The stability of the matrix of mono- and multi-species biofilms, formed by periodontal pocket microflora, in static and dynamic conditions of the environment in vitro and their antibiotic resistance
Vitebsk State Order of Peoples’ Friendship Medical University, Vitebsk, Republic of Belarus
Vestnik VGMU. 2017;16(5):136-144.
Objectives. To study the stability of the matrix of mono- and multi-species biofilms formed by periodontal pocket microflora in the static and dynamic conditions of the environment in vitro to enzymes, antiseptics, biological fluids and their antibiotic resistance.
Material and methods. Microbial biofilms of 10 clinical isolates, obtained from patients with chronic periodontitis as well as 15 isolates isolated in purulent-inflammatory diseases were studied. The ability to destroy the suspension of the exopolymer matrix of mono- and multi-species biofilms in static and dynamic conditions by the following enzymes and antiseptics was determined: alpha-DNase, hyaluronidase type I, trypsin, proteinase K, alpha-amylase pancreatic, acetylcysteine, 0,05% and 2% chlorhexedin, dimethylsulfoxide 25% hydrogen peroxide 3%. The antibiotic resistance of plankton forms of Streptococcus spp and also in the composition of biofilm to antibiotics has been determined.
Results. When studying the effect of antiseptics and enzymes on the biofilms matrix, it has been found that 25% dimethylsulfoxide and proteinase K were the most capable of destroying the exopolymer matrix. According to the data obtained, the matrix of monobiofilm Streptococcus spp (p <0,05) was more sensitive to most enzymes and antiseptics. The sensitivity of multi-species biofilms corresponds to the sensitivity of the most stable matrix. The dynamic environmental conditions affect the biofilm formation processes, increasing its resistance and stability to antiseptics and enzymes. When studying the sensitivity of Streptococcus spp. in the composition of biofilm to antibacterial drugs, a decrease in the sensitivity of bacteria to all the investigated antibodies with regard to plankton forms was revealed. The correlation between the biofilm weight and the level of resistance to antibacterial drugs was found. Thus microorganisms the ability of which to form a biofilm is higher are more resistant to antibacterial drugs.
Key words: biofilm, chronic periodontitis, antibiotics, antiseptics, enzymes, oral fluid.
1. Trakhtenberg ISh, Rubshteyn AP, Lemkina LM, Korobov VP, Morozov IA A. Formation of biomembranulas of staphilococci on the surface of titanium and a titanium with carbon diamondlike Membranula and action on them a low-molecular cationic peptide of a varnerin. Perspektiv Materialy. 2013;(4):39-44. (In Russ.)
2. An YH, Friedman RJ, Draughn RA, Smith EA, Nicholson JH, John JF. Rapid quantification of staphylococci adhered to titanium surfaces using image analyzed epifluorescence microscopy. J Microbiol Methods. 1995 Nov;24(1):29-40. doi: http://dx.doi.org/10.1016/0167-7012(95)00051-8
3. Sadovskaya I, Vinogradov E, Flahaut S, Kogan G, Jabbouri S. Extracellular carbohydrate-containing polymers of a model biofilm-producing strain, Staphylococcus epidermidis RP62A. Infect Immun. 2005 May;73(5):3007-17.
4. Kolchanova NE, Okulich VK, Shilin VE. Definition of formation of microbial biomembranula bacteria of a periodontal pocket and its fastness to chemical and biological objects. Immunopatologiia Allergologiia Infektologiia. 2015;(3):56-61. (In Russ.)
Information about authors:
Kalchanava N.E. – postgraduate of the Chair of Clinical Microbiology, Vitebsk State Order of Peoples’ Friendship Medical University.