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Pure and hydroxylated C 60 induce different types of cell death. (A-D) U251 cells were incubated without (control) or with 1000 lg/ml C 60 (OH) n for 24 h (A, C) or with 1 lg/ml nano-C 60 for 6 h (B, D). The number of cells staining with annexin V-FITC and/or PI (A, B), or those with fragmented DNA (PI low sub-G 1 fraction) (C, D), was assessed by flow cytometry. (E) The cytotoxicity of nano-C 60 (1 lg/ml) and C 60 (OH) n (1000 lg/ml) toward U251 cells was investigated after 24-h incubation in the absence or presence of the pan-caspase inhibitor z-VAD-fmk (0.5lM). The values in A-D are means ± SD from three separate experiments, while those in E are mean ± SD of triplicates and are representative of three experiments (*p < 0.05).
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The mechanisms underlying the cytotoxic action of pure fullerene suspension (nano-C60) and water-soluble polyhydroxylated fullerene [C60(OH)n] were investigated. Crystal violet assay for cell viability demonstrated that nano-C60 was at least three orders of magnitude more toxic than C60(OH)n to mouse L929 fibrosarcoma, rat C6 glioma, and U251 human...
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... two distinct types of cell death, we used double staining with annexin V-FITC and PI. Annexin V binds to the phosphatidylserine that is typically exposed at the outer side of cell membrane during apoptosis, while PI only enters the cells with membrane damage that occurs in necrotic cell death. Therefore, normal, healthy cells are annexin ÿ /PI ÿ (Fig. 3A, 3B, lower left quadrant), apoptotic cells express phosphatidylserine, but have preserved membrane (E) The cytotoxicity of nano-C 60 (1 lg/ml) and C 60 (OH) n (1000 lg/ml) toward U251 cells was investigated after 24-h incubation in the absence or presence of the pan-caspase inhibitor z-VAD-fmk (0.5lM). The values in A-D are means ± SD from ...
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... in the absence or presence of the pan-caspase inhibitor z-VAD-fmk (0.5lM). The values in A-D are means ± SD from three separate experiments, while those in E are mean ± SD of triplicates and are representative of three experiments (*p < 0.05). cells with C 60 (OH) n led to a significant increase in the number of apoptotic, but not necrotic, cells (Fig. 3A), while nano-C 60 caused a massive increase in the number of necrotic, but not apoptotic, cells (Fig. 3B). The analysis of cellular DNA , as well as the interaction of nano-C 60 and C 60 (OH) n (F) in causing L929 cell death was assessed by Chou-Talalay analysis as described in the ''Materials and Methods'' section. The results in A-C ...
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... SD from three separate experiments, while those in E are mean ± SD of triplicates and are representative of three experiments (*p < 0.05). cells with C 60 (OH) n led to a significant increase in the number of apoptotic, but not necrotic, cells (Fig. 3A), while nano-C 60 caused a massive increase in the number of necrotic, but not apoptotic, cells (Fig. 3B). The analysis of cellular DNA , as well as the interaction of nano-C 60 and C 60 (OH) n (F) in causing L929 cell death was assessed by Chou-Talalay analysis as described in the ''Materials and Methods'' section. The results in A-C are presented as mean ± SD of triplicate observations and are representative of at least three ...
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... [CI > 1] or synergism [CI < 1]). content, which is reduced in apoptosis due to DNA fragmen- tation, was consistent with the data from annexin/PI staining. Namely, the number of hypodiploid, presumably apoptotic, cells with fragmented DNA (sub-G 1 fraction of PI-stained cells) markedly increased upon the treatment of U251 cells with fullerol (Fig. 3C), while pristine fullerene was without effect (Fig. 3D). Similar results were obtained with C6 and L929 cells (data not shown). The ability of nano-C 60 to cause cell membrane damage indicative of necrosis was also con- firmed by a significant increase in LDH release (159 ± 15%, n ¼ 3, p < 0.05), which was not observed after C 60 (OH) n ...
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... reduced in apoptosis due to DNA fragmen- tation, was consistent with the data from annexin/PI staining. Namely, the number of hypodiploid, presumably apoptotic, cells with fragmented DNA (sub-G 1 fraction of PI-stained cells) markedly increased upon the treatment of U251 cells with fullerol (Fig. 3C), while pristine fullerene was without effect (Fig. 3D). Similar results were obtained with C6 and L929 cells (data not shown). The ability of nano-C 60 to cause cell membrane damage indicative of necrosis was also con- firmed by a significant increase in LDH release (159 ± 15%, n ¼ 3, p < 0.05), which was not observed after C 60 (OH) n treatment. Due to a difference in the kinetics of ...
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... ability of nano-C 60 to cause cell membrane damage indicative of necrosis was also con- firmed by a significant increase in LDH release (159 ± 15%, n ¼ 3, p < 0.05), which was not observed after C 60 (OH) n treatment. Due to a difference in the kinetics of cytotoxic action of C 60 (OH) n and nano-C 60 , the assessment of apoptosis/ necrosis presented in Figures 3A and 3C and Figures 3B and 3D was performed after 24 and 6 h, respectively. It should be noted, however, that the appearance of annexin þ cells in nano- C 60 -treated cultures could not be observed even at earlier time points (0.5-4 h; data not shown), which excluded the possibility that some early apoptotic events were overlooked. ...
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... we used z-VAD-fmk, a pharmacological inhibitor of caspase activation and subsequent apoptotic cell death, to evaluate the role of apoptosis in fullerene-mediated cytotoxic- ity. In accordance with the data presented in Figures 3A-3D, z-VAD-fmk exerted a significant protective effect in C 60 (OH) n - treated U251 cultures, but completely failed to rescue U251 cells from the toxicity of nano-C 60 (Fig. 3E). Therefore, pure C 60 and C 60 (OH) n apparently employ distinct cytotoxic mechanisms resulting in the preferential induction of cas- pase-independent necrosis and caspase-dependent apoptosis, respectively. ...
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... caspase activation and subsequent apoptotic cell death, to evaluate the role of apoptosis in fullerene-mediated cytotoxic- ity. In accordance with the data presented in Figures 3A-3D, z-VAD-fmk exerted a significant protective effect in C 60 (OH) n - treated U251 cultures, but completely failed to rescue U251 cells from the toxicity of nano-C 60 (Fig. 3E). Therefore, pure C 60 and C 60 (OH) n apparently employ distinct cytotoxic mechanisms resulting in the preferential induction of cas- pase-independent necrosis and caspase-dependent apoptosis, ...
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Citations
... Specifically, this study observed decreased toxicity of hydroxylated fullerene compared to the cytotoxic effects of fullerene aggregates in human skin (HDP) and liver carcinoma (HepG2) cells [11]. Similarly, it was observed that hydroxylation decreases the toxic potential of fullerene in mouse L929 fibrosarcoma, rat C6 glioma, and U251 human glioma cell lines [12]. Additionally, hydroxylated fullerene induced apoptotic changes in the investigated cell lines, while fullerene C 60 induced necrotic cell death. ...
... Additionally, hydroxylated fullerene induced apoptotic changes in the investigated cell lines, while fullerene C 60 induced necrotic cell death. The distinct effects of pristine and modified fullerene originate from the different nanoparticle interactions with the intracellular metabolic pathways [11,12]. ...
In order to maximally reduce the toxicity of fullerenol (the first derivative of C60, FD-C60), and increase its biomedical efficiency, the second derivative SD-C60 (3HFWC, Hyper-Harmonized Hydroxylated Fullerene Water Complex) was created. Several different methods were applied in the comparative characterization of FD-C60 and SD-C60 with the same OH groups in their core. FD-C60 as an individual structure was about 1.3 nm in size, while SD-C60 as an individual structure was 10–30 nm in size. Based on ten physicochemical methods and techniques, FD-C60 and SD-C60 were found to be two different substances in terms of size, structure, and physicochemical properties; FD-C60, at 100 °C, had endothermic characteristics, while SD-C60, at 133 °C, had exothermic characteristics; FD-C60 did not have water layers, while SD-C60 had water layers; the zeta potential of FD-C60 was −25.85 mV, while it was −43.29 mV for SD-C60. SD-C60 is a promising substance for use in cosmetics and pharmaceuticals.
... Specifically, this study observed decreased toxicity of hydroxylated fullerene compared to cytotoxic effect of fullerene aggregates to human skin (HDP) and liver carcinoma (HepG2) cells [11. Similarly, it was observed that hydroxylation decreases toxic potential of fullerene on mouse L929 fibrosarcoma, rat C6 glioma, and U251 human glioma cell lines [12]. Additionally, hydroxylated fullerene induced apoptotic changes on investigated cells lines, while fullerene C60 induced necrotic cell death. ...
... Additionally, hydroxylated fullerene induced apoptotic changes on investigated cells lines, while fullerene C60 induced necrotic cell death. Distinct effects of pristine and modified fullerene originate from the different nanoparticles' interaction with the intracellular metabolic pathways [11,12]. ...
(1) Background: Since fullerene C60 is insoluble in water and is toxic, it is not applicable in biomedicine and cosmetics. For this reason, the first derivative FD-C60 (fullerenol) was synthesized, which solved the problem of solubility in water and partially reduced toxicity. In order to maximally reduce toxicity and increase biomedical efficiency, the second derivative SD-C60 (3HFWC- Hyper-Harmonized-Hydroxylated Fullerene Water Complex), was created. (2) Methods: In order to obtain the most reliable data, several different methods are applicable in the characterization of FD-C60 and SD-C60 with water layers (NIR, FTIR, 13C-NMR, 1H-NMR, TGA/DTA, XRD, AFM/MFM etc.). (3) Results: FD-C60 as an individual structure is about 1.3 nm in size, while SD-C60 as an individual structure is 1030 nm in size. The aforementioned methods showed that SD-C60 contains water layers, which explains the difference between those two structures in the physicochemical properties, and their effects on biomolecules. (4) Conclusions: Based on the above methods and techniques, FD-C60 and SD-C60 are two different substances in terms of size, structure, and physicochemical properties; FD-C60 at 100C has endothermic characteristics and SD-C60 at 133C has exothermic characteristics, FD-C60 possesses water molecules only from humidity, while SD-C60 has water molecules from both humidity and extra water molecules in layers, the zeta potential of FD-C60 is -25.85 mV, while it is -43.29 mV for SD-C60.
... Specifically, in this study, the decreased toxicity of FD-C 60 compared with cytotoxic effect of fullerene aggregates to human skin (HDP) and liver carcinoma (HepG2) cells was observed [15]. Similarly, it was observed that hydroxylation decreases the toxic potential of fullerene on mouse L929 fibrosarcoma, rat C6 glioma, and U251 human glioma cell lines [16]. Additionally, FD-C 60 induced apoptotic changes on investigated cells lines, while fullerene C 60 induced necrotic cell death [16]. ...
... Similarly, it was observed that hydroxylation decreases the toxic potential of fullerene on mouse L929 fibrosarcoma, rat C6 glioma, and U251 human glioma cell lines [16]. Additionally, FD-C 60 induced apoptotic changes on investigated cells lines, while fullerene C 60 induced necrotic cell death [16]. The distinct effects of pristine and modified fullerene originate from the different nanoparticles' interaction with the intracellular metabolic pathways [16,17]. ...
... Additionally, FD-C 60 induced apoptotic changes on investigated cells lines, while fullerene C 60 induced necrotic cell death [16]. The distinct effects of pristine and modified fullerene originate from the different nanoparticles' interaction with the intracellular metabolic pathways [16,17]. ...
The human body contains 60–70% water, depending on age. As a body fluid, it is not only a medium in which physical and chemical processes take place, but it is also one of the active mediators. Water is the richest substance with non-covalent hydrogen bonds. Water molecules, by themselves (in vacuum), are diamagnetic but when organized into clusters, they become diamagnetic or paramagnetic. Also, biomolecules (DNA, collagen, clathrin, and other proteins) have non-covalent hydrogen bonds in their structure. The interaction, as well as signal transmission, between water and biomolecules is achieved through the vibrations of covalent and non-covalent hydrogen bonds, which determine the state and dynamics of conformational changes in biomolecules. Disruptive conformational changes in biomolecules, cells, and tissues lead to their dysfunctionality, so they are a frequent cause of many disorders and diseases. For example, the rearrangement of hydrogen bonding due to mitochondrial disease mutation in cytochrome bc1 disturbs heme bH redox potential and spin state. In order to prevent and repair the dysfunctional conformational changes, a liquid substance was developed based on the second derivative of the C60 molecule (SD-C60), which has classical and quantum properties. The characterization of SD-C60 by UV-VIS-NIR, FTIR, TEM, and AFM/MFM was performed and it is shown that SD-C60 water layers generate vibrations with near-zero phase dispersion which are transmitted through Fibonacci’s water chains to biomolecules. In comparison with previously published SD-C60 derivate (3HFWC, size until 10 nm, and 1–5 water layers), the improved formulation (3HFWC-W, size 10–25 nm, and 6–9 water layers) showed multiplied cytotoxic activity against melanoma cell lines of different aggressiveness. Apart from this, the mode of action was preserved and based on an induction of senescence rather than cell death. Importantly, high selectivity towards malignant phenotypes was detected. Observed effects can be ascribed to a machinery of hydrogen bonds, which are generated in SD-C60 and transmitted through water to biomolecules. This approach may open a new field in science and healthcare—a “water-based nanomedicine”.
... For this reason, the nanofilm at a concentration of 100 mg/L was used in further research. It seems that the structure of fullerenes determines their toxicity, and carbon soot C 60 / 70 with a predominance of C 70 , even at low concentrations, induces the necrosis of mouse fibroblast cells L929, rat glioma C6, human glioma U251 [26] and human lung cancer A549 [27]. Cell death after exposure to high doses of C 60 fullerene may be associated with ROS-dependent cell membrane damage, as previously described for melanoma cells B16 [28]. ...
Simple Summary
Understanding the relationship between hepatocellular carcinoma recurrence and the process of cellular phenotypic transformation is crucial for developing effective therapy. The main problem is that marginal cancer cells with strong migratory activity remain in the post-resection tumour bed. The degraded extracellular matrix of the liver combined with the presence of cytokines causes the abnormal transduction of signals into the cell and, consequently, cell migration through the vascular basement membrane. We have previously shown that growth factors induce the transformation of epithelial cells to a mesenchymal phenotype. In this study, for the first time, the effect of fullerene surfaces on the invasion of HepG2 and SNU-449 cells with epithelial and mesenchymal phenotypes was compared. It is predicted that fullerene C60, as a material that inhibits the secretion of the transforming growth factor, may reduce the invasion of mesenchymal cells and not affect epithelial cells by reducing the expression of extracellular matrix proteases. Our study aimed to determine the effect of the fullerene surface on reducing cell invasion by inducing mesenchymal–epithelial transition. Epithelial cells concentrated at the primary site of the tumours are an easier target for radioembolisation therapy.
Abstract
The epithelial–mesenchymal transition (EMT) is a process in which epithelial cells acquire the ability to actively migrate via a change to the mesenchymal phenotype. This mechanism occurs in an environment rich in cytokines and reactive oxygen species but poor in nutrients. The aim of this study was to demonstrate that the use of a fullerene C60 nanofilm can inhibit liver cancer cell invasion by restoring their non-aggressive, epithelial phenotype. We employed epithelial and mesenchymal HepG2 and SNU-449 liver cancer cells and non-cancerous mesenchymal HFF2 cells in this work. We used enzyme-linked immunosorbent assays (ELISAs) to determine the content of glutathione and transforming growth factor (TGF) in cells. We measured the total antioxidant capacity with a commercially available kit. We assessed cell invasion based on changes in morphology, the scratch test and the Boyden chamber invasion. In addition, we measured the effect of C60 nanofilm on restoring the epithelial phenotype at the protein level with protein membranes, Western blotting and mass spectrometry. C60 nanofilm downregulated TGF and increased glutathione expression in SNU-449 cells. When grown on C60 nanofilm, invasive cells showed enhanced intercellular connectivity; reduced three-dimensional invasion; and reduced the expression of key invasion markers, namely MMP-1, MMP-9, TIMP-1, TIMP-2 and TIMP-4. Mass spectrometry showed that among the 96 altered proteins in HepG2 cells grown on C60 nanofilm, 41 proteins are involved in EMT and EMT-modulating processes such as autophagy, inflammation and oxidative stress. The C60 nanofilm inhibited autophagy, showed antioxidant and anti-inflammatory properties, increased glucose transport and regulated the β-catenin/keratin/Smad4/snail+slug and MMP signalling pathways. In conclusion, the C60 nanofilm induces a hybrid mesenchymal–epithelial phenotype and could be used in the prevention of postoperative recurrences.
... Specifically, in this study decreased toxicity of FD-C60 compared to cytotoxic effect of fullerene aggregates to human skin (HDP) and liver carcinoma (HepG2) cells was observed [15]. Similarly, it was observed that hydroxylation decreases toxic potential of fullerene on mouse L929 fibrosarcoma, rat C6 glioma, and U251 human glioma cell lines [16]. Additionally, FD-C60 induced apoptotic changes on investigated cells lines, while fullerene C60 induced necrotic cell death [16]. ...
... Similarly, it was observed that hydroxylation decreases toxic potential of fullerene on mouse L929 fibrosarcoma, rat C6 glioma, and U251 human glioma cell lines [16]. Additionally, FD-C60 induced apoptotic changes on investigated cells lines, while fullerene C60 induced necrotic cell death [16]. Distinct effects of pristine and modified fullerene originate from the different nanoparticles' interaction with the intracellular metabolic pathways [16,17]. ...
... Additionally, FD-C60 induced apoptotic changes on investigated cells lines, while fullerene C60 induced necrotic cell death [16]. Distinct effects of pristine and modified fullerene originate from the different nanoparticles' interaction with the intracellular metabolic pathways [16,17]. ...
The human body contains 60-70% of water, depending on age. As a body fluid, it is not only a medium in which physical and chemical processes take place, but it is also one of the active mediators. Water is the richest substance with non-covalent hydrogen bonds. Water molecules, by themselves (in vacuum), are diamagnetic, but when organized into clusters, become diamagnetic or paramagnetic. Also, biomolecules (DNA, collagen, clathrin, and other proteins) have non-covalent hydrogen bonds in their structure. The interaction, as well as signal transmission, between water and biomolecules is achieved through the vibrations of covalent and non-covalent hydrogen bonds, which determine the state and dynamics of conformational changes of biomolecules. Disruptive conformational changes of biomolecules, cells, and tissues lead to their dysfunctionality, so they are a frequent cause of many disorders and diseases. For example, the rearrangement of hydrogen bonding due to mitochondrial disease mutation in cytochrome bc1 disturbs heme bH redox potential and spin state. In order to prevent and repair the dysfunctional conformational changes, a liquid substance was developed based on the second derivative of the C60 molecule (SD-C60), which has classical and quantum properties. Characterization of SD-C60 by UV-VIS-NIR, FTIR, TEM, and AFM/MFM was done and it is shown that SD-C60 water layers generate vibrations with near-zero phase dispersion which are transmitted through the Fibonacci’s water chains to biomolecules. In comparison to previously published SD-C60 derivate (3HFWC, size until 10 nm, and 1-5 water layers), improved formulation (3HFWC-W, size 10-25 nm, and 6-9 water layers) showed multiplied cytotoxic activity against melanoma cell lines of different aggressiveness. Apart from this, the mode of action was preserved and based on induction of senescence rather than cell death. Importantly, high selectivity toward malignant phenotype was detected. Observed effects can be ascribed to a machinery of hydrogen bonds, which are generated in SD-C60 and transmitted through water to biomolecules. This approach may open a new field in science and healthcare - a “water-based nanomedicine”.
... Specifically, exposures to Cu extracts led to a significant reduction of cell density as well as to cells' shrinkage, confirming the previously cytotoxic effect, ultimately leading to increased cell death rates, as opposed to the activity of untreated DMEM, Ti, and glass reference extracts, that had no effect on L929 mouse fibroblasts' morphology. It is well recognised that the L929 cells becomes round in shape after death [78]. Accordingly, exposure of L929 cells to both pure DLC and Ag extracts did not alter, neither confluence nor morphology/shape, also confirming their safe cytotoxic profile as evidenced by the Alamar blue assay. ...
Silver enriched diamond-like carbon (Ag/DLC) coatings are gaining a reputation for medical applications as Ag doping improves the intrinsic biocompatibility and antimicrobial features of DLC, and higher Ag amounts improve its hemocompatibility and growth inhabitation of tumours etc. However, higher Ag amounts in DLC coatings can also lead to increased cytotoxic effects. This work presents the co-sputter deposition of Ag-enriched DLC coatings with 17, 40, and 65 at.% Ag in a DLC matrix. The structural and mechanical properties of the coatings are studied with field emission scanning electron microscopy, electron dispersive X-ray, X-ray diffraction, contact surface profiler, and nanoindentation. The biocompatibility of 72 h and 168 h leached extracts of Ag/DLC coatings are assessed through Ag ion release rates in Dulbecco’s modified eagle medium using inductively coupled plasma optical emission spectroscopy and their corresponding biocompatibility profiles are mapped against the L929 mouse fibroblast cell line. EDX mapping shows a throughout distribution of Ag in DLC matrix instead of localization distributions and clustering. The XRD diffraction peaks reflect the amorphous-like behaviour of 17 at.% Ag/DLC and prominent Ag crystallinity for 40 and 65 at.% Ag/DLC coatings. The leached Ag ions concentration of 8 ppm demonstrated by the 65 at.% Ag/DLC coating completely diminishes its biocompatibility. The results suggest that 17 at.% Ag enriched DLC coatings are a safe candidate for biomedical application and retain biocompatibility when treated with extracts prepared over prolonged ion leaching periods of 168 h, when compared to 40 and 65 at.% Ag/DLC coatings. Similarly, the 17 at.% Ag/DLC coatings show less compromise on mechanical properties, which decline by over 50% for 40 and 65 at.% Ag/DLC coatings. This work has extended the typical range of Ag for depositing safe and biocompatible DLC coatings for medical applications.
... Studies showed that when compared to untreated mice, fullerene-treated mice have a higher survival rate and improved memory and learning abilities [207]. There were various other studies which showed these scavenging properties of fullerene and its derivative compounds such as polyhydrates and polycarboxylate having cellular protectivity, can cross blood-brain barrier and neural cell damage prevention [208,209]. The major cause of many acute and chronic disorders is oxidative stress, and study has shown that cancer cells have increased oxidative stress with higher ROS, thereby stimulating proliferation of cells, genetic instability and cellular mutation. ...
Porous carbon is a versatile material. They have been used for ages and have significant industrial applications in adsorption and catalysis. The field of biomedical science is evolving day by day which leads to a new generation of innovation and research. Nanotechnology has many industrial applications which may include construction material, military goods, nanomachining of nano-rods, and nanowires. Nanotechnology is the new tool of the twenty-first century for the scientific community. Porous carbon is a part of molecular nanotechnology that contributes to human health monitoring. In this chapter, we are going to discuss the application of porous carbon material in the field of biomedical sciences mainly in carbon nanostructure, graphene-based sensor materials, and magnetically activated porous carbon. We will also focus on recent advancements by using 3D printing for building biomedical devices using porous carbon as a bio-ink.
... However, the DLC coatings deposited with an increasing bias of − 40, − 80, and − 120 V with hardness values of 8.8, 14.9, and 20.3 GPa respectively, lose their optical density compared to negative controls and the soft DLC coating deposited with zero bias. Optical density is an indirect indicator to estimate the biocompatibility on cell adhesion criteria [84] and higher optical densities refer to more adherent (living) cells as the dead cells are detached [85] from the surface and float in the media. The potential of single wavelength (570 nm) measurement as an indirect indicator of both cell morphology and adhesion, associated with MTT assay, is also reported in the literature [84]. ...
... Fig. 7 presents the morphologies of L929 cells after 72 h interaction with 168 h leached extracts of DLC coatings and control materials. It is established that the living cells maintain a linear morphology while the dead cells become round [85]. The representative images, captured with a 10X lens microscope, show that the L929 cells have linear cellular morphologies and full cover the DLC coating surfaces in DMEM (Fig. 7A). ...
Biomedical applications of diamond-like carbon (DLC) coatings are continuously increasing due to their superior mechanical properties, low friction coefficient, antiwear characteristics, and biocompatibility. The mechanical and tribological characteristics of DLC coatings have been comprehensively investigated on various substrate systems as a function of their deposition parameter dependant features for various biomaterial applications. However, the relationship between biocompatibility and resulting hardness of DLC coatings as a function of their bias voltage driven intrinsic features like sp2 and sp3 bonds remains largely unexplored. In this work, a series of DLC coatings are prepared as a function of varying bias voltage from 0 to −120 V, and characterised for their atomic structure, physical, and mechanical properties, and biocompatibility. The contact angle and surface roughness of the DLC coatings reduce while hardness increases from 7.8 to 20.3 GPa with increasing bias voltage from 0 to −120 V. A relatively soft DLC coating is shown to retain excellent biocompatibility which is approximately 38% higher than the harder DLC coatings following exposure of their leached extracts to L929 mouse fibroblast cells. This work demonstrates the complex interdependence of biocompatibility and hardness of DLC coatings and the outcomes will support correct material selection with an appropriate balance of these key properties for specific biomedical applications such as load-carrying and non-load carrying devices.
... On the other hand, smallsize C60 fullerene has higher toxicity potency and leads to inhibition of DNA polymerase [4]. It should be noted that fullerene C60 and its derivatives may also have a toxic effect in some circumstances, whereas C60 can also protect the cells from the condition of oxidative damage or pathological statesThe most of reported data on fullerene bioactivity in respect to brain tissue cells were obtained with functionalized C60 derivates in both in vivo and in vitro studies [5][6][7]. C60 fullerene develops extremely wide range of bioactivity in dependence of a dose and the state of surface [8][9][10] Cytotoxic effect of several functionalized C60 fullerene is recognized as a function of its prooxidant effect [6,11,12]. Various anticancer effects of several fullerene forms were established including DNA methylation [13], cell cycle arrest [14], antiangiogenic effect [15] and stress-induced apoptosis [16]. ...
... It should be noted that fullerene C60 and its derivatives may also have a toxic effect in some circumstances, whereas C60 can also protect the cells from the condition of oxidative damage or pathological statesThe most of reported data on fullerene bioactivity in respect to brain tissue cells were obtained with functionalized C60 derivates in both in vivo and in vitro studies [5][6][7]. C60 fullerene develops extremely wide range of bioactivity in dependence of a dose and the state of surface [8][9][10] Cytotoxic effect of several functionalized C60 fullerene is recognized as a function of its prooxidant effect [6,11,12]. Various anticancer effects of several fullerene forms were established including DNA methylation [13], cell cycle arrest [14], antiangiogenic effect [15] and stress-induced apoptosis [16]. Besides, neuroprotective effect of pristine C60 fullerene on autophagy flux and apoptosis was demonstrated in rat brain stressed with hyperglycemia [17]. ...
... Distinct cytotoxic mechanisms were observed in respect with anticancer effect of pristine C60 fullerene and watersoluble polyhydroxylated fullerene. Despite necrotic effect of pristine C60 nanocrystals, hydroxylated fullerene C60 may induce various apoptotic events, including DNA fragmentation, ROS-independent cell death with characteristics of apoptosis and loss of the specific shape of cellular membrane [6]. Therefore, C60 nanoparticles exhibit extremely various bioactivity depending on the dose, the manners for solubilization and the rate of hydratation of C60 nanoparticles, which are critic for cytotoxic and cytoprotective features of C60 fullerene and its derivates. ...
Glioblastoma, kemo ve radyoterapiye karşı dirençli, en agresif beyin kanseri tiplerinden biridir. C60 fulleren türevi nanopartiküller, çeşitli modellerde antikanser aktivite amacı ile geliştirilmektedir. Birçok kemoterapi ajanının aksine, bu fulleren çeşitli konsantrasyonlarda toksik değildir. C60 fulleren, birçok biyomedikal uygulama için umut verici bir adaydır. Bu nedenle, suda çözünür hydrated C60 fullerene'in (HyC60Fn) insan glioblastoma U373 hücresinde PARP, Beclin1, LC3 ve GFAP ekspresyonu üzerindeki etkileri araştırılmıştır. Hücre canlılığı ve göçü, sırasıyla MTT ve yara iyileşmesi testi ile belirlendi. PARP, Beclin1 ve LC3 ekspresyonu western blot ile ve GFAP ise immünositokimya ile tespit edildi. 0.5 – 2.0 µM doz aralığındaki HyC60Fn, doza bağlı bir şekilde hücre canlılığını azalttığı belirlendi. Ayrıca, HyC60Fn 1.0 ve 2.0 µM dozları, glioblastoma hücre göçünü belirgin şekilde bastırmıştır. Mekanizma olarak, HyC60Fn'nin otofaji belirteçleri olarak Beclin-1'i ve LC3-II/LC3-I ekspresyon oranını belirgin şekilde yukarı regüle ettiği belirlendi. Ayrıca, suda çözünür HyC60Fn’nin PARP fragmanı ve bu durumun doğal sonuç olarak glioblastoma U373 hücrelerinde parthanatos aktive ettiği belirlendi. Mevcut sonuçlar, HyC60Fn'nin, glioblastoma hücrelerinde şiddetli otofaji akışı ve parthanatos kombinasyonu yoluyla anti-tümör etkisini başlatabildiğini göstermektedir. Bu nedenle HyC60Fn, glioblastoma hücrelerinin reaktivitesini ve programlanmış hücre ölümünü modüle ederek en azından kısmen hücre ölüm mekanizmasını etkiler. Bulgularımız, HyC60Fn 'in umut verici bir kanser karşıtı terapötik olabileceğini ve bu konuda daha fazla çalışmanın gerekli olduğunu göstermektedir.
... The generation of ROS by GO-FA/Ce6 was subsequently evaluated by fluorescence spectroscopy using dihydrorhodamine 123 (DHR123). This dye is not fluorescent, but after oxidation by ROS, it is transformed into fluorescent rhodamine 123, with a high red emission [50,51]. As shown in Figure 3b, the fluorescence emission of DHR123 was almost negligible in water or after the addition of 10 μg•mL −1 of GO-FA/Ce6, proving that the conjugate did not change the fluorescence of the dye. ...
... In order to exploit the photothermal and photodynamic properties for therapeutic applications, we evaluated the cellular uptake of GO-FA/Ce6 by MCF-7 cells using confocal microscopy, by tracking the fluorescence of Ce6 on GO at different time points (0, 4, The generation of ROS by GO-FA/Ce6 was subsequently evaluated by fluorescence spectroscopy using dihydrorhodamine 123 (DHR123). This dye is not fluorescent, but after oxidation by ROS, it is transformed into fluorescent rhodamine 123, with a high red emission [50,51]. As shown in Figure 3b, the fluorescence emission of DHR123 was almost negligible in water or after the addition of 10 µg·mL −1 of GO-FA/Ce6, proving that the conjugate did not change the fluorescence of the dye. ...
Graphene oxide (GO) is one of the most studied nanomaterials in many fields, including the biomedical field. Most of the nanomaterials developed for drug delivery and phototherapies are based on noncovalent approaches that lead to an unspecific release of physisorbed molecules in complex biological environments. Therefore, preparing covalently functionalized GO using straightforward and versatile methods is highly valuable. Phototherapies, including photothermal therapy (PTT) and photodynamic therapy (PDT), have shown great potential as effective therapeutic approaches against cancer. To overcome the limits of a single method, the combination of PTT and PDT can lead to a combined effect with a higher therapeutic efficiency. In this work, we prepare a folic acid (FA) and chlorin e6 (Ce6) double-functionalized GO for combined targeted PTT/PDT. This conjugate can penetrate rapidly into cancer cells and macrophages. A combined effect of PTT and PDT is observed, leading to a higher killing efficiency toward different types of cells involved in cancer and other diseases. Our work provides a simple protocol to prepare multifunctional platforms for the treatment of various diseases.
