Abstract
Human gingival mesenchymal stem cells (hGMSCs) and endothelial committed hGMSCs (e-hGMSCs) have considerable potential to serve as an in vitro model to replicate the inflammation sustained by Porphyromonas gingivalis in periodontal and cardiovascular diseases. The present study aimed to investigate the effect of ascorbic acid (AA) on the inflammatory reverting action of lipopolysaccharide (LPS-G) on the cell metabolic activity, inflammation pathway and reactive oxygen species (ROS) generation in hGMSCs and e-hGMSCs. Cells were treated with LPS-G (5 μg mL−1) or AA (50 μg mL−1) and analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay, immunofluorescence and Western blot methods. The rate of cell metabolic activity was decreased significantly in LPS-G-treated groups, while groups co-treated with LPS-G and AA showed a logarithmic cell metabolic activity rate similar to untreated cells. AA treatment attenuated the inflammatory effect of LPS-G by reducing the expression of TLR4/MyD88/NFκB/NLRP3/Caspase-1/IL-1β, as demonstrated by Western blot analysis and immunofluorescence acquisition. LPS-G-induced cells displayed an increase in ROS production, while AA co-treated cells showed a protective effect. In summary, our work suggests that AA attenuated LPS-G-mediated inflammation and ROS generation in hGMSCs and e-hGMSCs via suppressing the NFκB/Caspase-1/IL-1β pathway. These findings indicate that AA may be considered as a potential factor involved in the modulation of the inflammatory pathway triggered by LPS-G in an vitro cellular model.
Highlights
After treating Human gingival mesenchymal stem cells (hGMSCs) with the osteogenic differentiation medium, the formation of calcium deposits revealed by alizarin red staining was observable (Figure 1B, right panel)
These results indicate that hGMSCs can differentiate into both lineages
This study showed that in cells treated with ascorbic acid, high production of reactive oxygen species (ROS) is present in hGMSCS and e-hGMSCs stimulated with LPS-G, while the ROS levels are reduced in cells co-treated with AA and LPS-G
Summary
Ascorbic acid (AA) is well known to perform a key role in the maintenance of tissue integrity, scavenging free radicals, and has demonstrated immunomodulatory properties during chronic inflammatory diseases. AA showed a deceleration in the progression of tissue loss and induced regeneration by stimulating the progenitor cells of the periodontal ligament commitment [1]. Periodontal disease (PD) can be defined as a complex oral disease and its treatment represents a challenging condition for clinicians [2]. Periodontal disease involves chronic inflammation sustained by various types of bacteria that accumulate in dental plaque and cause localized inflammation by producing various
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