Персона: Скобельцин, Алексей Сергеевич
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Инженерно-физический институт биомедицины
Цель ИФИБ и стратегия развития – это подготовка высококвалифицированных кадров на базе передовых исследований и разработок новых перспективных методов и материалов в области инженерно-физической биомедицины. Занятие лидерских позиций в биомедицинских технологиях XXI века и внедрение их в образовательный процесс, что отвечает решению практикоориентированной задачи мирового уровня – диагностике и терапии на клеточном уровне социально-значимых заболеваний человека.
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- ПубликацияОткрытый доступEffect of Photodynamic Therapy with the Photosensitizer Methylene Blue on Cerebral Endotheliocytes In Vitro(2024) Makarov, V. I.; Skobeltsin, A. S.; Averchuk, A. S.; Loschenov, V. B.; Макаров, Владимир Игоревич; Скобельцин, Алексей Сергеевич; Лощенов, Виктор БорисовичBackground: Microvessels in tumor tissue play a crucial role in meeting the metabolic needs of transformed cells, controlling the entry of xenobiotics into tumor tissue, and regulating local inflammation that promotes metastasis. Methylene blue has photosensitizing properties and can also affect dysfunctional mitochondria. Methods: The study was performed on the primary culture of CECs. The cells underwent photodynamic treatment through 660 nm laser irradiation at a power density of 300 mW/cm2. The MTT, TMRE, and TUNEL assays were used to assess the survival, redox metabolism, mitochondrial activity, and apoptosis of CECs. Additionally, the metabolic activity of cells was evaluated using FLIM by measuring the fluorescence lifetime of NADH and FAD. Results: When CECs were incubated with MB, there was an increase in mitochondrial activity that was dependent on the concentration of MB. Additionally, mitochondrial activity increased when the CECs were exposed to 660 nm laser irradiation at an energy dose of up to 5 J/cm2. Following PDT, a slight shift towards oxidative phosphorylation was observed. Conclusions: In vitro application of MB accumulation or laser irradiation causes a shift in the redox status of CECs towards increased reducing activity, without causing any cell damage. However, the combined action of PS and laser radiation has the opposite effect on the redox status of cells, resulting in an increase in the oxidized form of FAD.
- ПубликацияТолько метаданныеSpectroscopic measurement of methylene blue distribution in organs and tissues of hamadryas baboons during oral administration(2021) Pominova, D. V.; Ryabova, A. V.; Rudenko, N. S.; Kulik, O. G.; Kozlikina, E. I.; Efendiev, K. T.; Skobeltsin, A. S.; Loschenov, V. B.; Эфендиев, Канамат Темботович; Скобельцин, Алексей Сергеевич; Лощенов, Виктор Борисович© 2021 by the authors. Licensee MDPI, Basel, Switzerland.New research on Methylene Blue (MB), carried out in 2020, shows that it can be an effective antiviral drug as part of COVID-19 treatment. According to the research findings, MB has potential as a direct antiviral drug for the prevention and treatment of COVID-19 in the first stages of the disease. However, the MB accumulation by various types of tissues, as well as by immune cells, has not been previously studied. Therefore, the objective of this study was to obtain spectral data on the interstitial distribution of the administered drug in endothelial tissues in primates. The data on interstitial MB distribution obtained by spectroscopic measurement at both macro-and microlevels during oral administration to Hamadryas baboon individuals demonstrate that MB accumulates in mucous membranes of gastrointestinal tract and the tissues of the respiratory, cardiovascular, immune, and nervous systems. Additionally, it was found that MB was present in lung and brain myeloid cells in significant concentrations, which makes it potentially useful for protection from autoimmune response (cytokine storm) and as a tool for the correction of immunocompetent cells’ functional state during laser irradiation. Since the cytokine storm starts from monocytic cells during SARS-CoV-2 cellular damage and since tumor-associated macrophages can significantly alter tumor metabolism, accumulation of MB in these cells provides a reason to conclude that the immune response correction in COVID-19 patients and change in macrophages phenotype can be achieved by deactivation of inflammatory macrophages in tissues with MB using laser radiation of red spectral range.
- ПубликацияТолько метаданныеThe use of methylene blue to control the tumor oxygenation level(2024) Pominova, D.; Ryabova, A.; Skobeltsin, A.; Markova, I.; Поминова, Дарья Вячеславовна; Рябова, Анастасия Владимировна; Скобельцин, Алексей СергеевичHypoxia is a characteristic feature of many tumors. It promotes tumor proliferation, metastasis, and invasion and can reduce the effectiveness of many types of cancer treatment. The aim of this study was to investigate the pharmacokinetics of methylene blue (MB) and its impact on the tumor oxygenation level at mouse Lewis lung carcinoma (LLC) model using spectroscopic methods. The pharmacokinetics of MB were studied qualitatively and quantitatively using video fluorescence imaging and fluorescence spectroscopy. The degree of hemoglobin oxygenation in vivo was examined by calculating hemoglobin optical absorption from the measured diffuse reflectance spectra. The distribution of MB fluorescence and the lifetime of NADH were analyzed using laser scanning microscopy and fluorescence lifetime imaging microscopy (FLIM) to assess cellular metabolism. After intravenous administration of MB at 10ў??20 mg/kg, it quickly transitioned in the tumor to a colorless leucomethylene blue, with maximum accumulation in the tumor occurring after 5ў??10 minutes. A concentration of 10 mg/kg resulted in a relative increase of the tumor oxygenation level for small tumors (volume 50ў??75 mm3) and normal tissue 120 minutes after the introduction of MB. A shift in tumor metabolism towards oxidative phosphorylation (according to the lifetime of the NADH coenzyme) was measured using FLIM method after intravenous administration of 10 mg/kg of MB. Intravenous administration of MB at 20 mg/kg results in a long-term decrease in oxygenation, which persisted for at least 120 minutes after the administration and did not return to its initial level. Administration of MB at 10 mg/kg shown to increase tumor oxygenation level, potentially leading to more effective antitumor therapy. However, at higher doses (20 mg/kg), MB may cause long-term decrease in oxygenation.
- ПубликацияТолько метаданныеDetection of Changes in Macrophage Polarization as a Result of 5-Aminolevulinic Acid Photodynamic Therapy Using Fluorescence-Lifetime Imaging Microscopy(2022) Ryabova, A.; Skobeltsin, A.; Markova, I.; Pominova, D.; Loschenov, V.; Рябова, Анастасия Владимировна; Скобельцин, Алексей Сергеевич; Поминова, Дарья Вячеславовна; Лощенов, Виктор БорисовичNon-invasive detection of polarized macrophages in tumors is an urgent task in terms of combined antitumor therapy. By analyzing the fluorescence lifetime of the metabolic cofactors—the reduced form of nicotinamide adenine dinucleotide (NADH) and flavins—differences in cellular metabolism of normal tissue, tumor, inflammatory and anti-inflammatory macrophages were demonstrated. In this work we studied changes in the polarization of macrophages obtained from THP-1 monocytes in response to photodynamic therapy with 5-aminolevulinic acid (ALA–PDT). Moderate ALA–PDT in vitro led to changes in M0 macrophages metabolism towards M1 polarization, wherein M1 and M2 macrophages died and were replaced by non-polarized cells. The interstitial distribution of polarized macrophages after ALA–PDT was studied in a mouse tumor model of grafted Lewis lung carcinoma. In response to ALA–PDT, there was an increase in the inflammatory macrophages fraction in the tumor node. Metabolic fluorescence-lifetime imaging microscopy (FLIM) was performed for macrophages in vitro and for tumor cryosections. It was shown that analysis of phasor diagrams for the NADH, flavins, and 5-ALA-induced protoporphyrin IX (PpIX) fluorescence lifetime helps to determine the change in metabolism in response to different modes of PDT at the cellular and tissue levels. These data can be used for post-surgery tissue inspection.
- ПубликацияТолько метаданныеPhotodynamic therapy with 5-ALA induced PpIX effect on macrophages polarization(2022) Ryabova, A. V.; Pominova, D. V.; Skobeltcin, A. S.; Romanishkin, I. D.; Loschenov, V. B.; Рябова, Анастасия Владимировна; Поминова, Дарья Вячеславовна; Скобельцин, Алексей Сергеевич; Лощенов, Виктор БорисовичIn this work, we have carried out the assessment of changes in the polarization of macrophages in response to photodynamic treatment according to the changes in the fluorescence lifetime of respiratory chain enzymes. The analysis of the time-resolved fluorescence of metabolic signatures and PpIX makes it possible to determine the cell metabolism types. © 2022 IEEE.
- ПубликацияТолько метаданныеPreliminary low-dose photodynamic exposure to skin cancer with chlorin e6 photosensitizer(2022) Shiryaev, A. A.; Solonina, I. L.; Fatyanova, A. S.; Reshetov, I. V.; Efendiev, K. T.; Alekseeva, P. M.; Skobeltsin, A. S.; Loschenov, V. B.; Эфендиев, Канамат Темботович; Скобельцин, Алексей Сергеевич; Лощенов, Виктор Борисович© 2022Background: The study aimed to investigate the chlorin e6 photosensitizer distribution in the tumor and tumor border (5 mm) during low-dose photodynamic treatment and to increase the effectiveness of the therapy for skin neoplasms. Methods: Sensitized boundaries of neoplasms were evaluated by video fluorescence imaging. The study of changes in the chlorin e6 distribution before/after photodynamic therapy and in the process of low-dose photodynamic exposure was carried out by the method of spectral fluorescence diagnostics. Results: All 19 patients with basal-cell skin cancer had a contrast of chlorin e6 accumulation compared to normal tissues. 3 hours after intravenous administration of the photosensitizer at a dose of 1 mg/kg, the chlorin e6 concentration was: in normal tissues - 0.18 mg/kg, in the tumor - 1.26 mg/kg, in the tumor border - 0.63 mg/kg. In most cases, the fluorescence indices of chlorin e6 in tumor tissues after low-dose photodynamic therapy increased and exceeded the values before light exposure. Conclusion: Low-dose photodynamic therapy seems to be an optimal method for treating select skin neoplasms, which does not cause severe pain in patients during the light exposure and allows for local increase of the photosensitizer concentration in tumor tissues. This method of photodynamic therapy can improve the effectiveness of thе treatment.
- ПубликацияОткрытый доступDEVELOPMENT OF A METHOD FOR ASSESSING THE DEPTH OF PENETRATION OF ETHOSOMES WITH METHYLENE BLUE INTO THE SKIN DURING APPLICATION AND PHOTODYNAMIC EXPOSURE РАЗРАБОТКА МЕТОДА ОЦЕНКИ ГЛУБИНЫ ПРОНИКНОВЕНИЯ ЭТОСОМ С МЕТИЛЕНОВЫМ СИНИМ В КОЖУ ПРИ АППЛИКАЦИОННОМ ПРИМЕНЕНИИ И ФОТОДИНАМИЧЕСКИМ ВОЗДЕИСТВИИ(2022) LoginovaA. G.; Никитенко, Г. В.; Tikhonowski, G. V.; Skobeltsin, A. S.; Loschenov, V. B.; Войтова, А. В.; Тихоновский, Глеб Валерьевич; Скобельцин, Алексей Сергеевич; Лощенов, Виктор Борисович
- ПубликацияТолько метаданныеPHOTO AND SPECTRAL FLUORESCENCE ANALYSIS OF THE SPINAL CORD INJURY AREA IN ANIMAL MODELS ФОТО И СПЕКТРАЛЬНЫИ ФЛУОРЕСЦЕНТНЫИ АНАЛИЗ ОБЛАСТИ ТРАВМЫ СПИННОГО МОЗГА НА ЖИВОТНЫХ МОДЕЛЯХ(2023) Udeneev, A. M.; Kalyagina, N. A.; Skobeltsin, A. S.; Loschenov, V. B.; Калягина, Нина Анатольевна; Скобельцин, Алексей Сергеевич; Лощенов, Виктор БорисовичThe purpose of the work is to follow the dynamics of changes in fluorescent signals in the near-surface layers of tissue of injured areas of the back of laboratory animals, which will allow, by indirect evidence, to evaluate the information content of fluorescence diagnosis for subsequent possible diagnostic monitoring of photodynamic therapy of the spinal cord. The model animals were Wistar rats. Two types of contusions were modeled: pneumo-contusion and contusion by a falling load. Methylene blue and indocyanine green were used as photosensitizers. Fluorescence measurements were carried out by imaging and spectrometric methods. A stroboscopic fluorescence imager with an excitation wavelength of 630 nm was used to acquire fluorescence images. The LESA-01-BIOSPEC spectrometer with a He-Ne laser excitation allowed to obtain spectra. It was shown that both methods make it possible to estimate the fluorescence value of methylene blue and indocyanine green in the tissues under study. Moreover, the photographic method also allows to obtain the spatial distribution of fluorescence. The general trend found in the data is a more intense and uniform fluorescence of the dorsal region of rats with methylene blue and a less intense, but more contrasting distribution of indocyanine green. The presented methods are non-invasive, which makes them attractive for diagnostic use. However, due to the shallow depth of signal reception, the condition of the spine can be determined only indirectly, by the condition of the near-surface layers of tissue that accumulate the photosensitizer.
- ПубликацияОткрытый доступSPECTROSCOPIC STUDY OF METHYLENE BLUE PHOTOPHYSICAL PROPERTIES IN BIOLOGICAL MEDIA СПЕКТРОСКОПИЧЕСКОЕ ИССЛЕДОВАНИЕ ФОТОФИЗИЧЕСКИХ СВОИСТВ МЕТИЛЕНОВОГО СИНЕГО В БИОЛОГИЧЕСКИХ СРЕДАХ(2023) Pominova, D. V.; Ryabova, A. V.; Markova, I. V.; Akhlyustina, E. V.; Skobeltsin, A. S.; Поминова, Дарья Вячеславовна; Рябова, Анастасия Владимировна; Ахлюстина, Екатерина Витальевна; Скобельцин, Алексей СергеевичA spectroscopic study of the photophysical properties of methylene blue (MB) in aqueous solutions was carried out. Absorption and fluorescence spectra as well as fluorescence lifetime were recorded. The concentration dependence of the intensity and shape of the spectra allowed establishing the ranges of MB concentrations for in vitro and in vivo studies at which aggregation is not observed (up to 0.01 mM, which corresponds to 3.2 mg/kg). Studies of photodegradation in biological media showed that photobleaching of more than 80% in plasma and culture media is observed already at a dose of 5 J/cm2 , while in water at this concentration and dose photobleaching is not yet observed, and at a dose of 50 J/cm2 photobleaching of MB is about 30%. It was found that in media containing proteins and having an alkaline pH, photobleaching occurs significantly faster than in neutral aqueous media. The ionic strength of the solution has no effect on the photobleaching rate. Such photobleaching is caused by the photodegradation of MB rather than the transition to the leucoform.The efficiency of singlet oxygen generation and photodynamic activity were evaluated in vitro. In the investigated range of MB concentrations, the efficiency of singlet oxygen generation is rather low, because positively charged MB binds to negatively charged cell membranes, which leads to a change in the type of photodynamic reaction. The emergence of other reactive oxygen species (ROS), different from singlet oxygen, in cells has been demonstrated. The generation of ROS and the low quantum yield of singlet oxygen generation indicate the tendency of MB to provide the type I photosensitization mechanism (electron transfer with the formation of semi-reduced and semi-oxidized MB+ radicals) rather than to the type II mechanism (energy transfer to oxygen with the formation of singlet oxygen) in biological media and in vivo.
- ПубликацияТолько метаданныеThe Combined Use of Methylene blue and Chlorin E6 Photosensitizers for Photodynamic Therapy and Correction of the Tumor Microenvironement(2024) Pominova, D. V.; Ryabova, A. V.; Skobeltsin, A. S.; Markova, I. V.; Поминова, Дарья Вячеславовна; Рябова, Анастасия Владимировна; Скобельцин, Алексей Сергеевич