Anti-inflammatory role of TPCA-1 encapsulated nanosomes in porcine chondrocytes against TNF-α stimulation
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In this study, we evaluated the hypothesis that immunonanosomes carrying the drug [5-(p-Fluorophenyl)-2-ureido]thiophene-3-carboxamide (TPCA-1) will help in reducing nuclear factor-kappaB (NF-κB)-associated inflammation in porcine chondrocytes against tumor necrosis factor-alpha (TNF-α)-induced stress. The nanosomes were tagged with monoclonal anti-type II collagen (MabCII) antibody to specifically target the exposed type II collagen in cartilage matrix. TPCA-1 at a concentration of 10 µM significantly reduced expression of the matrix-degrading enzyme, Matrix metalloproteinase-13 (MMP-13) and blocked the p65 nuclear translocation. In comparison to the TPCA-1 solution alone, the TPCA-1 nanosomes were found to be more effective in reducing the cellular toxicity, oxidative stress and inflammation in chondrocytes treated with TNF-α. In addition, TPCA-1 nanosomes were more effective in reducing the gene expression of hypoxia-inducible factor-2alpha (HIF-2α) that in turn is associated with the regulation of MMP-13 gene. TPCA-1 nanosomes significantly reduced expression of both these genes. The data also showed that TPCA-1 did not attenuate the down-regulated gene expression levels of anabolic genes aggrecan (ACAN) and collagen type II alpha (COL2A1). In conclusion, this study showed that TPCA-1 nanosomes carrying a dose of 10 µM TPCA-1 can effectively increase the survival of cultured porcine chondrocytes against TNF-α-induced stress. The findings of this study could be used to develop nanosome-based drug delivery systems (DDSs) for animal model of OA. Moreover, the approach presented here can be further utilized in other studies for targeted delivery of the drug of interest at a cellular level.
KeywordsNanosomes Inflammation Chondrocytes TNF-α TPCA-1 p65 translocation
FB and HC: designed and performed the experiments. FB and HC: performed the statistical analysis. FB and HC: wrote the manuscript. HC and KH: critically reviewed the manuscript. All authors read and approved the final manuscript.
This work was supported by grants from the Arthritis Foundation (Discovery award; H. Cho) and Oxnard Foundation (Medical Research; H. Cho). This research also supported by a VA Merit Review award and VA Research Career Scientist Award (K. Hasty) from the Department of Veterans Affairs.
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Conflict of interest
The authors declare that they have no competing interests.
- Maguire O, Collins C, O’Loughlin K, Miecznikowski J, Minderman H (2011) Quantifying nuclear p65 as a parameter for NF-kappaB activation: correlation between imagestream cytometry, microscopy, and Western blot. Cytometry A 79:461–469. https://doi.org/10.1002/cyto.a.21068 CrossRefPubMedPubMedCentralGoogle Scholar
- Podolin PL et al (2005) Attenuation of murine collagen-induced arthritis by a novel, potent, selective small molecule inhibitor of IkappaB Kinase 2, TPCA-1 (2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamide), occurs via reduction of proinflammatory cytokines and antigen-induced T cell proliferation. J Pharmacol Exp Ther 312:373–381. https://doi.org/10.1124/jpet.104.074484 CrossRefPubMedGoogle Scholar
- Rivard J, James A, Wegner G, Reagan K (2014) Modulating the tumor necrosis factor-α induced inflammatory response in human colonic epithelial cells by inhibiting NF-κB signaling (CCR5P.258). J Immunol 192:181.112Google Scholar
- TenBroek EM, Yunker L, Nies MF, Bendele AM (2016) Randomized controlled studies on the efficacy of antiarthritic agents in inhibiting cartilage degeneration and pain associated with progression of osteoarthritis in the rat. Arthritis Res Ther 18:24. https://doi.org/10.1186/s13075-016-0921-5 CrossRefPubMedPubMedCentralGoogle Scholar
- Tilstra JS, Gaddy DF, Zhao J, Dave SH, Niedernhofer LJ, Plevy SE, Robbins PD (2014) Pharmacologic IKK/NF-kappaB inhibition causes antigen presenting cells to undergo TNFalpha dependent ROS-mediated programmed cell death. Sci Rep 4:3631. https://doi.org/10.1038/srep03631 CrossRefPubMedPubMedCentralGoogle Scholar