Bioresonance and the Cell —Interactions and Beneficial Effects
Bioresonance and the Cell —Interactions and Beneficial Effects
Prof. Dr. rer. nat. habil. Peter C. Dartsch, graduate biochemist, Germany
Background
Since its foundation in 2002, Dartsch Scientific GmbH has focused exclusively on animal-free preclinical research and development using organ-specific cell culture models. In addition to standard procedures, the company has also developed numerous proprietary test procedures that are successfully used in a wide variety of areas to investigate active substances and methods. These possibilities will be illustrated in the lecture using examples.
In this textual summary, only the cell biological tests we have carried out with the BICOM bioresonance device and their results will be discussed. All the cell biological test systems used are recognized in the scientific and academic medical world and have already been described by us in detail in numerous publications.
What is cell culture?
Cell culture is the cultivation of animal or plant cells in a nutrient medium outside the organism. In addition to the origin (species and organ), a basic distinction is made between immortalized (immortal) cell lines, i.e. cells of a tissue type that can reproduce indefinitely in the course of their cultivation and are very stable in their properties, and primary cells. The latter are freshly isolated cells from organs that only have a limited lifespan in culture and then often die off or their properties change considerably with increasing in vitro age compared to the original cell and can no longer be used for further investigations. Cell cultures are widely used in basic biological and medical research, the development of therapeutic proteins and cell therapies as well as in the manufacture and production of vaccines and monoclonal antibodies.
Bioresonance device BICOM optima mobile and exposure of the cells
Dartsch Scientific GmbH was first commissioned in October 2017 to use current cell-biological methods to investigate whether the use of the BICOM bioresonance device can demonstrate beneficial effects in cultivated connective tissue cells. Only after several trials over a period of several months it was possible to determine beneficial effects. This is due to the complexity of the method and the sensitivity of the measuring device, as even the smallest details had to be taken into account and first optimized during practical application with cell cultures.
We were provided with a BICOM optima mobile loan device by REGUMED Regulative Medizintechnik GmbH, 0-82152 Planegg. In addition, the BICOM power applicator GST71 (optima) was used in the final optimization stage, which was housed in a mini incubator at 37 °C for the duration of the exposure. The "Pathogens Ai" program sequence was used as the basis. All
three sub-programs were set to a duration of 30 min, so that one exposure cycle lasted 90 min. This cycle ran twice in succession during the exposure of the cell cultures, so that the cells were exposed for a total of 180 min. The sample cup at the top right always
contained the same culture medium for the experiments as well as cell samples that had been pretreated under the same conditions.
We are of course aware that the duration of exposure on humans should not exceed one hour. However, the primary underlying question here was whether the BICOM bioresonance method can achieve a positive cell reaction at all.
4. Studies with connective tissue fibroblasts
Connective tissue fibroblasts of the cell line L-929 (Leibniz Institute; DSMZ - German Collection of Microorganisms and Cell Cultures, Braunschweig) were used over a period of several months. The cells were routinely cultivated in a specific culture medium (RPM! 1640 with 10 % growth mixture and 1 % gentamycin) in an incubator at 37 °C and an atmosphere of 5 % CO2 and 95 % air and at least 90 % humidity.
4.1 Cell vitality
For the experiments on cell vitality, the cells from mass cultures were sown in 24 central wells of two 96-hole culture plates per test batch and incubated for 24 hours until the cells were fully attached. As reference cells for the sample cup of the BICOM bioresonance device, the cells were seeded at the same density on round glass coverslips and also pre incubated for 24 hours. The first culture plate was placed directly on the power applicator and exposed for a total of 180 minutes with the program sequence "Pathogens Ai". The second culture plate was incubated as a reference. Afterwards both culture plates were incubated for further 21 hours until evaluation. Finally, cell vitality was measured by the color change of a specific dye. A total of 3 independent experiments were carried out.
Result: Exposure of the cells to the program sequence "Pathogens Ai" for 180 minutes resulted in a clear improvement in cell vitality compared to the control cells in all measured samples (Fig. 1). The calculation of cell vitality using the BICOM optima mobil across all measured values resulted in a statistically significant promotion of 38 ± 15 % (mean ± standard deviation for n = 3; p 0.01, Wilcoxon-Mann-Whitney test) compared to the control.
Fig. 1: Graphical representation of the absolute measured values for cell vitality after exposure to the BICOM optima mobil compared to the untreated control. The individual pairs of measured values in the 96-well culture plates are plotted as well as the respective mean values as a dashed line. The higher the measured value, the better the vitality of the cells. It can be clearly seen that the mean value for the exposed cells is significantly higher than the mean value for the untreated cells.
4.2 Cell regeneration/wound healing
Cell regenerative processes restore the functionality of damaged tissue in vivo. If an injury has occurred, the tissue can be closed and strengthened in the so-called granulation phase through the immigration and division of the predominant cell type in the tissue concerned. This phase in particular was 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 culture inserts 3 well made of silicone. The compartments of the inserts are separated from each other by a 500 p.m thick silicone bar and are bordered on the outside by a silicone bar. Due to the special adhesion area, an insert adheres firmly to the bottom of a culture dish and thus forms a defined cell-free area.
After reaching confluence (cells lie close together) within 48 hours of cell seeding, the inserts were removed with tweezers to obtain sharply demarcated cell-free areas between the compartments. The cells could then migrate into these cell-free areas and close the cell-free area by increased cell division. The first culture dish was placed directly on the power applicator and exposed to the "Pathogens Ai" program sequence for a total of 180 minutes. The second culture dish was incubated without treatment as a control. At the end of the exposure time, both culture plates were incubated for further 21 hours until analysis. The cells were then
fixed, stained, air-dried and the width of the remaining cell-free area was measured under the microscope. A total of 3 independent experiments were carried out.
Result: As shown in Fig. 2, exposure of the cultured connective tissue cells to the BICOM optima mobil and the program sequence "Pathogens Ai" resulted in faster migration and division of the cells by almost 25 %. As a result, the cell-free space was closed significantly faster than in the untreated control.
Fig. 2: Photomicrographs of cultured connective tissue cells after a total of 24 hours of incubation. The cells in the right-hand image (B) were exposed to the BICOM optima mobil and the "Pathogens Ai" program sequence for 180 minutes immediately after the silicone frames were
removed. The left image (A) shows the untreated control culture without exposure. It is clearly visible that the cell-free space on the right-hand image is significantly smaller than on the left-hand image.
4.3 Summary and conclusions
The use of the BICOM optima mobil device caused a pronounced and statistically significant stimulation of cell vitality in the cultivated connective tissue cells, which can lead to higher physical performance and thus to improved well-being in humans. In addition, it also caused a shortened cell regeneration time by stimulating cell migration and cell division. Especially during sporting exertion (or even overloading) this can play a very important role.
5. Studies with functional neutrophils 5.1 Function of neutrophils
In most mammals, neutrophils are the most common type of granulocyte, i.e. a specific type of white blood cell. They play a dual role, firstly as phagocytes (= scavenger cells) and secondly as inflammation-mediating cells. By swimming in the circulating blood, they form a cellular defense against invading microbial pathogens as an essential part of the innate immune system. During inflammation, the neutrophils migrate from the blood into the tissue and produce reactive oxygen species (= superoxide anion radicals) in a so-called oxidative or respiratory burst. Although these radicals fundamentally play an important role in intercellular signal transmission, an excess of radicals in the tissue (= local oxidative stress) can no longer be controlled by the body's own enzymes (e.g. superoxide dismutase or catalase) and lead to further undesirable cell and tissue damage. As a result, successful healing is usually considerably delayed or in the worst case even called into question.
5.2 Carrying out the experiment
Human promyelocytes (cell line HL-60; Leibniz Institute; DSMZ German Collection of Microorganisms and Cell Cultures, Braunschweig) were routinely used for the experiments. The cells were routinely cultured in a specific culture medium (DMEM/Ham's F12 1:1 with 10 % growth mixture and 1 % gentamycin) in an incubator at 37 °C and an atmosphere of 5 % CO2 and 95 % air as well as almost 90 % humidity. Under culture conditions with 1.5 vol% dimethyl sulfoxide in the culture medium the cells were differentiated within a five-day period into so-called functional neutrophils, i.e. cells that can produce a local excess of radicals in an oxidative burst after stimulation by a chemical agent (phorbol ester).
During the differentiation process, the cells were treated with the program sequence "Pathogens Ai" for 180 minutes on three consecutive days. The respective controls were incubated for the same time without exposure. On day 5 of differentiation, the cells were washed by centrifugation several times and resuspended again and taken up in phosphate buffer with 10 mM glucose. The functional neutrophils in the reaction mixture were stimulated to generate superoxide anion radicals by adding the phorbol ester. The radicals led to cleavage of the dye, which was also added to the experimental mixture. The amount of oxygen radicals present in the reaction mixture was directly proportional to the color change. In addition, the basal metabolic activity of the cells in the reaction mixture was determined in the same way without the stimulation of an oxidative burst. The color change was recorded at specific wavelengths using an elisa reader. A total of five independent experiments were carried out.
Result: The measurement of the basal cell metabolism of functional neutrophils without triggering an oxidative burst revealed an inhibition after exposure to the BICOM bioresonance device in every experiment compared to the untreated controls. If the mean value ± standard deviation is calculated from the five individual tests, a statistically significant inhibition of basal cell metabolism of 12 ± 5 % is obtained (p 0.01; Wilcoxon-Mann-Whitney test).
The inhibition of the formation of reactive oxygen radicals in the induced oxidative burst was consistent with the inhibition of the basal metabolism of functional neutrophils. Here, the inhibition across all five individual experiments was as high as 18 ± 4 % (mean value ± standard deviation; n = 5). Compared to the untreated control, this inhibition of radical formation was also statistically significant (p 0.01; Wilcoxon-Mann-Whitney test).
5.3 Summary and conclusions
In the animal-free studies carried out with inflammation-mediating cells (functional neutrophils), the BICOM bioresonance device with the program "Pathogens Ai" has shown that it can inhibit both the basal metabolism of these cells and their formation of reactive oxygen radicals.
Even if it is difficult to draw a direct conclusion from the test results to an entire organism, the results show the potential of the bioresonance device with this program combination to reduce local oxidative stress in the tissue in vivo and thus neutralize — at least partially — an important reaction cascade of a (chronic) inflammatory process, thereby improving the wellbeing of the user. In addition, a considerably stronger effect of the bioresonance device would undesirably reduce the potential of the cells circulating in the blood to defend against microbial pathogens as an innate immune defense.
6. Supplementary notes
The results of the study, which are only briefly described here, can also be found 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 oxidative burst of inflammation-mediating cells. Biomed J Sci & Tech Res 33: 25616-25620.