Diagnostic value of microbiome biomarkers of the periodonite microbiome in patients with the association of chronic periodontitis and diabetes mellitus type 2.

The place of high-tech methods of molecular biology in clinical laboratory diagnostics of various diseases and the development of a system of biomarkers as an important component of diagnostic research is currently attracting the closest attention of the scientific community. In this paper, an attempt is made to use high-tech metagenomic analysis to solve problems that arise due to the high frequency of association of periodontal diseases with systemic pathology, in particular, with type 2 diabetes mellitus. The aim of the study was to determine the taxonomic and metabolic features of the microbiome of periodontal tissues in periodontal diseases associated with type 2 diabetes mellitus, as a model of the ratio of local and systemic effects of periodontal pathogenic bacteria. The study included 16S shotgun sequencing of bacterial DNA as part of biological material from periodontal pockets/dentoalveolar furrows of 46 people - 15 patients with chronic periodontitis associated with type 2 diabetes mellitus, 15 patients with chronic periodontitis unrelated to systemic pathology, as well as 16 healthy people in the control group, followed by bioinformatic processing of the data obtained. The obtained data allowed us to establish the taxonomic features of the periodontal microbiome in the association of chronic periodontitis with type 2 diabetes mellitus, which included the predominance of representatives of the families Prevotellaceae and Spirochaetaceae in its composition. The features of metabolic processes in periodontal tissues with the participation of the microbiome were also revealed, which consisted in an increase in the exchange of cysteine and methionine against the background of a decrease in the metabolism of pyrimidine, methane, sphingolipids, and the synthesis of fatty acids, which are of diagnostic value in assessing the condition of patients with type 2 diabetes mellitus.

[Colonization activity of conditionally pathogenic microorganisms and features of biofilm formation on the surface of thermoplastic polymer material]. / Kolonizatsionnaya aktivnost' uslovno-patogennykh mikroorganizmov i osobennosti obrazovaniya bioplenki na poverkhnosti stomatologicheskogo termoplastichnogo polimernogo materiala.

THE PURPOSE OF THE STUDY: Was to assess the microbial colonization and biofilm-forming activity of conditionally pathogenic microorganisms in vitro to samples of acrylic-free thermoplastic polymer material and to assess the possibility of its use for the manufacture of removable tooth splinting structures by injection method. MATERIALS AND METHODS: The reference strains S. aureus, S. epidermidis, S. pyogenes, E. faecalis, E. coli, and C. albicans. Biofilm-forming activity was assessed by the level of ethanol extraction of a 0.1% aqueous solution of gentian violet by measuring on a PowerWave X microplate reader (USA). RESULTS: A low degree of severity of microbial colonization and sufficient colonization resistance to the formation of a microbial biofilm of E. coli (0.374±0.056 cu opt. Pl.), S. aureus (0.272±0.039 cu. Opt. Pl.), S. epidermdis (0.299±0.028 cu opt. Pl.), S. ryogenes (0.399±0.069 cu opt. Pl.), E. faecalis (0.401±0.089 cu opt. Pl.). Moreover, C. albicans strains form a more pronounced biofilm (0.425±0.104 cu opt. Pl.) in comparison with other strains studied in the experiment. CONCLUSION: Samples from a thermoplastic polymer based on polyoxymethylene have satisfactory biomedical characteristics and can be used at the stages of dental orthopedic treatment of patients without oral candidiasis.

[Experimental study of a bacterial adhesion to titanium plates used for osteosynthesis of facial bones]. / Rezul'taty izucheniya adgezii mikroorganizmov k titanovym plastinam, ispol'zuemym pri osteosinteze kostei litsevogo skeleta, v eksperimente.

BACKGROUND: The adhesion ability of microorganisms to the surface of titanium miniplates and screws is one of factors in the development of purulent-inflammatory complications in the postoperative period. One of the solutions to this problem is the use of an ion-plasma coating of mini-plates and screws. OBJECTIVE: The aim of the study was to prove a decrease adhesion level of pathogenic and conditionally pathogenic bacterium to samples of titanium mini-plates with ion-plasma coating, compared with samples without coating. MATERIAL AND METHODS: Present opinion about microbial adhesion, how the most important stage of development microbial colonization of abiotic surfaces, was been basis of this study. The strains of aerobic and facultative anaerobic pathogens, as well as the non-spore-forming (non-clostridial) obligate-anaerobic pathogens were used in the research. During the experiments the standard method for determining residual adhesion was applied. The data obtained were processed by the «Biostat 9.0¼ software package. For the data received by the methods of variational parametric and nonparametric statistics we used the Mann-Whitney test, taking into account the average value, error, and the number of observations (significance of differences at p<0.05). RESULTS: As a result, the differences between the indices residual adhesion of strains aerobic and facultative anaerobic pathogens on the samples with and without a protective coating were statistically reliable, however, the adhesion of the individual species varied. The same differences between the indices residual adhesion were obtained among the strains of obligate anaerobic pathogens. The low level of adhesion, which was observed in the case of using the protective coating, is a very promising way of development, since these microorganisms are dangerous as the may lead to the development of purulent-inflammatory complications and the rejection of implants. CONCLUSION: The decreased level of adhesion is a factor which reduces the microbial load during the postoperative period.

[Features and significance of adhesion of bacteria and fungi of the oral cavity as the initial stage of the formation of a microbial biofilm on dental polymer materials]. / Osobennosti i znachenie adgezii bakterii i gribov polosti rta kak etapa formirovaniya mikrobnoi bioplenki na stomatologicheskikh polimernykh materialakh.

OBJECTIVE: Characteristics of the adhesion of yeast fungi and oral bacteria of various types in vitro to samples of polymeric materials for fixed structures of dental prostheses, obtained using various technologies: adjective digital 3D printing and traditional methods. MATERIAL AND METHODS: Conducted model experiments on the adhesion of bacterial (including the main periodontopic-pathogenic species - P. gingivalis, P. intermedia, etc.) and fungal pathogens (C. albicans) to standard samples of polymer materials NextDent C & B Micro Filled Hybrid («NextDent¼, Netherlands), Detax Freeprint temp UV («Detax¼, Germany), obtained by digital additive 3D printing technology, and Luxatemp Automix Plus («DMG¼, Germany) and Acrytemp («Zhermack¼, Italy) - by the traditional method as a control. Removal of adhering microbes from the material was carried out using an ultrasound machine (exposure time 10 minutes, power 60 kHz). RESULTS: The dependence of the degree of microbial adhesion on the nature of the material and processing technology (3D printing, milling) was established. The materials of NextDent C & B Micro Filled Hybrid and Detax Freeprint temp UV showed high resistance to adhesion of clinical isolates of periodontal pathogenic bacteria and C. albicans fungi. CONCLUSION: The lowest adhesion values for periodontopathogenic species and C. albicans fungi were detected when using samples of materials obtained by 3D printing: NextDent C & B Micro Filled Hybrid and Detax Freeprint temp UV compared to Luxatemp Automix Plus and Acrytemp polymers.

[Control of primary adhesion of microorganisms and formation of biofilms on stomatological materials used for transdental implantation in dental stabilizing operations.]

Transdental implantation is an effective way to restore the lost biomechanical properties of a resected or amputated tooth. The choice of structural materials for the implant is the most important stage of treatment, in relation to the adhesion of aggressive microbiota to them. To characterize the adhesion of microorganisms of periodontopathogenic and cariogenic groups in vitro to experimental samples of zirconium dioxide and titanium alloys using cultural and electronic microscopic methods of adhesion evaluation as the first stage of biofilm formation. Samples for the experiment were prepared in the form of a tablet of standard form, on which the test strains were applied in an amount of 106CFU/ml. After shaking the unattached cells with ultrasound, they were seeded into dense nutrient medium to determine their number. In total, 14 strains of perodontopathogenic and cariogenic groups (including 3 reference strains) were used in the experiment. Scanning electron microscopy was used to visualize the adhesion of microorganisms. The results of the adhesion test to titanium nickelide and zirconium dioxide showed a significant reduction in adhesion for all microorganism species. In all variants (with all strains) the adhesion values to titanium nickelide and zirconium dioxide were statistically significantly lower than when using samples from a traditional titanium alloy. In scanning electron microscopy, single cells of test strains of perodontopathogenic microorganisms were determined on zirconium dioxide samples, while a considerable number of cells and the initial phase of biofilm formation were observed on the compared structural materials. Zirconium dioxide can be considered as an optimal choice material for the manufacture of transdental implants, which, in terms of its technological characteristics and low adhesion characteristics of microbes, is superior to the traditionally used titanium alloys.

[EXPERIMENTAL RATIONALE OF ENDODONTIC THERAPY OF CHRONIC FORMS OF PULPITIS- AND PERIODONTITIS USING PHOTO-ACTIVATED DISINFECTION AND ULTRASOUND TREATMENT].

AIM: Laboratory rationale of application of a complex effect of ultrasound treatment, and photo-activated disinfection in experiments in vitro using strains of anaerobic and microaerophilic microorganisms isolated from patients with chronic forms of pulpitis and periodontitis. MATERIALS AND METHODS: Microbiologic study of root canal was carried out using a bacteriologic method (2 species of obligatory-anaerobic and 2 -microaerophilic streptococci). Evaluation of the effect of ultrasound treatment in a complexwith a system of photo-activated disinfection on microflora of root canal was carried out using a bioreactor. Study of biofilms of the root canal system of a tooth in patients with chronic forms of pulpitis (52 individuals) and periodontitis (16 individuals) were carried out using scanning electron microscopy. RESULTS: The presence of an ample microbial bio- film in the ostiums of dentinal canaliculi and obturation of dentinal canaliculi in certain cases was established. Features of formation of growth curves for bacterial populations of oral microflora (Porphyromonasgingivalis, Fusobacterium nucleatum, Streptococcussanguinis, Streptococcus mutans) were described in the experimental part of the study as a result of a complex effect of photody- namic activation and ultrasound compared with control lacking those. CONCLUSION: The data obtained allow to conclude that complex effect significantly suppresses the growth or microbial populations of oral bacteria.

[Gingival fluid cytokine profile in patients after dental implantation and by periimplantitis].

The study included 32 patients with complications which have developed 3 months to 14 years after dental implants placement. ELISA method was used to assess cytokines content in various sites of tooth alignment in patients with Herpesviridae and periodontopathogenics bacteria associated periimplantitis. The results prove the necessity for Herpesviridae and selected bacteria monitoring after dental implantation.

[Expression of cytokines in dentogingival sulcus in patients after dental implantation and during development of peri-implantitis].

AIM: Determination of cytokine content in various areas of dentition in patients with peri-implantitis associated with parodontopathogenic bacteria species of I and II order. MATERIALS AND METHODS: 32 patients with complications that developed in 3 months to 14 years after installation of intraosteal dental implants were examined. Content of cytokines in various areas of dentition was determined by using solid phase enzyme immunoassay in patients with developed peri-implantitis associated with parodontopathogenic bacteria species of I and II order. Multiplex polymerase chain reaction was used for determination of parodontopathogenic bacteria marker DNA. RESULTS: Marker DNA of I order parodontopathogenic bacteria - Aggregatibacter (Actinobacillus) actinomycetemcomitans, Tannerella forsythia (Bacteroides forsythus), Porphyromonas gingivalis in peri-implantation tissues during implant rejection was detected with 34.4 - 75% frequency while II order (Prevotella intermedia, Treponema denticola, Parvimonas micros (Peptostreptococcus micros), Fusobacterium nucleatum/periodonticum etc.) - with significantly lower frequency. Total concentration of IL- 1beta, IL-4, IL-6, IL-8, TNFalpha, IL- 17A and INFgamma in contents of pathological pocket in the area of implants and the levels of each of them were significantly higher than in the contents of parodontal pockets, areas with stable implants and gingival fluid from areas with healthy teeth. Total interleukin content in the contents of pathological pockets in the area of rejected implants was significantly higher than in other studied areas. In the exudate of parodontal pockets of the preserved teeth it was 2.4 times lower, in the area of stable implants - 4.6 times lower, and in the areas with healthy teeth - 4.8 times lower than with rejected implants (p<0.05). CONCLUSION: The results of the study conducted allow to make a conclusion regard- ing reasonability of monitoring during dental implantation of parodontopathogenic microorganism strains and local cytokine response of the host organism with the objective of rational prophylaxis and therapy of inflammatory complications.

[Effect of antibacterial and immunomodulating therapy on cytokine status in the infectious-inflammatory complications of dental surgical interventions].

In patients with infectious-inflammatory complications after surgery in the oral cavity (cystectomy, dental implants, sinus lifting and removal of third molars) in the dynamics studied the content of pro-inflammatory cytokines (IL-1ß, IL-6 and TNF-α). A significant change in their content in blood serum, and their contents increased in the early postoperative period with normalization of cytokine levels within 2 weeks after antibiotical and immunomodulatory treatment.

[Comprehensive microbiological and molecular-biological control of using laser technologies in ambulatory surgical practice].

Results of comparative use of different type lasers for surgical treatment of patients with medium severity level of periodontitis were presented: group 1--erbium laser "Waterlaser Millennium" (41 people); group 2--diode laser "LS-0,97 - IRE-Polyus" (25 people); group 3--carbon dioxide laser "СО2-laser LST - 20/01 Lantset" (21 people). Patients of the comparative group undergone a traditional surgical curettage (30 people). A PCR-diagnostic was made to all patients that were sent to ambulatory surgical treatment. It revealed carriage of 1 to 4 types of periodontium pathogenic bacteria that present ed threat from the point of view of infectious complications development in the postoperative period. After using laser technologies an increase of periodontium pathogenic species eradication frequency was registered (А. actinomycetemcomitans, T. forsithia, P. gingivalis, P. intermedia, T. denticola): up to 60, 58 and 60%--for erbium, diode and carbon dioxide correspondingly.