RU-2861555-C1 - METHOD FOR INACTIVATION OF STAPHYLOCOCCUS AUREUS, KLEBSIELLA PNEUMONIAE, PSEUDOMONAS AERUGINOSA STRAINS UNDER EXPERIMENTAL CONDITIONS IN VITRO USING ANTIMICROBIAL PHOTODYNAMIC THERAPY WITH PHOTOSENSITISER IN AEROSOL FORM

Abstract

FIELD: medicine; pulmonology; microbiology. SUBSTANCE: invention can be used for the inactivation of Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa strains under experimental conditions in vitro. The method includes the use of antimicrobial photodynamic therapy using a photosensitiser in aerosol form. In the nebuliser chamber, 5 ml of 0.5% Photoditazine are diluted in 5 ml of a combined preparation of 7% NaCl solution and 0.1% sodium hyaluronate solution, activated using a multifunctional LED photoactivator with monochromatic light with a wavelength of 660 nm and an exposure dose of 1000 J/cm 2 , then nebulisation is carried out using a tube connected to the nebuliser chamber and to the Petri dish lid, the aerosol dose is 0.45 ml. EFFECT: reducing the absolute number of colony-forming units of bacterial conglomerates of Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa. 1 cl, 1 dwg, 1 tbl

Inventors

• Tsejmakh Aleksandr Evgenevich
• Tsejmakh Irina Yakovlevna
• Prokopev Vasilij Valerevich
• Timonin Andrej Viktorovich

Dates

Publication Date

20260505

Application Date

20250731

Claims (1)

1. A method for inactivating strains of Staphylococcus aureus, Klebsiella pneumonia, Pseudomonas aeruginosa under experimental conditions in vitro, carried out by applying antimicrobial photodynamic therapy using a photosensitizer in aerosol form, characterized in that 5 ml of 0.5% Photoditazin are diluted in 5 ml of a combined preparation of a 7% NaCl solution and a 0.1% sodium hyaluronate solution in a nebulizer container, activated using a multifunctional LED photoactivator with monochromatic light with a wavelength of 660 nm and an exposure dose of 1000 J/ cm2 , then nebulization is carried out using a tube connected to the nebulizer chamber and to a Petri dish lid, the aerosol dose is 0.45 ml.

Description

The invention relates to the field of medicine, namely to pulmonology and microbiology, and can be used to inactivate surface-attached microorganisms that form bacterial biofilms. According to modern concepts, a biofilm is a structurally organized community of microorganisms enclosed within an exopolymer matrix synthesized by the bacteria themselves and attached to biological or inorganic surfaces (Glushanova N. A., Blinov A. I., Alekseeva N. B. Bacterial biofilms in human infectious pathology. MvK. 2015. Special issue No. 2. URL: https://cyberlenirika.ru/artcheloveka; Ilyina T. S., Romanova Yu. M. Bacterial biofilms: role in chronic infectious processes and the search for means to combat them. Molecular genetics, microbiology and virology. 2021; 39 (2): 14-24). The formation of bacterial biofilms is associated with the resistance of a significant portion of the microorganisms that form these structures to antibiotics due to the slowdown or inability of medicinal compounds to penetrate the surface membrane and components of the polysaccharide matrix deep into the biofilm, slowdown in binding to bacterial cells and mechanisms of bacterial inactivation, slowdown in the rate of division and growth of bacteria, and the emergence of phenotypically resistant variants of bacteria within biofilms. Bacterial biofilm formation plays an important role in reducing the effectiveness of antimicrobial therapy and worsening respiratory outcomes in respiratory diseases, including cystic fibrosis, non-cystic fibrosis bronchiectasis, chronic obstructive pulmonary disease, and ventilator-associated pneumonia (Boisvert AA, Cheng MP, Sheppard DC, Nguyen D. Microbial Biofilms in Pulmonary and Critical Care Diseases. Ann Am Thorac Soc. 2016;13(9): 1615–1623; Ding L, Wang J, Cai S, Smyth H, Cui Z. Pulmonary biofilm-based chronic infections and inhaled treatment strategies. Int J Pharm. 2021; 604:120768). Most bacterial pathogens associated with severe course and adverse outcomes of these diseases have the ability to adhere and form biofilms on the respiratory mucosa, including Staphylococcus aureus, Klebsiella pneumonia, and Pseudomonas aeruginosa. Photodynamic inactivation of microorganisms is considered in a number of studies as a promising method for eradicating antibiotic-resistant microbial isolates and biofilms. The use of an exogenous photosensitizer or irradiation of microbial cells already supplied with endogenous photosensitizers makes photodynamic therapy a convenient tool for treating infections. The main disadvantage of photodynamic therapy is the need to deliver light radiation directly to the site of photosensitizer concentration, which makes the treatment of lower respiratory tract infections difficult (Rapacka-Zdo'nczyk A, Wo'zniak A, Michalska K. et al. Factors Determining the Susceptibility of Bacteria to Antibacterial Photodynamic Inactivation. Front. Med. 2021; 8:642609. doi: 10.3389/fmed.2021.642609). There are known methods for inactivating bacterial microorganisms and combating the formation of bacterial biofilms in the form of using hydrogen peroxide ( H2O2 ) in non-thermal technology for decontamination of food products; for deactivation of whole- cell bacteria by H2O2 in the development of bacterial vaccines; to combat bacterial biofilms during disinfection (Rosenzweig Z, Garcia J, Thompson GL, Perez LJ. Inactivation of bacteria using synergistic hydrogen peroxide with split-dose nanosecond pulsed electric field exposures. PLoS One. 2024; 19(11): e0311232; Fan Y, Mu Y, Lu L, Tian Y, Yuan F, Zhou B, Yu C, Wang Z, Li X, Lei S, Xu Y, Wu D, Yang L. Hydrogen peroxide-inactivated bacteria induces potent humoral and cellular immune responses and releases nucleic acids. Int Immunopharmacol. 2019; Labas MD, Zalazar CS, Cassano AE Reaction kinetics of bacteria disinfection employing hydrogen peroxide. 38: 78-87). These methods are based on the damage of proteins, DNA, and lipids of the bacterial cell wall by active oxygen radicals. These methods can only be used extracorporeally, since H2O2 can be harmful when exposed dermally or ophthalmologically, ingested, used for wound irrigation, by injection, or by inhalation, depending on the concentration of the solution and the type of exposure (Nelson AL, Porter L. Hydrogen Peroxide Toxicity. 2023 Aug 8. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025 Jan-. PMID: 36256749). A method for photodynamic inactivation of Mycobacterium tuberculosis by chlorin E6 (in the medicinal form of radochlorin) in vitro is known (Bredikhin D.A., Nikonov S.D., Cherednichenko A.G., Petrenko T.I. Photodynamic inactivation of Mycobacterium tuberculosis by radochlorin in vitro. Tuberculosis and lung diseases. 2018. Vol. 96, No. 1. pp. 5-10). In an experimental study conducted by the authors, a dose-dependent photoinactivation of the pathogen was established with light energy with a wavelength of 662 nm, while the effect of light radiation to activate the photosensi