--- slug: "jjm-167-final-version-10-10-2021" typ: "dartsch" quelle: "01 Wissenschaft/Studien Dartsch/JJM 167 final version_10.10.2021.pdf" chars: 28021 ---

JJM 167 final version_10.10.2021

Jpn J Med 2021, 4:1 495

Japan Journal of Medicine 2021; 4(1): 495- 500. doi: 10.31488/JJM. 167

Research article

Tesla Oscillator – Investigations on Its Beneficial Cell Effects Peter C. Dartsch Dartsch Scientific GmbH, Institut für zellbiologische Testsysteme, Auf der Vosshardt 25, D-49419 Wagenfeld, Germany * Corresponding author: Prof. Dr. Peter C. Dartsch, Dartsch Scientific GmbH, Institut für zellbiologische Testsysteme, Auf der Vosshardt 25, D-49419 Wagenfeld, Germany.

Received: June 08, 2021; Accepted: June 14, 2021; Published: June 16, 2021

Abstract Background. High-frequency treatment with Tesla currents is a medical science that is ancient and has been forgotten over time. The Tesla Oscillator is based on the basic theory of Nikola Tesla and Georges Lakhovsky, but is a real technical advancement. It is stated by the manufacturer to be able to inform, structure or energize water which activates the body's own energy fields and stimulates the self-healing power. Materials and Methods. In the present study, current cell biological test methods were used to investigate whether the Tesla experiment set, which includes the Tesla Oscillator and the handheld probe, is able to compensate an excess of endogenously generated radicals and to stimulate cell regeneration. Human promyelocytes (HL-60) were differentiated to functional neutrophils and were exposed to the frequencies of the Tesla Oscillator for a single time period up to 30 min. Then, superoxide anion generation was measured in the course of an induced oxidative burst. Connective tissue fibroblasts (L-929) were used to examine cell regeneration by closing a cell-free space due to migration and proliferation after exposure to the frequencies of the Tesla Oscillator for a single time period up to 30 min. Results. The results clearly demonstrate that the effects of a reduced radical generation after the exposure to the Tesla Oscillator was strictly time-dependent with a maximum of a nearly 40 % reduction after an exposure time of 30 min. The decrease in radical generation was statistically significant for exposure times ≥ 15 min. In addition, also the morphological examination of cell regeneration showed a clear time- dependent im-provement in the closure of the cell-free space after a single exposure of the cells to the frequencies of the Tesla Oscillator. The maximum stimulation of cell regeneration in comparison to the untreated control was measured after the 30 min exposure time and was nearly 30 % for exposure of the cells and 25 % for exposure of the culture medium. The stimulation was significant for exposure times ≥ 15 min. No significant difference was found between direct exposure of the cells or exposure of the culture medium which was used in the experiments afterwards. Conclusions. The use of the Tesla experiment set might improve and maintain personal health, vitality and well-being by an activation of cell regeneration and a reduced generation of superoxide anion radicals in complex wound healing or inflammatory processes. Keywords: Tesla Oscillator, tesla experiment set, oxidative burst, inflammation, functional neutrophil, connective tissue fibroblast, HL-60; L-929, cell regeneration, cell culture

Introduction The Tesla Oscillator is based on the basic theory of Nikola High-frequency treatment is a medical science that is ancient Tesla and Georges Lakhovsky, but it is no replica of the multi- and has been forgotten over time [1,2]. The so-called Tesla wave oscillator according to Lakhovsky [2], but a real technical currents were created in 1880 and were then implemented in advancement. Its manufacturer, Wassermatrix AG, CH-6343 high-frequency devices by various manufacturers. This thera- Rotkreuz, Switzerland, states that the Tesla Oscillator is able to peutic approach was further developed by Jacques-Arsène d’Ar- inform, structure or energize water by using the handheld probe. sonval in 1892 and used as a full-body treatment with high-fre- The water energized in this way is then supplied to the body by quency Tesla currents [3]. A further development was achieved drinking. This activates the body's own energy fields and stimu- by Kowarschick in Vienna [4]. lates the self-healing power. In this context, it must be taken into

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account that the human being consists of at least 70 % of water, with the green antenna rod pointing downwards in a table stand. so that a direct energizing of the body water is also possible. Treated cell cultures or test tubes made of polypropylene with According to the manufacturer, many users of the device report the cells or water to be informed were placed in a circle directly amazing improvements in vitality, health and well-being. around the antenna rod (Figure 1). The corresponding controls In the present study, current cell biological test methods were were located for the duration of the exposure in another room used to investigate whether the Tesla experiment set, which in- at least 10 meters away. The exposure was conducted at room cludes the Tesla Oscillator and the handheld probe, is able to temperature for t = 0 (untreated control), 5, 15 and 30 min. compensate an excess of endogenously generated radicals and Radical generation by functional neutrophils thus prevent oxidative stress with a delayed healing process. For Cells were cultivated as suspended mass cultures in special example, a local oxidative stress by reactive oxygen species in culture flasks with a ventilated lid (25 cm2 growth area; TPP, the tissue plays an important role in inflammatory processes [5- Switzerland) so that an atmospheric gas exchange can be inhibit- 8] or cell regenerative processes in the course of wound healing ed by turning the lid into another position. Thus, any pH changes [9,10] or after physical exercise or even overload [11-13]. It was in the bioassay during exposure to the frequencies of the Tesla also investigated whether the Tesla Oscillator has a direct benefi- Oscillator could be excluded. cial influence on cell regeneration, either by energizing the cells On day 5 of differentiation, cells were exposed to the frequen- themselves or by energizing the aqueous culture medium and cies of the Tesla Oscillator by the hand probe as described above causing a stimulation of cell migration and proliferation. and incubated for another 16 h in the incubator. The flasks were Material and Methods carefully wrapped in several layers of aluminium foil to avoid Tesla oscillator any unwanted interactions between the exposed cells and the unexposed controls. A complete Tesla experiment set including the Tesla Oscilla- tor and a handheld probe was kindly provided by Wassermatrix Finally, the cells of each experimental series were washed AG, CH-6343 Rotkreuz, Switzerland, for the duration of the ex- by several centrifugation steps (6 min at 190 x g) and repeated periments. The manufacturer states that the Tesla Oscillator is washings in phosphate buffered saline with calcium and magne- able to inform, structure or energize water by using the handheld sium. Cells were resuspended in phosphate buffered saline with probe. The water energized in this way is then supplied to the calcium and magnesium containing 10 mM glucose. 60 µl of body by drinking. This activates the body's own energy fields the cell suspension aliquots were pipetted to a reaction mixture and stimulates the self-healing power. In this context, it must be containing the tetrazolium dye WST-1 (Sigma-Aldrich, Deisen- taken into account that the human being consists of at least 70 hofen, Germany) and phorbol-12-myristate-13-acetate for the % of water, so that a direct energizing of the body water is also induction of an oxidative burst. The reactive superoxide anion possible. radicals in the reaction mixture caused the cleavage and color change of the dye. The amount of superoxide anion radicals Cell culture present in the reaction mixture was directly related to this color Human promyelocytes (cell line HL-60; ACC-3; ECACC change. The optical density was recorded at various time points 98070106; Leibniz-Institut; DSMZ German Collection for Mi- up to 45 min as a differential measurement ∆OD = 450 – 690 nm croorganisms and Cell Cultures, Braunschweig, Germany) were by an Elisareader (BioTek ELx 808 with software Gen 5 version cultivated over a period of several months. The cells were rou- 3.00) and calculated with Microsoft Excel 2016. tinely cultivated in RPMI 1640 medium supplemented with 10 Regeneration of connective tissue fibroblasts % growth supplement and 0.5 % gentamycin. Cells were seeded at a density of 100,000 cells/ml into the The non-adherent cells were routinely cultivated as suspended four cell culture compartments of silicone Culture-Insert 4 Well mass cultures and were regularly subcultured twice a week with (ibidi, Gräfelfing, Germany). The four compartments are sep- fresh culture medium. Cultures were incubated at 37 °C in an arated by a 500 μm thick silicone wall. Due to their especial- atmosphere of 5 % CO2 and 95 % air at almost 100 % humidity. ly designed surface, the inserts stick to the surface firmly and By addition of 1.5 % dimethylsulfoxide to the culture medium, completely prevent any cell attachment and growth under their cells were differentiated over a period of 6 days into functional silicone walls. When the silicone frame is removed, a cell-free neutrophils, which are capable of generating superoxide anion space (artificial wound) with sharp edges is left. The cell-free radicals after addition of phorbol-12-myristate-13-acetate (Sig- space can then be closed by cell migration and proliferation. ma-Aldrich, Deisenhofen, Germany) [14-18]. 48 h after seeding, the cell layers have become confluent and Connective tissue fibroblasts (cell line L-929; ACC-2; Leib- the silicone frames were removed. Immediately after removing niz Institute DSMZ - German Collection of Microorganisms and the silicone frame, cell cultures were exposed once for t = 0 (un- Cell Cultures, Braunschweig, Germany) were routinely grown treated control), 5, 15 and 30 min to the frequencies of the Tesla in RPMI 1640 with 10 % growth supplement and 0.5 % gen- Oscillator and then incubated for 20 h. In an additional series tamycin and incubated in an incubator at 37 °C in a humidified of experiments, not the cell cultures themselves, but the aque- atmosphere of 5 % CO2 and 95 % air. All cell culture reagents ous culture medium was exposed to the frequencies of the Tesla were purchased from Pan-Biotech, Aidenbach, Germany. experiment set for t = 0 (untreated control), 5, 15 and 30 min Experimental design for cell treatment and then pipetted to the cells before another 20 h incubation was The hand probe was clamped vertically outside the incubator performed. All cell samples were shielded by multiple wrapping

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Figure 1. Experimental setup for the exposure of functional neutrophils to investigate the formation of endogenously generated superoxide anion radicals (left picture), of connective tissue fibroblasts to investigate cell regeneration (middle picture) and by energizing the culture medium used for cell regeneration of connective tissue fibroblasts (right picture).

Figure 2. Representation of the time-dependent endogenous radical formation of cell cultures treated with the Tesla Oscillator. The decrease in radical generation is statistically significant for exposure times ≥ 15 min (p ≤ 0.01; Wilcoxon-Mann-Whitney test). The untreated control was set as “100%”. The data represent the mean value ± standard error of the mean of three independent experiments with duplicates

in aluminum foil and placed at different locations within the min, the generation of free radicals was reduced by 11.2 ± 5.5% incubator to avoid any unwanted interactions between exposed (mean value ± standard error of the mean) in comparison to the and unexposed cells. Finally, cells were fixed with methanol untreated control. After an exposure time of 30 min with the Tes- and stained with Giemsa’s azur eosin methylene blue solution la experiment set the reduced generation of radicals was 37.7 ± (Merck, Darmstadt, Germany) and air-dried. The width of the 7.7% (mean value ± standard error of the mean). The decrease remaining cell-free space was measured at 12 different locations in radical generation was statistically significant (p ≤ 0.01) for of each sample. Three independent experiments were conduct- exposure times ≥ 15 min. ed. Also the morphological examination of cell regeneration with Statistical analysis connective tissue fibroblasts showed a clear improvement in the Statistical analysis was done by using the non-parametric, closure of the cell-free space after a single exposure of the cells two-tailed Wilcoxon-Mann-Whitney test. to the frequencies of the Tesla experiment set (Figure 3). The evaluation of the cell-free space resulted in a more differentiated Results view on the time-dependent beneficial effect (Figure 4). There As shown in Figure 2, the reduced generation of endogenous was no difference between the direct exposure of the cell cul- radicals after exposure to the Tesla experiment set was strict- tures or by exposure of the culture medium which was then add- ly time-dependent. Even after a short exposure time of only 5 ed for the cell regeneration test.

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Figure 3. Representative micrographs of fixed and stained cell cultures demonstrating the closure of the cell-free space within 20 h. (A) Untreated control; (B) Cell culture which was directly exposed to the Tesla oscillator; (C) Cell culture with culture medium which was exposed to the Tesla Oscillator. Note the reduced cell-free space after both treatments. Olympus IX-50 inverted microscope equipped with an Olympus planachromate 10x and an Olympus E-10 digital camera with 4 megapixel resolution at bright field illumination.

Figure 4. Effect of the Tesla Oscillator on the regeneration of connective tissue fi-broblasts. The stimulation of cell regeneration with increasing exposure time is clearly visible and is statistically significant for exposure times ≥ 15 min (p ≤ 0.01; Wilcoxon-Mann-Whitney test). The untreated control was set as “100%”. Note that there is no difference between a direct exposure of the cells or an exposure of the culture medium which was then added to the cells. Data represent mean value ± standard error of the mean of three independent experiments.

The maximum stimulation of cell regeneration in comparison accepted in preclinical research. However, one might argue that to the untreated control was measured after the 30 min exposure cell cultures do not represent the human body with its complex time and was 26.8 ± 4.5 % (mean value ± standard error of the metabolic pathways and facets, but it should be considered that mean) for exposure of the cells and 25.1 ± 6.1 % (mean value ± cell cultures can shed light on selected aspects of living matter standard error of the mean) for exposure of the culture medium. and are a useful tool to avoid animal experiments. The stimulation was statistically significant for exposure times ≥ 15 min (p ≤ 0.01). Neutrophils are the most abundant type of granulocytes which make up about 60 % of all white blood cells in humans and are Discussion normally found in the bloodstream. They form an essential part Cell cultures are frequently used for a better understanding of of the innate immune system [21] and, therefore, play a key role the mechanisms that underlie cell activity in vivo. This includes in the front-line defense against invading microbial pathogens. differentiation, migration, proliferation, and Metabolism [19,20]. However, neutrophils are also one of the first responders of in- For over a century, two-dimensional cell cultures as used in this flammation and migrate from the blood into the inflamed tissue present study, have been used as in vitro models and are well and generate highly reactive superoxide anion radicals in the

Japan Journal of Medicine Jpn J Med 2021, 4:1 499 course of an oxidative or respiratory burst [5,22]. cies and their detoxification in healing skin wounds. J Investigat The present study with functional neutrophils has shown that Dermatol Symp Proc. 2006; 11:106-111. the frequencies of the Tesla experiment set are able to reduce the 11. Apor P, Rádi A. [Physical exercise, oxidative stress and damage]. generation of unwanted superoxide anion radicals by functional Orvosi Hetilap. 2006; 147:1025-1031. neutrophils in a time-dependent manner after only one single 12. Vollaard NB, Cooper CE, Shearman JP. Exercise-induced oxida- application of the Tesla Oscillator. Thus, a (chronic) inflamma- tive stress in overload training and tapering. Med Sci Sports Exer- tory process in the tissue might be improved by the inhibition of cise. 2006; 38:1335-1341. radical generation in the inflamed tissue. 13. Larsen MK, Matchkov VV. Hypertension and physical exercise: Somehow synergistically is the beneficial effect of the fre- The role of oxidative stress. Medicina. 2016; 52:19-27. quencies of the Tesla Oscillator on cell regeneration which is 14. Babior BM. NADPH oxidase: An update. Blood. 1999; 93:1464- stimulated in a time-dependent manner. In this present test sys- 1476. tem, especially the granulation phase is simulated which is char- 15. Tan AS, Berridge MV. Superoxide produced by activated neutro- acterized by migration and proliferation of the cells in order to phils efficiently reduces the tetrazolium salt WST-1 to produce close the cell-free space (= wounded area) in vivo. However, a a soluble formazan: a simple colorimetric assay for measuring complicated wound healing process is also related to local oxi- respiratory burst activation and for screening anti-inflammatory dative stress [23-25] which is reduced by the frequencies at the agents. J Immunol Meth. 2000; 238:59-68. same time. 16. Teufelhofer O, Weiss RM, Parzefall W, et al. Promyelocytic HL60 cells express NADPH oxidase and are excellent targets in a rapid The results clearly demonstrate that the frequencies of the spectrophotometric microplate assay for extracellular superoxide. Tesla Oscillator are able to inform, structure or energize the wa- Toxicol Sci. 2003; 76:376-383. ter of the aqeous culture medium surrounding the cells. There-

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To cite this article: Peter C. Dartsch. Tesla Oscillator – Investigations on Its Beneficial Cell Effects. Japan Journal of Medicine. 2021; 4:1.

© 2021 Peter C. Dartsch.

Japan Journal of Medicine