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FRAUENPOWER! => ~~~ FRAUENPOWER! ~~~ => Topic started by: YanTing on June 06, 2019, 02:01:59 AM
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Biological Actions, Electrical Conductance and Silicon-Containing Microparticles of Arsenicum Album Prepared in Plastic and Glass Vials
Homeopathy 2019; 108(01): 012-023
Biological Actions, Electrical Conductance and Silicon-Containing Microparticles of Arsenicum Album Prepared in Plastic and Glass Vials
Luciane Costa Dalboni et al. (15 authors total)
Universidade Paulista – UNIP, São Paulo, São Paulo, Brazil
Funding We thank the Brazilian Association of Homeopathic Pharmacists (Associação Brasileira de Farmacêuticos Homeopatas—ABFH) for funding the study and CAPES-PROSUP (number 1678566) for the PhD grant.
Abstract
Introduction:
According to the “silica hypothesis” formulated to explain homeopathy, the information of starting materials would be transferred to cells by silica nanoparticles detached from the glassware walls by serial dilution and agitation through epitaxy. We compared the biological activity, electrical current and silicon microparticle content (by means of scanning electron microscopy/energy-dispersive X-ray spectroscopy) of high dilutions (HDs) of arsenic prepared in plastic and glass vials to investigate the role of silica in their biological effects in vitro.
Materials and Methods:
Co-cultures of macrophages and yeast (Saccharomyces cerevisiae) were treated with different HDs of arsenic prepared in plastic and glass vials. Macrophage morphology, phagocytosis index, nitric oxide (NO), and cytokine production were evaluated.
Results:
Measurable amounts of silicon microparticles were detected only in the HDs prepared in glass vials, but ultra-centrifugation eliminated them. Specific and non-specific results were observed. Non-specific pro-inflammatory effects were seen in all dilutions prepared in plastic vials, including elevation of pro-inflammatory cytokines, NO and macrophage phagocytic index. Only the 200th centesimal dilution of arsenic produced specific decrease in interleukin-6 production in macrophages, and it was independent of the vial type or the presence of microparticles of silica in the medicine samples. The nature of the vials had an impact on the electric flow in the respective fluids.
Conclusion:
The non-specific, pro-inflammatory effects might be attributed to organic residuals detached from the vials' plastic walls during manipulation. Instead, specific silica-independent effects of the homeopathic medicine can be attributed to the decrease of interleukin-6 after treatment with the 200th centesimal dilution of arsenic.
Free full text:
https://www.thieme-connect.com/products/ejournals/html/10.1055/s-0038-1670675
Excerpts:
Introduction
“Nanoparticles (NPs) of starting materials and contaminants were recently detected in homeopathic high dilutions (HDs) above Avogadro's number.[1] [2] Since homeopathic medicines are traditionally prepared using glass vials, it was postulated that contaminant silica NPs detaching from vials during the step of agitation might represent a critical component involved in the biological effects of these medicines observed experimentally.[1] [2] [3] [4] [5] [6] The understanding of their real effects (specific or non-specific) is crucial to characterize the mechanisms involved.
“It is known that nanosilica can form stable three-dimensional structures using DNA, proteins or living cells as template (epitaxial growth).[7] [8] [9] [10] [11] [12] [13] This phenomenon led to a microstructural hypothesis to account for the action of homeopathic medicines. Briefly, nanosilica would behave as non-specific amplifier or vehicle for the specific (structural or electromagnetic) information contained in starting materials, subsequently identified and processed by biological systems.[14] [15] [16] Hypothetically, initial interaction and adsorption of starting material with silica NPs in low homeopathic dilutions would create “nanoseeds” that would be preserved in the preparation of the subsequent dilutions.[6] [17] [18] [19] Nanoparticles might also be associated with changes in electrical conductivity, as was shown for the homeopathic medicine Ferrum metalicum,[20] although other factors can also alter the conductivity of homeopathic aqueous solutions.[21]
“Few studies have sought to assess the influence of glass (silica) on the effects of homeopathic HDs. In the pioneer study carried out by Boiron et al,[22] HDs prepared in paraffin-covered glass vials had smaller effect compared with the ones prepared in standard glass vials. One single study investigated trace elements in plastic and glass vials used to prepare and store homeopathic HDs.[23] The results showed absence of silica in the propylene vials and increase in the concentration of trace elements, mainly silica, lithium and sodium, when bi-distilled water was transferred from propylene to glass vials. The largest difference in trace element concentration occurred in the step from mother tincture to the first centesimal dilution (1cH); the later steps to 30cH were not associated with increase in trace element concentration. To the best of our knowledge, no study investigated differences in biological effects between HDs prepared in plastic and glass vials.”
“The experimental design and choice of the medicine were based on the literature[24] and the model chosen, macrophages in co-culture with microorganisms, was previously used in homeopathic fundamental research.[25] Macrophages are myeloid cells specialized for phagocytosis of micro-organisms, such as yeasts, and stand out for their ability to respond epigenetically to small environmental stimuli, mainly by production of cytokines/chemokines[26] [27] which makes these cells adequate biological sensors in HD research.”
“Arsenic raw material containing 99.984% and 0.99984 mol/mol2 of arsenic trioxide, as declared by the supplier, was acquired as a fine powder. The homeopathic manufacturing process followed the statements of Brazilian Homeopathic Pharmacopoeia[28] and started with the separation of lactose powder in three equal parts. One-third of the lactose was placed in a porcelain vessel to avoid As2O3 loss. Following this, As2O3 was added and triturated, meeting the centesimal scale (1 part in 99 parts of inert vehicle), and the complex (Arsenicum album plus lactose) was submitted to two independent and vigorous cycles of crushing and scraping for 20 minutes. After this first cycle, the second and third parts of lactose were added to the porcelain vessel, and submitted to the second and third cycles of crushing and scraping, for 20 minutes each, respectively.”
“The Arsenicum album 3cH was used as matrix to the following dynamizations, always according to Brazilian Homeopathic Pharmacopoeia.[28] HDs were prepared using standard type II amber glass or PET (polyethylene terephthalate, R18/400, Apace, São Paulo, Brazil) vials. The plastic vials were sterilized by microwave heating, three 5-minute shocks at high intensity.”
“High dilutions were prepared as follows: centesimal serial dilutions (6 for 6cH and 200 for 200cH) were made using different flasks and performing 100 strong vertical motions (agitation) for each dilution. All dilutions were prepared from Ars 3cH using 70% hydro-alcoholic solution (p/p)—which is equivalent to 77% (v/v)—in the following steps...Vertical agitation (‘potentization’) was performed in each step by means of a mechanical arm used specifically for this purpose (Denise, AUTIC, São Paulo, Brazil). Theoretically, the concentration of Ars 6cH and Ars 200cH was 1.3×10^-11 M and Ars 1.3×10^-366 M, respectively.”
“Discussion
“The results of the present study show plastic-related non-specific effects and very specific effects on IL production, which were independent of microparticles containing silicon but probably dependent on the smallest nanostructures, which could include water and the silica itself. In this sense, dilutions of arsenic prepared in plastic vials exhibited pro-inflammatory effects attributed to putative organic residuals detached from the vials' walls during disinfection and agitation, since these effects were not seen in glass-prepared medicines...”
“Macrophage spreading, phagocytic activity and increase in IL-1a, IL-6, MCP-1, and VEGF in the culture medium were found only in samples treated with medicines prepared in plastic vials—Ars 6cHP and Ars 200cHP, but not in Ars 6cHG or Ars 200cHG. These findings suggest that plastic-derived soluble contaminants present in higher potencies might be somehow related to the increase in macrophage activity.
“Available evidence indicates that PET bottles subjected to high temperature (40–60°C) do not exhibit cytotoxic activity,[30] but biomaterials containing PET might modify the macrophage activity in vitro.[31] [32] In any case, plastic components have relevant biological actions, this fact being of interest for homeopathic pharmaceutics and, thus, deserving of more thorough study. For instance, one study compared homeopathic medicine Gelsemium sempervirens 6c, 30c, and 200c to agitated vehicle controls (6c, 30c, and 200c) and non-agitated solvent prepared in vials with natural cork stoppers and with silicone stoppers. All the vials with cork stoppers exhibited more than 4×10^8 NPs/mL, significantly more than with the non-agitated silicone stoppers. On ultraviolet–visible spectroscopy, the agitated control 30c exhibited significantly higher absorbance, while the non-agitated silicone stopper controls had significantly lower mean absorbance than all the other samples.[33]
“These findings suggest that the agitation inherent to homeopathic remedy manufacture released trace materials from natural cork stoppers that could interfere with various properties of HDs, and eventually also with their biological activity.[33] Also, potentized water exhibited a residual effect on macrophage spreading, which might be attributed to a non-specific action of micro- and/or nano-bubbles.[34] A direct action of medicines on yeast is unlikely in the present case because, according to the literature, treatment with Ars in decimal dilutions 8x and 16x did not induce any change in S. cerevisiae growth.[35]
“The decrease in IL-6 production might be considered as the main specific biological effect of Arsenicum album 200cH, since it was observed not only in this potency but in all situations: Ars 200cHG, Ars 200cHGC, and Ars 200cHP. Therefore, these specific effects were not related to the identification of silicon-containing microparticles and agree with the effects of Ars 200cH on leukocytes previously described.[24]”
“The correlation between electric current flow through the liquids and the type of the vial used corroborates the hypothesis in which metal particles in suspension could improve conductivity[1] [2] [21] [41] and possible organic contaminants from plastic could impair it.”
“In short, the results of the present study indicate lack of correlation between the presence of silicon-containing microparticles and biological (specific or non-specific) effects of the tested HDs. On these grounds, one might not infer that silicon-containing microparticles could carry the information of homeopathic medicines to biological systems. However, smaller particles, as NPs,[1] [2] [4] [5] [6] [22] [32] [33] might explain the divergence. Moreover, in this model, NPs of the original substance could not be observed, maybe due to the limitations of the microscope used. Some studies showing these particles and their putative hormetic effects are well described in the literature.[41] [42] [43]
“Another hypothesis remains to be checked in the future: that the capacity of water itself acts as information carrier, based on electromagnetic properties derived from its dipole behavior.[44] [45] [46] [47] [48] Alternatively, the participation of both elements together—water and particles, mixed in complex nano-structures that are almost impossible to separate in nature—could elicit some biological effect.[49]
“Many studies using refined methods are still needed to establish which hypotheses account for the mechanism of action of homeopathy, since several rationales represent reasonable possibilities.[50] [51]”