Comparative Studies of the Structural and Physicochemical Properties of the First Fullerene Derivative FD-C60 (Fullerenol) and Second Fullerene Derivate SD-C60 (3HFWC).

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.

The presymptomatic treatment with 3HFWC nanosubstance decreased plaque load in 5XFAD mouse model of Alzheimer's disease.

INTRODUCTION: In the present study, we assessed the effects of the hyper-harmonized-hydroxylated fullerene-water complex (3HFWC) on Alzheimer's disease (AD) neuropathological hallmarks in 5XFAD mice, an AD animal model. METHODS: The 3-week-old 5XFAD mice were exposed to 3HFWC water solution ad libitum for 3 months in the presymptomatic phase of pathology. The functional effects of the treatment were confirmed through near-infrared spectroscopy (NIRS) analysis through machine learning (ML) using artificial neural networks (ANNs) to classify the control and 3HFWC-treated brain tissue samples. The effects of 3HFWC treatment on amyloid-ß (Aß) accumulation, plaque formation, gliosis, and synaptic plasticity in cortical and hippocampal tissue were assessed. RESULTS: The 3HFWC treatment significantly decreased the amyloid-ß plaque load in specific parts of the cerebral cortex. At the same time, 3HFWC treatment did not induce the activation of glia (astrocytes and microglia) nor did it negatively affect synaptic protein markers (GAP-43, synaptophysin, and PSD-95). CONCLUSION: The obtained results point to the potential of 3HFWC, when applied in the presymptomatic phase of AD, to interfere with amyloid plaque formation without inducing AD-related pathological processes such as neuroinflammation, gliosis, and synaptic vulnerability.

The Second Derivative of Fullerene C60 (SD-C60) and Biomolecular Machinery of Hydrogen Bonds: Water-Based Nanomedicine.

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".

Physicochemical Properties of Gold Nanoparticles for Skin Care Creams.

Gold nanoparticles (AuNPs) have now been used in skin care creams for several years, with marketed anti-aging, moisturizing, and regenerative properties. Information on the harmful effects of these nanoparticles is lacking, a concern for the use of AuNPs as cosmetic ingredients. Testing AuNPs without the medium of a cosmetic product is a typical method for obtaining this information, which is mainly dependent on their size, shape, surface charge, and dose. As these properties depend on the surrounding medium, nanoparticles should be characterized in a skin cream without extraction from the cream's complex medium as it may alter their physicochemical properties. The current study compares the sizes, morphology, and surface changes of produced dried AuNPs with a polyvinylpyrrolidone (PVP) stabilizer and AuNPs embedded in a cosmetic cream using a variety of characterization techniques (TEM, SEM, DLS, zeta potential, BET, UV-vis). The results show no observable differences in their shapes and sizes (spherical and irregular, average size of 28 nm) while their surface charges changed in the cream, indicating no major modification of their primary sizes, morphology, and the corresponding functional properties. They were present as individually dispersed nanoparticles and as groups or clusters of physically separated primary nanoparticles in both dry form and cream medium, showing suitable stability. Examination of AuNPs in a cosmetic cream is challenging due to the required conditions of various characterization techniques but necessary for obtaining a clear understanding of the AuNPs' properties in cosmetic products as the surrounding medium is a critical factor for determining their beneficial or harmful effects in cosmetic products.

Melanoma Cell Reprogramming and Awakening of Antitumor Immunity as a Fingerprint of Hyper-Harmonized Hydroxylated Fullerene Water Complex (3HFWC) and Hyperpolarized Light Application In Vivo.

In our recent study, we showed that in vitro treatment of melanoma cells with hyperpolarized light (HPL) as well as with the second derivative of fullerene, hyper-harmonized hydroxylated fullerene water complex (3HFWC) reduced viability of cells by decreasing their proliferative capacity and inducing senescence and reprogramming towards a normal, melanocytic phenotype. Therefore, we wanted to determine whether these effects persisted in vivo in the syngeneic mouse melanoma model with a combined treatment of HPL irradiation and 3HFWC per os. Our results demonstrated the potent antitumor effects of 3HFWC nanosubstance assisted by HPL irradiation. These effects were primarily driven by the stimulation of melanoma cell growth arrest, the establishment of a senescent phenotype, and melanocytic differentiation on the one hand, and the awakening of the antitumor immune response on the other. In addition, the combined treatment reduced the protumorigenic activity of immune cells by depleting T regulatory cells, myeloid-derived suppressors, and M2 macrophages. The support of the 3HFWC substance by HPL irradiation may be the axis of the new approach design based on tumor cell reprogramming synchronized with the mobilization of the host's protective immune response.

Combined Action of Hyper-Harmonized Hydroxylated Fullerene Water Complex and Hyperpolarized Light Leads to Melanoma Cell Reprogramming In Vitro.

(1) Background: Their unique structure and electron deficiency have brought fullerenes into the focus of research in many fields, including medicine. The hyper-harmonized hydroxylated fullerene water complex (3HFWC) formulation has solved the limitations of the poor solubility and bioavailability of fullerenes. To achieve better antitumor activity, 3HFWC was combined with short-term irradiation of cells with hyperpolarized light (HPL) generated by the application of a nanophotonic fullerene filter in a Bioptron® device. The benefits of HPL were confirmed in the microcirculation, wound healing and immunological function. (2) Methods: B16, B16-F10 and A375 melanoma cells were exposed to a wide spectrum of 3HFWC doses and to a single short-term HPL irradiation. (3) Results: Apart from the differences in the redox status and level of invasiveness, the effects of the treatments were quite similar. Decreased viability, morphological alteration, signs of melanocytic differentiation and cellular senescence were observed upon the successful internalization of the nanoquantum substance. (4) Conclusions: Overall, 3HFWC/HPL promoted melanoma cell reprogramming toward a normal phenotype.

Mechanisms of skin moisturization with hyperharmonized hydroxyl modified fullerene substance.

BACKGROUND: Hyper Harmonized Hydroxyl Modified Fullerene Substance (3HFWC+ ) establishes hydrogen bonds with the surrounding water molecules and organizes them in clusters with the liquid crystalline state, similar to the properties of water surrounding the biomolecules. AIMS: To investigate the moisturizing properties of hyperharmonized fullerenol-3HFWC+ as an emulsion O/W ingredient on the skin. PATIENTS/METHODS: We have analyzed the reflexion of the blue light from the skin, in vivo, with different levels of moisturization and compared the influence of three groups of cosmetic products (with various active ingredients, 3HFWC+ or water in same percentage in the "vehiculum") on skin moisturization by measuring paramagnetic/diamagnetic properties by Optomagnetic Imaging Spectroscopy. RESULTS: Regenerating Cream and Body Lotion have shown statistically significant increase of diamagnetic features predominantly in all 3 types of creams, while the Hand Cream formulations have not shown statistically significant results after 4 weeks of treatment. Anti-aging cream with 3HFWC+ demonstrated statistically significant increase of paramagnetic properties of the skin (p+ /p- from 0.90 to 0.62) and showed beneficial effects on information transfer and water content in stratum corneum. CONCLUSION: Although it does not act as a classical moisturizing agent on the skin, 3HFWC+ can increase moisturization as a result of hydrogen bonds established among 3HFWC+ and water or biomolecules, liquid crystalline state of water in 3HFWC+ resembling the properties of water surrounding the biomolecules, and finally, liquid crystalline state of water in the products.

Influence of hyper-harmonized fullerene water complex on collagen quality and skin function.

BACKGROUND: Fullerene water complex establishes the optimal order and function of biomolecules in natural, biophysical way by transducing the signal through water hydrogen bonds to biomolecules. OBJECTIVES: This paper considers the effects of the patented hyper-harmonized-hydroxylated fullerene water complex (3HFWC) on biophysical properties of the skin collagen molecules. METHOD: Optomagnetic imaging spectroscopy (OMIS) has been used for the analysis of the biophysical skin properties (diamagnetic/paramagnetic) after applying three groups of different cosmetic products. Tested cosmetic products were prepared by replacing the active ingredients with 3HFWC or with water in four commercial products. The original commercial creams and their vehiculums with water added served as control groups. Data were statistically analyzed using paired t test in R software. RESULTS: t Test gave statistically significant results for all of the products with 3HFWC, while within the control group, only body lotion and hand cream did show statistically significant results (P < 0.05). Significant improvements in abundance and quality of collagen in the dermis were achieved with body lotion with 3HFWC (p+ /p- ranged from 0.82 to 0.97). While body lotion vehiculum made collagen-water interaction more stable (p+ /p- ranged from 0.3 to 0.55), hand cream with 3HFWC made it more dynamic (p+ /p- ranged from 0.63 to 0.49). Body lotion vehiculum improved the compactness of the dermis (p+ /p- ranged from 0.2 to 1.03), as well as commercial hand cream (p+ /p- ranged from 0.28 to 0.85). CONCLUSION: Compared to the control groups, cosmetic products with 3HFWC demonstrated positive effects on the biophysical properties of the skin. Increased paramagnetic properties are linked to more unpaired electrons, their faster movement, and, finally, better signal transduction. Thus, products with 3HFWC could enable faster regeneration of collagen and prompt skin reaction to the negative environmental influences.

Fetus sound stimulation: cilia memristor effect of signal transduction.

BACKGROUND: This experimental study evaluates fetal middle cerebral artery (MCA) circulation after the defined prenatal acoustical stimulation (PAS) and the role of cilia in hearing and memory and could explain signal transduction and memory according to cilia optical-acoustical properties. METHODS: PAS was performed twice on 119 no-risk term pregnancies. We analyzed fetal MCA circulation before, after first and second PAS. RESULTS: Analysis of the Pulsatility index basic (PIB) and before PAS and Pulsatility index reactive after the first PAS (PIR 1) shows high statistical difference, representing high influence on the brain circulation. Analysis of PIB and Pulsatility index reactive after the second PAS (PIR 2) shows no statistical difference. Cilia as nanoscale structure possess magnetic flux linkage that depends on the amount of charge that has passed between two-terminal variable resistors of cilia. Microtubule resistance, as a function of the current through and voltage across the structure, leads to appearance of cilia memory with the "memristor" property. CONCLUSION: Acoustical and optical cilia properties play crucial role in hearing and memory processes. We suggest that fetuses are getting used to sound, developing a kind of memory patterns, considering acoustical and electromagnetically waves and involving cilia and microtubules and try to explain signal transduction.

Nanomagnetic behavior of fullerene thin films in Earth magnetic field in dark and under polarization light influences.

In this paper magnetic fields intensity of C60 thin films of 60 nm and 100 nm thickness under the influence of polarization lights are presented. Two proton magnetometers were used for measurements. Significant change of magnetic field intensity in range from 2.5 nT to 12.3 nT is identified as a difference of dark and polarization lights of 60 nm and 100 nm thin films thickness, respectively. Specific power density of polarization light was 40 mW/cm2. Based on 200 measurement data average value of difference between magnetic intensity of C60 thin films, with 60 nm and 100 nm thickness, after influence of polarization light, were 3.9 nT and 9.9 nT respectively.