Oral hydrolyzed type 2 collagen protects against the OA of obesity and mitigates obese gut microbiome dysbiosis

Abstract

The aim of this study was to analyze the anti-inflammatory effect of VA692, in comparison with celecoxib. In addition, we would establish if the new compound presents any advantages over other COX-2 inhibitors already available in therapeutic use. By iTRAQ methodology, we quantitatively analyzed the different expressed profiles in T/C-28a2 cell line treated with the studied drugs in presence of IL-1b. Methods: Human T/C-28a2 chondrocytes cell line were generated by Goldring group. Human articular cartilage was obtained from femoral heads of five OA patients. Cells were incubated with VA692 and celecoxib (1, 0.5 and 0.2mM) with interleukin (IL)-1b (5ng/ml) for 48hrs. The expression of inflammatory cytokines and anti-oxidant enzymes was evaluated by quantitative qRT-PCR, prostaglandins (PG)-E 2 release by ELISA. Apoptosis levels and ROS production were evaluated by flow cytometry. Proteins extracted by T/C-28a2 cell line was employed to carry out western blot analysis and the iTRAQ methodology. Statistical analysis was performed by ANOVA and Bonferroni multiple comparison tests. Results: IL-1b-stimulated chondrocytes showed a significant increase (p<0.001) of COX-2, IL-1b, IL-6, IL-8, superoxide dismutase (SOD)-2 and catalase (CAT) gene expression, as well as of PGE 2 levels in OA chondrocyte and in T/C-28a2 cell line. The tested drugs significantly counteracted the effect of IL-1b, with a better modulation by VA692 1mM in T/C-28a2 (p<0.01 for COX-2, IL-1b, IL-8, CAT; p<0.001 for IL-6, SOD-2 and PGE 2). Regarding apoptosis and mitochondrial superoxide anion production, the new drug was able to significantly reduce (p<0.05) their increase induced by IL-1b (p<0.05). Proteomic analysis led to the identification of 797 proteins in T/C-28a2 cell line, 123 of which were significantly modulated by VA692 in presence of IL-1b (p<0.001), while 34 were modulated by IL-1b alone (p<0.05). 21 proteins were commonly modulated by the two groups, indicating that 101 proteins were regulated by VA692 in a specific manner. Among the proteins downregulated by VA692, some with structural function were detected, responsible for cytoskeleton reorganization, as well as chaperones (heat shock proteins) and glycolitic enzymes. Proteins involved in calcium metabolism and in ribosome biogenesis resulted up-regulated instead, as well as SOD-2 as confirmed by western blot analysis. Conclusions: Our data demonstrated the anti-inflammatory effect of VA692, suggesting also its anti-apoptotic and anti-oxidant role. The proteomic analysis demonstrated that VA692 induced not only an antiinflammatory regulation in chondrocytes but, interestingly, seemed to regulate their anabolic response. On the basis of our results, we can hypothesize that VA692 could present any advantages over other COX-2 inhibitors already available for therapeutic use. However, further in vitro and in vivo experiments are necessary to elucidate and confirm the importance of this pharmacological compound before its use in the therapeutic approach of joint diseases.

β-Elemene Suppresses Obesity-Induced Imbalance in the Microbiota-Gut-Brain Axis

“…The relative order abundance of Coriobacteriales, which belong to the phylum Actinobacteria, was decreased by HFD feeding and reversed by β-elemene intervention. Similar results were also indicated by Soniwala et al [40], who found that obesity reduces the order largely and hydrolyzed type 2 collagen showed excellent effects on alleviating the tendency and mitigating obese gut microbiome dysbiosis. At the genus level, an increasing tendency of Akkermansia was observed after β-elemene treatment.…”

Development of a Mobile App to Monitor the Effectiveness of a Hydrolyzed Cartilage Matrix Supplement on Joint Discomfort: Real-World Study

“…The fact that participants were not restricted to any diet or lifestyle highlights the relevance of HCM supplementation in a real-world setting where outside factors are not controlled (contrary to strictly controlled clinical trial designs). This design allowed us to investigate if the benefits of HCM observed in an OA mouse model extend to a human heterogeneous population [ 13 ].…”

“…The first preclinical report on HCM supplementation in obese mice with posttraumatic OA demonstrated cartilage protection and a significant suppression of key proinflammatory microbial strains in the gut, suggesting that HCM might indirectly protect cartilage by modulating the gut microbiome [ 13 ]. Future randomized controlled trial (RCT) protocols are warranted to assess extended clinical outcomes, capturing biomechanical function, biomarkers of inflammation, or imaging techniques to get more insights into the mode of action of HCM.…”

“…The effect of the washout was directly noticeable in the HCM group with a gradual increase of the joint pain score over the 4-week period, while remaining constant in the placebo group, suggesting a direct mechanistic correlation between supplementation and benefit, such as an anti-inflammatory effect. CS has been reported to have anti-inflammatory activity [ 24 , 25 ], and so has HCM in a preclinical study [ 13 ], which could explain the attenuation of the experienced benefit in the washout period. However, joint discomfort was still lower than in the placebo group at the end of the washout period, suggesting that other, more long-lasting mechanisms, might contribute to the pain-alleviating effect of HCM.…”

“…The hydrolyzed cartilage matrix (HCM) containing collagen and chondroitin sulfate (CS) has shown positive effects on joint protection in a mouse model for obesity-accelerated posttraumatic osteoarthritis (OA) [ 13 ]. However, the effects of this natural matrix are yet to be tested in the general population.…”