2.8. In Vitro Magnetic Hyperthermia

A549 and HGF cells were seeded and exposed to the same concentration of IOMNPs as described in Section 2.7. Upon the washing and trypsinization steps, the cells were dispersed in 1 mL of cell culture media. Two equal aliquots of 400 µL each were separated. The cells were gently centrifuged for 10 min at 100 g and then 200 µL of cell culture media was removed from each aliquot. Therefore, one aliquot (IOMNP loaded cells dispersed in 200 µL of cell culture) was exposed for 30 min to an Alternating Magnetic Field (AMF), while the other aliquot was immersed in a water bath maintained at 37 °C during the MH experiments to serve as a control. Three different intensities of the AMF (30, 45, and 60 kA/m) were evaluated. After the exposure to the AMF, cells were seeded in 96-well plates and the cell mortality was evaluated after 24 h using both AB and NR assays. Cell mortality was calculated as the relative ratio between cell loaded IOMNPs exposed to AMF and the control. The experiments were performed with six biological replicates, each including 3–6 technical replicates.