TNFα-Induced Inflammation Model—Evaluation of Concentration and Passage-Dependent Effects on Bovine Chondrocytes
Abstract
:1. Introduction
2. Results
2.1. Anabolic Markers
2.2. Catabolic Markers
2.3. Inflammation
2.4. Apoptosis
2.5. Functional Analysis of Glycosaminoglycans, Nitric Oxide, and Multiplex Inflammation Assay
3. Discussion
4. Materials and Methods
4.1. Experimental Design
4.2. Isolation and In Vitro Expansion of Chondrocytes
4.3. TNFα Chondrocyte Inflammation Model Setup
4.4. RNA Extraction, Reverse Transcription, and Gene Expression Analysis
4.5. Multiplex ELISA
4.6. Nitric Oxide (NO), Glycosaminoglycan (GAG), and Viability Analysis
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Allen, K.D.; Thoma, L.M.; Golightly, Y.M. Epidemiology of osteoarthritis. Osteoarthr. Cartil. 2022, 30, 184–195. [Google Scholar] [CrossRef] [PubMed]
- Ossendorff, R.; Thimm, D.; Wirtz, D.C.; Schildberg, F.A. Methods of Conservative Intra-Articular Treatment for Osteoarthritis of the Hip and Knee. Dtsch. Arztebl. Int. 2023, 120, 575–581. [Google Scholar] [CrossRef] [PubMed]
- Jin, Z.; Wang, D.; Zhang, H.; Liang, J.; Feng, X.; Zhao, J.; Sun, L. Incidence trend of five common musculoskeletal disorders from 1990 to 2017 at the global, regional and national level: Results from the global burden of disease study 2017. Ann. Rheum. Dis. 2020, 79, 1014–1022. [Google Scholar] [CrossRef] [PubMed]
- Hunter, D.J.; Schofield, D.; Callander, E. The individual and socioeconomic impact of osteoarthritis. Nat. Rev. Rheumatol. 2014, 10, 437–441. [Google Scholar] [CrossRef]
- Zheng, L.; Zhang, Z.; Sheng, P.; Mobasheri, A. The role of metabolism in chondrocyte dysfunction and the progression of osteoarthritis. Ageing Res. Rev. 2021, 66, 101249. [Google Scholar] [CrossRef]
- Kapoor, M.; Martel-Pelletier, J.; Lajeunesse, D.; Pelletier, J.-P.; Fahmi, H. Role of proinflammatory cytokines in the pathophysiology of osteoarthritis. Nat. Rev. Rheumatol. 2011, 7, 33–42. [Google Scholar] [CrossRef] [PubMed]
- Sokolove, J.; Lepus, C.M. Role of inflammation in the pathogenesis of osteoarthritis: Latest findings and interpretations. Ther. Adv. Musculoskelet. Dis. 2013, 5, 77–94. [Google Scholar] [CrossRef]
- Saklatvala, J. Tumour necrosis factor alpha stimulates resorption and inhibits synthesis of proteoglycan in cartilage. Nature 1986, 322, 547–549. [Google Scholar] [CrossRef]
- Chisari, E.; Yaghmour, K.M.; Khan, W.S. The effects of TNF-alpha inhibition on cartilage: A systematic review of preclinical studies. Osteoarthr. Cartil. 2020, 28, 708–718. [Google Scholar] [CrossRef]
- Xu, M.; Ji, Y. Immunoregulation of synovial macrophages for the treatment of osteoarthritis. Open Life Sci. 2023, 18, 20220567. [Google Scholar] [CrossRef]
- Frondoza, C.G.; Sohrabi, A.; Polotsky, A.; Phan, P.V.; Hungerford, D.S.; Lindmark, L. An in vitro screening assay for inhibitors of proinflammatory mediators in herbal extracts using human synoviocyte cultures. In Vitro Cell. Dev. Biol. Anim. 2004, 40, 95–101. [Google Scholar] [CrossRef] [PubMed]
- Hu, G.; Zhao, X.; Wang, C.; Geng, Y.; Zhao, J.; Xu, J.; Zuo, B.; Zhao, C.; Wang, C.; Zhang, X. MicroRNA-145 attenuates TNF-α-driven cartilage matrix degradation in osteoarthritis via direct suppression of MKK4. Cell Death Dis. 2017, 8, e3140. [Google Scholar] [CrossRef]
- Mohanraj, B.; Huang, A.H.; Yeger-McKeever, M.J.; Schmidt, M.J.; Dodge, G.R.; Mauck, R.L. Chondrocyte and mesenchymal stem cell derived engineered cartilage exhibits differential sensitivity to pro-inflammatory cytokines. J. Orthop. Res.® 2018, 36, 2901–2910. [Google Scholar] [CrossRef]
- Kobayashi, M.; Squires, G.R.; Mousa, A.; Tanzer, M.; Zukor, D.J.; Antoniou, J.; Feige, U.; Poole, A.R. Role of interleukin-1 and tumor necrosis factor alpha in matrix degradation of human osteoarthritic cartilage. Arthritis Rheum. 2005, 52, 128–135. [Google Scholar] [CrossRef]
- Kuyinu, E.L.; Narayanan, G.; Nair, L.S.; Laurencin, C.T. Animal models of osteoarthritis: Classification, update, and measurement of outcomes. J. Orthop. Surg. Res. 2016, 11, 19. [Google Scholar] [CrossRef]
- Teeple, E.; Jay, G.D.; Elsaid, K.A.; Fleming, B.C. Animal models of osteoarthritis: Challenges of model selection and analysis. AAPS J. 2013, 15, 438–446. [Google Scholar] [CrossRef] [PubMed]
- Robinson, W.H.; Lepus, C.M.; Wang, Q.; Raghu, H.; Mao, R.; Lindstrom, T.M.; Sokolove, J. Low-grade inflammation as a key mediator of the pathogenesis of osteoarthritis. Nat. Rev. Rheumatol. 2016, 12, 580–592. [Google Scholar] [CrossRef] [PubMed]
- Ragni, E.; de Luca, P.; Valli, F.; Zagra, L.; de Girolamo, L. Inflammatory Treatment Used to Mimic Osteoarthritis and Patients’ Synovial Fluid Have Divergent Molecular Impact on Chondrocytes In Vitro. Int. J. Mol. Sci. 2023, 24, 2625. [Google Scholar] [CrossRef]
- Mattiuzzo, E.; Faggian, A.; Venerando, R.; Benetti, A.; Belluzzi, E.; Abatangelo, G.; Ruggieri, P.; Brun, P. In Vitro Effects of Low Doses of β-Caryophyllene, Ascorbic Acid and d-Glucosamine on Human Chondrocyte Viability and Inflammation. Pharmaceuticals 2021, 14, 286. [Google Scholar] [CrossRef]
- Gao, Y.; Liu, S.; Huang, J.; Guo, W.; Chen, J.; Zhang, L.; Zhao, B.; Peng, J.; Wang, A.; Wang, Y.; et al. The ECM-cell interaction of cartilage extracellular matrix on chondrocytes. Biomed Res. Int. 2014, 2014, 648459. [Google Scholar] [CrossRef]
- Benya, P.D.; Shaffer, J.D. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell 1982, 30, 215–224. [Google Scholar] [CrossRef] [PubMed]
- Zhu, D.; Trinh, P.; Liu, E.; Yang, F. Cell-Cell Interactions Enhance Cartilage Zonal Development in 3D Gradient Hydrogels. ACS Biomater. Sci. Eng. 2023, 9, 831–843. [Google Scholar] [CrossRef] [PubMed]
- Woods, A.; Wang, G.; Beier, F. Regulation of chondrocyte differentiation by the actin cytoskeleton and adhesive interactions. J. Cell. Physiol. 2007, 213, 1–8. [Google Scholar] [CrossRef] [PubMed]
- Al-Maslamani, N.A.; Oldershaw, R.; Tew, S.; Curran, J.; D’Hooghe, P.; Yamamoto, K.; Horn, H.F. Chondrocyte De-Differentiation: Biophysical Cues to Nuclear Alterations. Cells 2022, 11, 4011. [Google Scholar] [CrossRef]
- He, Y.; Li, Z.; Alexander, P.G.; Ocasio-Nieves, B.D.; Yocum, L.; Lin, H.; Tuan, R.S. Pathogenesis of Osteoarthritis: Risk Factors, Regulatory Pathways in Chondrocytes, and Experimental Models. Biology 2020, 9, 194. [Google Scholar] [CrossRef]
- Bradley, J.R. TNF-mediated inflammatory disease. J. Pathol. 2008, 214, 149–160. [Google Scholar] [CrossRef]
- Cope, P.J.; Ourradi, K.; Li, Y.; Sharif, M. Models of osteoarthritis: The good, the bad and the promising. Osteoarthr. Cartil. 2019, 27, 230–239. [Google Scholar] [CrossRef] [PubMed]
- Sophia Fox, A.J.; Bedi, A.; Rodeo, S.A. The basic science of articular cartilage: Structure, composition, and function. Sports Health 2009, 1, 461–468. [Google Scholar] [CrossRef]
- Tilwani, R.K.; Vessillier, S.; Pingguan-Murphy, B.; Lee, D.A.; Bader, D.L.; Chowdhury, T.T. Oxygen tension modulates the effects of TNFα in compressed chondrocytes. Inflamm. Res. 2017, 66, 49–58. [Google Scholar] [CrossRef]
- Ossendorff, R.; Grede, L.; Scheidt, S.; Strauss, A.C.; Burger, C.; Wirtz, D.C.; Salzmann, G.M.; Schildberg, F.A. Comparison of Minced Cartilage Implantation with Autologous Chondrocyte Transplantation in an In Vitro Inflammation Model. Cells 2024, 13, 546. [Google Scholar] [CrossRef]
- Liu, T.; Zhang, L.; Joo, D.; Sun, S.-C. NF-κB signaling in inflammation. Signal Transduct. Target. Ther. 2017, 2, 17023. [Google Scholar] [CrossRef] [PubMed]
- Sharma, J.N.; Al-Omran, A.; Parvathy, S.S. Role of nitric oxide in inflammatory diseases. Inflammopharmacology 2007, 15, 252–259. [Google Scholar] [CrossRef]
- Aizawa, T.; Kon, T.; Einhorn, T.A.; Gerstenfeld, L.C. Induction of apoptosis in chondrocytes by tumor necrosis factor-alpha. J. Orthop. Res.® 2001, 19, 785–796. [Google Scholar] [CrossRef] [PubMed]
- Schuerwegh, A.J.; Dombrecht, E.J.; Stevens, W.J.; van Offel, J.F.; Bridts, C.H.; de Clerck, L.S. Influence of pro-inflammatory (IL-1 alpha, IL-6, TNF-alpha, IFN-gamma) and anti-inflammatory (IL-4) cytokines on chondrocyte function. Osteoarthr. Cartil. 2003, 11, 681–687. [Google Scholar] [CrossRef]
- Wang, N.; Grad, S.; Stoddart, M.J.; Niemeyer, P.; Südkamp, N.P.; Pestka, J.; Alini, M.; Chen, J.; Salzmann, G.M. Bioreactor-Induced Chondrocyte Maturation Is Dependent on Cell Passage and Onset of Loading. Cartilage 2013, 4, 165–176. [Google Scholar] [CrossRef]
- Lampropoulou-Adamidou, K.; Lelovas, P.; Karadimas, E.V.; Liakou, C.; Triantafillopoulos, I.K.; Dontas, I.; Papaioannou, N.A. Useful animal models for the research of osteoarthritis. Eur. J. Orthop. Surg. Traumatol. Orthop. Traumatol. 2014, 24, 263–271. [Google Scholar] [CrossRef]
- Vincent, T.L.; Williams, R.O.; Maciewicz, R.; Silman, A.; Garside, P. Mapping pathogenesis of arthritis through small animal models. Rheumatology 2012, 51, 1931–1941. [Google Scholar] [CrossRef] [PubMed]
- Riboh, J.C.; Cvetanovich, G.L.; Cole, B.J.; Yanke, A.B. Comparative efficacy of cartilage repair procedures in the knee: A network meta-analysis. Knee Surg. Sports Traumatol. Arthrosc. 2017, 25, 3786–3799. [Google Scholar] [CrossRef]
- Niemeyer, P.; Hanus, M.; Belickas, J.; László, T.; Gudas, R.; Fiodorovas, M.; Cebatorius, A.; Pastucha, M.; Hoza, P.; Magos, K.; et al. Treatment of Large Cartilage Defects in the Knee by Hydrogel-Based Autologous Chondrocyte Implantation: Two-Year Results of a Prospective, Multicenter, Single-Arm Phase III Trial. Cartilage 2022, 13, 19476035221085146. [Google Scholar] [CrossRef]
- Niemeyer, P.; Feucht, M.J.; Fritz, J.; Albrecht, D.; Spahn, G.; Angele, P. Cartilage repair surgery for full-thickness defects of the knee in Germany: Indications and epidemiological data from the German Cartilage Registry (KnorpelRegister DGOU). Arch. Orthop. Trauma Surg. 2016, 136, 891–897. [Google Scholar] [CrossRef]
- Angele, P.; Fritz, J.; Albrecht, D.; Koh, J.; Zellner, J. Defect type, localization and marker gene expression determines early adverse events of matrix-associated autologous chondrocyte implantation. Injury 2015, 46 (Suppl. 4), S2–S9. [Google Scholar] [CrossRef] [PubMed]
- Grad, S.; Gogolewski, S.; Alini, M.; Wimmer, M.A. Effects of simple and complex motion patterns on gene expression of chondrocytes seeded in 3D scaffolds. Tissue Eng. 2006, 12, 3171–3179. [Google Scholar] [CrossRef] [PubMed]
- Lee, C.R.; Grad, S.; Maclean, J.J.; Iatridis, J.C.; Alini, M. Effect of mechanical loading on mRNA levels of common endogenous controls in articular chondrocytes and intervertebral disk. Anal. Biochem. 2005, 341, 372–375. [Google Scholar] [CrossRef]
- Tang, H.; Panemangalore, R.; Yarde, M.; Zhang, L.; Cvijic, M.E. 384-Well Multiplexed Luminex Cytokine Assays for Lead Optimization. J. Biomol. Screen. 2016, 21, 548–555. [Google Scholar] [CrossRef] [PubMed]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Ossendorff, R.; Wang, S.; Kurth, S.; Jaenisch, M.; Assaf, E.; Strauss, A.C.; Bertheloot, D.; Welle, K.; Burger, C.; Wirtz, D.C.; et al. TNFα-Induced Inflammation Model—Evaluation of Concentration and Passage-Dependent Effects on Bovine Chondrocytes. Int. J. Mol. Sci. 2024, 25, 9136. https://doi.org/10.3390/ijms25179136
Ossendorff R, Wang S, Kurth S, Jaenisch M, Assaf E, Strauss AC, Bertheloot D, Welle K, Burger C, Wirtz DC, et al. TNFα-Induced Inflammation Model—Evaluation of Concentration and Passage-Dependent Effects on Bovine Chondrocytes. International Journal of Molecular Sciences. 2024; 25(17):9136. https://doi.org/10.3390/ijms25179136
Chicago/Turabian StyleOssendorff, Robert, Su Wang, Sarah Kurth, Max Jaenisch, Elio Assaf, Andreas C. Strauss, Damien Bertheloot, Kristian Welle, Christof Burger, Dieter C. Wirtz, and et al. 2024. "TNFα-Induced Inflammation Model—Evaluation of Concentration and Passage-Dependent Effects on Bovine Chondrocytes" International Journal of Molecular Sciences 25, no. 17: 9136. https://doi.org/10.3390/ijms25179136