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

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

Ascorbic Acid: A New Player of Epigenetic Regulation in LPS-gingivalis Treated Human Periodontal Ligament Stem Cells

Periodontitis is usually sustained from microorganism of oral cavity, like Porphyromonas gingivalis (P. gingivalis). Periodontal disease is an infectious disease that afflicts a large number of people. Researches are investigating on the mesenchymal stem cells (MSCs) response to inflammatory events in combination with antioxidant substances. In particular, ascorbic acid (AA) increased cell proliferation, upregulated the cells pluripotency marker expression, provide a protection from inflammation, and induced the regeneration of periodontal ligament tissue. The purpose of the present research was to investigate the effects of AA in primary culture of human periodontal ligament stem cells (hPDLSCs) exposed to P. gingivalis lipopolysaccharide (LPS-G). The effect of AA on hPDLSCs exposed to LPS-G was determined through the cell proliferation assay. The molecules involved in the inflammatory pathway and epigenetic regulation have been identified using immunofluorescence and Western blot analyses. miR-210 level was quantified by qRT-PCR, and the ROS generation was finally studied. Cells co-treated with LPS-G and AA showed a restoration in terms of cell proliferation. The expression of NFκB, MyD88, and p300 was upregulated in LPS-G exposed cells, while the expression was attenuated in the co-treatment with AA. DNMT1 expression is attenuated in the cells exposed to the inflammatory stimulus. The level of miR-210 was reduced in stimulated cells, while the expression was evident in the hPDLSCs co-treated with LPS-G and AA. In conclusion, the AA could enhance a protective effect in in vitro periodontitis model, downregulating the inflammatory pathway and ROS generation and modulating the miR-210 level