MicroRNA 210 Mediates VEGF Upregulation in Human Periodontal Ligament Stem Cells Cultured on 3DHydroxyapatite Ceramic Scaffold

The aim of the present research was the evaluation of the behavior of human periodontal ligament stem cells (hPDLSCs), cultured in presence of Endobon® Xenograft Granules (G), a fully deproteinated hydroxyapatite ceramic scaffold derived from cancellous bovine bone. hPDLSCs were seeded with and without G for 24 h to 1 week. The cell growth, morphological features, adhesiveness, differentiation ability, modulation of miR-210 and Vascular Endothelial Growth Factor (VEGF) secretion were analyzed by means of MTT assay, Scanning Electron Microscopy (SEM), Confocal Laser Scanning Microscopy (CLSM), Alizarin Red S assay, RT-PCR and ELISA test, respectively. hPDLSCs grown on the biomaterial showed the ability to form focal adhesion on the substrate, as demonstrated by vinculin expression. These data were supported by SEM analysis showing that an adhesiveness process associated to cell growth occurs between cells and biomaterials. The osteogenic differentiation, evaluated by morphological, biochemical, and RT-PCR analysis, was pronounced in the hPDLSCs grown in the three-dimensional inorganic bovine bone substitute in the presence of osteoinductive conditions. In addition, an upregulation of miR-210 and VEGF was evident in cells cultured in presence of the biomaterial. Our results inspire us to consider granules not only an adequate biocompatible three-dimensional biomaterial, but also an effective inductor of miR-210 and VEGF; in fact, the involvement of miR-210 in VEGF secretion could offer a novel regulatory system in the early steps of the bone-regeneration process.

Download Full-text

VEGF/VEGF-R/RUNX2 Upregulation in Human Periodontal Ligament Stem Cells Seeded on Dual Acid Etched Titanium Disk

Materials ◽

10.3390/ma13030706 ◽

2020 ◽

Vol 13 (3) ◽

pp. 706 ◽

Cited By ~ 2

Author(s):

Francesca Diomede ◽

Guya Diletta Marconi ◽

Marcos F. X. B. Cavalcanti ◽

Jacopo Pizzicannella ◽

Sante Donato Pierdomenico ◽

...

Keyword(s):

Stem Cells ◽

Cell Adhesion ◽

Confocal Laser Scanning Microscopy ◽

Periodontal Ligament ◽

Laser Scanning ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Rt Pcr ◽

Periodontal Ligament Stem Cells ◽

Titanium Surfaces

In restorative dentistry, the main implants characteristic is the ability to promote the osseointegration process as the result of interaction between angiogenesis and osteogenesis events. On the other hand, implants cytocompatibility remains a necessary feature for the success of surgery. The purpose of the current study was to investigate the interaction between human periodontal stem cells and two different types of titanium surfaces, to verify their cytocompatibility and cell adhesion ability, and to detect osteogenic and angiogenic markers, trough cell viability assay (MTT), Confocal Laser Scanning Microscopy (CLSM), scanning electron microscopy (SEM), and gene expression (RT-PCR). The titanium surfaces, machined (CTRL) and dual acid etched (TEST), tested in culture with human periodontal ligament stem cells (hPDLSCs), were previously treated in two different ways, in order to evaluate the effects of CTRL and TEST and define the best implant surface. Furthermore, the average surface roughness (Ra) of both titanium surfaces, CTRL and TEST, has been assessed through atomic force microscopy (AFM). The vascular endothelial growth factor (VEGF) and Runt-related transcription factor 2 (RUNX2) expressions have been analyzed by RT-PCR, WB analysis, and confocal laser scanning microscopy. Data evidenced that the different morphology and topography of the TEST disk increased cell growth, cell adhesion, improved osteogenic and angiogenic events, as well osseointegration process. For this reason, the TEST surface was more biocompatible than the CTRL disk surface.

Download Full-text

Enhanced VEGF/VEGF-R and RUNX2 Expression in Human Periodontal Ligament Stem Cells Cultured on Sandblasted/Etched Titanium Disk

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2020.00315 ◽

2020 ◽

Vol 8 ◽

Cited By ~ 2

Author(s):

Guya Diletta Marconi ◽

Francesca Diomede ◽

Jacopo Pizzicannella ◽

Luigia Fonticoli ◽

Ilaria Merciaro ◽

...

Keyword(s):

Stem Cells ◽

Periodontal Ligament ◽

Periodontal Ligament Stem Cells ◽

Runx2 Expression ◽

Titanium Disk ◽

Cells Cultured

Download Full-text

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

BioMolecular Concepts ◽

10.1515/bmc-2020-0016 ◽

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.

Download Full-text

Human periodontal ligament stem cells cultured onto cortico-cancellous scaffold drive bone regenerative process

10.22203/ecm.v032a12 ◽

2016 ◽

Vol 32 ◽

pp. 181-201 ◽

Cited By ~ 28

Author(s):

F Diomede ◽

◽

N Zini ◽

V Gatta ◽

S Fulle ◽

...

Keyword(s):

Stem Cells ◽

Periodontal Ligament ◽

Regenerative Process ◽

Periodontal Ligament Stem Cells ◽

Cells Cultured

Download Full-text

In vivo periodontal tissue regeneration by periodontal ligament stem cells and endothelial cells in three-dimensional cell sheet constructs

Journal of Periodontal Research ◽

10.1111/jre.12405 ◽

2016 ◽

Vol 52 (3) ◽

pp. 408-418 ◽

Cited By ~ 14

Author(s):

C. P. Panduwawala ◽

X. Zhan ◽

W. L. Dissanayaka ◽

L. P. Samaranayake ◽

L. Jin ◽

...

Keyword(s):

Stem Cells ◽

Tissue Regeneration ◽

Periodontal Ligament ◽

Cell Sheet ◽

Three Dimensional ◽

Periodontal Tissue ◽

Periodontal Ligament Stem Cells ◽

Periodontal Tissue Regeneration ◽

Dimensional Cell

Download Full-text

Osteogenic Potential of Periodontal Ligament Stem Cells Cultured in Osteogenic and Regular Growth Media: Confocal and Scanning Electron Microscope Study

The Journal of Contemporary Dental Practice ◽

10.5005/jp-journals-10024-2822 ◽

2020 ◽

Vol 21 (7) ◽

pp. 776-780

Author(s):

Areej H Mukhtar ◽

Montaser N Alqutub

Keyword(s):

Stem Cells ◽

Periodontal Ligament ◽

Electron Microscope Study ◽

Regular Growth ◽

Osteogenic Potential ◽

Growth Media ◽

Periodontal Ligament Stem Cells ◽

Scanning Electron Microscope Study ◽

Scanning Electron ◽

Cells Cultured

Download Full-text

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

Mediators of Inflammation ◽

10.1155/2016/8467849 ◽

2016 ◽

Vol 2016 ◽

pp. 1-12 ◽

Cited By ~ 10

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.

Download Full-text

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

Applied Sciences ◽

10.3390/app11136040 ◽

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.

Download Full-text