Exploring the Sequential Cellular Events of Phagocytosis Triggered by Godanti Bhasma in Mammalian Cells

Phagocytosis is a vital cellular mechanism through which cells engulf and degrade foreign particles, pathogens, or debris, playing a key role in immune defense and the maintenance of tissue homeostasis. Disruptions in this process are associated with various diseases. To explore the complex events involved in the phagocytosis pathway, advanced smart particles and effective monitoring techniques are essential. Godanti Bhasma (GB), a traditional Indian medicine composed of bioactive calcium sulfate particles, is rapidly internalized by phagocytosis in mammalian cells, inducing significant cytoplasmic vacuolation. The key stages of GB-induced phagocytosis were evaluated here using flow cytometry (FC), live-cell imaging, and specific staining techniques. Flow cytometric analysis demonstrated the formation of phagocytic cup-like structures associated with particle internalization. Live-cell imaging enabled real-time observation of phagocytic processes, including particle uptake, vacuole formation, degradation of engulfed materials, and vacuolar turnover. Staining with neutral red and acridine orange was employed to assess vacuolar acidification. Interestingly, treatment with the lysosomal inhibitor BFA1 in GB-treated cells did not lead to vacuolation, as evidenced by the lack of neutral red uptake, emphasizing the requirement for an acidic environment for vacuolation to occur. These findings underscore the potential of GB-induced phagocytosis as a model to elucidate the sequential cellular events involved in this process, which is critical for understanding host-pathogen interactions, intracellular trafficking, and developing innovative therapeutic strategies for disorders related to phagocytosis.