Clinical Studies

Studies on Arthritis

Abstract
Osteoarthritis (OA) is the most common disorder of the musculoskeletal system and is a consequence of mechanical and biological events that destabilize tissue homeostasis in articular joints. Controlling chondrocyte death and apoptosis, function, response to anabolic and catabolic stimuli, matrix synthesis or degradation and inflammation is the most important target of potential chondroprotective treatment, aimed to retard or stabilize the progression of OA. Although many drugs or substances have been recently introduced for the treatment of OA, the majority of them relieve pain and increase function, but do not modify the complex pathological processes that occur in these tissues. Pulsed electromagnetic fields (PEMFs) have a number of well-documented physiological effects on cells and tissues including the upregulation of gene expression of members of the transforming growth factor b super family, the increase in glycosaminoglycan levels, and an antiinflammatory action. Therefore, there is a strong rationale supporting the in vivo use of biophysical stimulation with PEMFs for the treatment of OA. In the present paper some recent experimental in vitro and in vivo data on the effect of PEMFs on articular cartilage were reviewed. These data strongly support the clinical use of PEMFs in OA patients.

Summary
Objective: Hartley guinea pigs spontaneously develop arthritis that bears morphological, biochemical, and immunohistochemical similarities to human osteoarthritis. It is characterized by the appearance of superficial fibrillation by 12 months of age and severe cartilage lesions and eburnation by 18 months of age. This study examines the effect of treatment with a pulsed electromagnetic field (PEMF) upon the morphological progression of osteoarthritis in this animal model.
Design: Hartley guinea pigs were exposed to a specific PEMF for 1 h/day for 6 months, beginning at 12 months of age. Control animals were treated identically, but without PEMF exposure. Tibial articular cartilage was examined with histological / histochemical grading of the severity of arthritis, by immunohistochemistry for cartilage neoepitopes, 3B3(−) and BC-13, reflecting enzymatic cleavage of aggrecan, and by immunoreactivity to collagenase (MMP-13) and stromelysin (MMP-3). Immunoreactivity to TGFβ, interleukin (IL)-1β, and IL receptor antagonist protein (IRAP) antibodies was examined to suggest possible mechanisms of PEMF activity.
Results: PEMF treatment preserves the morphology of articular cartilage and retards the development of osteoarthritic lesions. This observation is supported by a reduction in the cartilage neoepitopes, 3B3(−) and BC-13, and suppression of the matrix-degrading enzymes,collagenase and stromelysin. Cells immunopositive to IL-1 are decreased in number, while IRAP-positive cells are increased in response totreatment. PEMF treatment markedly increases the number of cells immunopositive to TGFβ.
Conclusions: Treatment with PEMF appears to be disease-modifying in this model of osteoarthritis. Since TGFβ is believed to upregulate gene expression for aggrecan, downregulate matrix metalloprotease and IL-1 activity, and upregulate inhibitors of matrix metalloprotease,the stimulation of TGFβ may be a mechanism through which PEMF favorably affects cartilage homeostasis.

Summary
Objective: To investigate the role of pulsed electromagnetic field (PEMF) exposure parameters (exposure length, magnetic field peak amplitude, pulse frequency) in the regulation of proteoglycan (PG) synthesis of bovine articular cartilage explants.
Methods: Bovine articular cartilage explants were exposed to a PEMF (75 Hz; 2 mT) for different time periods: 1, 4, 9, 24 h. Then, cartilage explants were exposed for 24 h to PEMFs of different magnetic field peak amplitudes (0.5, 1, 1.5, 2 mT) and different frequencies (2, 37, 75, 110 Hz). PG synthesis of control and exposed explants was determined by Na2-35SO4 incorporation.
Results: PEMF exposure significantly increased PG synthesis ranging from 12% at 4 h to 17% at 24 h of exposure. At all the magnetic field peak amplitude values, a significant PG synthesis increase was measured in PEMF-exposed explants compared to controls, with maximal effect at 1.5 mT. No effect of pulse frequency was observed on PG synthesis stimulation.
Conclusions: The results of this study show the range of exposure length, PEMF amplitude, pulse frequency which can stimulate cartilage PG synthesis, and suggest optimal exposure parameters which may be useful for cartilage repair in in vivo experiments and clinical application.

Summary
Background Osteo-arthritis, a painful joint disorder involving degenerative changes of the articular cartilage and subchondral bone, often results in progressive functional impairment and disability. One particular modality used by physiotherapists that shows very promising results in reducing the joint damage and pain found in osteo-arthritis is pulsed
electromagnetic fields.
Objective The present objective was to examine the
rationale for, and the potential efficacy of, applying pulsed electromagnetic fields for reducing joint pain and other related symptoms of osteo-arthritis.
Methods The related English language literature was
extensively reviewed to examine whether changes in pain might be expected from the application of pulsed electromagnetic fields to an osteo-arthritic joint, and why.
Results The basic and clinical research in this field, while somewhat limited, supports the insightful application of pulsed electromagnetic fields to ameliorate pain and disability due to osteo-arthritis.
Conclusion Further basic and clinical research to validate the use of pulsed electromagnetic fields in facilitating function and possibly in facilitating joint reparative processes in osteo-arthritis, as well the lessening of osteo-arthritic joint pain and joint dysfunction is indicated.

Indian Journal of Aerospace Medicine : 48(2) 2004

Rotational Field Quantum Magnetic Resonance (RFQMR) in the Treatment of Osteo Arthritis of the Knee Joint

Studies on Cancer

NMR technology has dramatically contributed to the revolution of image diagnostic. NMR apparatuses use combinations of microwaves over a homogeneous strong (1 Tesla) static magnetic field. We had previously shown that low intensity (0.3–66 mT) static magnetic fields deeply affect apoptosis in a Ca2+ dependent fashion (Fanelli et al., 1999 FASEBJ., 13;95–102). The rationale of the present study is to examine whether exposure to the static magnetic fields of NMR can affect apoptosis induced on reporter tumor cells of haematopoietic origin. The impressive result was the strong increase (1.8–2.5 fold) of damage-induced apoptosis by NMR. This potentiation is due to cytosolic Ca2+ overload consequent to NMR-promoted Ca2+ influx, since it is prevented by intracellular (BAPTA-AM) and extracellular (EGTA) Ca2+ chelation or by inhibition of plasma membrane L-type Ca2+ channels. Three-days follow up of treated cultures shows that NMR decrease long term cell survival, thus increasing the efficiency of cytocidal treatments. Importantly, mononuclear white blood cells are not sensitised to apoptosis by NMR, showing that NMR may increase the differential cytotoxicity of antitumor drugs on tumor vs normal cells. This strong, differential potentiating effect of NMR on tumor cell apoptosis may have important implications, being in fact a possible adjuvant for antitumor therapies.

We investigated the effects of pulsed magnetic stimulation on tumor development processes and immune functions in mice. A circular coil (inner diameter¼15 mm, outer diameter¼75 mm) was used in the experiments. Stimulus conditions were pulse width¼238 ms, peak magnetic field¼0.25 T
(at the center of the coil), frequency¼25 pulses/s, 1000 pulses/sample/day and magnetically induced eddy currents in mice¼0.79–1.54 A/m2. In an animal study, B16-BL6 melanoma model mice were exposed to the pulsed magnetic stimulation for 16 days from the day of injection of cancer cells. A tumor growth study revealed a significant tumor weight decrease in the stimulated group (54%
of the sham group). In a cellular study, B16-BL6 cells were also exposed to the magnetic field (1000 pulses/sample, and eddy currents at the bottom of the dish¼2.36–2.90 A/m2); however, the magnetically induced eddy currents had no effect on cell viabilities. Cytokine production in mouse spleens was measured to analyze the immunomodulatory effect after the pulsed magnetic stimulation.

Tumor necrosis factor (TNF-a) production in mouse spleens was significantly activated after the exposure of the stimulus condition described above. These results showed the first evidence of the antitumor effect and immunomodulatory effects brought about by the application of repetitive magnetic stimulation and also suggested the possible relationship between anti-tumor effects and the increase of TNF-a levels caused by pulsed magnetic stimulation.

Extremely low frequency (ELF) pulsed-gradient magnetic field (with the maximum intensity of
0.6–2.0 T, gradient of 10–100 T · M_1, pulse width of 20–200 ms and frequency of 0.16–1.34 Hz treatment of mice can inhibit murine malignant tumour growth, as seen from analyses at
different hierarchical levels, from organism, organ, to tissue, and down to cell and macromolecules.
Such magnetic fields induce apoptosis of cancer cells, and arrest neoangiogenesis,
preventing a supply developing to the tumour. The growth of sarcomas might be amenable to such new method of treatment.

Contact inhibition of division: Involvement of the electrical transmembrane potential

Cellular potentials of normal and cancerous fibroblasts and hepatocytes.   

Deficits in elevating membrane potential of rat fibrosarcoma cells after cell contact

Most cancer cells are known to have lower resting cellular potentials than do their normal counterparts. This study investigates how these potentials establish themselves during growth and cellular contact in tissue culture. Normal quail embryonic fibroblasts and quail fibrosarcoma (QT-35) and normal rat kidney cells and rat fibrosarcoma (from rat fibroblasts chemically transformed by nitroquinoline oxide) were recorded intracellularly using high-impedance micropipets. In high-density high-contact cultures, both quail and rat cancer cells had lower potentials than did normal cells (-20.7 compared to -40.1 mV for quail and -30.7 compared to -61.9 mV for rat). In low-density mitotically synchronous cultures, the rat cells were recorded every 4 hr for 96 hr. Starting at a low density, normal cell membrane potential is maintained at a low level through subsequent cell divisions. Without any additional change in cell density, the potential suddenly elevates to a high level. The membrane potential of cancer cells is by contrast unrelated either to cell density or to time. Cancer cells maintained an intermediate potential from low to very high densities and never elevated their potential to high values. The failure of cancer cells to reach high potentials may be linked to their uncontrolled cell division.

Calcium ion and the membrane potential of tumor cells

Binggeli R, Weinstein RC, Stevenson D Cancer Biochemisry Biophysics 1994 Oct;14(3):201-10.

Calcium ion affects ion permeability and membrane potential among many other aspects of cell function. Initial effects of increasing extracellular calcium upon membrane potential were studied in a quail fibrosarcoma (QT35) where calcium had a dose dependent effect, and normal quail fibroblasts, where there was little effect. Comparisons were then made in six different human hepatocellular carcinomas (Tong, HepG2, Hep3B, PLC/PRF/5, Mahlavu, and HA22T) in response to smaller changes in concentration. There were insignificant changes in membrane potential in two cell lines and significant elevations in four. Cytolysis by natural killer cells also declined in rough proportion to the increase in membrane potential. The less differentiated hepatocellular carcinoma cells have both higher baseline membrane potentials and a greater potential increase to increased calcium. By contrast, more highly differentiated tumor cells had paradoxically smaller membrane potentials and along with normal cells had small potential responses to calcium increases.

Summary
Low-energy, pulsed electromagnetic fields (PEMFs) have reversed therapeutically resistant pathologic processes in the musculo-skeletal system. Their development as a non-thermal therapeutic agent is based on 30 years of study of the electro-biological properties of connective tissues. Specific energy characteristics in applied PEMFs produce selected biological effects by modifying synthetic and other behavioral patterns of target cells; some mechanisms of action are defined. The technology appears safe and effective in clinical treatment of un-united fractures, avascular necrosis of bone, and chronic, refractory tendinitis. An expanding, rational use in biomedical science is predicted.