{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T23:20:35Z","timestamp":1791328835357,"version":"4.3.1"},"reference-count":44,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2024,8,23]],"date-time":"2024-08-23T00:00:00Z","timestamp":1724371200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Deutsche Arthrosehilfe e.V., the BONFOR-Forschungskommission der Medizinischen Fakult\u00e4t Bonn, the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)","award":["EXC2151\u2014390873048"],"award-info":[{"award-number":["EXC2151\u2014390873048"]}]},{"name":"Deutsche Arthrosehilfe e.V., the BONFOR-Forschungskommission der Medizinischen Fakult\u00e4t Bonn, the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)","award":["62214"],"award-info":[{"award-number":["62214"]}]},{"name":"John Templeton Foundation","award":["EXC2151\u2014390873048"],"award-info":[{"award-number":["EXC2151\u2014390873048"]}]},{"name":"John Templeton Foundation","award":["62214"],"award-info":[{"award-number":["62214"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Inflammation models are widely used in the in vitro investigation of new therapeutic approaches for osteoarthritis. TNF\u03b1 (tumor necrosis factor alpha) plays an important role in the inflammatory process. Current inflammation models lack uniformity and make comparisons difficult. Therefore, this study aimed to systematically investigate whether the effects of TNF\u03b1 are concentration-dependent and whether chondrocyte expansion has an effect on the inflammatory model. Bovine chondrocytes were enzymatically isolated, expanded to passages 1\u20133, and transferred into a 3D pellet culture. Chondrocyte pellets were stimulated with recombinant bovine TNF\u03b1 at different concentrations for 48 h to induce inflammation. Gene expression of anabolic (collagen 2, aggrecan, cartilage oligomeric protein (COMP)), catabolic (matrix metalloproteinases (MMP3, MMP13)), dedifferentiation (collagen 1) markers, inflammation markers (interleukin-6 (IL-6), nuclear factor kappa B (NFkB), cyclooxygenase-2 (COX), prostaglandin-E-synthase-2 (PTGES2)), and the apoptosis marker caspase 3 was determined. At the protein level, concentrations of IL-6, nitric oxide (NO), and sulfated glycosaminoglycans (GAG) were evaluated. Statistical analysis was performed using the independent t-test, and significance was defined as p &lt; 0.05. In general, TNF\u03b1 caused a decrease in anabolic markers and an increase in the expression of catabolic and inflammatory markers. There was a concentration-dependent threshold of 10 ng\/mL to induce significant inflammatory effects. Most of the markers analyzed showed TNF\u03b1 concentration-dependent effects (COMP, PRG4, AGN, Col1, MMP3, and NFkB). There was a statistical influence of selected gene expression markers from different passages on the TNF\u03b1 chondrocyte inflammation model, including Col2, MMP13, IL-6, NFkB, COX2, and PTGES2. Considering the expression of collagen 2 and MMP3, passage 3 chondrocytes showed a higher sensitivity to TNF\u03b1 stimulation compared to passages 1 and 2. On the other hand, MMP13, IL-6, NFkB, and caspase 3 gene expression were lower in P3 chondrocytes compared to the other passages. On the protein level, inflammatory effects showed a similar pattern, with cytokine effects starting at 10 ng\/mL and differences between the passages. TNF\u03b1 had a detrimental effect on cartilage, with a clear threshold observed at 10 ng\/mL. Although TNF\u03b1 effects showed concentration-dependent patterns, this was not consistent for all markers. The selected passage showed a clear influence, especially on inflammation markers. Further experiments were warranted to explore the effects of TNF\u03b1 concentration and passage in long-term stimulation.<\/jats:p>","DOI":"10.3390\/ijms25179136","type":"journal-article","created":{"date-parts":[[2024,8,26]],"date-time":"2024-08-26T05:16:24Z","timestamp":1724649384000},"page":"9136","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["TNF\u03b1-Induced Inflammation Model\u2014Evaluation of Concentration and Passage-Dependent Effects on Bovine Chondrocytes"],"prefix":"10.3390","volume":"25","author":[{"given":"Robert","family":"Ossendorff","sequence":"first","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3362-2061","authenticated-orcid":false,"given":"Su","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Kurth","sequence":"additional","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5998-0224","authenticated-orcid":false,"given":"Max","family":"Jaenisch","sequence":"additional","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elio","family":"Assaf","sequence":"additional","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7777-3253","authenticated-orcid":false,"given":"Andreas C.","family":"Strauss","sequence":"additional","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9495-706X","authenticated-orcid":false,"given":"Damien","family":"Bertheloot","sequence":"additional","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9340-9881","authenticated-orcid":false,"given":"Kristian","family":"Welle","sequence":"additional","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3431-4721","authenticated-orcid":false,"given":"Christof","family":"Burger","sequence":"additional","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6189-5114","authenticated-orcid":false,"given":"Dieter C.","family":"Wirtz","sequence":"additional","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0797-1945","authenticated-orcid":false,"given":"Frank A.","family":"Schildberg","sequence":"additional","affiliation":[{"name":"Department of Orthopedics and Trauma Surgery, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/j.joca.2021.04.020","article-title":"Epidemiology of osteoarthritis","volume":"30","author":"Allen","year":"2022","journal-title":"Osteoarthr. Cartil."},{"key":"ref_2","first-page":"575","article-title":"Methods of Conservative Intra-Articular Treatment for Osteoarthritis of the Hip and Knee","volume":"120","author":"Ossendorff","year":"2023","journal-title":"Dtsch. Arztebl. Int."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1014","DOI":"10.1136\/annrheumdis-2020-217050","article-title":"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","volume":"79","author":"Jin","year":"2020","journal-title":"Ann. Rheum. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1038\/nrrheum.2014.44","article-title":"The individual and socioeconomic impact of osteoarthritis","volume":"10","author":"Hunter","year":"2014","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"101249","DOI":"10.1016\/j.arr.2020.101249","article-title":"The role of metabolism in chondrocyte dysfunction and the progression of osteoarthritis","volume":"66","author":"Zheng","year":"2021","journal-title":"Ageing Res. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1038\/nrrheum.2010.196","article-title":"Role of proinflammatory cytokines in the pathophysiology of osteoarthritis","volume":"7","author":"Kapoor","year":"2011","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1177\/1759720X12467868","article-title":"Role of inflammation in the pathogenesis of osteoarthritis: Latest findings and interpretations","volume":"5","author":"Sokolove","year":"2013","journal-title":"Ther. Adv. Musculoskelet. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1038\/322547a0","article-title":"Tumour necrosis factor alpha stimulates resorption and inhibits synthesis of proteoglycan in cartilage","volume":"322","author":"Saklatvala","year":"1986","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1016\/j.joca.2019.09.008","article-title":"The effects of TNF-alpha inhibition on cartilage: A systematic review of preclinical studies","volume":"28","author":"Chisari","year":"2020","journal-title":"Osteoarthr. Cartil."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"20220567","DOI":"10.1515\/biol-2022-0567","article-title":"Immunoregulation of synovial macrophages for the treatment of osteoarthritis","volume":"18","author":"Xu","year":"2023","journal-title":"Open Life Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1290\/1543-706X(2004)040<0095:AIVSAF>2.0.CO;2","article-title":"An in vitro screening assay for inhibitors of proinflammatory mediators in herbal extracts using human synoviocyte cultures","volume":"40","author":"Frondoza","year":"2004","journal-title":"In Vitro Cell. Dev. Biol. Anim."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e3140","DOI":"10.1038\/cddis.2017.522","article-title":"MicroRNA-145 attenuates TNF-\u03b1-driven cartilage matrix degradation in osteoarthritis via direct suppression of MKK4","volume":"8","author":"Hu","year":"2017","journal-title":"Cell Death Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2901","DOI":"10.1002\/jor.24061","article-title":"Chondrocyte and mesenchymal stem cell derived engineered cartilage exhibits differential sensitivity to pro-inflammatory cytokines","volume":"36","author":"Mohanraj","year":"2018","journal-title":"J. Orthop. Res.\u00ae"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1002\/art.20776","article-title":"Role of interleukin-1 and tumor necrosis factor alpha in matrix degradation of human osteoarthritic cartilage","volume":"52","author":"Kobayashi","year":"2005","journal-title":"Arthritis Rheum."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1186\/s13018-016-0346-5","article-title":"Animal models of osteoarthritis: Classification, update, and measurement of outcomes","volume":"11","author":"Kuyinu","year":"2016","journal-title":"J. Orthop. Surg. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1208\/s12248-013-9454-x","article-title":"Animal models of osteoarthritis: Challenges of model selection and analysis","volume":"15","author":"Teeple","year":"2013","journal-title":"AAPS J."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"580","DOI":"10.1038\/nrrheum.2016.136","article-title":"Low-grade inflammation as a key mediator of the pathogenesis of osteoarthritis","volume":"12","author":"Robinson","year":"2016","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Ragni, E., de Luca, P., Valli, F., Zagra, L., and de Girolamo, L. (2023). Inflammatory Treatment Used to Mimic Osteoarthritis and Patients\u2019 Synovial Fluid Have Divergent Molecular Impact on Chondrocytes In Vitro. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24032625"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Mattiuzzo, E., Faggian, A., Venerando, R., Benetti, A., Belluzzi, E., Abatangelo, G., Ruggieri, P., and Brun, P. (2021). In Vitro Effects of Low Doses of \u03b2-Caryophyllene, Ascorbic Acid and d-Glucosamine on Human Chondrocyte Viability and Inflammation. Pharmaceuticals, 14.","DOI":"10.3390\/ph14030286"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Gao, Y., Liu, S., Huang, J., Guo, W., Chen, J., Zhang, L., Zhao, B., Peng, J., Wang, A., and Wang, Y. (2014). The ECM-cell interaction of cartilage extracellular matrix on chondrocytes. Biomed Res. Int., 2014.","DOI":"10.1155\/2014\/648459"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/0092-8674(82)90027-7","article-title":"Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels","volume":"30","author":"Benya","year":"1982","journal-title":"Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"831","DOI":"10.1021\/acsbiomaterials.2c00469","article-title":"Cell-Cell Interactions Enhance Cartilage Zonal Development in 3D Gradient Hydrogels","volume":"9","author":"Zhu","year":"2023","journal-title":"ACS Biomater. Sci. Eng."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1002\/jcp.21110","article-title":"Regulation of chondrocyte differentiation by the actin cytoskeleton and adhesive interactions","volume":"213","author":"Woods","year":"2007","journal-title":"J. Cell. Physiol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Al-Maslamani, N.A., Oldershaw, R., Tew, S., Curran, J., D\u2019Hooghe, P., Yamamoto, K., and Horn, H.F. (2022). Chondrocyte De-Differentiation: Biophysical Cues to Nuclear Alterations. Cells, 11.","DOI":"10.3390\/cells11244011"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"He, Y., Li, Z., Alexander, P.G., Ocasio-Nieves, B.D., Yocum, L., Lin, H., and Tuan, R.S. (2020). Pathogenesis of Osteoarthritis: Risk Factors, Regulatory Pathways in Chondrocytes, and Experimental Models. Biology, 9.","DOI":"10.3390\/biology9080194"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1002\/path.2287","article-title":"TNF-mediated inflammatory disease","volume":"214","author":"Bradley","year":"2008","journal-title":"J. Pathol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1016\/j.joca.2018.09.016","article-title":"Models of osteoarthritis: The good, the bad and the promising","volume":"27","author":"Cope","year":"2019","journal-title":"Osteoarthr. Cartil."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1177\/1941738109350438","article-title":"The basic science of articular cartilage: Structure, composition, and function","volume":"1","author":"Bedi","year":"2009","journal-title":"Sports Health"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1007\/s00011-016-0991-5","article-title":"Oxygen tension modulates the effects of TNF\u03b1 in compressed chondrocytes","volume":"66","author":"Tilwani","year":"2017","journal-title":"Inflamm. Res."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Ossendorff, R., Grede, L., Scheidt, S., Strauss, A.C., Burger, C., Wirtz, D.C., Salzmann, G.M., and Schildberg, F.A. (2024). Comparison of Minced Cartilage Implantation with Autologous Chondrocyte Transplantation in an In Vitro Inflammation Model. Cells, 13.","DOI":"10.3390\/cells13060546"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"17023","DOI":"10.1038\/sigtrans.2017.23","article-title":"NF-\u03baB signaling in inflammation","volume":"2","author":"Liu","year":"2017","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1007\/s10787-007-0013-x","article-title":"Role of nitric oxide in inflammatory diseases","volume":"15","author":"Sharma","year":"2007","journal-title":"Inflammopharmacology"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1016\/S0736-0266(00)00078-4","article-title":"Induction of apoptosis in chondrocytes by tumor necrosis factor-alpha","volume":"19","author":"Aizawa","year":"2001","journal-title":"J. Orthop. Res.\u00ae"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1016\/S1063-4584(03)00156-0","article-title":"Influence of pro-inflammatory (IL-1 alpha, IL-6, TNF-alpha, IFN-gamma) and anti-inflammatory (IL-4) cytokines on chondrocyte function","volume":"11","author":"Schuerwegh","year":"2003","journal-title":"Osteoarthr. Cartil."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1177\/1947603512471345","article-title":"Bioreactor-Induced Chondrocyte Maturation Is Dependent on Cell Passage and Onset of Loading","volume":"4","author":"Wang","year":"2013","journal-title":"Cartilage"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1007\/s00590-013-1205-2","article-title":"Useful animal models for the research of osteoarthritis","volume":"24","author":"Lelovas","year":"2014","journal-title":"Eur. J. Orthop. Surg. Traumatol. Orthop. Traumatol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1931","DOI":"10.1093\/rheumatology\/kes035","article-title":"Mapping pathogenesis of arthritis through small animal models","volume":"51","author":"Vincent","year":"2012","journal-title":"Rheumatology"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3786","DOI":"10.1007\/s00167-016-4300-1","article-title":"Comparative efficacy of cartilage repair procedures in the knee: A network meta-analysis","volume":"25","author":"Riboh","year":"2017","journal-title":"Knee Surg. Sports Traumatol. Arthrosc."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"19476035221085146","DOI":"10.1177\/19476035221085146","article-title":"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","volume":"13","author":"Niemeyer","year":"2022","journal-title":"Cartilage"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"891","DOI":"10.1007\/s00402-016-2453-5","article-title":"Cartilage repair surgery for full-thickness defects of the knee in Germany: Indications and epidemiological data from the German Cartilage Registry (KnorpelRegister DGOU)","volume":"136","author":"Niemeyer","year":"2016","journal-title":"Arch. Orthop. Trauma Surg."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"S2","DOI":"10.1016\/S0020-1383(15)30012-7","article-title":"Defect type, localization and marker gene expression determines early adverse events of matrix-associated autologous chondrocyte implantation","volume":"46","author":"Angele","year":"2015","journal-title":"Injury"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3171","DOI":"10.1089\/ten.2006.12.3171","article-title":"Effects of simple and complex motion patterns on gene expression of chondrocytes seeded in 3D scaffolds","volume":"12","author":"Grad","year":"2006","journal-title":"Tissue Eng."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1016\/j.ab.2004.10.005","article-title":"Effect of mechanical loading on mRNA levels of common endogenous controls in articular chondrocytes and intervertebral disk","volume":"341","author":"Lee","year":"2005","journal-title":"Anal. Biochem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1177\/1087057116644164","article-title":"384-Well Multiplexed Luminex Cytokine Assays for Lead Optimization","volume":"21","author":"Tang","year":"2016","journal-title":"J. Biomol. Screen."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/17\/9136\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:41:55Z","timestamp":1760110915000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/17\/9136"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,8,23]]},"references-count":44,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2024,9]]}},"alternative-id":["ijms25179136"],"URL":"https:\/\/doi.org\/10.3390\/ijms25179136","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,8,23]]}}}