DETERMINATION OF BIOFILM FORMING ACTIVITY OF MICROORGANISMS ON SYNTHETIC POLYMERIC MATERIALS
Authors: Godovalov A. P. , Stepanov M. S. , Yakovlev M. V. , Kobzarenko E. E. , Batog K. A. 
Published: February 2020

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.