BSP in osteogenic-conditioned medium. AZM enhanced the osteogenic differentiation of PDLSCs that were stimulated with TNF-α. Western blot analysis showed that β-catenin, phosphorated p65, and phosphorylated IκB-α protein expression decreased in PDLSCs treated with AZM. In addition, pretreatment of PDLSCs with AZM (10 μg/ml, 20 μg/ml) prevented TNF-α-induced apoptosis by decreasing caspase-8 and caspase-3 expression. Conclusions. Our results showed that AZM promotes PDLSCs osteogenic differentiation in an inflammatory microenvironment by inhibiting the WNT and NF-κB signaling pathways and by suppressing TNF-α-induced apoptosis. This suggests that AZM has potential as a clinical therapeutic for periodontitis.

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