Could FA-PG-SPIONs act as a hyperthermia sensitizing agent? An in vitro study
Graphical abstract
Introduction
Cancer is a major public health problem in the world. According to the most recent statistics published by the American Cancer Society in 2014, cervical cancer is a very common malignant tumor in women, and remains one of the leading malignancies in women worldwide (Siegel et al., 2014). Among cancer types, cervical cancer is the fourth leading cause of death in women; and in order to treat cervical cancer, different methods such as radiotherapy, surgery, and chemotherapy are used. Using relatively new methods such as hyperthermia for cancer treatment has been increasingly expanding during the past few decades. In clinical procedures, hyperthermia is usually referred to as heating the tissue to a temperature range from 41 to 45 °C (Beik et al., 2016b). In this temperature range, only tumors are destroyed due to irregular poor blood flow in the tumors and their lower thermal tolerance compared to normal tissues (Huang et al., 2008). Therefore, because of less dissipation of heat in tumors than normal tissues, this temperature difference causes heat sensitivity (Chatterjee et al., 2011). The results of hyperthermia are to denature the protein and loosen the walls of the cell membrane therefore causing irreversible damage to the cells (Day et al., 2011, Huang et al., 2008). Tumor tissues in this condition appear to be more sensitive to thermal killing and prevent the thermal damage repair. Therefore, this temperature difference causes heat sensitivity (Chatterjee et al., 2011). There are several methods such as radio frequency, microwaves, ultrasound and shortwave diathermy and hot water bath (Hall and Giaccia, 2006, Norouzi et al., 2018) which induce hyperthermia. Among the mentioned methods, the simplest and most reliable method to induce heat is hot water bath with cells cultured in vitro (Hall and Giaccia, 2006). On the other hand, recently, magnetic nanoparticles such as iron oxide nanoparticles have been increasingly utilized in medical applications such as targeted drug delivery, magnetic resonance imaging (MRI), magnetic separation, drug delivery, treatment-inducing hyperthermia; and for the biomedical purposes such as hyperthermia, targeted drug delivery and MRI contrast agent (Emtiazi et al., 2017, Gupta et al., 2014, Jahandar et al., 2015, Mohammadi et al., 2015, Mostaghasi et al., 2018, Pankhurst et al., 2003, Sun et al., 2008, Zarrabi et al., 2011, Zarrabi et al., 2014). Moreover, the vessel wall is different in a cancerous cell compared to normal cell particularly because cancer cells are irregular in shape, leaky, defective (lack of basal membrane, endothelial cells poorly aligned) and dilated. It is predicted that most nanoparticles accumulate within cancer cells due to lack of lymphatic drainage in cancerous cells, a phenomenon called enhanced permeation and retention (EPR) effects. According to the EPR concept, it is predicted that most nanoparticles accumulate within cancer cells compared to normal cells (Maeda, 2001, Zwicke et al., 2012). In addition, recent researches have shown that exposing iron oxide nanoparticles to hyperthermia enables it to produce enhanced cytotoxicity of the nanoparticle-labeled cells (Beik et al., 2016a, Beik et al., 2016c, Ogden et al., 2009, Rodríguez-Luccioni et al., 2011, Shakeri-Zadeh et al., 2015b). Furthermore, hyperthermia-induced cell death depends on the duration of hyperthermia and temperature used for treatment (Roti Roti, 2008). Meanwhile, the penetration rate of nanoparticles is effective in inducing hyperthermia effectiveness, and there are several ways to increase the penetration of nanoparticles including the application of folic acid, peptides and aptamer which are generally referred to as targeting agents (Sun et al., 2015).
By targeting the tumor, it would be possible to eliminate many unwanted side effects of the current cancer therapies (Ghaznavi et al., 2017, Mirrahimi et al., 2017, Neshastehriz et al., 2017, Samadian et al., 2016, Shakeri-Zadeh et al., 2015a, Shakeri-Zadeh et al., 2014). Folic acid is a B vitamin, being essential for the body and playing a crucial role in cell proliferation. Interestingly, abundant receptors exist for vitamin B on the surface of special cancer cell lines such as head and neck, cervix, uterine, and ovarian (Sudimack and Lee, 2000, Toffoli et al., 1997).
Cancer cells express more folate receptors for their cellular levels due to their faster growth compared to normal cells since cells require folate for their cell cycle and metabolism. Therefore, folate-receptor is a suitable targeting agent (Sun et al., 2015). In this regard, therapeutic agents for the treatment of cancer can be targeted intentionally towards cancerous cells by binding to this vitamin (Leamon and Reddy, 2004).
Coating these nanoparticles with a biocompatible polymer like hyperbranched polyglycerol not only could improve its hydrophilicity, but also facilitates the attachment of targeting agents on the surface of polymer. In this regard, folic acid, one of the best widely used targeting agents, which had specific receptors on the surface of some certain cancer cell types, has been widely recommended. Thus, it could improve the specificity of cancer tissue treatment through active targeting of therapeutic agents.
Numerous articles show that there is a need for studying the enhancement effect of hyperthermia such as microwave and ultrasound in combination with different types of nanoparticles. However, the effect of hyperthermia with hot water bath that utilizes heat at 43 °C for 20, 40, 60 min combined with targeted/ non-targeted iron oxide nanoparticle on human cervical cancer cell line HeLa has not been investigated. Therefore, in the present in vitro investigation, the cytotoxic effect of SPIONs hyperthermia treatment with a hot water bath has been determined. FA-PG-SPIONs and PG-SPIONs with different concentrations () have been taken into consideration in order to determine the effect of iron oxide nanoparticles in the enhancement of hyperthermia in cancer cells. This could introduce the targeted nanoparticles as a hyperthermia-sensitizing agent.
Section snippets
Cell line
HeLa cell line was provided as a gift by Central Laboratory of Isfahan University of Medical Sciences, and it was procured from Cell Bank of Pasteur Institute of Iran. It was cultured in Dulbecco's Modified Eagle's Medium (DMEM, GIBCO, USA) supplemented with 10% FBS (GIBCO, USA), 100 U/ml of penicillin and 100 mg/ml of streptomycin (Sigma, USA). HeLa cells were incubated and subcultivated twice a week in T-25 tissue culture flasks (T25 flasks, SPL) as a monolayer with a density of 25 × 104
Cell characteristics
Human cervical carcinoma cell line HeLa grew as a monolayer on 24 well-plate, and an approximate doubling time of 25.47 ± 2.00 h for the cell population was obtained.
Nanoparticles synthesis and characterization
Coated and uncoated SPIONs were synthesized by thermal decomposition method. The described synthesis method in this paper results in the production of nanoparticles with narrow size distribution, which is a critical point for biological applications. Furthermore, SPIONs coated with PG are used in hyperthermia for the first time in
Discussion
Combination of iron oxide nanoparticle with hyperthermia therapy can be utilized in the field of cancer therapy due to the significant increase in cancer cell death using this combination.
The current study was planned to assess the cytotoxic effects of FA-PG-SPIONs and PG-SPIONs alone and in combination with hyperthermia on HeLa cell line.
This experiment can be assessed in four ways:
- 1.
The role of time durations in hyperthermia in increasing the cytotoxic effects
- 2.
The role of different
Conclusion
In summary, our results indicate that targeted iron oxide nanoparticles (FA-PG-SPIONs) in combination with hyperthermia can be introduced as a sensitizer agent when easily enter into cancer cells thus increasing the hyperthermia-induced cell death. Furthermore, it was revealed that the effect of cell exposure time to hyperthermia conditions is nanoparticle's dose dependance. Nevertheless, further study and researches in vitro (for other cell lines) and especially in vivo are needed to confirm
Declaration of interest
The authors report no conflicts of interest.
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