biophBiomedical PhotonicsBiomedical Photonics2413-9432Non-profit partnership for development of domestic photodynamic therapy and photodiagnosis10.24931/2413-9432-2025-14-3-4-13bioph-729Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESИсследование фотодинамической активности метиленового синего на суспензиях эритроцитов in vitroStudy of methylene blue photodynamic activity on erythrocyte suspensions in vitroМарковаИ. В.MarkovaI. V.

Москва

Moscow

РябоваА. В.RyabovaA. V.

Moscow

РоманишкинИ. Д.RomanishkinI. D.

Moscow

ПоминоваД. В.PominovaD. V.

Moscow

pominovadv@gmail.comИнститут общей физики им. А. М. Прохорова Российской академии наук; Национальный исследовательский ядерный университет «МИФИ»РоссияProkhorov General Physics Institute of Russian Academy of Sciences; National Research Nuclear University MEPhI (Moscow Engineering Physics Institute)Russian FederationИнститут общей физики им. А. М. Прохорова Российской академии наукРоссияProkhorov General Physics Institute of Russian Academy of SciencesRussian Federation202517102025143413Copyright © Маркова И.В., Рябова А.В., Романишкин И.Д., Поминова Д.В., 20252025Маркова И.В., Рябова А.В., Романишкин И.Д., Поминова Д.В.Markova I.V., Ryabova A.V., Romanishkin I.D., Pominova D.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.pdt-journal.com/jour/article/view/729

В работе исследована фотодинамическая активность (по скорости утилизации молекулярного кислорода при облучении) метиленового синего (МС) в суспензиях эритроцитов in vitro. Методами спектроскопии и конфокальной микроскопии с флуоресцентными сенсорами на синглетный кислород и другие активные формы кислорода показано, что при увеличении концентрации МС (10–100 мг/кг) молярная фотодинамическая активность снижается. Установлено, что 5-10% от добавленного к эритроцитам МС прочно связывается с мембранами эритроцитов, а генерация синглетного кислорода (¹O₂) подавляется в пользу реакций I типа (образование H₂O₂, O₂•⁻, •OH). Еще порядка 40% от добавленного к эритроцитам МС переходит в бесцветную лейкоформу, однако при фотодинамическом воздействии окисляется обратно до МС. Максимальный квантовый выход генерации ¹O₂ (φΔ ) в суспензиях эритроцитов составил 0,014 для концентрации МС 10 мг/кг, что на порядок ниже значений для МС в органических растворителях и для фотосенсибилизатора сравнения фотосенс (φΔ = 0,38). Взаимодействие с эритроцитами (агрегация, восстановление в лейкоформу, конкуренция за кислород) объясняет снижение эффективности МС в физиологических условиях по сравнению с органическими растворителями. Полученные результаты важны с точки зрения оптимизации системного применения МС в фотодинамической терапии.

In this paper we studied the photodynamic activity (the rate of molecular oxygen utilization during irradiation) of methylene blue (MB) in erythrocyte suspensions in vitro. Using spectroscopy and confocal microscopy with fluorescent sensors for singlet oxygen and other active oxygen species, it was shown that with an increase in the MB concentration (10–100 mg/kg), the molar photodynamic activity decreases. It was found that 5–10% of the MB added to erythrocytes tightly binds to the erythrocyte membranes, and the generation of singlet oxygen (¹O₂) is suppressed in favor of type I reactions (formation of H₂O₂, O₂•⁻, •OH). Another 40% of the MB added to erythrocytes is converted into a colorless leuco form, but is reoxidized back to MB under photodynamic exposure. The maximum relative quantum yield of ¹O₂ generation (φΔ ) among those measured in erythrocyte suspensions was 0.014 for a 10 mg/kg MB concentration, which is an order of magnitude lower than the values for MB in organic solvents and for the aluminum sulfonated phthalocyanine comparison photosensitizer (PS) (φΔ = 0.38). Interaction with erythrocytes (aggregation, reduction to the leuco form, competition for oxygen) explains the decrease in the MB efficiency under physiological conditions compared to organic solvents. The obtained results are important from the point of view of optimizing the systemic use of MB in photodynamic therapy.

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The authors declare that there are no conflicts of interest present.