Abstract
Background: The problem of bacterial colonization of implants used in medical practice continues to be relevant regardless of the material of the implant. Particular attention deserves polymeric implants, which are prepared ex tempore from polymethyl methacrylate, for example - duting orthopedic surgical interventions (so-called "bone cement"). The protection of such implants by antibiotic impregnation is subjected to multiple criticisms, therefore, as an alternative to antibiotics, lytic bacteriophages with a number of unique advantages can be used - however, no experimental studies have been published on the possibility of impregnating bacteriophages into polymethyl methacrylate and their antibacterial activity assessment under such conditions.Aims: to evaluate the possibility of physical placement of bacteriophages in polymethylmethacrylate and to characterize the lytic antibacterial effect of two different strains of bacteriophages when impregnated into polymer carrier ex tempore during the polymerization process in in vitro model.Materials and methods: First stage - Atomic force microscopy (AFM) of polymethyl methacrylate samples for medical purposes was used to determine the presence and size of caverns in polymethyl methacrylate after completion of its polymerization at various reaction temperatures (+6…+25°C and +18…+50°C).The second stage was performed in vitro and included an impregnation of two different bacteriophage strains (phage ph20 active against S. aureus and ph57 active against Ps. aeruginosa) into polymethyl methacrylate during the polymerization process, followed by determination of their antibacterial activity.Results: ACM showed the possibility of bacteriophages placement in the cavities of polymethyl methacrylate - the median of the section and the depth of cavities on the outer surface of the polymer sample polymerized at +18…+50°C were 100.0 and 40.0 nm, respectively, and on the surface of the transverse cleavage of the sample - 120.0 and 100.0 nm, respectively, which statistically did not differ from the geometric dimensions of the caverns of the sample polymerized at a temperature of +6…+25°C.The study of antibacterial activity showed that the ph20 bacteriophage impregnated in polymethyl methacrylate at +6…+25°C lost its effective titer within the first six days after the start of the experiment, while the phage ph57 retained an effective titer for at least 13 days.Conclusion: the study confirmed the possibility of bacteriophages impregnation into medical grade polymethyl methacrylate, maintaining the effective titer of the bacteriophage during phage emission into the external environment, which opens the way for the possible application of this method of bacteriophage delivery in clinical practice. It is also assumed that certain bacteriophages are susceptible to aggressive influences from the chemical components of "bone cement" and / or polymerization reaction products, which requires strict selection of bacteriophage strains that could be suitable for this method of delivery.
Highlights
The problem of bacterial colonization of implants used in medical practice continues to be relevant regardless of the material of the implant
Particular attention should be given to polymeric implants which are produced “ex tempore” from polymethyl methacrylate, for example, during orthopedic surgical interventions. The protection of such implants by antibiotic impregnation is subjected to multiple criticisms; as an alternative to antibiotics, lytic bacteriophages with a number of unique advantages can be used ― no experimental studies have been published on the possibility of impregnating bacteriophages into polymethyl methacrylate and their antibacterial activity assessment under such conditions
ACM showed the possibility of bacteriophages placement in the cavities of polymethyl methacrylate ― the median of the section and the depth of cavities on the outer surface of the polymer sample polymerized at +18...+50°C were 100.0 and 40.0 nm, respectively, and on the surface of the transverse cleavage of the sample ― 120.0 and 100.0 nm, respectively, which statistically did not differ from the geometric dimensions of the caverns of the sample polymerized at a temperature of +6...+25°C
Summary
Отдельного внимания заслуживают имплантируемые в организм человека полимерные имплантаты, которые изготовляют ex tempore (по мере надобности) из полиметилметакрилата, например при ортопедических хирургических вмешательствах (так называемый костный цемент). Цель исследования ― изучить возможность физического размещения бактериофагов в полиметилметакрилате и в модели in vitro охарактеризовать литический антибактериальный эффект двух различных штаммов бактериофагов при их импрегнации в изготавливаемый ex tempore полимерный носитель на этапе полимеризации. Вторым этапом in vitro было проведена импрегнация двух различных штаммов бактериофагов (ph, активного в отношении Staphylococcus aureus, и ph, активного в отношении Pseudomonas aeruginosa) в полиметилметакрилат на этапе полимеризации с последующим определением их антибактериальной активности. В исследовании была подтверждена возможность импрегнации бактериофагов в полиметилметакрилат медицинского назначения с поддержанием эффективного титра бактериофага при его эмиссии во внешнюю среду, что открывает пути возможного применения такого способа доставки бактериофагов в клинической практике. (Для цитирования: Самохин А.Г., Козлова Ю.Н., Корнеев Д.В., Таранов О.С., Фёдоров Е.А., Павлов В.В., Тикунова Н.В.
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