Homeopathy in Experimental Cancer Models: A Systematic Review.

BACKGROUND: Complementary and alternative medicine, including homeopathy, is widely used to improve well-being among cancer patients and reduce adverse effects of conventional treatment. In contrast, there are few studies on the use of homeopathic medicines to treat the disease itself. Yet, evidence of possible effectiveness of homeopathic high dilutions in experimental cancer models has been published during the past 20 years. AIM: The aim of the study was to perform a systematic review of fundamental research studies on homeopathic high dilutions in cancer. METHODS: Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline, we conducted a literature search in the database PubMed for original publications, from 2000 to 2018 and in English, on in vitro and in vivo experimental cancer models testing homeopathic high dilutions. RESULTS: Twenty-three articles met the inclusion criteria-14 in vitro, eight in vivo, and one in vitro plus in vivo experimental models. Most studies were from India. Research prominently focused on cytotoxic effects involving apoptotic mechanisms. Intrinsic aspects of homeopathy should be considered in experimental designs to emphasize the specificity of such effects. CONCLUSION: Fundamental research of homeopathy in cancer is still at an early stage and has mainly been performed by a few groups of investigators. The results point to an interference of well-selected homeopathic medicines with cell cycle and apoptotic mechanisms in cancer cells. However, these findings still need independent reproduction.

Effects of Homeopathic Phosphorus on Encephalitozoon cuniculi-Infected Macrophages In-Vitro.

INTRODUCTION: Encephalitozoon cuniculi (E. cuniculi), a fungus that acts as an intracellular pathogen, causes a marked neurological syndrome in many host species and is a zoonotic concern. Although no well-established treatment for this syndrome is known, previous successful clinical experience using homeopathic phosphorus has been described in which symptom remission with no mortality occurred in 40/42 animals by means of unknown immunological mechanisms. The latter observation was the main motivation for this study. OBJECTIVE: To verify, in an in-vitro model, if macrophages infected with E. cuniculi can change in function after treatment with different potencies of phosphorus. MATERIALS AND METHODS: RAW 264.7 macrophages were infected with E. cuniculi in-vitro and treated with various homeopathic potencies of phosphorus. The vehicle was used as a control solution (0.06% succussed ethanol). After 1 and 24 hours, the following parameters were analyzed: parasite internalization (by the Calcofluor staining method), lysosome activity (by the acridine orange method), cytokine/chemokine production (by the MAGPIX system), and cell ultrastructure. Automatic image analysis was used when applicable, and the experiments were performed in triplicate. RESULTS: Treatment with vehicle alone increased interleukin (IL)-6, tumor necrosis factor alpha and monocyte chemotactic protein -1 production (p ≤ 0.05) and reduced the number of internalized parasites (p ≤ 0.001). A progressive and time-dependent increase in RANTES (regulated on activation, normal T-cell expressed and secreted) and lysosome activity (p ≤ 0.002) was observed only after treatment with the highest potency of phosphorus (Phos 200cH), together with decreased apoptosis rate, intense parasite digestion, and the presence of non-internalized spores. CONCLUSIONS: Phos 200 cH has a modulatory action on the activity of infected macrophages, especially a specific increase in RANTES, a key element in the prognosis of E. cuniculi-infected and of immunosuppressed patients bearing infections.

Biological Actions, Electrical Conductance and Silicon-Containing Microparticles of Arsenicum Album Prepared in Plastic and Glass Vials.

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.

High-diluted thymulin on Ehrlich tumor growth in mice and the importance of tumor microenvironment

Introduction: The aim of the present study was to describe different biological aspects of Ehrlich tumor in mice, such as body weight evolution, tumor growth rate, histological organization and systemic immune response after treatment with high-diluted thymulin (10-9 M, named 5CH). Methods: Tumor assessment was focused on macro- and microscopic aspects; parameters included occurrence of necrosis, embolism and tumor development, in addition to quantitative analysis of apoptosis (caspase-3), cell proliferation (Ki-67) and angiogenesis (vascular endothelial growth factor - VEGF) by means of specific immunohistochemistry markers. Spleen cell populations were evaluated by flow cytometry analysis. Results: Mice treated with thymulin 5CH exhibited changes in the tumor microenvironment, such as reduced micro-embolism incidence and cytokeratin expression, with increased caspase-3 expression in the tumor cells. These findings indicate some apoptotic activity by the tumor cells induced by the treatment, even though no reduction of the macroscopic tumor mass occurred. No changes in the systemic immune response were detected, as the balance among spleen cell populations remained unchanged. Conclusions: The results indicate that treatment of mice bearing Ehrlich tumor with thymulin 5CH induces some specific changes in the tumor environment. However, it did not influence systemic immunity parameters. Adjuvant use of thymulin 5CH in oncological clinical practice is still a matter of discussion. (AU)

Homeopathic treatments modify inflammation but not behavioral response to influenza antigen challenge in BALB/c mice.

BACKGROUND: Influenza affects thousands of people worldwide every year, motivating the development of new therapies. In this work, the effects of two homeopathic preparations (influenza biotherapies and thymulin) were chosen following two different rationales: isotherapy and endo-isotherapy models. The homeopathic effects were evaluated individually considering the inflammatory and behavioral responses against influenza virus antigen were studied in BALB/c mice. METHODS: Male adult mice were treated orally and blindly for 21 days with highly diluted influenza virus or with thymulin, and were divided in two sets of experiments. The first series of experiments aimed to describe their behavior, using an open field (OF) device. In the second series, mice were challenged subcutaneously with influenza hemagglutinin antigen (7 µg/200 µl) at day 21. At day 42, behavior and inflammation response were evaluated. RESULTS: No behavioral changes were seen in OF tests at any time point after treatments. Flow cytometry and morphometry revealed significant changes in T and B cell balance after influenza antigen challenge, varying according to treatment. CONCLUSION: The results show that both homeopathic treatments induced subtle changes in acquired immune anti-viral response regulation. A deeper understanding of the mechanism could elucidate their possible use in influenza epidemiological situations.

The use of animal models in homeopathic research--a review of 2010-2014 PubMed indexed papers.

BACKGROUND: In the 1990s, a study was performed on the effects of highly diluted thyroxine on frog metamorphosis. This model represented one of the most discussed examples of the biological effects of high dilutions over the next two decades. In 2010, another critical conceptual review of the use of animal models in homeopathy and high-dilution research was published. The main contribution of these studies was the elucidation of the biological features and phenomenology of the effects of high dilutions on living systems, representing an important step forward in our understanding of the mechanisms of action of homeopathic medicines. METHODS: We performed a further review of this line of investigation using the same methods. Fifty-three articles that were indexed in the PubMed database and used 12 different animal species were systematically evaluated. Only a fraction of the studies (29/53) reported herein were performed with "ultra high" dilutions. The other studies were performed with dilutions in ranges below 10(-23) (14/53 articles) or commercial complexes (10/53 articles). RESULTS: Only two articles reported negative results; both used in vivo protocols to test commercial complexes, one in fish and one in bees. The quality of the employed techniques improved in 2010-2014 compared with the studies that were reviewed previously in 2010, with the inclusion of more ethically refined protocols, including in vitro primary cell cultures and ex vivo studies (10/53 articles), often with three or more replicates and analyses of epigenetic mechanisms that were previously unknown in 2010. CONCLUSION: In our updated review of the past 5 years, we found further demonstrations of the biological effects of homeopathy using more refined animal models and in vitro techniques.

Modulation of inflammation response to murine cutaneous Leishmaniasis by homeopathic medicines: Antimonium crudum 30cH.

BACKGROUND: Leishmaniasis is a zoonotic disease caused by protozoan parasites of the mononuclear phagocytic system. The modulation activity of these cells can interfere in the host/parasite relationship and influences the prognosis. METHODS: We evaluated the effects of the homeopathic preparation Antimonium crudum 30cH on experimental infection induced by Leishmania (L.) amazonensis. Male Balb/c mice were inoculated with 2 × 10(6)Leishmania (L.) amazonensis promastigotes into the footpad and, after 48 h (acute phase) or 60 days (chronic phase), cell population of lymphocytes and phagocytes present in the peritoneal washing fluid and spleen were analyzed by flow cytometry and histopathology, with histometry of the subcutaneous primary lesion, local lymph node and spleen. Immunohistochemistry was performed to quantify CD3 (T lymphocyte), CD45RA (B lymphocyte) and CD11b (phagocytes) positive cells. RESULTS: In treated mice, during the acute phase, there was significant increase of the macroscopic lesion, associated to inflammatory edema, as well increase in the number of free amastigotes and B lymphocytes inside the lesion. Increase of B lymphocytes (predominantly B-2 cells) was also seen in the local lymph node, spleen and peritoneum. In the chronic phase, the inflammatory process in the infection focus was reduced, with reduced phagocyte migration and peritoneal increase of B-1a cells (precursors of B-2 immunoglobulin producers cells) and T CD8+ cells. CONCLUSION: The treatment of mice with Antimonium crudum 30cH induced a predominantly B cell pattern of immune response in Leishmania (L.) amazonensis experimental infection, alongside the increase of free amastigote forms number in the infection site. The clinical significance of this study is discussed, further studies are suggested.

Modulation of inflammation response to murine cutaneous Leishmaniosis by homeopathic medicines: thymulin 5cH.

BACKGROUND: In previous studies, we observed that thymulin 5cH could modulate BCG (Bacillus Calmette-Guerin) induced chronic inflammation by increasing peritoneal B1 stem cells differentiation into phagocytes and improving phagocytosis efficiency. METHODS: We used the same protocol to study the effects of thymulin 5cH in the experimental murine Leishmaniasis, in order to elucidate some aspects of the parasite-host relation under this homeopathic treatment. Male Balb/c mice were orally treated with thymulin 5cH or vehicle during 60 days, after the subcutaneous inoculation of 2 × 10(6) units of Leishmania (L.) amazonensis into the footpad. Washied inflammatory cell suspension from peritoneal cavity, spleen, local lymph node and infected subcutaneous tissue were harvested after 2 and 60 days from infection to quantify the inflammation cells by flow cytometry and histometry methods. RESULTS: After a transitory increase of peritoneal T reg cells, treated mice presented, chronically, increase in the peritoneal and spleen B1 cells percentage (p = 0.0001) in relation to other cell types; more organized and exuberant inflammation response in the infection site, and decrease in the number of parasites per field inside the primary lesion (p = 0.05). No difference was seen in local lymph node histology. CONCLUSIONS: Thymulin 5cH is able to improve B1 cell activation and Leishmania (L) amazonensis phagocytosis efficiency in mice, similarly to that observed previously in BCG experimental infection.

Immunomodulation of Homeopathic Thymulin 5CH in a BCG-Induced Granuloma Model.

The present study analyzed the immune modulation mechanisms of thymulin 5CH in a granuloma experimental model. Male adult Balb/c mice were inoculated with BCG into the footpad to induce granuloma, which was quantitatively evaluated. The phenotypic characterization of phagocyte, T- and B-lymphocyte populations in the peritoneum, and local lymph node was done by flow cytometry. During all experimental periods, thymulin 5CH and vehicle (control) were given ad libitum to mice, diluted into the drinking water (1.6 × 10(-17) M). After 7 days from inoculation, thymulin-treated mice presented reduction in the number of epithelioid cytokeratine-positive cells (P = 0.0001) in the lesion, in relation to young phagocytes. After 21 days, the differentiation of B1 peritoneal stem cells into phagocytes reached the peak, being higher in thymulin-treated mice (P = 0.0001). Simultaneously, the score of infected phagocytes in the lesion decreased (P = 0.001), and the number of B1-derived phagocytes, CD4+ and CD8+ T lymphocytes in the local lymph node increased in relation to control (P = 0.0001). No difference was seen on the CD25+ Treg cells. The results show that thymulin 5CH treatment is able to improve the granuloma inflammatory process and the infection remission, by modulating local and systemic phagocyte differentiation.

Rats Born to Mothers Treated with Dexamethasone 15 cH Present Changes in Modulation of Inflammatory Process.

As little information about the effect of ultra high dilutions of glucocorticoid in reproduction is available in the literature, pregnant female Wistar rats (N = 12) were blindly subcutaneously treated during all gestational and lactation period with: dexamethasone 4 mg/kg diluted into dexamethasone 15 cH (mixed); or dexamethasone 4 mg/kg diluted in water; or dexamethasone 15 cH, or vehicle. Parental generation had body weight, food and water consumption monitored. The F1 generation was monitored regarding to newborn development. No birth occurred in both groups treated with dexamethasone 4 mg/kg. After 60 days from birth, 12 male F1 rats were randomly selected from each remaining group and inoculated subcutaneously with 1% carrageenan into the footpad, for evaluation of inflammatory performance. Edema and histopathology of the footpad were evaluated, using specific staining methods, immunohistochemistry and digital histomorphometry. Mothers treated with mixed dexamethasone presented reduced water consumption. F1 rats born to dexamethasone 15 cH treated females presented significant increase in mast cell degranulation, decrease in monocyte percentage, increase in CD18+ PMN cells, and early expression of ED2 protein, in relation to control. The results show that the exposure of parental generation to highly diluted dexamethasone interferes in inflammation modulation in the F1 generation.

Efeitos da timulina 5cH na evolução do granuloma e da diferenciação de células B1: um modelo experimental para entender seus mecanismos biológicos / Effects of thymulin 5cH in granuloma evolution and B1 cell differentiation: an experimental model to understand its biological mechanisms

Em estudos prévios, foi verificado que a timulina (um hormônio tímico), quando preparada em potências homeopáticas 5cH, tem a propriedade de melhorar a performance produtiva de frangos infectados com reovirus, bem como modular o desenvolvimento do tumor de Ehrlich e de lesões granulomatosas em camundongos por mecanismos imunomediados. O objetivo deste trabalho foi estudar os mecanismos imunonodulatórios da timulina 5cH em um modelo de granuloma experimental, pela injeção subcutânea de BCG em camundongos, focando o envolvimento das células B1 e do zinco no processo. Três grupos de camundongos Balb/c machos (grupo A tratado com timulina 5cH, grupo B tratado com timulina 5cH incubada com Chelex ® - um quelante de zinco ? e grupo C, controle, tratado com veículo) foram inoculados com BCG no coxim plantar esquerdo e o granuloma subcutâneo e baço foram colhidos para análise histomorfométrica, após 7, 14 e 21 dias. Colorações de Ziehl-Neelsen, hematoxilina eosina e azul da Prússia foram também utilizadas. A citometria de fluxo também foi usada nos mesmos tempos para caracterizar e quantificar células peritoneais. Células positivas para CD11b (fagócitos ativados, células B1), CD 19 (B1 e B2), CD23 (negativas em B1, positivas em B2) e CD5 (B1a) foram analisadas em um aparelho FACS Calibur (BD). A análise estatística foi Kruskal-Wallis/Dunn para avaliações não paramétricas e ANOVA/Tuckey-Krammer para avaliações paramétricas. O método X2 foi usado para avaliar a proporção entre células na citometria. Valores de p?0.05 foram considerados estatisticamente significantes. Camundongos tratados com timulina 5cH apresentaram alta atividade de macrófagos e aumento na área folicular esplênica após 7 dias da inoculação do BCG. O aumento do tamanho macroscópico da lesão e a redução da infecção local foram vistos após 21 dias. Nesse período, a citometria demonstrou aumento na proporção de fagócitos derivados de células B1 peritoneais em camundongos tratados com timulina 5cH, independentemente da incubação ou não com Chelex®, a qual bloqueou apenas os efeitos da timulina sobre células B2 no peritôneo e reduziu significativamente os níveis de manganês na preparação medicamentosa. Conclui-se que a timulina 5cH modula o granuloma induzido por BCG através de mais de um mecanismo, especialmente da diferenciação de células B1 peritoneais em fagócitos.(AU)

In previous studies, we found that thymulin (a thymic hormone), when prepared in homeopathic 5cH potency, had the property to improve the productive performance of broiler chickens infected with reovirus, as well as modulate the development of Ehrlich tumor and granuloma inflammatory lesions in mice by immune-mediated mechanisms. The aim of the present work was to study the immunomodulatory mechanisms of thymulin 5cH in a granuloma experimental model, by subcutaneous inoculation of BCG in mice, focusing the B-1 cells and zinc involvement in this process. Three groups of male Balb/c SPF mice (group A treated with thymulin 5cH, group B treated with thymulin 5cH incubated in Chelex ® - a zinc chelant - and group C, control, treated with vehicle) were inoculated with BCG in the left footpad and subcutaneous granuloma and spleen were harvested for histomorphometry analysis, after 7, 14 and 21 days. Ziehl-Neelsen, HE and Prussia Blue staining methods were used. Flow cytometry was also used in the same times to characterize and quantify peritoneal cells. Positive cells for CD11b (activated phagocytes, B-1 cells), CD19 (B-1 and B-2 cells), CD23 (negative B-1 cells, positive B2 cells) and CD5 (B-1a cells) were analyzed in a FACS Calibur (BD) device. Statistical analysis was performed using Kruskal - Wallis / Dunn for nonparametric evaluations and ANOVA / Tuckey-Krammer for the parametric ones. The X² method was used to evaluate the cell count in flow cytometry. P values ? 0.05 were considered statistically significant. Mice treated with thymulin 5cH presented higher macrophage activity and increase in the follicular area were seen in spleen after 7 days. Increase in gross lesion diameter and decrease in local BCG infection were seen after 21 days. At this time, the flow cytometry demonstrated the increase in peritoneal phagocytes derived from B-1 cells in thymulin 5cH treated mice, independently of Chelex ® incubation. The incubation of thymulin 5cH with Chelex ® blocked its effects only upon the number of B2 cells in the peritoneum and reduces Mn levels in the medicine solution. We conclude that thymulin 5CH modulates the BCG-induced granuloma through more than one mechanism, especially by peritoneal B1 cell differentiation into phagocytes.(AU)

Avaliação da resposta imune e comportamental de camundongos BALB/c após tratamento com soluções homeopáticas de H3N2 / Evaluation of BALB/c mice behavior and relations to the immune response after treatment with H3N2 homeopathic solutions

In Brazil, homeopathic medicines are prepared according to the Homeopathic Pharmacopeia, regulated by ANVISA. Among several categories of medicines, there is the biotherapic group, which is prepared from etiologic agents. In this study, we developed a biotherapic from influenza A virus, aiming the influenza infection prevention. Influenza is a disease that affects thousands of people worldwide every year, with an important economic impact, what motivates the development of new low cost therapies. The H3N2 biotherapic developed in this study was administered to Balb/c mice to evaluate their immune response to viral specific antigens and behavior (homeopathic proving). Sixty-two 4 weeks old Balb/c mice were divided into five experimental groups (n=14 per group), after approval by the Ethics Committee of Animal Use (Protocol DFBCICB 037) and stimulated daily, blindly, with 1% (v/v) different homeopathic medicines, for a maximum period of 42 days. The tested medicines were: biotherapic 30x prepared from inactivated influenza A virus; biotherapic 30x prepared with infectious influenza A virus; and thymulin 5cH, a thymus hormone. The two control groups were treated with water 30x and nothing (baseline group). After 21 days of treatment, half of the animals from each group was challenged subcutaneously with the viral hemagglutinin antigen (7 g / 200 L) and monitored by 21 days further, to evaluate the humoral immune response and general behavior, using an open device. The remaining animals were evaluated by the same behavioral tests at the end of the first 21 days, as an attempt to define the proving features. After euthanasia, all animals were autopsied and the spleen, lungs, heart and mediastine lymph nodes were weighed. Histometry of the spleen follicles was also made. Histopathological and behavioral analyses showed absence of behavioral effects, however, there was increase of spleen lymphoid follicles diameter in immunized animals treated with thymulin and with the biotherapic prepared from infectious influenza A, when compared to the control group. This experiment is being repeated using flow cytometry to complete the analysis and confirm the results.(AU)

Medicamentos homeopáticos são preparados de acordo com a farmacotécnica homeopática regulamentada pela ANVISA. Dentre as várias categorias destes medicamentos, destaca-se o grupo dos bioterápicos, medicamentos que são preparados a partir do próprio agente etiológico. No presente estudo, foi desenvolvido um bioterápico a partir do vírus influenza A, visando a profilaxia da gripe. A gripe é uma doença que atinge milhares de pessoas anualmente em todo o mundo e o desenvolvimento de novas terapias para esta doença vem sendo estimulado com frequência. O bioterápico desenvolvido foi administrado a camundongos do tipo Balb/c para avaliação da resposta imune e comportamental. Para tanto, sessenta e dois camundongos Balb/c com 4 semanas de vida foram separados em cinco grupos experimentais, após aprovação pelo Comitê de Ética de Uso de Animais (Protocolo DFBCICB 037) e estimulados diariamente, de maneira cega, por diferentes soluções homeopáticas, na concentração de 1% (V/V), durante um prazo máximo de 42 dias. Três medicamentos homeopáticos foram testados: bioterápico contendo o vírus influenza A inativado 30DH; bioterápico contendo o vírus influenza A íntegro 30DH; timulina 5CH. Um grupo controle foi tratado com água 30x e o outro não recebeu tratamento. Após 21 dias de tratamento, metade dos animais de cada grupo (31 animais) foi desafiada, por via subcutânea, com o antígeno viral hemaglutinina na concentração de 7 g/ 200L e acompanhados por mais 21 dias para avaliação da resposta imune humoral e do comportamento, pela técnica do campo aberto. Os animais restantes foram submetidos aos mesmos testes ao final dos primeiros 21 dias de tratamento, antes do desafio antigênico. Após a eutanásia, todos os animais foram necropsiados e o baço, o pulmão, o coração e o linfonodo mediastínico foram colhidos para análise de peso e histometria do baço. As análises histopatológica e comportamental mostraram a ausência de efeitos patogenéticos perceptíveis neste modelo experimental, mas houve aumento da reatividade dos folículos linfóides do baço nos animais desafiados antigenicamente e tratados com bioterápico de influenza A íntegro e timulina, em relação ao grupo controle. Este experimento está sendo repetido(AU)

Inflammation in rats born to mothers treated with dexamethasone 15cH during pregnancy: an immunohistochemical study / Inflammation in rats born to mothers treated with dexamethasone 15cH during pregnancy: an immunohistochemical study

In previous studies, we observed that rats born to mothers treated with dexamethasone 15CH (10-33M) had a higher level of mast cell degranulation and greater arteriolar dilation after the exposure of an inflammatory stimulus, suggesting the possibility of vertical transmission of the effects of ultra-diluted substances between mother and offspring. In this study, a more detailed assessment of the cellular events in acute inflammation was made using techniques of immunohistochemistry. The identification of adhesion molecules expression was made by the markers: anti-CD54 (ICAM-1) and anti-CD18 (?2-Integrin). The identification of inflammatory cells was performed by the markers anti-MAC387 (mononuclear cells) and anti-CD163 (active macrophages). Polymorphonuclear cells were identified by hematoxylin-eosin staining. The number of labeled cells per was recorded, except for the anti-CD54 marker, whose intensity of staining on the endothelial cells was defined by scores assigned by two independent observers. The results point toward to an up regulation of the whole inflammatory process in rats born to mothers treated with dexamethasone 15CH during pregnancy. This conclusion is justified by the following statistically significant (p?0.05) findings: a) bigger mast cell degranulation and increased of arteriolar diameter; b) increased migration of polymorphonuclear cells in relation to the mononuclear cells; c) earlier expression of CD163 in monocytes, d) higher level of adhesion molecules expression.(AU)

Inflammatory Process Modulation by Homeopathic Arnica montana 6CH: The Role of Individual Variation.

The effects of Arnica montana 6cH on the individual modulation of acute inflammation kinetics in rats were evaluated. Adult male Wistar rats were inoculated with 1% carrageenan into the footpad and treated with Arnica montana 6cH, dexamethasone (4.0 mg/kg; positive control) or 5% hydroalcoholic solution (negative control), per os, each 15 minutes, between 30 and 180 minutes after the irritant inoculation. Histopathological and immunohistochemistry procedures were done in order to get a panel of inflammatory positive cells for CD3 (T lymphocytes), CD45RA (B lymphocytes), CD18 (beta 2 integrin), CD163 (ED2 protein), CD54 (ICAM-1), and MAC 387 (monocytes and macrophages). The statistical treatment of data included a posteriori classification of animals from each group (N = 20) in two subgroups presenting spontaneous precocious or late oedema. Animals that presented precocious oedema were less responsible to Arnica montana 6cH in relation to hemodynamic changes. Instead, rats that exhibited late oedema presented less intense oedema (P = .01), lower percentage of mast cell degranulation (P = .0001), and increase in lymphatic vessels diameter (P = .05). The data suggest an individually qualitative adjustment of inflammatory vascular events by Arnica montana 6cH.

Inflammation in rats born to mothers treated with dexamethasone 15cH during pregnancy: an immunohistochemical study / Inflammation in rats born to mothers treated with dexamethasone 15cH during pregnancy: an immunohistochemical study

In previous studies, we observed that rats born to mothers treated with dexamethasone 15CH (10-33M) had a higher level of mast cell degranulation and greater arteriolar dilation after the exposure of an inflammatory stimulus, suggesting the possibility of vertical transmission of the effects of ultra-diluted substances between mother and offspring. In this study, a more detailed assessment of the cellular events in acute inflammation was made using techniques of immunohistochemistry. The identification of adhesion molecules expression was made by the markers: anti-CD54 (ICAM-1) and anti-CD18 (?2-Integrin). The identification of inflammatory cells was performed by the markers anti-MAC387 (mononuclear cells) and anti-CD163 (active macrophages). Polymorphonuclear cells were identified by hematoxylin-eosin staining. The number of labeled cells per was recorded, except for the anti-CD54 marker, whose intensity of staining on the endothelial cells was defined by scores assigned by two independent observers. The results point toward to an up regulation of the whole inflammatory process in rats born to mothers treated with dexamethasone 15CH during pregnancy. This conclusion is justified by the following statistically significant (p?0.05) findings: a) bigger mast cell degranulation and increased of arteriolar diameter; b) increased migration of polymorphonuclear cells in relation to the mononuclear cells; c) earlier expression of CD163 in monocytes, d) higher level of adhesion molecules expression.