This study aimed to evaluate the slow release and internalization of azacitidine-bentonite combination in THP-1 and K562 cell cultures in acute myeloid leukemia morphology.

The morphology of bentonite clay was assessed using two Scanning Electron Microscopes. The bentonite-azacytidine combination was assessed in THP-1 and K562 cell cultures via in vitro cell proliferation tests, proliferation with CCK-8, and drug release tests with dialysis membranes. Additionally, apoptosis and internalization were determined using the Annexin V-FITC Kit and fluorescence methods, respectively.

Our findings showed that azacytidine achieved complete and controlled release within 8 hours. Bentonite displayed significant antiproliferative effects at concentrations of 10, 25, 50, and 100 µg/ml in both cell lines. The combination of azacytidine and bentonite exhibited a synergistic effect in inhibiting cell proliferation, with significant decreases in cell viability in the 1 µM azacytidine + 10 µg/ml bentonite, 5 µM azacytidine + 10 µg/ml bentonite, and 10 µM azacytidine + 10 µg/ml bentonite groups compared to the controls. The drug release profile of the bentonite-azacytidine combination demonstrated slow release, with 50% released in the first two hours and approximately 90% released in the fourth hour, with prolonged release exceeding eight hours, potentially reducing side effects and increasing efficacy in target cells.

In conclusion, bentonite NPs exhibited significant potential as drug carriers for azacytidine in the treatment of leukemia and offered benefits such as improved solubility, bioavailability, controlled release, protection from harsh environments, and cost-effectiveness.