An in vitro evaluation of graphene oxide reduced by Ganoderma spp. in human breast cancer cells (MDA-MB-231)

Abstract

Background Recently, graphene and graphene-related materials have attracted much attention due their unique properties, such as their physical, chemical, and biocompatibility properties. This study aimed to determine the cytotoxic effects of graphene oxide (GO) that is reduced biologically using Ganoderma spp. mushroom extracts in MDA-MB-231 human breast cancer cells. Methods Herein, we describe a facile and green method for the reduction of GO using extracts of Ganoderma spp. as a reducing agent. GO was reduced without any hazardous chemicals in an aqueous solution, and the reduced GO was characterized using a range of analytical procedures. The Ganoderma extract (GE)-reduced GO (GE-rGO) was characterized by ultraviolet-visible absorption spectroscopy, X-ray diffraction, Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, dynamic light scattering, scanning electron microscopy, Raman spectroscopy, and atomic force microscopy. Furthermore, the toxicity of GE-rGO was evaluated using a sequence of assays such as cell viability, lactate dehydrogenase leakage, and reactive oxygen species generation in human breast cancer cells (MDA-MB-231). Results The preliminary characterization of reduction of GO was confirmed by the red-shifting of the absorption peak for GE-rGO to 265 nm from 230 nm. The size of GO and GE-rGO was found to be 1,880 and 3,200 nm, respectively. X-ray diffraction results confirmed that reduction processes of GO and the processes of removing intercalated water molecules and the oxide groups. The surface functionalities and chemical natures of GO and GE-rGO were confirmed using Fourier-transform infrared spectroscopy and X-ray photoelectron spectroscopy. The surface morphologies of the synthesized graphene were analyzed using high-resolution scanning electron microscopy. Raman spectroscopy revealed single- and multilayer properties of GE-rGO. Atomic force microscopy images provided evidence for the formation of graphene. Furthermore, the effect of GO and GE-rGO was examined using a series of assays, such as cell viability, membrane integrity, and reactive oxygen species generation, which are key molecules involved in apoptosis. The results obtained from cell viability and lactate dehydrogenase assay suggest that GO and GE-rGO cause dose-dependent toxicity in the cells. Interestingly, it was found that biologically derived GE-rGO is more toxic to cancer cells than GO. Conclusion We describe a simple, green, nontoxic, and cost-effective approach to producing graphene using mushroom extract as a reducing and stabilizing agent. The proposed method could enable synthesis of graphene with potential biological and biomedical applications such as in cancer and angiogenic disorders. To our knowledge, this is the first report using mushroom extract as a reducing agent for the synthesis of graphene. Mushroom...

Combination of graphene oxide–silver nanoparticle nanocomposites and cisplatin enhances apoptosis and autophagy in human cervical cancer cells

“…23,32 Briefly, rGO was obtained from the reaction of C-phycocyanin with GO. In a typical reduction experiment, 10 mL C-phycocyanin (10 mg) was added to 90 mL 1 mg/mL aqueous GO; the mixture was stirred at 30°C for 12 hours.…”

“…53 Also, recently we observed that both GO and AgNPs were capable of inducing ROS in several cancer cell lines. 21,22,32 Therefore, we were interested to investigate whether Cis, rGO-AgNPs, or a combination of Cis and rGO-AgNPs induced a dramatic effect on ROS-induced cell toxicity. To study the oxidative impact of Cis and rGOAgNPs on cellular oxidation-reduction (redox) state, we measured cellular ROS levels after exposing HeLa cells to Cis, rGO-AgNPs, or a combination of both for 24 hours.…”

“…19 AgNPs, GO, and rGO can induce the leakage of LDH into the media and significantly induce cytotoxicity in various types of cancer cell lines. 22,24,32 Next, we examined the effect of Cis, rGO-AgNPs, or a combination of both in LDH-induced cell toxicity. To study LDH leakage, we measured the LDH level after exposing HeLa cells to Cis, rGO-AgNPs, or a combination of both for 24 hours.…”

Reduction of graphene oxide by resveratrol: a novel and simple biological method for the synthesis of an effective anticancer nanotherapeutic molecule

“…8,9 Owing to its excellent characteristics, graphene can be potentially used in electronic devices such as flexible thin-film transistors, touch panels, and solar cells. [5][6][7]10 Furthermore, graphene has been used in various biomedical applications such as bio-sensing, 11 antibacterial composites, [12][13][14] drug delivery, 15 and tissue scaffolds.…”

“…6,7,26 The most commonly used chemical-reducing agents are hydrous hydrazine, hydrazine monohydrate, sodium borohydride, and hydrogen sulfide, which are highly toxic and harmful to living organisms and the environment. 6,9,[26][27][28] To overcome the aggregation and solubility problems, several polymers or surfactants have been used such as poly(sodium-4-styrenesulfonate), 29 alkaline conditions, 30 poly(N-vinyl-2-pyrrolidone), 31 and poly(allylamine). 32 Recently, biological molecules have been used to reduce GO; thus, Salas et al 33 have reported "green" reduction of GO via bacterial respiration.…”