Aluminium Phthalocyanine Nanoparticles Application for Fluorescent Diagnostics and Photodynamic Therapy in Dentistry

Early diagnosis of tooth-enamel microcracks is of great importance in modern dentistry for caries prevention. It is known that the accumulation of the bacteria in the enamel microcracks can be reason of caries. A promising substance for early diagnostics of the accumulation of pathogenic microflora on the tooth enamel surface is aluminum phthalocyanine (AlPc). It could be observed that AlPc does not fluoresce in the nanoparticles form but in the monomeric molecular form it does. This allows identification of local pathological microflora accumulation within microcracks of the tooth enamel because AlPc nanoparticles (nAlPc) can be activated by pathological microflora. This paper describes the nAlPc application for fluorescent diagnostics (FD) of the enamel surface in vitro. To reduce the time from the beginning of interaction of nAlPc with the microflora to the appearance of nAlPc fluorescence, Protelan MST-35 surfactant was used as an additional activator. For FD in dentistry a model compound with nAlPc, Protelan MST-35 and with the complementary components was prepared. The following components were used as complementary: the methylparaben, carbopol, carboxymethyl cellulose, titanium dioxide, sodium phosphate, sodium saccharin and sorbitol, which are commonly used in toothpastes. Human teeth were used for the investigation of the interaction between nAlPc colloid and the model compound with nAlPc for the detection of microcracks and the areas of accumulation of pathogenic microflora on the enamel surface. Statistical processing of the experimental results showed the effectiveness of the surfactant usage for the additional activation of nAlPc and a reduction in the FD time. The application of nAlPc as a marker will make it possible to detect microcracks of the enamel tooth surface at the earliest stages and the areas of the pathogenic microflora accumulation, which can lead to the development of a caries. Keywords: fluorescent diagnostics, nanoparticles, aluminum phthalocyanine, Protelan MST-35, surfactants, enamel microcracks, tooth enamel

Активация наночастиц фталоцианина алюминия для локальной флуоресцентной спектроскопии в стоматологии

Early diagnosis of caries and tooth enamel microcracks is of great importance for preventing the destruction of healthy tooth enamel. Inorder to detect microcracks in the enamel and pathogenic microflora foci that can cause caries, nanoform of aluminum phthalocyanine (AlPc) can be used as a marker. In a colloidal solution, the nanoparticles do not fluoresce, unlike their molecular form. To convert the particle into its molecular form, it is necessary to have a solvent or specific environment (bacteria, macrophages, etc.). That is why the hydrophobic nanoparticles of aluminum phthalocyanine (nAlPc) can act as markers for detecting hidden pathogenic microflora during fluorescent diagnostics. Further reduction of the diagnosis time and increase the efficiency can be achieved by using biologically compatible surfactants as additional activators of nAlPc.In order to carry out local fluorescence spectroscopy of enamel microcracks and pathogenic microflora foci on the enamel surface, a model compound containing surfactants, auxiliary components and nAlPc colloid at a concentration of 10 mg/l was prepared.Studies on the interaction of the model compound with nAlPc and Protelan MST-35 with tooth enamel ex vivo have shown this surfactant to be a promising auxiliary activator of the nanoparticles, allowing conducting local fluorescence spectroscopy of the tooth enamel surface 3 min after application. In addition, statistical processing of the results showed the effectiveness of using the model compound for local fluorescence spectroscopy of the enamel surface in order to detect the enamel microcracks and the pathogenic microflora accumulation foci that can lead to the development of a cariogenic process.Для выявления микротрещин эмали и очагов скопления патогенной микрофлоры, которые могут стать причиной развития кариеса, в качестве маркера используется фталоцианин алюминия (AlPc) в виде наночастиц. В коллоидном растворе наночастицы не обладают собственной флуоресценцией, в отличие от молекулярной формы. Для перевода частицы в молекулярную форму необходимо присутствие растворителя или специфического окружения (бактерии, макрофаги и др.). Поэтому гидрофобные наночастицы фталоцианина алюминия (nAlPc) могут выступать в качестве маркера для обнаружения скрытых очагов скопления патогенной микрофлоры во время проведения флуоресцентной диагностики. Для сокращения времени диагностики и увеличения эффективности в качестве дополнительных активаторов nAlPc могут быть использованы биологически совместимые поверхностно-активные вещества (ПАВ).Для проведения локальной флуоресцентной спектроскопии микротрещин эмали и очагов скопления патогенной микрофлоры на поверхности эмали была приготовлена модельная смесь, содержащая ПАВ, вспомогательные компоненты и коллоида nAlPc в концентрации 10 мг/л.Исследования по взаимодействию модельной смеси с nAlPc и протеланом с эмалью зубов ex vivo показали перспективность использования этого ПАВ для дополнительной активации наночастиц, что позволяет проводить локальную флуоресцентную спектроскопию поверхности эмали зубов через 3 мин после нанесения. Также статистическая обработка результатов показала эффективность использования модельной смеси для локальной флуоресцентной спектроскопии поверхности эмали для выявления микротрещин эмали и очагов скопления патогенной микрофлоры, которая может привести к развитию кариесогенного процесса

Взаимосвязь спектроскопических и структурных свойств j-агрегатов индоцианина зеленого

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.Индоцианин зеленый (ICG), находясь в растворе, способен образовывать стабильные структуры наночастиц или коллоидный раствор, изменяя при этом свои спектроскопические свойства. В работе различными методами, основанными на светорассеянии, были исследованы степень агрегации и средний размер наночастиц в зависимости от концентрации коллоидного раствора наночастиц индоцианина зеленого (ICG NPs) в форме J-агрегатов. Размер наночастиц представляет собой важный параметр с точки зрения клинического применения, так как техника внутривенного введения препаратов, с целью избежания тромбозов микрососудов и эмболии, предусматривает лекарственные формы с включениями, в виде отдельных молекул или их кластеров, не превышающими в диаметре 500 нм. С другой стороны, наночастицы размером менее 30 нм длительно циркулируют в организме и могут проникать в клетки здоровой ткани. В ходе исследований, было установлено, что увеличение концентрации ICG NPs в растворе ведет к увеличению среднего размера спонтанно формируемых J-агрегатов, что в свою очередь ведет к уменьшению коэффициента поглощения в агрегатах. Предположительно, нелинейная зависимость поглощения J-агрегата от его размера, может быть объяснен формированием центров поглощения на поверхности J-агрегата в виде подвижных поверхностных молекул. Был определен пороговый диапазон концентрации молекул ICG, при котором происходит переход от агрегации с увеличением размера с медленным прибавлением молекул J-агрегата ICG в высоту, но с быстрым прибавлением в ширину

ИССЛЕДОВАНИЕ РАСПРЕДЕЛЕНИЯ ХЛОРИНОВЫХ ФОТОСЕНСИБИЛИЗАТОРОВ В ДВУХМЕРНЫХ И ТРЕХМЕРНЫХ КЛЕТОЧНЫХ КУЛЬТУРАХ

The aim of this study was to analyze the distribution of chlorin type photosensitizers (PS): chlorin e6 (Ce6) and dimethyl ester of chlorin e6 (DME), in human  adenocarcinoma HT29 monolayer and multicellular spheroid cell cultures. There is an assumption,  that the chemical modification of Ce6 molecules  causes a change  of intracellular location and the enhanced photosensitizing activity. Indeed, photodynamic therapy  on monolayer cell culture with DME showed two times higher photokilling ability comparing with that of non-modified analogue.Ce6 and  DME biodistribution  processes  in tumor  tissue  were studied  on multicellular tumor  spheroids  model. Total amount  of DME in multicellular  tumor  spheroids  exceeded 1,3 times  accumulation  of Ce6. According  to  fluorescence  microscopy  studies, Ce6 and  DME distribution patterns in the spheroids bulk were similar. Application of 2D and 3D tumor models for the analysis of photosensitizer distribution may allow predicting the photosensitizer biodistribution  features for photodynamic therapy in vivo.Цель  данного  исследования   заключалась  в  оценке  распределения  хлориновых  фотосенсибилизаторов  –  хлорина  е6    (Ce6) и  его диметилового  эфира  (DME) – в клетках НТ29 аденокарциномы человека  в моделях  монослоя  клеток и мультиклеточных сфероидах. Согласно полученным данным, DME накапливается  в 2,3 раза  более  эффективно в клетках монослоя  по сравнению  с Ce6. Высказано предположение,  что химическая модификация молекул Ce6 приводит к изменению внутриклеточной локализации, а также к увеличению фотосенсибилизирующей активности. Так, проведение фотодинамической терапии на клетках монослоя с использованием DME показало увеличение способности к фотоповреждению  более чем в 2 раза по сравнению с его немодифицированным аналогом.Процессы распределения DME и Ce6 в опухолевой  ткани были изучены на модели  мультиклеточных опухолевых сфероидов.  Общее количество  DME в опухолевых сфероидах  превышало  накопление  Ce6 в 1,3 раза.  Согласно данным  флуоресцентной  микроскопии, характер  распределения Ce6 и DME в сфероидах  не отличается. Использование  двухи трёхмерных моделей  для анализа  процессов связывания фотосенсибилизаторов может позволить прогнозировать особенности процессов распределения фотосенсибилизаторов при проведении фотодинамической терапии in vivo

МЕТОДЫ ФОТОНИКИ ДЛЯ ОЦЕНКИ КАЧЕСТВА ПРИЖИВЛЕНИЯ КОЖНЫХ ТРАНСПЛАНТАТОВ

 In this review, based on more than 70 articles of Russian and foreign authors, methods of skin engraftment monitoring are discussed. Main processes occurring in skin on cellular and subcellular levels at different stages of engraftment are considered. Optical methods which allow performing non-invasive analysis of blood vessels, collagen concentration and form of cellular respiration (by NADH fluorescence) are described. Comparative analysis of nuclear and optical methods for engraftment monitoring highly developed and widespread nowadays is presented. The advantages of optical methods includes multifunctionality, usability and clarity of results, safety and low cost. In contrast to X-ray CT, MRI and ultrasound, optical methods can be used in monitoring mode. One of the promising directions for improving quality of engraftment due to antibacterial effect, photodynamic therapy, is described in details. The use of crystalline organic nanophotosensitizers (particularly aluminum phthalocyanine) is shown to be the most promising. The main distinctive feature of its application is that nanoparticles injected into wound surface or contact area of tissue graft are not photoactive until the moment the inflammation starts. The development of method for assessing skin condition by spectroscopic properties of tissue components (using fluorescent dyes and photosensitizers in molecular and nanoforms), which allows analyzing physiological state of skin (degree and rate of engraftment or rejection) and controlling certain biochemical and physiological parameters of a tissue graft or an entire area of affected skin is shown to be crucial.Настоящий обзор, основанный более чем на 70 проанализированных статьях российских и зарубежных авторов, посвящен методам мониторинга состояния трансплантируемых участков кожи. В обзоре рассмотрены основные процессы, происходящие в коже на клеточном и субклеточном уровнях на разных этапах приживления трансплантата. Описаны оптические методы, позволяющие проводить неинвазивный анализ состояния кровеносных сосудов, концентрации коллагена, типа клеточного дыхания (по флуоресценции NADH). Приведена сравнительная таблица ядерных и оптических методов мониторинга состояния приживления трансплантата, наиболее широко развитых и использующихся на сегодняшний день. Отмечено, что преимущество оптических методов заключается в многофункциональности, простоте использования и интерпретации результатов, безопасности, низкой стоимости, а также в возможности использования в режиме мониторинга, в отличие от рентгено-компьютерной томографии, МРТ и УЗИ. Подробно рассмотрено одно из перспективных направлений, используемых для улучшения качества приживления трансплантатов преимущественно за счет антимикробного эффекта – фотодинамическая терапия. Отмечено, что перспективным является использование для фотодинамической терапии кристаллических наночастиц органических фотосенсибилизаторов, в частности фталоцианина алюминия. Основная отличительная особенность их использования заключается в том, что наночастицы, введенные в область раневой поверхности или в контактную область приживляемой ткани или трансплантата, не проявляют фотоактивности до момента начала воспаления. Сделан вывод, что актуальной является задача по разработке метода оценки состояния кожного покрова по спектроскопическим свойствам компонентов тканей (с использованием флуоресцентных красителей и фотосенсибилизаторов в молекулярной и наноформах), который позволит анализировать физиологическое состояние кожного покрова (степень и скорость приживления или отторжения), а также контролировать некоторые биохимические и физиологические параметры трансплантата или всей области поражения кожи

Assessment of the efficiency of energy- and resource-saving technologies in the model of open innovation

© The Authors, published by EDP Sciences. The relevance of this work is determined by the fact that the issues of resource-saving technologies implementation in the model of open innovation have not been fully addressed yet and require further study and systematization of the determining factors, which is especially important on the back of the transition to a new technological pattern and the use of the emerging technological opportunity windows. The solution of the problems mentioned will reveal new opportunities for qualitative and quantitative growth of production systems by improving the innovation targeting in the field of resource saving and energy efficiency. The purpose of the article is to identify the functional dependence between the Networked Readiness Index and the emerging technological opportunity windows in order to improve the efficiency of resource-saving technologies in open innovation and production systems. The main research methods underlying the article include the method of description used to identify trends in the Networked Readiness Index across the globe, the correlation analysis method used to identify a close relationship between the Networked Readiness Index and indicators of state support granted to the innovations in production, and the regression analysis method used to build a regression model of the dependence between the resource-saving system and production indicators. The article touches upon the aspects of improving the resource-saving system efficiency in the framework of the open innovation model in the field of production. The correlation between the Networked Readiness Index and the indicators of the state institutions quality characterizing the level of innovation support provided to the industry across the world countries has been revealed, and the functional connection between the use of waste at petrochemical plants and the petrochemical products shipped has been proved using the emerging economies as an example. The materials of the article can be used in the development of strategies and programs aimed to improve the resource-saving system efficiency in petrochemical companies of developing countries, taking into account the emerging technological opportunity windows and technology readiness of the production for innovative transformations

Assessment of the efficiency of energy- and resource-saving technologies in the model of open innovation

© The Authors, published by EDP Sciences. The relevance of this work is determined by the fact that the issues of resource-saving technologies implementation in the model of open innovation have not been fully addressed yet and require further study and systematization of the determining factors, which is especially important on the back of the transition to a new technological pattern and the use of the emerging technological opportunity windows. The solution of the problems mentioned will reveal new opportunities for qualitative and quantitative growth of production systems by improving the innovation targeting in the field of resource saving and energy efficiency. The purpose of the article is to identify the functional dependence between the Networked Readiness Index and the emerging technological opportunity windows in order to improve the efficiency of resource-saving technologies in open innovation and production systems. The main research methods underlying the article include the method of description used to identify trends in the Networked Readiness Index across the globe, the correlation analysis method used to identify a close relationship between the Networked Readiness Index and indicators of state support granted to the innovations in production, and the regression analysis method used to build a regression model of the dependence between the resource-saving system and production indicators. The article touches upon the aspects of improving the resource-saving system efficiency in the framework of the open innovation model in the field of production. The correlation between the Networked Readiness Index and the indicators of the state institutions quality characterizing the level of innovation support provided to the industry across the world countries has been revealed, and the functional connection between the use of waste at petrochemical plants and the petrochemical products shipped has been proved using the emerging economies as an example. The materials of the article can be used in the development of strategies and programs aimed to improve the resource-saving system efficiency in petrochemical companies of developing countries, taking into account the emerging technological opportunity windows and technology readiness of the production for innovative transformations

INVESTIGATION OF CHLORIN PHOTOSENSITIZERS DISTRIBUTION IN MONOLAYER AND SPHEROID CELL CULTURES

The aim of this study was to analyze the distribution of chlorin type photosensitizers (PS): chlorin e6 (Ce6) and dimethyl ester of chlorin e6 (DME), in human  adenocarcinoma HT29 monolayer and multicellular spheroid cell cultures. There is an assumption,  that the chemical modification of Ce6 molecules  causes a change  of intracellular location and the enhanced photosensitizing activity. Indeed, photodynamic therapy  on monolayer cell culture with DME showed two times higher photokilling ability comparing with that of non-modified analogue.Ce6 and  DME biodistribution  processes  in tumor  tissue  were studied  on multicellular tumor  spheroids  model. Total amount  of DME in multicellular  tumor  spheroids  exceeded 1,3 times  accumulation  of Ce6. According  to  fluorescence  microscopy  studies, Ce6 and  DME distribution patterns in the spheroids bulk were similar. Application of 2D and 3D tumor models for the analysis of photosensitizer distribution may allow predicting the photosensitizer biodistribution  features for photodynamic therapy in vivo

PHOTONIC METHODS FOR QUALITY EVALUATION OF SKIN ENGRAFTMENT

 In this review, based on more than 70 articles of Russian and foreign authors, methods of skin engraftment monitoring are discussed. Main processes occurring in skin on cellular and subcellular levels at different stages of engraftment are considered. Optical methods which allow performing non-invasive analysis of blood vessels, collagen concentration and form of cellular respiration (by NADH fluorescence) are described. Comparative analysis of nuclear and optical methods for engraftment monitoring highly developed and widespread nowadays is presented. The advantages of optical methods includes multifunctionality, usability and clarity of results, safety and low cost. In contrast to X-ray CT, MRI and ultrasound, optical methods can be used in monitoring mode. One of the promising directions for improving quality of engraftment due to antibacterial effect, photodynamic therapy, is described in details. The use of crystalline organic nanophotosensitizers (particularly aluminum phthalocyanine) is shown to be the most promising. The main distinctive feature of its application is that nanoparticles injected into wound surface or contact area of tissue graft are not photoactive until the moment the inflammation starts. The development of method for assessing skin condition by spectroscopic properties of tissue components (using fluorescent dyes and photosensitizers in molecular and nanoforms), which allows analyzing physiological state of skin (degree and rate of engraftment or rejection) and controlling certain biochemical and physiological parameters of a tissue graft or an entire area of affected skin is shown to be crucial.</p

Assessment of the efficiency of energy-and resource-saving technologies in the model of open innovation

The relevance of this work is determined by the fact that the issues of resource-saving technologies implementation in the model of open innovation have not been fully addressed yet and require further study and systematization of the determining factors, which is especially important on the back of the transition to a new technological pattern and the use of the emerging technological opportunity windows. The solution of the problems mentioned will reveal new opportunities for qualitative and quantitative growth of production systems by improving the innovation targeting in the field of resource saving and energy efficiency. The purpose of the article is to identify the functional dependence between the Networked Readiness Index and the emerging technological opportunity windows in order to improve the efficiency of resource-saving technologies in open innovation and production systems. The main research methods underlying the article include the method of description used to identify trends in the Networked Readiness Index across the globe, the correlation analysis method used to identify a close relationship between the Networked Readiness Index and indicators of state support granted to the innovations in production, and the regression analysis method used to build a regression model of the dependence between the resource-saving system and production indicators. The article touches upon the aspects of improving the resource-saving system efficiency in the framework of the open innovation model in the field of production. The correlation between the Networked Readiness Index and the indicators of the state institutions quality characterizing the level of innovation support provided to the industry across the world countries has been revealed, and the functional connection between the use of waste at petrochemical plants and the petrochemical products shipped has been proved using the emerging economies as an example. The materials of the article can be used in the development of strategies and programs aimed to improve the resource-saving system efficiency in petrochemical companies of developing countries, taking into account the emerging technological opportunity windows and technology readiness of the production for innovative transformations