Safety Evaluation of Arsenicum album 6C in Cell Lines and BALB/c Mice Through Toxicity, Proliferation and Apoptosis Studies

Studies on safety evaluation, efficacy and molecular mechanisms of homeopathic medicines are limited. Despite the fact that homeopathic medicines are utilized all over the globe, the toxicity evaluation of individual drugs like Arsenicum album are still unknown. Hence, to shed light on the above facts, we evaluated the cytotoxicity of Arsenicum album 6C in vitro by MTT assay on RAW 264.7 cells and acute oral toxicity in BALB/c mice as per OECD 423 guideline. The cells tolerated up to 10% concentration of the medicine in culture for a maximum of 40 min. Wound healing and MTT assay showed that Arsenicum album 6C did not interfere with the normal rate of proliferation of BHK-21 cells. Moreover, DAPI staining exhibited normal nuclear morphology and ethidium bromide- acridine orange (EtBr-AO) cocktail fluorescence demonstrated absence of apoptosis in Arsenicum album 6C treated cells. Acute oral toxicity study in BALB/c mice with single dose 50, 300 and 2000 mg/kg Arsenicum album 6C did not show lethality in female BALB/c mice and the median lethal dose (LD 50 ) of the drug corresponded to Category 5 or unclassified (LD 50 cut off is ∞) of Globally Harmonized Classification System. The histology of lung, liver, spleen and kidney of treated mice also appeared normal. As this drug elicited no adverse effects on cell proliferation, nuclear morphology and apoptotic states of cells both in vitro and in vivo , it may be used as therapeutic agent against diseases but molecular mechanism of action needs to be elucidated further.