Controlled release of Ag⁺ ions to human cancer cells selectively neutralized with silver nanoparticles of different sizes produced by a green synthesis method

We present a facile green method to synthesize silver nanoparticles (AgNPs) with different sizes and uniform size distributions using an extract of Rhododendron mucronulatum flowers. Here, AgNPs were utilized to modify the release of Ag⁺ ions and selectively annihilate human cancer cells. AgNPs were prepared at ambient temperature (RT-AgNPs) and in higher-temperature water baths at 60 °C (W60-AgNPs) and 70 °C (W70-AgNPs) to modify the diameter and uniformity of the AgNPs. The W60-AgNPs were further treated with acid to modify their size and surface state (named AW60-AgNPs). The water-bath technique enabled an alteration of the relative rates of initial nucleation and growth of nanoparticles in a homogeneous environment, promoting the creation of small AgNPs with a uniform diameter distribution. The acid treatment eroded the surface particles to reduce their diameter and simultaneously introduced functional groups. The Ag⁺-ion release efficiency of the prepared nanoparticles and the toxicity of Ag⁺ ions on cancer cells were examined. Among the prepared samples, the AW60-AgNPs released the greatest amount of Ag⁺ ions and produced the largest amount of reactive oxygen species (O₂̇⁻, ˙OH, and H₂O₂₎. All of the AgNP samples were applied to tumor cells (K562 and HT1080) and healthy cells (LO2) in vitro. In the presence of K562 cells, AW60-AgNPs showed the most efficient dose- and time-dependent early and late apoptosis as well as reduced viability. A crystal violet assay showed that the synthesized particles preferentially attack HT1080 cancer cells rather than LO2 normal cells. We found that Ag⁺ ions release efficiency and selectivity varied with particle size, and that DNA damage in tumor cells was due to the formation of Ag⁺-coordinated complexes within DNA base pairs.