Anti-Tumoral Potential of Ultra-Diluted Viscum album Preparations in Murine Melanoma Cells: Insights from In Vitro Studies

Background: Cytotoxic effects of Viscum album mother tinctures (VAMT) have been demonstrated on different tumoral cell lineages. A metabolomics approach has highlighted the influence of the host tree on the biological properties of VAMT. This study investigated the effects of different VAMT potencies on murine melanoma cells (B16F10 lineage) through in vitro assays. Methods: VAMTs were prepared using fresh plant material harvested during the European summer from two host trees: Quercus robur (VAMTQ) and Abies alba (VAMTA). Homeopathic protocols were employed to prepare the potencies 3X, 12X, 30X, 6C, 12C, 200C, 2LM, 3LM, and 5LM, which were then evaluated on B16F10 cells. Biological effects were assessed using MTT assays (mitochondrial activity), crystal violet staining (cell viability), flow cytometry (cell death mechanisms, reactive oxygen species [ROS] generation, and cytokine release), and optical microscopy (cell morphology via Giemsa staining). Succussed water, ethanol, and carboplatin were used as controls. Results: Both VAMTs significantly reduced tumoral metabolism ( p < 0.001), with the highest cytotoxic values observed after 1 hour of incubation. Morphological changes indicative of karyolysis and pyknosis were noted, along with an increase in ROS and inflammatory cytokine levels at 1 and 24 hours post-incubation, respectively. The 12X and 5LM potencies derived from VAMTQ induced alterations in cell replication and oxidative stress, accompanied by a significant reduction in MCP-1 cytokine release ( p < 0.0001). In contrast, the 200C potency of both VAMTs exhibited the highest inflammatory profile, triggering cytotoxic effects alongside elevated ROS production and inflammatory cytokine production after 24 hours of incubation. Conclusion: The homeopathic 12X and 5LM potencies showed modulation of cellular metabolism and a promising anti-inflammatory effect, highlighting the potential anti-tumoral properties of Viscum album preparations in vitro .