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In this study, we evaluated the immunomodulatory effects of xanthan gum (XG) in RAW264.7 macrophages and the underlying molecular mechanisms. We used scanning electron microscopy (SEM) to analyze the morphology of XG-treated RAW264.7 cells with and without lipopolysaccharide (LPS) stimulation and investigated the subsequent effects on nitric oxide (NO), interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor (TNF-α), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) levels in LPS-activated mouse RAW264.7 macrophages. We also analyzed the binding affinity of XG to Toll-like receptor 4 (TLR4) with surface plasmon resonance (SPR) analysis and observed that XG decreased NO, IL-6 and TNF-α secretion into the culture medium and iNOS and COX-2 protein levels induced by LPS. This study reveals a two-way immunomodulatory effect of XG on inflammatory mediators in RAW264.7 macrophages that may involve the TLR4 signal pathway, providing a pharmacological basis for the use of XG in the control of inflammatory disorders. [ABSTRACT FROM AUTHOR]

The current study explored the efficacy of an intra-articular (IA) injection of allogeneic adipose tissue–derived stem cells (ADSCs) combined with xanthan gum (XG) in a rat osteoarthritis (OA) model. We confirmed that XG significantly inproved proliferation of ADSCs in a dose dependent manner in vitro . The rat OA model was induced by an anterior cruciate ligament transection (ACLT), and at 4 weeks after surgery, rats were divided into four groups: the XG-ADSCs group, the ADSCs group, the XG group and the phosphate-buffered saline (PBS) group. A single dose of 1 × 10 6 allogeneic ADSCs suspended in 1% XG, ADSCs suspended in PBS, 1% XG alone or PBS alone was injected into the OA joint of rats in the respective treatment groups. Rats were sacrificed at 8 weeks after surgery. Treatment outcomes were evaluated by weight-bearing control of the hind limbs, gross morphological analysis, histological analysis and specific staining of articular cartilage, and measurement of inflammatory factors in synovial fluid. For the rats in the XG-ADSC-s and ADSCs-treated groups, the weight-bearing percentage of the right hind limb was significantly increased compared to that in the PBS group and was sustained over 4 weeks. However, the positive effect in the XG-ADSCs group was significantly greater than that in the ADSCs group. For the rats in the XG group, the efficacy decreased during the third week after surgery. The articular cartilage was relatively normal in the XG-ADSCs group, and moderate degeneration was observed in the ADSCs and XG groups. ADSCs and XG-ADSC treatments significantly decreased the concentrations of IL-1β, TNF-α, MMP-3 and MMP-13 in synovial fluid; however, the attenuating effect of the XG-ADSCs treatment was significantly enhanced compared with that of the ADSCs treatment alone. These results indicate that a single IA injection of allogeneic ADSCs combined with XG efficiently attenuated OA progression with a therapeutic effect that was significantly greater than that of either ADSCs or XG alone. IA injection of XG-ADSCs might be an effective treatment for OA in humans. [ABSTRACT FROM AUTHOR]

We have previously reported the application of low molecular weight XG(LM-XG), with molecular weights ranging from 1 × 10 6 Da to 1.5 × 10 6 Da for treating osteoarthritis. In this study, we investigated the anti-apoptotic activity of LM-XG under oxidative stress conditions, activated by hydrogen peroxide (H 2 O 2 )-treated chondrocytes in vitro . Chondrocytes were pretreated with various doses of LM-XG (0, 10, 100, 500, or 1000 μg/mL) or 1000 μg/mL sodium hyaluronate for 12 h, and then exposed to 0.5 mmol/L H 2 O 2 for another 12 h. After treatment, chondrocyte viability was evaluated using a cell counting kit-8; DNA fragmentation was detected using Hoechst33258 staining; the percentage of DNA fragmentation was evaluated using the diphenylamine DNA assay kit; the apoptosis rate was evaluated using flow cytometry; chondrocyte ultra-microscopic morphology was observed using transmission electron microscopy; intracellular reactive oxygen species levels were observed and quantified using 2,7-dichlorofuorescin diacetate, mitochondrial permeability transition analysis was performed using MitoTracker Red CMXRos and 4′,6-diamidino-2-phenylindole staining; and finally, caspase-3 activity was detected by western blot. The results showed that, compared with H 2 O 2 -treated chondrocytes, LM-XG improved cell viability, decreased the percentage of DNA fragmentation, reduced the apoptosis rate, decreased the levels of intracellular reactive oxygen species and mitochondrial permeability transition, reverted the morphological damage, and downregulated cleaved caspase-3 levels. These results demonstrate that LM-XG has anti-apoptotic activity in H 2 O 2 -treated chondrocytes. [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA) is one of the most common chronic diseases and characterized by degradation of articular cartilage. We have previously reported xanthan gum (XG) injection preparation with high molecular weights ( Mw ) in ranging from 3 × 10 6 Da to 5 × 10 6 Da (HM-XG) could enhance the viscosity of synovial fluid, protect joint cartilage in rabbit, and the therapeutical effect has no significance difference with an existing clinical medication (sodium hyaluronate, SH) at the same injection frequency (once weekly for 5 weeks). Herein, we prepared a XG injection preparation with a low M w (LM-XG) in ranging from 1 × 10 6 Da to 1.5 × 10 6 Da, and evaluated the therapeutical effect for OA therapy at once every 2 weeks for 5 weeks with an SH at once weekly for 5 weeks as reference. The model of OA was induced using anterior cruciate ligament transection (ACLT) in a rabbit in vivo and also using sodium nitroprusside (SNP) in cell culture in vitro . The results showed that LW-XG could also protect cartilage from damage, decrease the concentration of nitric oxide (NO) in synovial fluid and reverse the amplification of the knee joint width similar to HM-XG as our previously reported. At the cellular level, LW-XG promotes proliferation while decreases apoptosis of chondrocytes. Mechanistically at the molecular level, these effects are elicited via down-regulation of the protein levels of caspase-3 and bax and up-regulation of the protein levels of bcl-2 in cartilage in both in vivo and in vitro. These results showed that LW-XG maybe become an excellent candidate long-acting drug for treating OA. [ABSTRACT FROM AUTHOR]