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Methylene blue (MB) is a promising photosensitizer (PS) for the treatment of pathological neoplasms, since it has both photodynamic activity (under laser irradiation) and redox and catalytic properties (in the absence of light). In the framework of this work, using spectroscopic methods, the effect of intravenous administration of MB on tissue oxygenation of hemoglobin in small animals in vivo in tumor and normal tissues was analyzed. The influence of MB on cell metabolism was analyzed. It has been shown that the use of MB promotes an increase in oxygen consumption by the tumor, and also leads to a shift in metabolism towards oxidative phosphorylation. It was shown that the use of MB contributes to an increase in oxygen consumption by the tumor, and also leads to a shift in metabolism towards oxidative phosphorylation.

A wide range of literature sources report on the potential benefits of transdermal drug delivery. Among these advantages, the following are distinguished – minimal injury, reduction of side effects, and prevention of degradation or metabolism in the gastrointestinal tract or liver. However, transdermal delivery of most molecules often excludes due to the barrier function of the skin, which prevents the penetration of exogenous substances. To overcome this barrier and increase skin absorption, ethosomal complexes use, by means penetration into the deep layers of the skin and/or systemic circulation is possible. This work devotes to the development of a non-invasive method for assessing the depth of penetration by ethosomes with methylene blue (MB) into the skin during application and photodynamic exposure. MB as photosensitizer (PS) was chosen, since there are a sufficient number of publications on its positive effect on the restoration of the cell’s respiratory chain of various organs and therefore the restoration of their metabolism. Besides MB has proven to be an effective PS, destructed pathogenic microbes and viruses, including SARS-CoV-2. However, for more effective Covid-19 therapy and antibiotic-resistant microbial diseases, the penetration of MB into the vascular system of the epidermis or mucous tissue is required. Nowadays, the existing methods for assessing the penetration depth of PS are time consuming and require the use of animal skin or model samples. The LESA-01 BIOSPEC system with specially designed optical adapters that allow assessing the drug fluorescence intensity on skin surface and at a depth of up to 2 mm in the investigation was used.

Introduction. Today, antimicrobial photodynamic therapy (a-PDT) becomes more and more popular, because of it pronounced bactericidal activity, anti-inflammatory effect. At the same time, no animal model studies have been conducted on morphological changes in cells after exposure to PDT on venous ulcers (VU) when using different types of photosensitizers (PS). The problem of comparing morphological changes in tissues when using a-PDT vs conventional PDT have not yet been resolved.Aim. Evaluation of the effectiveness of PDT and APDT in a comprehensive examination of trophic ulcers in an in vivo experiment and compare them with standard methods for the treatment of VLU.Materials and мethods. A series of experiments was conducted on 21 rabbits, separated into 3 equal groups of 7 rabbits each. Venous trophic ulcer was originally modeled for all rabbits. To obtain a VU, we performed an additional ligation of v. femoralis. The control group received standard therapy for VU. The PDT group had PDT with Photosens. The a-PDT group underwent a-PDT using Cholosens. Every 3 days, picture of local inflammation, regeneration rate and ulcer volume were determined. A morphological study of VLU was carried out on the first, 9th and 15th days.Results. The a-PDT group, day 15: 100% wound epithelization. Control group, day 21st: The volume of wounds decreased on average by 50%. The PDT group: 100% wound epithelization. The morphological study indicated a positive trend in the a-PDT group compared with PDT and control groups, which resulted in a decrease in the total mass of necrotic detritus, a change in the quantitative and qualitative composition of inflammatory infiltrate.Conclusion. The data obtained indicate that the use of a-PDT is recommended in treatment of VLU. Both PDT and a-PDT methods showed better results in comparison with standard therapy.

Indocyanine green (ICG), when in free form in a liquid, can form stable nanoparticle structures or colloidal solution, while changing its spectroscopic properties. In the work, the aggregation degree and the average size of nanoparticles depending on the concentration of a colloidal solution of indocyanine green (ICG NPs) in the form of J-aggregates were investigated by various methods based on light scattering. The size of nanoparticles is an important parameter from the point of view of clinical application, because the technique of intravenous administration of drugs, in order to avoid microvascular thrombosis and embolism, provides dosage forms with inclusions of individual molecules or their clusters, not exceeding 500 nm diameter. In turn, small nanoparticles less than 30 nm lead to prolonged circulation of the drug in the body with an increased possibility of permeation into cells of healthy tissue. In the course of studies, it was found that an increase in the concentration of ICG NPs in the solution leads to an increase in the average size of spontaneously formed J-aggregates, which, in turn, leads to a decrease in the absorption coefficient in the aggregates. Presumably, this phenomenon, i.e. the established nonlinear dependence of the J-aggregate absorption on its size, can be explained by the formation of absorption centers on the J-aggregate surface in the form of mobile surface molecules. The threshold range of ICG molecule concentration was determined, at which there is a transition from aggregation with an increase in size with a slow addition of ICG J-aggregate molecules in height to a rapid addition in width.

Nanoparticles are used as drug carriers to increase the selectivity and effectiveness of therapy, as well as for combined therapy that utilizes different effects. Iron oxide nanoparticles are promising in this aspect. Due to magnetic properties, they can be used as a contrast agent for magnetic resonance imaging. Also, iron oxide nanoparticles could be coated with a photosensitizer for photodynamic therapy and their laser or magnetic heating can be used for phototherapy. Local enhancement of the electromagnetic field near iron oxide nanoparticles can increase the fluorescence intensity of photosensitizers and the efficiency of singlet oxygen generation. This paper presents the results of a study of iron oxide nanoparticles focused on the photophysical aspects of the formation of “hot spots” under laser irradiation. The photoinduced effects of iron oxide nanoparticles observed in in vitro experiments lead to the rupture of lysosomes. Theoretical modeling showed that the heating of iron oxide nanoparticles with a radius of 35 nm under the action of laser radiation is about 89°C and 19°C for wavelengths of 458 and 561 nm, respectively. Local field enhancement occurs in pairs of nanoparticles of various sizes and strongly depends on the distance between them. The maximum gain is achieved at small distances between nanoparticles. For a dimer of nanoparticles with radii of 10 and 35 nm at a distance of 1 nm, an enhancement factor of two orders of magnitude was obtained. The investigated phenomenon of «hot spots» is in demand for precision therapy, because the photo-induced processes occur at small distances between nanoparticles, in areas of their high accumulation.

Today, photodynamic therapy is one of the most promising minimally invasive methods of treatment of various diseases, including cancer. The main limitation of this method is the insufficient penetration into the tissue of laser radiation used to activate photosensitizer molecules, which makes it difficult to carry out therapy in the treatment of large or deep-seated tumors. In this regard, there is a great interest in the development of new strategies for photodynamic therapy using infrared radiation for excitation, the wavelengths of which fall into the “transparency window” of biological tissues. In this work, it was proposed to use upconversion NaGdF4 :Yb:Er nanoparticles (UCNP), which absorb infrared excitation and serve as a donor that transfers energy to the photosensitizer. Photosens and phthalosens were chosen as the most promising photosensitizers for the study. The aim of this work was to study the energy transfer processes between upconversion nanoparticles doped with rare-earth ions and photosensitizer molecules. in order to excite photosensitizers with IR radiation and carry out photodynamic therapy of deep-seated neoplasms. Using spectroscopic and time-resolved methods, it has been demonstrated that there is an efficient energy transfer between upconversion particles and photosensitizers phthalosens and photosens. The calculated efficiency of energy transfer by the Foerster mechanism was 41% for the UCNP + photosens system and 69% for the UCNP + phthalosens system. It has been experimentally and theoretically proved that there is a binding of photosensitizer molecules with UCNP by means of surfactants, leading to a reduction in the distance between them, due to which effective nonradiative energy transfer is realized. The generation of singlet oxygen by the phthalosens photosensitizer upon excitation by means of energy transfer from UCNP, excited at 980 nm wavelength of, has been demonstrated.

The main problem in the photodynamic therapy (PDT) of tumors is insufficient light exposure to tissue or the appearance of undesirable surface effects. The reason is the irregular distribution of the absorbed light dose by depth. The influence of the spot diameter on the relative fluence rate in the near-surface layer of the cervical tissue was studied by Monte Carlo simulation. Photodynamic exposure with chlorine-type photosensitizer (PS) was carried out on the tissue model with laser 660 nm at the same power density with a change in spot diameter from 5 to 15 mm and radiation energy density from 100 to 300 J cm−2. The distributions of the fluorescence indices of the PS and the hemoglobin oxygenation degree by depth were obtained. The dependence of PS photobleaching on the energy density was established at the same power density and different spot diameters. The developed method increased the efficiency of PDT by delivering a sufficient energy density of laser radiation to the entire tumor tissue by depth without thermal damage, that allowed minimizing side effects and prevent possible growth and recurrence of the disease.

Neurosurgery of intracranial tumors, especially of glial origin, is a non-trivial task due to their infiltrative growth. In recent years, optical methods of intraoperative navigation have been actively used in neurosurgery. However, one of the most widely used approaches based on the selective accumulation of fluorescent contrast medium (5-ALA-induced protoporphyrin IX) by the tumor cannot be applied to a significant number of tumors due to its low accumulation. On the contrary, Raman spectroscopy, which allows analyzing the molecular composition of tissues while preserving all the advantages of the method of fluorescence spectroscopy, does not require the use of an exogenous dye and may become a method of choice when composing a system for intraoperative navigation or optical biopsy. This work presents the first results of using the principal component method to classify Raman spectra of human glioblastoma with intermediate processing of spectra to minimize possible errors from the fluorescence of both endogenous fluorophores and photosensitizers used in fluorescence navigation. As a result, differences were found in the principal component space, corresponding to tissue samples with microcystic components, extensive areas of necrosis, and foci of fresh hemorrhages. It is shown that this approach can serve as the basis for constructing a system for automatic intraoperative tissue classification based on the analysis of Raman spectra.

The paper presents the results of a comparative study of methods of cluster analysis of optical intraoperative spectroscopy data during surgery of glial tumors with varying degree of malignancy. The analysis was carried out both for individual patients and for the entire dataset. The data were obtained using combined optical spectroscopy technique, which allowed simultaneous registration of diffuse reflectance spectra of broadband radiation in the 500–600 nm spectral range (for the analysis of tissue blood supply and the degree of hemoglobin oxygenation), fluorescence spectra of 5‑ALA induced protoporphyrin IX (Pp IX) (for analysis of the malignancy degree) and signal of diffusely reflected laser light used to excite Pp IX fluorescence (to take into account the scattering properties of tissues). To determine the threshold values of these parameters for the tumor, the infltration zone and the normal white matter, we searched for the natural clusters in the available intraoperative optical spectroscopy data and compared them with the results of the pathomorphology. It was shown that, among the considered clustering methods, EM‑algorithm and k‑means methods are optimal for the considered data set and can be used to build a decision support system (DSS) for spectroscopic intraoperative navigation in neurosurgery. Results of clustering relevant to thepathological studies were also obtained using the methods of spectral and agglomerative clustering. These methods can be used to postprocess combined spectroscopy data.

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.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.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.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.

In view of their potential in the treatment of oncological diseases, photosensitizers (PS) have been the object of active research for the last years. The applied therapeutic photodynamic method involves the activation of a PS by light at a specific wavelength. PS interacts with oxygen and catalyzes the production of singlet oxygen molecules or radical oxygen species which leads to tumor cell death. This work is devoted to the study of the effectiveness of the photodynamic action of new PSs based on polycationic derivatives of synthetic bacteriochlorin on human lung cancer cells A549. The results obtained show that these PS effectively bind to these cells, have high phototoxicity and low ‘dark’ cytotoxicity for them. Morphological and immunohistochemical studies show that photodynamic effect leads to induction of cell necrosis and apoptosis, as well as a sharp decrease in mitotic activity.

Laser hyperthermia is one of the promising methods for treatment of oncological diseases. For routine clinical use of hyperthermia, it is necessary to control the uniformity and localization of heat within the tumor. Local heating can be achieved by using special thermal agents, such as nanoparticles doped with rare-earth ions. Measurement of the temperature of the thermal agents will allow timely regulation of the applied laser radiation excitation power and optimization of the hyperthermia process.The paper presents the results of a study on the non-invasive determination of the YPO4 nanoparticles doped with Nd3+ temperature with sensitivity of 0.2% °С-1 in 30-60°С temperature range. The temperature of the nanoparticles was calculated from the Nd3+ luminescence spectra in the 800-1000 nm range under excitation into 4F5/2 energy state by 805 nm laser. A calibration procedure for recalculating the ratio of the luminescence intensities from the Stark sublevels of the 4F3/2 Nd3+ state into the values of the real NP temperature in accordance with the Boltzmann distribution is given. An algorithm for calculating luminescence intensities for individual Stark components is proposed. After calculating the intensities corresponding to each individual Stark component, all the intensities related to the transition from the upper and lower Stark sublevels of the 4F3/2 state are summed, and then their ratio is calculated. The resulting ratio is normalized to the value of the ratio at room temperature and, in accordance with the calibration dependence, is recalculated into the NP heating temperature. It was demonstrated that the investigated 1%Nd3+:YPO4 nanoparticles can be used as "primary” thermometers that do not require additional recalibration to evaluate the temperature in the range used for hyperthermia.

The mortality rate from various oncological diseases remains consistently extremely high, even in spite of recent breakthrough discoveries in the field of cancer therapy [1-3]. One of the most promising cancer treatments is photodynamic therapy (PDT) combined with fluorescence diagnostics (FD) [4, 5]. This study aims to evaluate the efficiency of the PDT procedure using an experimental LED device for photodynamic therapy and fluorescence diagnostics of neoplasms by the level of photobleaching of the photosensitizer protoporphyrin IX (PPIX) in optical phantoms of the skin. The results of spectral measurements showed that the LEDs is able to effectively induce oxidative stress in cancer cells, and due to the presence of a video system, it becomes possible to register the level of tissue fluorescence.

Photodynamic therapy (PDT) is widely implemented in clinical practice. This contributes to the development of new devices for conducting fluorescence diagnostics (FD) and PDT. One of the key parameters for the new devices is the ability to perform PDT and evaluate the fluorescence signal during PDT. Most PDT devices do not have the ability to perform PD and register a fluorescent signal, which is important for monitoring the quality of treatment. The purpose of this work is to develop a new device that will be able to conduct PD and PDT during the operation. Using a video system with software for it and an LED irradiator with several types of LEDs. The results show the effective use of this system and the ability to observe FD, as well as conduct.

Antibacterial photodynamic therapy (APDT) is a promising method of treating local infected foci, in particular, surgical and burn wounds, trophic and diabetic ulcers. Photodynamic inactivation (PDI) is able to effectively destroy bacterial cells without them developing resistance in response to treatment.This work was dedicated to the study of photophysical and antibacterial properties of new photosensitizers (PS) based on polycationic phthalocyanines and synthetic bacteriochlorins for photodynamic inactivation of P. aeruginosa bacteria and their biofilms. Gram-negative bacteria P. aeruginosa are often found in infected wounds, presumably in biofilm state and are characterized by rather low susceptibility to APDT, which is a problem. PS were studied for possible aggregation at various concentrations by means of absorption and fluorescence spectroscopy. The results of studies of the ZnPcChol

The aim of the study is to optimize the irradiation procedure during endobiliary photodynamic therapy for patients with cholangiocarcinoma. The studies were performed using fiber-optic light delivery tools in various configurations and using different irradiation power levels. Control of photosensitizer accumulation at the site of therapy was also applied. It is shown that the use of fiber-optic light delivery tools with cylindrical diffusers provides more effective and less painful photodynamic therapy. It is shown that after the injection of the photosensitizer, there is an optimal time for the therapy, when the maximum difference in the accumulation of the photosensitizer in the tumor is observed in comparison with the intact tissue. Keywords: endobiliary photodynamic therapy, fluorescent diagnostics, fiber-optic light delivery tools, cholangiocarcinoma

A new method of light dose monitoring during the photodynamic therapy of abdominal cavity organs was developed. To apply this method specially developed sensors were constructed. For these sensors the temporal and distance dependences of light doses were received and analyzed. It was shown that the light dose from the operating lamp is large enough in laser light field to be taken into account when planning photodynamic therapy. Keywords: PDT, light doses, other source of light in PDT, photodynamic therapy

A new type of the surgical instruments is being developed—sapphire scalpels with a possibility of the fluorescent diagnostics of the state of incised tissue near the cutting edge directly in the process of the surgical operation. The principle of the new system is based on the use of two isolated capillary channels in the volume of the sapphire scalpel for introducing of quartz waveguides. One of the waveguides is used for delivering the laser radiation directly to the blade and excitation of photoluminescence near the cutting edge. The second one is used for catching and transfer of photoluminescence to spectrometer. The scalpel employs rapid real‐time feedback analysis for on‐line diagnostics of tissues during surgery to remove a tumor.