Bioresonance and cells – interactions and beneficial effects

Prof. Peter C. Dartsch, Dartsch Scientific GmbH, Wagenfeld, Germany

1. Introduction

Since its foundation in 2002 in the immediate vicinity of the university town of Tübingen, Dartsch Scientific GmbH has been exclusively engaged in animal free research and development through the use of organ-specific cell cultures. In addition to standard procedures, the company has also developed numerous proprietary test procedures that are successfully used in very different areas for investigating active substances and methods. This brief summary is intended to describe the cell biological investigations we have performed using the BICOM bioresonance device and their results. The cell biological test systems we use are recognised in the scientific and conventional medical world and we have reported on them in numerous publications before.

2. Bioresonance device BICOM optima Mobile and exposure of the cells

Dartsch Scientific GmbH was first commissioned in October 2017 to use the latest cell biological methods to investigate whether beneficial effects can be demonstrated in cultivated connective tissue cells by using the BICOM bioresonance device. It took several experimental attempts over a period of several months to establish beneficial effects. This is due to the complexity of the method and the sensitivity of the measuring device, since even the smallest details had to be considered and first optimised in practical application with cell cultures. REGUMED Regulative Medizintechnik GmbH, D-82152 Planegg, Germany, kindly loaned us a BICOM optima mobil device. In addition, the final optimisation stage used the BICOM power applicator GST71 (optima), which was housed in a mini incubator at 37°C for the duration of the exposure. The program sequence “Pathogens Ai” was used as the basis.

All three sub-programs were set to last 30 minutes, so that one exposure cycle took 90 minutes. This cycle ran 2x consecutively during the exposure of the cell cultures, so that the cells were exposed for a total of 180 minutes. The sample cup at the top right always contained the same culture medium for the experiments along with cell samples that had been pre-treated under the same conditions.

We naturally realise that the duration of exposure in humans should not exceed one hour. However, the primary underlying question here was whether the BICOM bioresonance method can achieve a positive reaction in cells at all.

3. Investigations with connective tissue fibroblasts

Connective tissue fibroblasts of cell line L-929 (Leibniz Institute DSMZ, German Collection of Microorganisms and Cell Cultures, Braunschweig) were used over a period of several months. Cells were routinely grown in a specific culture medium in an incubator at 37 °C in an atmosphere of 5% CO2 and 95% air and near 100% humidity.

Cell vitality

For the cell viability experiments, cells were seeded from mass cultures into 24 central wells of each of two 96-hole culture dishes per set of experiments and incubated for 24 hours until the cells had become fully attached. The cells were seeded in the same density on round glass cover slips and also pre-incubated for 24 h as reference cells for the sample cup of the BICOM bioresonance device. The first culture dish was placed directly on the power applicator and exposed for a total of 180 minutes using the “Pathogens Ai” program sequence. The second culture dish was incubated as a reference. Subsequently, both culture dishes were incubated for another 21 hours until evaluation. Finally, cell vitality was measured by the change in colour of a specific dye. A total of 3 independent experiments were performed.

in comparison to the untreated control sample. The upper figure plots the individual pairs of measurements of the same wells in the two 96-hole culture dishes and the corresponding mean value as a dashed line. It can be clearly seen that, despite the fluctuations that a biological measurement system always entails, the mean value for the exposed cells is significantly higher than that for the untreated cells. The difference becomes even more evident when – as shown in the lower figure – the percentage difference between the exposed cells is plotted in comparison to the untreated control cells. The dashed line also represents the mean value for the exposed cells, which is significantly higher than that for the untreated cells. The dashed line also represents the mean value here.

Cell regeneration/wound healing

Cell regenerative processes restore the functionality of a damaged tissue in vivo. If an injury occurred previously, closure and tissue consolidation can be achieved in what is known as the granulation phase through cell migration and division of the predominant cell type in the tissue in question. This is the phase that is simulated in the test system used here.

The cells were seeded and incubated at a high density in the three individual compartments of so-called 3-well culture inserts made of silicone. The compartments of the inserts are separated from each other by a 500 µm thick silicone bar and bordered on the outside by a silicone bar. An insert will adhere firmly to the bottom of a culture dish due to the special adhesion area, thus forming a defined cell-free area.

After reaching confluence (cells lying very close to each other) within 48 hours after cell seeding, the inserts were pulled off with forceps to obtain sharply defined cell-free areas between the compartments. The cells could now migrate into these cell-free areas and close that cell-free area again through increased cell division. The first culture dish was placed directly on the power applicator and exposed for a total of 180 minutes using the “Pathogens Ai” program sequence. The second culture dish was incubated without treatment as a control sample. After the period of exposure, both culture dishes were incubated for another 21 hours until evaluation. The cells were then fixed, stained, and airdried, and the width of the remaining cell-free area was measured under the microscope.

A total of 3 independent experiments were performed.

Result:

As shown in Fig. 2, exposure of the cultured connective tissue cells to the BICOM optima mobil and the “Pathogens Ai” program sequence resulted in faster migration and division of the cells by almost 25 %. As a result, the cell-free space closed significantly faster than in the untreated control sample.

Fig. 2: Photomicrographs of cultured connective tissue cells after a total of 24 hours incubation. The cells in the image on the right (B) were exposed to the BICOM optima Mobile and the “Pathogens Ai” program sequence for 180 minutes directly after removal of the silicone frames. The image on the left (A) shows the control culture without exposure. It is clearly visible that the cell-free space in the image on the right is much smaller than in the one on the left.

Summary and conclusions

The application of the BICOM optima mobil device caused a pronounced and statistically significant stimulation of cell vitality in the cultivated connective tissue cells; this can lead to increased physical performance in humans and hence to an improved sense of well-be-ing. In addition, it also resulted in a shorter cell regeneration time by stimulating cell migration and cell division. This can play a very important role, especially when it comes to sporting strain (or even excessive strain).

4. Studies with functional neutrophils

Neutrophils are the most common type of granulocyte, a specific type of white blood cell, in most mammals. They play a dual role, firstly as phagocytes (= scavenger cells) and secondly as proinflammatory or inflammation-mediating cells. They form a cellular defence against invading microbial pathogens by floating in the circulating blood as an essential component of the innate immune system. In cases of inflammation, neutrophils migrate from the blood into the tissue and produce reactive oxygen species, predominantly superoxide anion radicals, in what is termed an oxidative or respiratory burst. Although these radicals basically play an important role in intercellular signal transmission, an excess of radicals in the tissue (local oxidative stress) can no longer be neutralised by the body’s own enzymes (for example superoxide dismutase or catalase) and lead to further undesirable cell and tissue damage. As a result, this generally delays the success of the cure considerably or, in the worst case, even jeopardises it.

Human promyelocytes (cell line HL-60; Leibniz Institute; DSMZ German Collection for Microorganisms and Cell Cultures, Braunschweig) were used for the experiments as a matter of routine. Cells were routinely grown in a specific culture medium in an incubator at 37 °C in an atmosphere of 5% CO2 and 95% air and near 100% humidity. The cells were differentiated into so-called functional neutrophils under culture conditions with 1.5 vol% dimethyl sulphoxide in the culture medium over a five-day period, i.e. cells that can produce a local excess of radicals in an oxidative burst after stimulation by a chemical agent (phorbolester) (Fig. 3).

Fig 3: Simulation of one aspect of the inflammatory response in tissue through endogenously generated oxygen radicals in functional neutrophils. The cells were treated with the “Pathogens Ai” program sequence for 180 minutes on each of three consecutive days during the differentiation process. The corresponding control samples were incubated for the same length of time without exposure. The cells were washed by repeated centrifugation and resuspension on day 5 of differentiation and incorporated into a phosphate buffer with 10 mM glucose. The functional neutrophils in the reaction mixture were stimulated to generate superoxide anion radicals by adding a phorbol ester. Here, the radicals caused a cleavage of the dye that was also added to the experimental batch. The amount of oxygen radicals present in the reaction mixture was directly proportional to the change in colour. In addition, the basal metabolic activity of the cells without being stimulated to an oxidative burst was determined in the same way in the reaction mixture. The change in colour was recorded at specific wavelengths using an Elisa Reader. A total of 5 independent experiments were performed.

Result

Measurement of the basal cellular metabolism of functional neutrophils without triggering an oxidative burst showed inhibition after exposure to the BICOM bioresonance device when compared with untreated control samples in each experiment performed. Calculating the mean value ± standard deviation from the 5 individual tests yields a statistically significant inhibition of basal cell metabolism amounting to 12 ± 5 %. The inhibition of the basal metabolism of functional neutrophils was in accordance with the inhibition of the formation of reactive oxygen radicals in the induced oxidative burst. Here, the inhibition over all 5 individual trials was as high as 18 ± 4 % (mean ± standard deviation). This inhibition of radical formation was statistically highly significant compared to the untreated control sample.

Summary and conclusions

In the animal-free studies conducted here with inflammation-mediating cells (functional neutrophils), the BICOM bioresonance device with the “Pathogens Ai” program sequence demonstrated that it can inhibit both basal metabolism and the formation of reactive oxygen radicals through these cells.

Even though it is difficult to draw a direct conclusion from the results of the investigations to a whole organism, the results do show the potential of the bioresonance device with this program combination to reduce local oxidative stress in tissue also in vivo and thus to neutralise – at least partially – an important reaction cascade of a (chronic) inflammatory process and hence to improve well-being. Furthermore, a significantly more powerful effect of the bioresonance device would result in an undesirable reduction in the efficiency of the potential of the cells circulating in the blood to defend against microbial pathogens as an innate immune defence.

5. Supplementary note

The results of the study, which are only briefly presented here, can also be read in detail in two scientific publications in international journals:

Dartsch PC (2021) Investigations on the beneficial effects of BICOM optima mobile bioresonance-device

on cultured connective tissue fibroblasts. J Biomed Sci Res 3 (1): 133.

Dartsch PC (2021) Effects of the BICOM optima mobile bioresonance device on cell metabolism and oxi-

dative burst of inflammation-mediating cells. Biomed J Sci & Tech Res 33: 25616-25620.

BRIEF CV

Prof. Peter C. Dartsch, Dartsch Germany

Prof. Dr. rer. nat. Peter C. Dartsch initially studied chemistry at the Technical University of Darmstadt and then biochemistry at the University of Tübingen. He first worked at the Medical Faculty of the University of Tübingen on the cultivation of organ-specific primary cells and the development and establishment of invivo cell culture models, when writing his diploma thesis (1985) and later during his doctorate (1989) and

habilitation in human physiology (1991). He became an associate professsor in 1997. In 2002 he left his position of deputy managing director of the Institute for Occupation and Social Medicine and retired from public service. He then established Dartsch Scientific GmbH as the managing director and sole shareholder, which to this day deals with animal-free cell biological test systems in preclinical research and development.

To date, he has written more than 150 scientific publications and book contributions, given 150 lectures and poster presentations and supervised more than 30 medical doctoral theses. He was an external member of the Medical Faculty of the University of Tübingen until his retirement in 2020, but is still active today as Managing Director of Dartsch Scientific GmbH.