In Vivo and in Vitro Demonstration of Gold Nanorod Aided Photothermal Presoftening of B16F10 Melanoma for Efficient Chemotherapy Using Doxorubicin Loaded Graphene Oxide

A combined photothermal therapy (PTT) and chemotherapy (chemo) were performed in vitro on B16F10 melanoma cells and in vivo using melanoma bearing C57BL/6 mice. The 785 nm (100 mW) irradiated gold nanorods (AuNRs) were used as the PT agent, and electrostatically conjugated Doxorubicin (Dox) to a nanocarrier graphene oxide (GO) worked as the chemotherapeutic. Selection of dosage was optimized from the individual viability studies, and finally a combined therapeutic (AuNR (100 ppm), GO (125, and 250 ppm), Dox (0.0058, and 0.00058 ppm)), was delivered in vitro. PTT, followed by chemo, sequentially, resulted in <10% viability, whereas simultaneous PTT with chemo resulted in a viability of similar to 40% for the melanoma cells. Flow cytometry indicated optical inhomogeneity in the cells that internalized GO, and AuNR; however, the Dox amount was identical within the cells treated with or without PTT. Confocal microscopy revealed that GO+Dox was internalized, and Dox was distributed uniformly within the cells irrespective of the treatment protocol. In vivo results in melanoma bearing C57BL/6 mice resembled the in vitro data closely. The tumor growth inhibition index was highest at 0.78 for the group receiving sequential treatment, followed by 0.61 for those receiving simultaneous treatment, where the control group had a score of 0. For the sequential treatment, presoftening of the cells with PTT, followed by the chemo resulted in significantly improved toxicity of the treatment, whereas simultaneous PTT with chemo results were dominated by the Dox alone.