Altered Osteogenic Differentiation in Mesenchymal Stem Cells Isolated from Compact Bone of Chicken Treated with Varying Doses of Lipopolysaccharides
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PROVIDER: S-EPMC10669906 | biostudies-literature | 2023 Nov
REPOSITORIES: biostudies-literature
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Project description:Lipopolysaccharides (LPS) are cell wall components of gram-negative bacteria and endotoxins that cause inflammation and sickness in vertebrate animals. Our objective to identify the plasma proteomic changes in an avian model of inflammation. Chickens were treated with either saline or LPS and the blood was collected 24 hours after injection. The pooled plasma samples were depleted of high abundant proteins, analyzed by Matrix assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) and liquid chromatography–tandem mass spectrometry (LC-MS/MS). MALDI-TOF analyses showed an increase in fibrinogen beta derived peptide and decrease in apolipoprotein AII- isoform X1 derived peptide in LPS samples. Label free quantitation of LC-MS/MS spectra revealed an increase in alpha-1 acid glycoprotein, chemokine-CCLI10, and cathelicidin-2 and decrease in interferon stimulated gene-12-2 in the LPS group. These differentially expressed proteins are associated with inflammation, cytokines, immunomodulation, and defense which are useful as candidate biomarkers of infection and inflammation.
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Project description:BackgroundPeriodontitis is a chronic infection initiated by oral bacterial and their virulence factors, yet the severity of periodontitis is largely determined by the dysregulated host immuno-inflammatory response. Baicalein is a flavonoid extracted from Scutellaria baicalensis with promising anti-inflammatory properties. This study aims to clarify the anti-inflammatory and osteogenic effects of baicalein in periodontal ligament cells (PDLCs) treated with lipopolysaccharides (LPS).MethodsHuman PDLCs were incubated with baicalein (0-100 μM) for 2 h prior to LPS challenge for 24 h. MTT analysis was adopted to assess the cytoxicity of baicalein. The mRNA and protein expression of inflammatory and osteogenic markers were measured by real-time polymerase chain reaction (PCR), western blot and enzyme-linked immunosorbent assay (ELISA) as appropriate. Alkaline phosphatase (ALP) and Alizarin red S (ARS) staining were performed to evaluate the osteogenic differentiation of PDLCs. The expression of Wnt/β-catenin and mitogen-activated protein kinase (MAPK) signaling related proteins was assessed by western blot.ResultsMTT results showed that baicalein up to 100 μM had no cytotoxicity on PDLCs. Baicalein significantly attenuated the inflammatory factors induced by LPS, including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), matrix metalloprotein-1 (MMP-1), MMP-2 and monocyte chemoattractant protein 1 (MCP-1) at both mRNA and protein level. Moreover, MAPK signaling (ERK, JNK and p38) was significantly inhibited by baicalein, which may account for the mitigated inflammatory response. Next, we found that baicalein effectively restored the osteogenic differentiation of LPS-treated PDLCs, as shown by the increased ALP and ARS staining. Accordingly, the protein and gene expression of osteogenic markers, namely runt-related transcription factor 2 (RUNX2), collagen-I, and osterix were markedly upregulated. Importantly, baicalein could function as the Wnt/β-catenin signaling activator, which may lead to the increased osteoblastic differentiation of PDLCs.ConclusionsWith the limitation of the study, we provide in vitro evidence that baicalein ameliorates inflammatory response and restores osteogenesis in PDLCs challenged with LPS, indicating its potential use as the host response modulator for the management of periodontitis.
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Project description:ObjectivesPeriodontitis is one of the most widespread inflammatory diseases; it causes tooth loss and is also associated with a variety of systemic diseases. Mesenchymal stem cells (MSCs) have been used to treat periodontitis. However, it is unknown whether bacterial toxins in the periodontal environment affect MSC-mediated periodontal regeneration. Porphyromonas gingivalis lipopolysaccharides (Pg-LPS) are key toxins for development of periodontitis. The purpose of the present study was to investigate effects of P. gingivalis LPS on biological properties of MSCs.Materials and methodsMesenchymal stem cells from bone marrow (BMMSCs) were treated with different concentrations of P. gingivalis LPS (0.1-10 ?g/ml), then its effects were evaluated on biological properties of BMMSCs including proliferation, apoptosis, osteogenic differentiation and capacities to inhibit activated T cells.ResultsLow concentration of P. gingivalis LPS (0.1 ?g/ml) accelerated MSC proliferation, osteogenic differentiation and capacities to inhibit activated T cells via up-regulation of nitric oxide. However, high concentration of P. gingivalis LPS (10 ?g/ml) reduced MSC proliferation, osteogenic differentiation and capacities to inhibit activated T cells.ConclusionsMesenchymal stem cells were functionally different following exposure to P. gingivalis LPS at the investigated concentrations. These findings suggest that MSC-mediated periodontal regeneration may be regulated by P. gingivalis LPS.
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