In vitro evaluation of the effects of Viscum album homeopathic preparations on murine melanoma cells

Viscum album extract (VA) is a complementary treatment in cancer, with in vitro and in vivo cytotoxic effects on several tumor types when applied in phytochemical doses. However, highly diluted ethanolic homeopathic preparations' effects and mechanisms need further study. Aims:To assess the in vitro effects of highly diluted VA from the subspecies V. album abietis and V. album album at different potency levels in different dilution ratios on murine melanoma cells. Methodology:The VA mother tinctures (MT)from Abies alba (MTA) and Quercus robur (MTQ) were prepared with summer and winter samples, harvested in Switzerland. They were submitted to homeopathic ethanolic maceration and a subsequent dynamization process. MTA, MTQ and the following respective potencies were tested in B16F10 murine cells: 3x, 12x, 30x, 6cH, 12cH, 200cH, 2LM, 3LM, and 5LM. Dynamized water, dynamized and non-dynamized ethanol, and carboplatin were used as control groups. The mitochondrial activity and cell viability analysis were performed at 1, 24, 48, and 72 hours by in vitro incubation. MTA and MTQ harvested in summer, as well as 12x, 200cH and 5LM potencies were also tested to cell apoptosis and necrosis markers, reactive oxygens species (ROS) production, inflammatory cytokines profile, cell morphology, and migratory capacity. Results and discussion: MTA and MTQ induced a decrease in cell metabolism and higher cytotoxicity within 1 hour, with significant morphological changes and increased production of ROS and inflammatory cytokines. Both homeopathic dilutions 12x and 5LM showed an influence on cell metabolism, cell replication, and oxidative stress modulation with inflammatory cytokines, mitosis, and migration pattern changes. On the other hand, Quercus robur and Abies alba 200cH showed increased on cytotoxicity and ROS levels, respectively. Conclusion:The in vitro effects of Viscum album homeopathic solutions in melanoma cells highlight the promising antitumoral potential and reinforce the need for further research to better understanding their mechanisms of action.

Carcinosinumproduces phenotypic changes of 4T1 tumor cells in vitro

Considering that there are few published studies that specifically address the exclusive use of Carcinosinumin different potencies and, most of them focused on genotypic and clinical effects, the present study was proposed to identify possible phenotypic changes, including viability, expression of HER-2 and metastatic abilities, using 4T1 cells in vitroas a model. Carcinosinum was tested in different homeopathic potencies (12cH; 30cH; 200cH) mechanically prepared using sterile pure water. The time space between preparing the potencies and using them was 24 hours.The final dilutions were inserted into the culture medium in a volume equal to 10%, at the time of cell seeding. The same succussed vehicle used to prepare the drugs (70% ethanol) diluted 1:100 in sterile pure water was used as control. All treated cells were cultured in 25 mL flasks, with cell density of 5 x 105cells/mL. After 24 hours of treatment, cells were analyzed for apoptosis index using Annexin V kit and the Countess® system. The morphology of 4T1 cells was monitored by staining cell smears with hematoxylin-eosin and Giemsa methods. HER-2 expression was assessed by immunocytochemistry and metalloproteinase activity was assessed by zymography. The determination of the cytokine profile was performed using Cytometric Bead Array (CBA). The samples were evaluated in quadruplicate and the data were analyzed by one-way ANOVA. Carcinosinum30cH presented the highest apoptotic index and reduction of MMP-9-Pro expression; Carcinosinum200cH produced the highest positivity for HER-2 and no specific effect was seen after the treatment with Carcinosinum12cH. No change in cytokine expression was seen among treatments. We conclude that Carcinosinum30cH and 200cH can change phenotypic features important totumor development in vitro. The clinical meaning of these data deserves further investigation.

Phenotypic Changes in Mammary Adenocarcinoma (4T1) cells In Vitro after Treatment with Carcinosinum.

OBJECTIVE: The present study aimed to identify possible phenotypic changes in 4T1 (murine mammary adenocarcinoma) cells in vitro, including viability, HER-2 (human epidermal growth factor receptor-type 2) expression, and metastatic potential, after treatment with Carcinosinum in different homeopathic dilutions (12cH, 30cH, 200cH) shaken mechanically in pure, sterile, water from a commercial stock dilution. METHODS: Treated cells were cultured in R10 medium, using 24-well plates, 105 cells per well, and treated with vehicle, Carcinosinum 12cH, 30cH or 200cH; untreated cells were used as the baseline control. After 24 hours of treatment, the percentage of apoptotic cells was analyzed by annexin V. Cell morphology was evaluated by microscopy after hematoxylin-eosin and Giemsa staining, whilst HER-2 expression was assessed using immunocytochemistry. The metastatic potential was determined by the expression and activity of the enzyme matrix metalloproteinase 9 (MMP-9) using zymography. The cytokine profile was established using the cytometric bead array method. RESULT: Treatment of 4T1 cells in vitro with Carcinosinum 30cH produced an increase in the number of annexin V-positive cells (apoptosis) and decreased expression of proactivated MMP-9. Cells treated with Carcinosinum 200cH presented hyper-expression of HER-2 on the plasma membrane, identified by immunocytochemistry. There were no differences in cytokine production among treatments. CONCLUSION: The data show promising results for Carcinosinum 30cH in vitro, but in vivo studies are also required to evaluate the role of tumor microenvironment in its effects.

Alterações dimensionais em pacientes submetidos à expansão rápida da maxila com aparelho tipo Haas / Evaluation of dimensional changes in patients undergoing rapid maxillary expansion appliance type with Haas

A atresia maxilar é uma alteração no sentido transversal,que pode redundar em uma mordida cruzada posterior unilateral oubilateral. Para correção da atresia do arco dentário superior, sãoutilizados aparelhos com intenção de expandir o arco de maneiraortopédica ou ortodôntica, obtendo assim uma compatibilidade transversalentre os arcos dentários. O objetivo deste estudo foi avaliar asalterações dimensionais da maxila em pacientes na fase de dentaduramista com atresia do arco superior, utilizando radiografias oclusaistomadas antes, após a expansão rápida da maxila e após a remoçãodo aparelho. Material e métodos: a amostra consistiu de 35 pacientesque utilizaram o aparelho tipo Haas convencional, na dentadura mista,de acordo com o protocolo padrão de instalação, ativação, contençãoe remoção dos aparelhos de expansão rápida da maxila. Resultado: otrabalho confirma que a abertura da sutura ocorre em maior quantidadena região anterior (4,3 mm) do que na posterior (3,74 mm), representandouma abertura triangular de 7° em média com a porção posterior,abrindo 87% da quantidade da abertura anterior. Na região posterior,os molares expandiram por volta de 5 mm, e a base óssea 3,7 rnrn,representando uma expansão óssea de 74% da expansão dentária.

The maxillary atresia is a alteration in the transversedimension that can result in a unilateral or bilateral crossbite. Forcorrection of atresia of the upper dental arch appliances with the intent toexpand the arch of orthopedic or orthodontic manner are used, obtainingacross-compatibility between the dental arches. The purpose of this studywas to evaluate the dimensional changes of the maxillary in patients inthe mixed dentition with atresia in the upper dental arch, using oclusalradiographs taken before, after rapid maxillary expansion and afterremoval of the appliance. Methods: the sample consisted of 35 patientswho used the appliance type conventional Haas, in the mixed dentition,according to the standard protocol for installation, activation, containmentand removal of the appliances rapid maxillary expansion. Results: theresults confirm that the suture opening occurs in greater quantities in theanterior (4.3 mm) than posterior to (3. 74 tnm), representing a triangularopening of 7 degrees on average with the posterior opening 87% of thequantity of anterior opening. The molars region expanded about 5 mm,and the base bone 3.7 mm, representing a 74% bone expansion of theexpansion teeth.