TY - JOUR
T1 - Quantification of cytokines and inflammatory mediators in a three-dimensional model of inflammatory arthritis
AU - Rai, Muhammad F.
AU - Sivaramakrishna Rachakonda, P.
AU - Manning, Kizzie
AU - Vorwerk, Brita
AU - Brunnberg, Leo
AU - Kohn, Barbara
AU - Schmidt, Michael F.G.
N1 - Funding Information:
We are thankful to Christiane Palissa and Anna Hermann for their technical support in cell culture and protein expression studies. We are grateful to Michael Sittinger (Tissue Engineering Laboratory, Department of Rheumatology, Charité-University Medicine, Berlin) for help in chondrocytes isolation. We also acknowledge the important technical support with alginate culturing of chondrocytes through Henning Madry and co-workers from Laboratory for Experimental Orthopaedics, Department of Orthopaedic Surgery, Saarland University Medical Centre, Homburg, Germany. We thank Gerd Schlenker and colleagues (Department of Animal and Environmental Hygiene, Free University Berlin) for help with the assay for endotoxins. We gratefully acknowledge the funding of this project (Schm 442/10-1) by Deutsche Forschungsgemeinschaft (DFG). M.F. Rai is recipient of a Ph.D. fellowship from German Academic Exchange Service (DAAD), Germany and Higher Education Commission (HEC) Govt. of Pakistan. This work is in partial fulfilment of his Ph.D. thesis at the FU Berlin, Germany.
PY - 2008/4
Y1 - 2008/4
N2 - Human recombinant IL-1β and TNFα have been previously used to induce a cytokine response in canine chondrocytes. In order to establish this functional relation in a homologous system in vitro, we have developed both 2D and 3D models of inflammatory arthritis using canine recombinant cytokines in canine articular chondrocytes. IL-1β and TNFα were cloned and subsequently expressed in Escherichia coli. The purified recombinant canine cytokines were used to simulate inflammation in vitro and the expression of typical inflammation markers such as proinflammatory cytokines (IL-1β, IL-6, IL-8, GM-CSF and TNFα), enzyme mediators (MMP-3 MMP-13, iNOS, COX-2) and their catabolites (NO, PGE2) was measured. High expression of proinflammatory cytokines, enzyme mediators and their catabolites was only observed in IL-1β/TNFα stimulated cells. We conclude that the canine IL-1β and TNFα generated in this study are biologically active and equally effective in the canine cell culture systems. Inducing an inflammatory pathway by canine exogenous cytokines in canine chondrocytes provides a useful tool for the study of canine inflammatory arthritis.
AB - Human recombinant IL-1β and TNFα have been previously used to induce a cytokine response in canine chondrocytes. In order to establish this functional relation in a homologous system in vitro, we have developed both 2D and 3D models of inflammatory arthritis using canine recombinant cytokines in canine articular chondrocytes. IL-1β and TNFα were cloned and subsequently expressed in Escherichia coli. The purified recombinant canine cytokines were used to simulate inflammation in vitro and the expression of typical inflammation markers such as proinflammatory cytokines (IL-1β, IL-6, IL-8, GM-CSF and TNFα), enzyme mediators (MMP-3 MMP-13, iNOS, COX-2) and their catabolites (NO, PGE2) was measured. High expression of proinflammatory cytokines, enzyme mediators and their catabolites was only observed in IL-1β/TNFα stimulated cells. We conclude that the canine IL-1β and TNFα generated in this study are biologically active and equally effective in the canine cell culture systems. Inducing an inflammatory pathway by canine exogenous cytokines in canine chondrocytes provides a useful tool for the study of canine inflammatory arthritis.
KW - Canine chondrocytes
KW - Cytokines
KW - Interleukin (IL)-1β
KW - Osteoarthritis
KW - Tumour necrosis factor (TNF)α
UR - http://www.scopus.com/inward/record.url?scp=41149083874&partnerID=8YFLogxK
U2 - 10.1016/j.cyto.2008.02.004
DO - 10.1016/j.cyto.2008.02.004
M3 - Article
C2 - 18343681
AN - SCOPUS:41149083874
SN - 1043-4666
VL - 42
SP - 8
EP - 17
JO - Cytokine
JF - Cytokine
IS - 1
ER -