Sensitization of glioblastoma cancer cells to radiotherapy and magnetic hyperthermia by targeted temozolomide-loaded magnetite tri-block copolymer nanoparticles as a nanotheranostic agent.

AIMS: Recently, the development of new strategies in the treatment and diagnosis of cancer cells such as thermo-radiation-sensitizer and theranostic agents have received a great deal of attention. In this work, folic acid-conjugated temozolomide-loaded SPION@PEG-PBA-PEG nanoparticles (TMZ-MNP-FA NPs) were proposed for use as magnetic resonance imaging (MRI) contrast agents and to enhance the cytotoxic effects of hyperthermia and radiotherapy. MAIN METHODS: Nanoparticles were synthesized by the Nano-precipitation method and their characteristics were determined by dynamic light scattering (DLS), scanning electron microscopy (SEM) and X-ray powder diffraction (XRD). To evaluate the thermo-radio-sensitization effects of NPs, C6 cells were treated with nanoparticles for 24 h and then exposed to 6-MV X-ray radiation. After radiotherapy, the cells were subjected to an alternating magnetic field (AMF) hyperthermia. The therapeutic potential was assessed using clonogenic assay, ROS generation measurement, flow cytometry assay, and qRT-PCR analysis. Also, the diagnostic properties of the nanoparticles were assessed by MRI. KEY FINDINGS: MRI scanning indicated that nanoparticles accumulated in C6 cells could be tracked by T2-weighted MR imaging. Colony formation assay proved that TMZ-MNP-FA NPs enhanced the anti-proliferation effects of AMF by 1.94-fold compared to AMF alone (P < 0.0001). Moreover, these NPs improved the radiation effects with a dose enhancement factor of 1.65. All results showed that the combination of carrier-based chemotherapy with hyperthermia and radiotherapy caused a higher anticancer efficacy than single- or two-modality treatments. SIGNIFICANCE: The nanoparticles advanced in this study can be proposed as the promising theranostic and thermo-radio-sensitizer platform for the diagnosis and tri-modal synergistic cancer therapy.

Physical and Biological Properties of 5-Fluorouracil Polymer-Coated Magnetite Nanographene Oxide as a New Thermosensitizer for Alternative Magnetic Hyperthermia and a Magnetic Resonance Imaging Contrast Agent: In Vitro and In Vivo Study.

This study reports a new procedure for utilizing 5-fluorouracil (5-Fu)-loaded polycaprolactone (PCL)/chitosan-covered magnetite nanographene oxide (5-Fu/SPION/NGO@PCL-LMWC) as a platform for synergistic thermo-chemotherapy. In fact, superparamagnetic iron oxide nanoparticles/nanographene oxide (SPION/NGO) nanoparticles can be coated with copolymers PCL/chitosan to attain better colloidal stability in the biological environment. Nanoparticles were synthesized and characterized for their size, surface charge, X-ray patterns, polymer content, and in vitro heat-triggered release. In vitro cytotoxic effects of nanoparticles on CT-26 cells were assessed with an MTT assay and real-time polymerase chain reaction. In vivo tumor growth inhibition was evaluated on an allograft mouse model of CT-26 cells. Tumor-bearing mice were injected with 5-Fu-loaded nanoparticles intravenously, and then, the targeted delivery was amplified using a magnetic field and finally exposed to an alternating magnetic field (AMF) (40 A/m, 13.56 MHz), during which the tumor site temperature increased to 43 °C. By using an infrared camera, we managed to heat the nanoparticles up to a constant temperature between 42.5 and 43.5 °C, with a tolerance ±0.03 °C. Finally, in vitro results showed that 5-Fu-loaded nanoparticles combined with AMF hyperthermia significantly reduced the plating efficiency of the cells (P < 0.01) and increased the Bax/Bcl-2 ratio (1.42 times, P < 0.01) compared with those achieved with each one alone. Furthermore, in vivo results demonstrated that the treatment of 5-Fu-loaded nanoparticles combined with the AMF diminished the growth of CT-26 tumor cells and increased the life span of the tumor-bearing mice (P < 0.001) by thermal energy deposition compared to that of the free 5-Fu drug. Also, the high level of accumulation of the nanoparticles within the tumor site was easily monitored with magnetic resonance imaging. It was concluded that the multifunctional magnetic nanoparticles could be used as a promising nanocarrier platform for achieving concurrent goals, drug delivery, magnetic targeting, thermal-sensitizing, cell death induction, and real-time monitoring of response to treatment.

Z-scan method to measure the nonlinear optical behavior of cells for evaluating the cytotoxic effects of chemotherapy and hyperthermia treatments.

The effects of new treatments must be investigated in vitro before using clinically or in vivo. The aim of this study was to introduce the Z-scan technique as a fast, accurate, inexpensive, and safe in vitro method to distinguish the cytotoxic effects of various treatments. C6 and OLN-93 cell lines were prepared and treated with Temozolomide (TMZ), radiofrequency hyperthermia (HT), and chemo-hyperthermia (HT+TMZ). The cytotoxic effects of different treatments on both cell lines were evaluated using colony formation assay and Z-scan method. The results of colony assay showed that the surviving fraction (SF) of C6 cells treated with TMZ, HT, and HT + TMZ were significantly decreased compared to the control group. Whereas, hyperthermia treatment had no significant effect on the SF of OLN-93 cells. The results of Z-scan technique indicated that the control group of C6 cells had the negative nonlinear refractive index (n2). Whereas, the C6 cells treated with HT, TMZ, and HT + TMZ had the positive n2 index. The sign of n2 index in the control and HT groups of OLN-93 cells was positive but treatment of cells with TMZ and HT + TMZ changed the sign of it. Moreover, with increasing the cytotoxic effects of different treatments, the SF value of both cell lines decreased and the magnitude of n2 index increased. The results of Z-scan technique were completely in line with the results of colony assay. Therefore, Z-scan method could distinguish the cytotoxic effects of various treatments by examining the nonlinear optical properties of the samples.

In vitro anti-cancer efficacy of multi-functionalized magnetite nanoparticles combining alternating magnetic hyperthermia in glioblastoma cancer cells.

Localized hyperthermia and the targeted release of the chemotherapy drug are one of the most challenging problems in chemo-hyperthermia therapy. In the present study, magnetite nanoparticles as a carrier of Temozolomide (TMZ) functionalized with folic acid-ligand (TMZ-MNP-FA) were designed and developed for targeted chemotherapy and radiofrequency hyperthermia of cancer cells. Nanoparticles were synthesized and characterized for hydrodynamic diameter, zeta potential, morphology, drug loading capacity, and in vitro RF-triggered release. Their cytotoxicity and efficacy as targeted drug delivery systems were evaluated in both cancer and normal cells and the therapeutic efficacy was analyzed on the C6 glioblastoma cancer cells. The C6 cells were treated with the nanoparticles and subjected to an alternating magnetic field (AMF) to reach a typical hyperthermia temperature of 43 °C. Then induction of apoptotic cells and the proliferation capacity of cancer cells were evaluated. The in vitro release studies exhibited that the drug release from TMZ-loaded magnetite nanoparticles was minimal at 37 °C but was noticeably boosted under an AMF irradiation. The developed targeted magnetite nanoparticles revealed higher cytotoxic effect and cellular uptake in folate-receptor overexpressing C6 cancer cells compared to OLN-93 normal cells. All results showed that combined magnetite chemo-hyperthermia (AMF + TMZ-MNP-FA) treatment was significantly more efficacious in cancer cells than hyperthermia, chemotherapy, or chemo-hyperthermia treatments (P < 0.0001). In conclusion, TMZ-MNP-FA had a key role to convert the externally delivered radiofrequency energy to heat in cancer cells. Additionally, localized hyperthermia triggered a TMZ release from the nanocarriers that resulted in cancer cell damage with synchronizing hyperthermia and chemotherapy.

Evaluation of nonlinear optical behavior of mouse colon cancer cell line CT26 in hyperthermia treatment.

Hyperthermia treatment can induce component changes on cell. This study explored the potential of Z-scan to improve accuracy in the identification of subtle differences in mouse colon cancer cell line CT26 during hyperthermia treatment. Twenty-one samples were subjected individually to treatment of hyperthermia at 41, 43, and 45 °C. Each hyperthermia treatment was done in six different time (15, 30, 45, 60, 75, and 90 min). Two optical setups were used to investigate the linear and nonlinear optical behavior of samples. Prior to the Z-scan technique, all samples were fixed with 1 mL of 5% paraformaldehyde. The linear optical setup indicated that extinction coefficient cannot monitor cell changes at different treatment regimes. But the nonlinear behavior of CT26 in all hyperthermia treatment regimens was different. By increasing the time and/or temperature of hyperthermia treatments, change in the sign of nonlinear refractive index from negative to positive occurred in earlier time intervals. This phenomenon was seen for 41, 43, and 45 °C in 75, 60, and 45 min, respectively. The results showed that the Z-scan technique is a reliable method with the potential to characterize cell changes during hyperthermia treatment regimes. Nonlinear refractive index can be used as a new index for evaluation of cell damage.

Evaluation of nonlinear optical differences between breast cancer cell lines SK-BR-3 and MCF-7; an in vitro study.

BACKGROUND: This study aimed to explore the potential of the Z-scan technique to improve accuracy in identifying SK-BR-3 and MCF-7 breast cancer cell lines. METHODS: Three in vitro samples were prepared for each breast cancer cell line. A closed-aperture Z-scan technique was used to measure the sign and magnitude of the nonlinear refractive index of each sample. Prior to the Z-scan, all samples were fixed with 1 mL of 5% paraformaldehyde. RESULTS: The sign of the nonlinear refractive indices of MCF-7 and SK-BR-3 breast cancer cell lines were negative and positive, respectively. The repeated Z-scan measurements for all samples of each cell line were similar. CONCLUSION: The results indicated that the proposed bio-optical method is a reliable method for characterizing differences in various breast cancer cell types. It is suggested that the nonlinear refractive index of cells be considered as an indicator for differentiating various breast cell lines from each other.