Microorganisms are able to form biofilms on surfaces of biotic and abiotic nature. In turn, in human biotopes there are optimal conditions for the implementation of biofilm-forming activity. Moreover, in medical practice, polymeric materials are often used for drainage or prosthetics, which can also be successfully colonized by bacteria. However, in laboratory practice, the formation of biofilms is usually evaluated on glass or polystyrene. The purpose of the study is to evaluate the methodological features of studying the biofilm-forming activity of microorganisms on the surface of synthetic polymeric materials. We used strains of Staphylococcus aureus ATCC 25923, Escherichia coli K-12, Candida albicans ATCC 10231, as well as synthetic polymeric materials - DentLight Flow light-curing composite material (nano-hybrid fluid composite; Russia), glass ionomer chemical curing Fuji 1 (Japan), cement for temporary fixation of orthopedic constructions TempBond NE (USA), acrylic, polyurethane and polyvinyl chloride. The formation of biofilms in flat-bottomed ELISA plates in this study was considered as a control group. If the polymer belonged to cold curing materials, sterile flat-bottomed tablets were used, the bottom of which was filled with a thin layer of plastic. After hardening of the plastic, biofilms were formed in the tablets. In the second series of experiments, hot cured materials cut into equal parts 5×5×1 mm in size were placed in the wells of a plate and again used to determine biofilm formation with subsequent coloring. To extract the dye, the pieces were transferred to a new plate to exclude the amount of film biomass formed on the walls of the plate wells. In both cases, cultivation was carried out at 37° C for 24-48 hours. The biomass of the film was stained with fuchsin. Statistical data processing was performed using t-Student criterion. For the threshold level of significance, the value p <0.05 was taken. It is established that the proposed options for determining biofilm forming ability are available and indicative. It was revealed that the same microorganisms have individual biofilm formation indicators for each polymer material. The light curing dental composite and polyvinyl chloride exhibit the more pronounced antiadhesive properties than cements and polyurethane. Up to date, most of the studies of biofilm formation have been carried out using glass or polystyrene, which, as a rule, are not used for the manufacture of prostheses, catheters, drains, etc., which makes it difficult to assess the true film-forming activity of microorganisms. The proposed methodological approaches, especially the second option for preparing testing samples, solve this problem. In general, the proposed approaches to testing biofilm-forming activity on polymers are very simple to implement and generally available. For an adequate study of the biofilms formation, it will be advisable to use polymer materials, directly used in medicine, rather than polystyrene tablets, the material of which is found exclusively in laboratory practice.
Microbial biofilms are heterogeneous, moving and constantly changing communities of microorganisms, often of various taxons. Approaches to study and assessment anti-biofilm drugs widely available today do not adequately assess their effects, while the results of studying the interaction of drugs with components of the film composition can provide them the right choice. The aim of investigation was to test a new method of morphological evaluation of biofilms. To form biofilms, we used an approach when the slide was placed at an angle of 30o-45o relatively to the Petri dish, and a suspension of test strains S. epidermidis in peptone broth was poured into the space between the Petri dish and the slide. A sterile cotton swab moistened with distilled water was placed next to the glass slide to create optimal humidity. The system was placed in a thermostat for 24 hours. The formed films were examined under a microscope using the DCM 310 video eyepiece and the Scope photo x86,3.1.312 program that allowed to conduct a complete morphometric study of the film: select layers, channels, cavities and make measurements, and then save the results on electronic media in jpg file format. Microscopy of the stained slides revealed that the biofilm has a layered structure. In each image obtained using a video eyepiece, it was possible to differentiate 4 layers. From the border of the two media to the inside: the fragmentation layer, the dense layer, the matrix substance layer, and the last one - the persistence layer. Channels of different diameters (from 10 to 24 microns) are observed across the entire thickness of the biofilm. Thus, used approach allows us to visualize and evaluate the structure of microbial biofilm, measure the thickness of layers and channel diameters. In addition, this method can be used to study the effect of antimicrobial drugs on bacterial films.
Aim. To study and compare the features of formation of biofilms S. aureus and S. epidermidis on synthetic materials from polyvinylchloride and polystyrene. Materials and methods. In the experiments, S. aureus and S. epidermidis strains as well as polymeric synthetic materials were used. Expression of biofilm formation of these strains on polystyrene and polyvinylchloride was studied. Biofilm formation was determined in sterile flat-bottomed tablets. Into a part of tablet holes, pieces of polyvinylchloride measuring 551 mm were placed. They were cultivated at the temperature 37 C for 2448 hours. Results. On polyvinylchloride, biofilm-forming ability of staphylococci was practically absent. S. aureus is more active in formation of biofilms as compared with S. epidermidis. As incubation period is increased, S. aureus elevates biofilm-forming activity but S. epidermidis does not. Conclusions. It was established that one and the same microorganisms are different in manifestation of biofilm-forming activity on polystyrene and polyvinylchloride that can be conditioned by various chemical structure of these materials.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.