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16MEDICAL SCIENCES
MICROBIOLOGICAL CONDITION OF THE ORAL CAVITY AFTER DENTAL FILLING WITH
VARIOUS COMPOSITE MATERIALS
Gafforova S.S.,
Basic doctoral student of the Department of Facultetic Therapeutic Dentistry,
Tashkent State Dental Institute
Gafforov S.A.
Head of the Department of Dentistry, Pediatric Dentistry and Orthodontics, Сenter for the development ofprofessional qualifications of medical workers,
Doctor of Medical Sciences, Professor
Abstract
It can hardly be denied that dental caries is the most common pathology among dental diseases of the oral activity. In this article, adhesive microbial colonization of the oral cavity to various filling materials is comparatively evaluated.
Keywords: microbiology of the oral cavity, dental caries and their treatment, dental filling materials, prevention.
I.Introduction
It is known that dental caries is the most common pathology among dental diseases of the oral cavity (OR). Speaking about the reasons for the development of the carious process, we can say that caries is a poly-etiological disease [8, 9, 20]. A number of authors believe that the main factor in the onset and development of caries are PR microorganisms [3, 16], and their role in this process has been proven by animal experiments [14, 15].
A number of authors argue that one of the main factors in the onset and development of caries, including secondary caries, is the adhesive activity of cario-genic PR microorganisms to the surface of filling materials and hard tissues of teeth, as a result of physico-chemical interaction between microorganisms, filling material and adsorbed saliva proteins are formed biofilms [1, 2, 4, 24, 7, 11, 19, 21, 25]. In addition, the quantity and quality of colonization by the bacterial flora depends on the type of tooth surfaces and filling materials [10, 18, 23]. Also, it was found that on fiberglass composite material and ceramics, it contains a large amount of agglutinins that stimulate the adhesion of S. mutans [17, 18]; others, it was found on glass-ion-omer cements (GIC), S. sobrinus showed the least adhesive activity to the composite filling material "Charisma" and GIC "Prodigy" [12, 22]; the role in the colonization of microflora with the degree of roughness of the surface of filling materials [5].
There is evidence that composites accumulate more microorganisms and bacteria in comparison with other filling materials. To solve this problem, the authors propose the development of a new high-quality and fine-particle inorganic filler and further improvement of the existing ones [13], another researcher recommends conducting studies in the long-term assessment of their antibacterial properties, both in experimental and clinical studies, to increase the effectiveness of such modified materials [6]; Despite this, it remains important to study and improve the properties of composite and other filling materials, which will further
minimize the risk of secondary caries. The above literature indicates the need for further study of the microbiological state of PR in patients during filling with various filling materials, most often used in modern dentistry.
II. Analysis
The aim of the study was to study the sensitivity of cariogenic microorganisms to chlorhexidine acetate (HCA) in the oral cavity and to assess adhesive micro-bial colonization regarding the domestic light-cured Firuze-Grand filling material for the restoration of anterior and posterior teeth.
The study involved 120 patients (78 women and 42 men), aged 18 to 60 years in the dental clinic of Ltd. "Sardor-Denta", and the outpatient clinic of therapeutic dentistry, where the department of "Faculty therapeutic dentistry" of the Tashkent Dental Institute is located. (Table №1). Research methods included clinical and laboratory assessment and examination of PR; study of the condition of filled teeth with caries, microbiological assessment of the condition of PR after filling in the period: 6 months, 12 months and 18 months. The filling materials used are light-cured composites: nanocompo-site - "Filtek Ultimate" (3M ESPE, USA) (2-group A); Latelux (Latus, Ukraine) (2-group B); "Restavrin" (Technodent, Russia) (2-group C) and 1-st A, 1-st B, 1st C groups - patients who used the domestic filling material Firuze-Grand (Dentals Pharma GmbH), which began to be produced since 2020 in the Zamin free economic zone of the Jizzakh region according to German technologies.
We collected complaints, anamnesis and clinical examination of patients. For each patient under study, medical records were filled out. The map recorded the dental formula deployed over the surfaces, the presence or absence of complaints, the state of the mucous membrane (RM) of the gingival papillae and marginal gums. To detect primary caries, we used the method of vital staining with a fuchsin solution using a caries marker (Voco), and the condition of previously placed fillings of foreign materials was assessed using clinical criteria
approved by the International Federation of Dentists KPU index, which characterizes the number of teeth af-FDI. At the stage of the clinical study, the intensity and fected by caries, filled and removed. prevalence of dental caries was studied according to the
Table №1.
The number of examined patients and filled teeth. Distribution by types of filling materials.
Group A (40 people and 1120 physiological teeth (100%)) Group B (40 people and 1120 physiological teeth (100%)) Group C (40 people and 1120 physiological teeth (100%))
1 2 1 2 1 2
Number of patients (total -120) 120 22/55% 18/45% 26/65% 14/35% 27/67,5% 13/32,5%
100% 22/18,3% 18/15% 26/21,7% 14/11,7% 27/22,5% 13/10,8%
Gender Men (42) 42/35% 8/20% 10/25% 12/30% 6/15% 3/7,5% 3/7,5%
8/19,4% 10/24% 12/28,5% 6/14,3% 3/7,2% 3/7,14%
Women (78) 78/65% 14/35% 8/20% 14/35% 8/20% 24/60% 10/25%
14/17,9% 8/10,25% 14/17,9% 8/10,25% 24/30,8% 10/12,8%
Age 18-45 age (52) 52/43,3% 7/17,5% 6/15% 14/35% 6/15% 12/30% 7/17,5%
7/13,5% 6/11,5% 14/27% 6/11,5% 12/23% 7/13,5%
46-60 age (36) 36/30% 3/7,5% 6/15% 8/20% 5/12,5% 10/25% 4/10%
3/8,3% 6/16,7% 8/22,2% 5/13,9% 10/27,8% 4/11,1%
61 age and elder (32) 32/26,7% 12/30% 6/15% 4/10% 3/7,5% 5/15% 2/5%
12/37,5% 6/18,75% 4/12,5% 3/9,4% 5/15,6% 2/6,25%
Quality of inspection of teeth = 3360/100% Healthy (2210) 2210/65,8 % 501/44,8 328/29,3% 386/34,5% 340/30,3% 345/30,8% 310/27,7%
501/22,6% 328/14,8% 386/17,4% 340/15,3% 345/15,6% 310/14,0%
Carious (1150) 1150/34,2 % 223/19,9% 228/20,3% 211/18,9% 203/18,1% 140/12,5% 145/12,9%
223/19,4% 228/19,8% 211/18,3% 203/17,6% 140/12,2% 145/12,6%
Quality examination of teeth according to the KPU index (2860/100%) Intact (476) 476/16,6% 128/11,4% 116/10,3% 88/7,8% 82/7,3% 27/2,4% 35/3,1%
128/26,9% 116/24,4% 88/18,5% 82/17,2% 27/5,7% 35/7,3%
Carious (730) 730/25,5% 144/12,85 % 122/10,9% 146/13% 158/14% 90/8% 70/6,2%
144/19,7% 122/16,71 % 146/20% 158/21,6% 90/12,3% 70/9,6%
Sealed (650) 650/22,7% 160/14,3% 143/12,8% 86/7,8% 77/6,9% 94/8,4% 90/8%
160/24,61 % 143/22,0% 86/13,23% 77/11,84% 94/14,46% 90/13,84%
Removed (1004) 1004/35,1 % 108/9,64% 125/11,16 % 208/18,57 % 214/19,10 % 220/19,64 % 129/11,51 %
108/10,75 % 125/12,45 % 208/20,71 % 214/21,31 % 220/21,91 % 129/12,84 %
The composition of the investigated filling material "Firuze-Grand" (Dentals Pharma GmbH): di-methacrylate oligomers (bis-GMA, TEGDMA, UDMA, bis-EMA, PEGDMA, AMD), a radiopaque combined filler made of modified aluminosilicate glass (0.02-2.0 ^m) and nanosized particles of silicon oxide (5-20 nm). The proportion of inorganic filler is -80%, the curing process takes place under the influence of the visible light spectrum, with a wavelength of 400-500 nm. The material has the properties of fluorescence and color stability, radiopacity, and for the purpose of aesthetic imitation of dental tissues, the material is produced in a range of shades according to the VITA shade scale and three degrees of transparency.
The culture media for the cultivation of bacteria were selected for optimal growth of specific types of microorganisms. Also, to study the antibacterial properties of chlorhexidine acetate (HCA) at concentrations of 0.5% and 1.9%, the sensitivity of clinical strains S. mutans, S. mitis / oralis, S. sanguinis, S. gordonii, L. salivarius and the reference strain C. albicans ATCC 10231.
To study the antimicrobial activity of HCA solutions and Firuze-Grand filling material containing AMD, the disk-diffusion method was used. Pilot studies of the sensitivity of microorganisms to the composite material were carried out on 160 samples of the Firuze-Grand filling material without the addition of an antiseptic and with the addition of HCA in 3 repetitions for each group. Museum reference strains Microtrol were used as test strains in pilot studies; American type culture collection; S. pneumoniae ATCC 6303; S. pyogenes ATCC 19615; E. faecalis ATCC 29212; H. influenzae ATCC 49766; E. coli ATCC 25922 and C. albicans ATCC 10231.
Several colonies of microorganisms, grown on an appropriate medium, were collected with a sterile bacteriological loop and suspended in sterile isotonic solution (0.85% p / reNaCe). The suspension was vortexed to a density of 0.5 units according to the McFarland turbidity standard, which corresponds to 1.5 x 108 L / ml. Experiments were performed on 8 surfaces for each group. We used clinical isolates of S. mutans, S. mitis / oralis, S. sanguinis, S. gordonii, L. salivarius strains
from patients with dental caries. Also, to study the adhesive activity of microorganisms, 16 polished samples of the composite material "Firuze-Grand" with the addition of HCA were used to determine the adhesion to the tested samples of the filling material of clinical isolates of S. and the museum strain C. albicans ATCC 10231.
To study the color parameters of the Firuze-Grand filling material with the addition of HCA in different concentrations, a Spectron-M color analyzer was used. Water absorption reflects the mass amount of water that is absorbed by a sample of filling material after 7 days of exposure in distilled water at 37 ° C.
For the manufacture of 12 samples of each composite material, premolars and molars of patients removed for orthodontic indications and due to complications of periodontal diseases were used. The extracted teeth were cleaned from soft tissues and dental deposits and placed in distilled water, stored at a temperature of 4 ° C. Each tooth was sawn with a milling cutter in the sagittal direction using a dental micromotor with a rotation speed of 1500-3000 rpm, with constant air-water cooling and humidification.
Results of determining the sensitivity of microorgai
On the exposed tooth surface, an adhesive bond was created with the Firuze-Grand composite material. The enamel and dentin surfaces were etched with 37% phosphoric acid for 15-20 seconds. To study the adhesion shear strength, 12 samples of each investigated group of the Firuze-Grand composite material were made, the shear strength was determined when the crosshead was moving at a speed of 5.0 mm / min.
The obtained results of the studies were processed, the arithmetic mean, standard deviation and error of the arithmetic mean were calculated, Student's t-test (t) and the probability of error (p) were determined. At p <0.05, the differences between the two samples were considered significant.
III. Discussion Analyzing the results obtained, it can be noted that the clinically isolated S. mutans strain from patients with dental caries showed moderate sensitivity to HCA solutions at concentrations of 0.5% - 11.8 ± 0.22 mm, 1.9% - 12.6 ± 0, 27 mm, with a probability of p <0.05. Similar results were obtained for the S. sanguinis strain, which showed sensitivity to HCA at a concentration of 0.5% - 11.8 ± 0.22 mm, and 1.9% - 12.8 ± 0.22 mm.
Table №2.
ms involved in the formation of caries to HCA solu-
M / o Zone of inhibition in mm (M ± m) at concentrations of HCA Criterion of Student t, p
0,5% (n=5) 1,9% (n=5)
S.mutans 11,8±0,22 12,6±0,27 t=2,46, p<0,05**
S.mitis/oralis 0 0 -
S.sanguinis 11,8±0,22 12,8±0,22 t=3,42, p<0,01**
S.gordonii 9,6±0,27 10,8±0,22 t=3,61, p<0,01**
L. salivarius 14,6±0,27 15,4±0,45 t=1,65, p>0,05**
C. albicans АТСС 10231 14,2±0,22 17,8±0,42 t=6,35, p <0,001**
Note: * 0-the absence of a visible zone of growth disk (d = 7); ** Student's criterion was determined by centration of HCA solutions 0.5% and 1.9%.
The zones of growth inhibition of the S. gordonii strain for both antiseptic concentrations were lower than those of S. sanguinis and S. mutans. Thus, the sensitivity to HCA at a concentration of 0.5% in S. gordo-nii was 9.6 ± 0.27 mm, and to an antiseptic at a concentration of 1.9% - 10.8 ± 0.22 mm.
The S. mitis / oralis strain proved to be resistant to the action of HCA at concentrations of 0.5% and 1.9%. However, in experiments using Firuze-Grand composite discs with AMD, this strain showed sensitivity to a sample containing 5.0% HCA, which inhibited the growth of bacteria in the 11.75 ± 0.48 mm zone. This may indicate an insufficient concentration of the HCA solution of 1.9% to inhibit the growth of S. mitis / oralis on a nutrient medium; when comparing the general results of the studies, the S. mitis / oralis strain turned out to be the least susceptible to antiseptic effects among all studied streptococci species.
i inhibition corresponds to the diameter of the filter paper comparing growth inhibition zones in relation to the con-
Unlike streptococci, the clinical strain L. salivar-ius and the museum strain C. albicans ATCC 10231 were highly sensitive to both concentrations of the antiseptic solution. Thus, the diameter of the growth inhibition zone when using filter paper disks with HCA 0.5% was 14.2 ± 0.22, and by 1.9% - 17.8 ± 0.42, with p <0.001.
A pilot study of the sensitivity of microorganisms to the composite material was carried out for control samples of the Firuze-Grand filling material without the addition of an antiseptic and test samples with the addition of HCA at concentrations of 0.1%, 0.5%. After inoculation of microorganisms on solid nutrient media in Petri dishes, the introduction of disks of filling material with HCA and incubation in a thermostat at 37 ° C, the values of the zones of inhibition of the growth of cultures of microorganisms were taken into account (Table №3).
Table №3.
Sensitivity of museum strains of microorganisms to the filling material "Firuze-Grand" with the addition of different concentrations of HCA
M/o Zone of inhibition in mm (M ± m) at a concentration of HCA Criterion of Student t , p
0,0%* (n=4) 0,1%* (n=4) 0,5%* (n=4) 5,0% (n=4)
S.pneumoniae ATCC 6303 0 0 10,50±0,33 11,75±0,29 t=3,09,p<0,05**
S.pyogenes ATCC 19615 0 0 12,25±0,29 14,75±0,30 t=6,31,p<0,05**
E.coli ATCC 25922 0 0 10,25±0,29 12,50±0,33 t=5,29,p<0,05**
E.faecalis ATCC 29212 0 6±2,31 10,25±0,29 12,75±0,30 t=1,75, p>0,05*** t=2,48
H.influenzae ATCC 49766 0 0 0 8,75±0,29 -
C.albicans ATCC 10231 0 0 10,38±0,28 17,50±0,29 t=17,50, p<0,001**
Note: * 0-disc diameter (d = 7); ** Student's test in relation to the concentration of HCA 0.5%; *** Student's t-test in relation to the concentration of HCA 0.1%.
Samples of the composite material "Firuze-Grand" with a concentration of HCA 0.1% had no antimicrobial effect on all studied strains of microorganisms, except for E. faecalis. But, at a concentration of 0.5% HCA in S. pneumoniae ATCC 6303 zones of growth inhibition were 10.50 ± 0.33 mm, with 5.0% - 11.75 ± 0.29 mm. The statistical significance of differences in the width of the zones of growth inhibition at these concentrations was noted (p <0.05).
The size of the zone of inhibition of growth of S. pyogenes with an antiseptic concentration of 0.5% was 12.25 ± 0.29 mm, and for samples with 5.0% HCA -14.75 ± 0.30 mm. Differences in the width of growth inhibition zones at these antiseptic concentrations are statistically significant (p <0.05), the linear correlation coefficient is 0.72.
The diameter of the growth inhibition zone of the E. coli strain ATCC 25922 when using samples of the composite material "Firuze-Grand" with a HCA concentration of 0.5% was 10.25 ± 0.29 mm, and with an antiseptic concentration of 5.0% - 12.50 ± 0.33 mm. The growth retardation zones for composite samples with a HCA concentration of 0.5% are 18% lower than for samples with an antiseptic concentration of 5.0%.
When comparing the values of the zones of inhibition of the E. faecalis strain with the concentration of HCA 0.1% and 0.5%, no statistically significant difference (p> 0.05) was obtained for the samples of the composite material with 0.5% HCA 10.25 ± 0,29 mm and samples with 5.0% AMD 12.75 ± 0.30 mm. The linear correlation coefficient was 0.66.
Positive results were obtained in determining the sensitivity of H. influenzae to samples of filling material with an antiseptic concentration of 5.0%, the size of the growth inhibition zone was 8.75 ± 0.29 m. The degree of sensitivity of the C. albicans strain to the action of HCA at a concentration of 0.5% in the composition of the composite material "Firuze-Grand" in the form
of the formation of a zone of growth inhibition was 10.38 ± 0.28 mm, and the size of the zone of growth inhibition in samples with 5.0% HCA was 17.50 ± 0.29 mm.
IV. Conclusion
The obtained positive results made it possible to use this method to study the antibacterial activity of the composite material "Firuze-Grand" with AMD, in relation to microorganisms involved in the development of carious lesions of hard dental tissues.
Thus, HCA solutions have a pronounced antiseptic effect on PR microorganisms, which are involved in the development of dental carious lesions, and can be recommended for introduction into composite filling materials to reduce the risk of complications in the form of secondary caries and pulp diseases.
On the basis of pilot studies, it was found that the addition of HCA to the composite material "Firuze-Grand" at concentrations of 0.5% and 5.0% leads to inhibition of the growth of museum strains of microorganisms. Moreover, in all cases, the linear correlation coefficient was below 1.0, which indicates the presence of a direct relationship between the concentration of the antiseptic and the sensitivity of the strains. In addition, clinical studies of the condition of filled teeth revealed differences in adhesive microbial colonization to various filling materials.
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