Preliminary study on the effect of inflamed TMJ synovial fluid on the intracellular calcium concentration and differential expression of iNOS and COX-2 in human immortalized chondrocyte C28/I2

  • Choi, Eun-Ah (Department of Orthodontics, College of Dentistry, Seoul National University) ;
  • Lee, Dong-Geun (Department of Orthodontics, Section of Dentistry, Seoul National University Bundang Hospital) ;
  • Chae, Chang-Hoon (Department of Oral and Maxillofacial Surgery, College of Medicine, Hallym University) ;
  • Chang, Young-Il (Department of Orthodontics, College of Dentistry, Seoul National University) ;
  • Park, Young-Ju (Department of Oral and Maxillofacial Surgery, College of Medicine, Hallym University) ;
  • Kim, Young-Kyun (Department of Oral & Maxillofacial Surgery, Section of Dentistry, Seoul National University Bundang Hospital)
  • Published : 2006.02.28

Abstract

Objective. The objective of this study was to examine the hypothesis that inflammatory synovial fluid from TMJ internal derangement initiates a transient increase in intracellular calcium concentration ([$Ca^{2+}$]i) in chondrocytes and the induced Ca2+ signaling affects iNOS/COX-2 gene expression patterns following exposure to inflamed synovial fluid. Materials and Methods. Two female adult patients with symptoms of TMD who agreed to participate in the study were selected for this study. Immortalized human juvenile costal chondrocyte C-28/I2 was grown to 80% confluency and synovial fluids from two patients were added respectively to culture media for 24 hours at the concentration of 100ng/10ml. Confocal laser scanning microscope (CLSM) was used to examine changes of intracellular calcium concentration ([$Ca^{2+}$]i). RT-PCR was performed to identify the expression profile of IL-1${\alpha}$, iNOS, COX-2. Results. Increased [$Ca^{2+}$]i was observed in chondrocytes subjected to inflamed synovial fluid compared to control cultures and in respective cultures exposed to inflamed synovial fluids from each patient, IL-1${\beta}$, COX-2 mRNA were detected. However, in neither case iNOS mRNA was expressed. IL-1${\alpha}$, COX-2, and iNOS mRNA were expressed in control culture. Conclusion. Our results show that immortalized chondrocytes cultured with inflamed synovial fluids from patients diagnosed as disc displacement without reduction and limitation in mouth opening showed increased calcium concentration and expression of COX-2 while inhibiting the production of iNOS, which in turn could adversely affect the chondrocytes in at least short term by hindering physiologic role of NO against inflammatory cascades. These findings suggest that inflamed synovial fluid may differentially regulate the transcriptomes of relevant inflammatory mediators, especially iNOS/COX-2 axis in chondrocytes through adjusting calcium transients.

Keywords

References

  1. Myers SL: Synovial fluid markers in osteoarthritis. Rheum Dis Clin North Am 1999;25(2):433-449 https://doi.org/10.1016/S0889-857X(05)70077-6
  2. Bresnihan B: Are synovial biopsies of diagnostic value? Arthritis Res Ther 2003;5(6):271-278 https://doi.org/10.1186/ar1003
  3. Schmidt HH, Nau H, Wittfoht W, Gerlach J, Prescher KE, Klein MM, Niroomand F, Bohme E: RArginine is a physiological precursor of endothelium-derived nitric oxide. Eur J Pharmacol 1988;13;154(2):213-216
  4. Sakurai H, Kohsaka H, Liu MF, Higashiyama H, Hirata Y, Kanno K, et al.: Nitric oxide production and inducible nitric oxide synthase expression in inflammatory arthritides. J Clin Invest 1995;96:2357-2363 https://doi.org/10.1172/JCI118292
  5. Takahashi T, Homma H, Nagai H, Seki H, Kondoh T, Yamazaki Y, Fukuda M: Specific expression of inducible nitric oxide synthase in the synovium of the diseased temporomandibular joint. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2003;95(2):174-181 https://doi.org/10.1067/moe.2003.45
  6. Homma H, Takahashi T, Seki H, Ohtani M, Kondoh T, Fukuda M: Immunohistochemical localization of inducible nitric oxide synthase in synovial tissue of human temporomandibular joints with internal derangement. Arch Oral Biol 2001;46:93-97 https://doi.org/10.1016/S0003-9969(00)00086-8
  7. Turini ME, DuBois RN: Cyclooxygenase-2: a therapeutic target. Annu Rev Med 2002;53:35-57. Review https://doi.org/10.1146/annurev.med.53.082901.103952
  8. Quinn JH, Kent JH, Moise A, Lukiw WJ: Cyclooxygenase-2 in synovial tissue and fluid of dysfunctional temporomandibular joints with internal derangement. J Oral Maxillofac Surg 2000;58(11):1229-1232; discussion 1232- 1233 https://doi.org/10.1053/joms.2000.16619
  9. Hardingham GE, Chawla S, Johnson CM, Bading H: Distinct functions of nuclear and cytoplasmic calcium in the control of gene expression. Nature 1997;16;385(6613): 260-2653
  10. Verschure PJ, Van Marle J, Van Noorden CJ, Van den Berg WB: The contribution of quantitative confocal laser scanning microscopy in cartilage research: chondrocyte insulinlike growth factor-1 receptors in health and pathology. Microsc Res Tech 1997;15;37(4):285-298. Review
  11. Dworkin SF, LeResche L: Research diagnostic criteria for temporomandibular disorders: review, criteria, examinations and specifications, critique. J Craniomandib Disord 1992:6:301-355
  12. Knight MM, Roberts SR, Lee DA, Bader DL: Live cell imaging using confocal microscopy induces intracellular calcium transients and cell death. Am J Physiol Cell Physiol 2003;284(4):C1083-1089 https://doi.org/10.1152/ajpcell.00276.2002
  13. Stichtenoth DO, Frolich JC: Nitric oxide and inflammatory joint diseases. Br J Rheumatol 1998;37(3):246-257. Review https://doi.org/10.1093/rheumatology/37.3.246
  14. Wahl SM, McCartney-Francis N, Chan J, Dionne R, Ta L, Orenstein JM: Nitric oxide in experimental joint inflammation. Benefit or detriment? Cells Tissues Organs 2003; 174(1-2):26-33 https://doi.org/10.1159/000071657
  15. MacIntyre I, Zaidi M, Alam AS, Datta HK, Moonga BS, Lidbury PS, Hecker M, Vane JR: Osteoclastic inhibition: an action of nitric oxide not mediated by cyclic GMP. Proc Natl Acad Sci USA 1991;1;88(7):2936-2940
  16. Stefanovic-Racic M, Morales TI, Taskiran D, McIntyre LA, Evans CH: The role of nitric oxide in proteoglycan turnover by bovine articular cartilage organ cultures. J Immunol 1996;1;156(3):1213-1220
  17. van der Veen RC: Nitric oxide and T helper cell immunity. Int Immunopharmacol. 2001;1(8):1491-1500 https://doi.org/10.1016/S1567-5769(01)00093-5
  18. Ralston SH, Todd D, Helfrich M, Benjamin N, Grabowski PS: Human osteoblast-like cells produce nitric oxide and express inducible nitric oxide synthase. Endocrinology 1994;135(1):330-336 https://doi.org/10.1210/en.135.1.330
  19. Goldring MB, Berenbaum F: The regulation of chondrocyte function by proinflammatory mediators: prostaglandins and nitric oxide. Clin Orthop Relat Res. 2004;(427 Suppl): S37-46. Review https://doi.org/10.1097/01.blo.0000143739.07632.7c
  20. Veihelmann A, Landes J, Hofbauer A, Dorger M, Refior HJ, Messmer K, Krombach F: Exacerbation of antigen-induced arthritis in inducible nitric oxide synthase-deficient mice. Arthritis Rheum 2001;44(6):1420-1427 https://doi.org/10.1002/1529-0131(200106)44:6<1420::AID-ART237>3.0.CO;2-K
  21. Clements KM, Price JS, Chambers MG, Visco DM, Poole AR, Mason RM: Gene deletion of either interleukin-1beta, interleukin-1beta-converting enzyme, inducible nitric oxide synthase, or stromelysin 1 accelerates the development of knee osteoarthritis in mice after surgical transection of the medial collateral ligament and partial medial meniscectomeniscectomy. Arthritis Rheum 2003;48(12):3452-3463 https://doi.org/10.1002/art.11355
  22. Palmer, R. M., M. S. Hickery, I. G. Charles, S. Moncada, and M. T. Bayliss:1993. Induction of nitric oxide synthase in human chondrocytes. Bio-chem Biophys Res Commun 193:398-405 https://doi.org/10.1006/bbrc.1993.1637
  23. Rediske, J., C. Koehne, B. Zhang, and M. Lotz: The inducible production of nitric oxide by articular cell types. Osteoarthritis and Cartilage 1994;2:199-206 https://doi.org/10.1016/S1063-4584(05)80069-X
  24. Geng Y, Lotz M: Increased intracellular Ca2+ selectively suppresses IL-1-induced NO production by reducing iNOS mRNA stability. J Cell Biol 1995;129(6):1651-1657 https://doi.org/10.1083/jcb.129.6.1651
  25. Geng Y, Blanco FJ, Cornelisson M, Lotz M: Regulation of cyclooxygenase-2 expression in normal human articular chondrocytes. J Immunol 1995;15;155(2):796-801
  26. Salvemini D, Misko TP, Masferrer J, Seibert K, Currie MG, Needleman P: Nitric oxide activates cyclooxygenase enzymes. Proc Natl Acad Sci USA 1993;90:7240-7244
  27. Needleman P, Manning PT: Interactions between the inducible cyclooxygenase (COX-2) and nitric oxide synthase (iNOS) pathways: implications for therapeutic intervention in osteoarthritis. Osteoarthritis Cartilage 1999;7(4):367-370 https://doi.org/10.1053/joca.1998.0237