The Influence of Nuclear Magnetic Resonance Therapy (NMRT) and Interleukin IL1-b Stimulation Cal 78 Chondrosarcoma Cells and C28/ I2 Chondrocytes

Introduction: For the last decade the Nuclear Magnetic Resonance (NMR) therapy has turned out a success in pain treatment of patients who suffer from osteoarthritis (OA) of knees or hands, low back pain and osteoporosis, respectively. While clinical outcome could be proved, less is known about the underlying mechanism by which NMRT modulates cellular processes leading to observed pain reduction. This study implements the analysis of potential signal transduction pathways involved in NMRT signal transfer in Cal-78 chondrosarcoma cells and NMRT influencing cell growth and viability of interleukin IL- 1β stimulated Cal-78 cells and C28/I2 chondrocytes. Changes in expression of inflammatory proteins like metalloproteinases MMP1, MMP3, MMP8, MMP9, MMP10 and MMP13, as well as interleukins IL6 and IL8 were evaluated. Methods: Basic pathway analysis was performed by the gene array technology ensued by the reporter-gene technique (Cignal™ Reporter Assay). Cell proliferation was tested by calcein- and eFlour 670 staining; the S-phase of the cells was determined by use of the BrdU assay. Changes in expression of the inflammatory proteins were evaluated by performing quantitative RT-PCR, enzyme immuno assays and luminex measurements. Results: The interpretation of the gene array combined with the results of the reporter gene assay roughly reveals indices of NMRT influencing the transforming growth factor (TGF)- β and Mitogen Activated Protein (MAP) kinase pathway. The cell growth and viability of both cell lines was not changed by NMRT although their inhibition caused by IL-1β was more pronounced in the absence of NMRT stimulation. Even though IL-1β provably stimulated the increase of the expression of MMPs and ILs, these effects seemed to be divergently regulated within the two cell lines. Interestingly at the level of RNA MMP13 and IL-8 are statistically down regulated by NMRT only in C28/I2 cells. Conclusions: Our findings concerning effects of IL-1β being less significant under NMRT than under control conditions, plus the reduced MMP-13 expression substantiate the postulated anti-inflammatory effect of NMRT relevant for pain relief. Clarification of how NMRT signaling is processed by targeted cells will broaden perspectives for clinical adaptations of NMRT by more precise and efficient application of this alternative therapeutically action.