The effect of two different polymeric-coated magnetite nano-graphene oxide as 5-fluorouracil carrier and radiofrequency hyperthermia on colon cancer in vitro

Introduction: Colon carcinoma is one of the most common form of cancer that affects the health and lives of people. 5-Fluorouracil (5-Fu) has become useful as synergistically therapy with other antitumor agents in a colon cancer therapy. Short half-life, wide distribution and different side effects of 5-Fu limit its medical applicability. In this study, we have investigated the drug release and the requisite time of hyperthermia of PCL-Chitosan coated magnetite nano-graphene oxide (SPION-NGO-PCL-Chitosan) and PLGA-coated magnetite nano- graphene oxide core (SPION-NGO-PLGA) as carriers of 5-Fu on CT26 colon cancer cells.   Materials and Methods: The in vitro 5-Fu release from nanoparticles was performed by the diffusion method. A weighed amount of nanoparticle was suspended in PBS and next transferred into a dialysis bag. The bag was placed into 12mL PBS at different temperature conditions (37 and 43°C) under pH=7.4 as a release medium. The released 5-Fu concentration was measured using a UV visible spectrophotometer analysis at 265 nm. In order to obtain the requisite time of radiofrequency hyperthermia using AMF at 43oC, 5×105 CT26 cells were seeded in T-25 flasks. After 24 hours, the cells were treated with 50.24 μg/mL of 5-FU-SPION-NGO-PLGA or 14.4 μg/mL 5-Fu-SPION-NGO-PCL-Chitosan nanoparticles (equivalent to about 1 μΜ of 5-Fu) for 24 hours. After 24h, the cells were washed twice with PBS and the culture medium was replaced with a fresh medium. Finally, the cells were irradiated with the alternative magnetic field (power of 50 W) (Basafan, Iran). Moreover, control samples without NPs treatment were prepared and irradiated with AMF. Temperature variations of the cells were evaluated with a digital infrared (IR) thermal camera (Testo 875-1i, Germany) during AMF irradiation. Results: The results show that 5-Fu was released from PLGA-coated NPs and PCL-chitosan- coated NPs to the extent of 41.36% and 75.84% within approximately 24 h at 37oC, respectively. Total release from 5-Fu-SPION-NGOPLGA and 5-Fu-SPION-NGO-PCL-Chitosan at 37oC over 7 days was 57% and 81%, respectively. The 5-Fu release within 4h was equivalent to 71% for PLGA-coated NPs and 97% for PCL-chitosan-coated NPs at 43°C. To evaluate the requisite time of hyperthermia at 43oC, treated flasks with nanoparticles were irradiated with AMF and monitored with IR camera. Finally, As observed, the increasing temperature profiles of the cells as a function of time. The mean temperature of the cells irradiated to AMF was increased to 43°C at 17, 6 and 5 minutes for control, SPION-NGO-PCL-chitosan, and SPION-NGO-PLGA groups, respectively. Conclusion: According to this study, SPION-NGO-PCL-chitosan NP and SPION-NGO-PLGA NP as 5-Fluorouracil carriers and AMF hyperthermia increased the effect of the hyperthermia which lead to lower dosages of drugs that could induce harmful side effects.