Gut microbiota dysbiosis links chronic apical periodontitis to liver fibrosis in nonalcoholic fatty liver disease: Insights from a mouse model.

AIM: In this study, we investigated the systemic implications of chronic apical periodontitis (CAP). CAP may contribute to the nonalcoholic fatty liver disease (NAFLD) progression through the gut microbiota and its metabolites, which are related to the degree of fibrosis. METHODOLOGY: Sixteen 7-week-old male apolipoprotein E knockout (apoE-/-) mice were randomly divided into two groups: the CAP and Con groups. A CAP model was established by sealing the first- and second-maxillary molars with bacterium-containing cotton balls. Apical lesions were evaluated by micro-CT. Histological evaluations of NAFLD were performed using second harmonic generation/two-photon excitation fluorescence (SHG/TPEF) assays. Additionally, we comprehensively analyzed the gut microbiota using 16S rRNA gene sequencing and explored metabolic profiles by liquid chromatography-mass spectrometry (LC-MS). Immunofluorescence analysis was used to examine the impact of CAP on tight junction proteins and mucin expression. Transcriptome assays have elucidated gene expression alterations in liver tissues. RESULTS: Micro-CT scans revealed an evident periapical bone loss in the CAP group, and the total collagen percentage was increased (Con, 0.0361 ± 0.00510%, CAP, 0.0589 ± 0.00731%, p < .05). 16S rRNA sequencing revealed reduced diversity and distinct taxonomic enrichment in the CAP group. Metabolomic assessments revealed that differentially enriched metabolites, including D-galactosamine, were enriched and that 16-hydroxyhexadecanoic acid and 3-methylindole were depleted in the CAP group. Immunofluorescence analyses revealed disruptions in tight junction proteins and mucin production, indicating intestinal barrier integrity disruption. Liver transcriptome analysis revealed upregulation of Lpin-1 expression in the CAP group. CONCLUSION: This study provides comprehensive evidence of the systemic effects of CAP on liver fibrosis in NAFLD patients by elucidating alterations in the gut microbiota composition and metabolism.

Integration of single-cell RNA sequencing of endothelial cells and proteomics to unravel the role of ICAM1-PTGS2 communication in apical periodontitis: A laboratory investigation.

AIM: Endothelial cells (EDs) play a key role in angiogenesis and are associated with granulomatous lesions in patients with chronic apical periodontitis (CAP). This study aimed to investigate the diversity of EDs using single-cell ribonucleic acid sequencing (scRNA-seq) and to evaluate the regulation of intercellular adhesion molecule 1 (ICAM1) on the ferroptosis-related protein, prostaglandin-endoperoxide synthase 2 (PTGS2), in CAP. METHODOLOGY: EDs from the uploaded scRNA-seq data of five CAP samples (GSE181688 and GSE197680) were categorized using distinct marker genes. The interactions between vein EDs (veinEndo) and other cell types were analysed using CellPhoneDB. Differentially expressed proteins in the proteomics of human umbilical vein EDs (HUVECs) and THP-1-derived macrophages infected with Porphyromonas gingivalis were compared with the differentially expressed genes (DEGs) of VeinEndo in scRNA-seq of CAP versus healthy control periodontal tissues. The protein-protein interaction of ICAM1-PTGS2 in macrophages and HUVECs was validated by adding recombinant ICAM1, ICAM1 inhibitor and PTGS2 inhibitor using real-time polymerase chain reaction (PCR), western blotting, and immunofluorescence staining. RESULTS: EDs in patients with CAP were divided into eight subclusters: five vein ED, capillaries, arterials and EC (PLA). There were 29 mutually upregulated DEGs and two mutually downregulated DEGs in vein cells in the scRNA-seq data, as well as differentially expressed proteins in the proteomics of HUVECs. Real-time PCR and immunofluorescence staining showed that ICAM1 and PTGS2 were highly expressed in CAP, infected HUVECs, and macrophages. Recombinant protein ICAM1 may improve PTGS2 expression, reactive oxygen species (ROS), and Fe2+ levels and decrease glutathione peroxidase 4 (GPX4) and SLC7A11 protein levels. ICAM1 inhibitor may inverse the above changes. CONCLUSIONS: scRNA-seq revealed the diversity of EDs in CAP and identified the possible regulation of ICAM1 by the ferroptosis-related protein, PTGS2, in infected HUVECs and macrophages, thus providing a basis for therapeutic approaches that target the inflammatory microenvironment of CAP.

Correlation between PD-1/PD-L1 and RANKL/OPG in chronic apical periodontitis model of Sprague-Dawley rats.

Chronic apical periodontitis (CAP) is characterized by inflammation and destruction of the apical periodontium that is of pulpal origin, appearing as an apical radiolucent area, and does not produce clinical symptoms. Little is known about whether the PD-1/PD-L1 ratio is associated with the balance between RANKL and OPG in CAP. The relationship between PD-1/PD-L1 and RANKL/OPG in CAP is investigated in this study. A CAP rat model was established using Sprague-Dawley rats. The pulp chambers were exposed to the oral cavity to allow bacterial contamination. The apical tissues of the bilateral mandibular first molars were analyzed for histological morphology using hematoxylin and eosin (H&E) staining. Immunohistochemistry and qRT-PCR were used to determine the expression of PD-1, PD-L1, OPG, and RANKL mRNA and proteins in periapical tissues and mandibular samples, respectively. The radiological images indicated a poorly defined low-density shadow and alveolar bone resorption after periodontitis induction. Histological analysis revealed an infiltration of inflammatory cells and alveolar bone resorption in the periapical tissues. Mandibular mRNA and periapical protein expression of PD-1, PD-L1, and RANKL was upregulated 7-28 days after periodontitis induction, while the expression of OPG was downregulated. No significant relationship was observed between PD-1/PD-L1 and RANKL/OPG at either mRNA or protein levels in CAP. There is an increased expression of PD-1, PD-L1, and RANKL and a decreased expression of OPG, indicating progression of CAP.

CRISPR/Cas13d targeting GZMA in PARs pathway regulates the function of osteoclasts in chronic apical periodontitis.

Chronic apical periodontitis is a prevalent oral disease characterized by bone loss, and its underlying mechanisms remain unclear. This study aimed to investigate the role and mechanism of the serine protease GZMA in osteoclasts during chronic apical periodontitis. To address this, we employed crRNA/Cas13d to inhibit GZMA expression and examined its impact on osteoclast behavior. Our findings revealed that GZMA plays a significant role in promoting osteoclast cell proliferation while inhibiting cell apoptosis. Additionally, the inhibition of GZMA led to a notable increase in miR-25-3p expression, which, in turn, downregulated the expression of TGF-ß. Consequently, the reduction in TGF-ß expression led to a decrease in PAR1 expression within the PARs pathway. These results suggest that GZMA might serve as a promising therapeutic target for the treatment of chronic apical periodontitis. Furthermore, our study highlights the potential of targeting GZMA using crRNA/Cas13d as a valuable approach for future therapeutic interventions.

Prevalence of oral infections in chronic kidney disease patients: A cross-sectional study.

OBJECTIVES: The purpose of this investigation was to analyze the prevalence of apical periodontitis (AP) and periodontal disease (periodontitis) (PD) in Chronic kidney disease (CKD) patients in relation to their treatment phase. SUBJECTS AND METHODS: In this cross-sectional study, 188 patients with CKD were divided into two groups: patients without dialysis (WD group, n = 53) and patients on dialysis (DP group, n = 135). Panoramic radiographs were used to diagnose AP. The presence of periodontal disease was evaluated radiographically assessing alveolar bone loss. Student's t-test, chi-squared test, and logistic regression analysis were used to determine the significance of differences between groups. RESULTS: In the WD group, 55% of patients had at least one tooth with AP, whereas in the DP group 67% had at least one tooth with AP (OR = 2.11; 95% CI = 1.09-4.08; p < 0.05). PD was more prevalent in the DP group (78%) than in the WD group (36%) (OR = 6.26; CI 95% = 3.13-12.52; p < 0.01). CONCLUSIONS: Oral infections are more prevalent in the advanced stages of CKD. The treatment of PD and AP should be incorporated in the treatment planning of patients with CKD.

Gut microbiota may mediate the impact of chronic apical periodontitis on atherosclerosis in apolipoprotein E-deficient mice.

AIM: There are growing evidences linking chronic apical periodontitis (CAP) to atherosclerosis. Gut microbiota is found to be involved in the development of atherosclerosis. Recent studies have shown that CAP could change the diversity and composition of the gut microbiota. It was therefore, we hypothesized that gut microbiota and its metabolites could mediate the impact of CAP on atherosclerosis. METHODOLOGY: Twenty-four 5-week-old lipoprotein E knockout (apoE-/- ) mice were randomly divided into four groups: the CAP group, Con group, Co-CAP (cohoused with CAP) and Co-Con (cohoused with Con) group. In the CAP group, sterile cotton wool containing P. gingivalis was placed into the exposed pulp chamber, followed by coronal resin-based composite restoration of the bilateral maxillary first and second molars. In the Con group, a sham operation was performed. Biweekly, mice in the CAP group were anaesthetised to check the sealing of coronal access. Meanwhile, the animals in the Con group were anaesthetised. The cohousing approach was used to introduce gut microbiota from the CAP and Con groups into the Co-CAP and Co-Con groups, respectively. Alterations in the abundance and diversity of the gut microbiota were detected using 16S rRNA sequencing, Oil-red O staining was used to demonstrate the extent of lesions, and serum levels of trimethylamine N-oxide (TMAO), and immunohistochemistry of flavin-containing monooxygenase 3 (FMO3) in liver were used to assess TMAO-related metabolic alterations. RESULTS: Alterations of alpha and beta diversity were shown both in the CAP and the Co-CAP groups. Moreover, the percentage of atherosclerotic lesion area increased in the CAP and Co-CAP groups (p < .05). Linear discriminant analysis effect size (LEfSe) at the family level found the increases of Lachnospiraceae and Ruminococcaceae (p < .05), which were positively correlated with serum TMAO levels (p < .05). In the redundancy analysis technique (RDA), serum levels of TMAO were positively associated with the atherosclerotic lesions. Co-occurrence analysis revealed that the relative abundances of Lachnospiraceae and Porphyromonadacae were positively correlated with both the percentage of lesion area and TMAO level (p < .05). CONCLUSION: Thus, within the limitations of this study, the data suggest that the gut microbiota can mediate the effects of CAP on atherosclerosis.

Hypercitrullination and anti-citrullinated protein antibodies in chronic apical periodontitis, a laboratory investigation. Does autoimmunity contribute to the pathogenesis?

AIM: The presence of Gram-negative anaerobic bacteria, in particular, Porphyromonas gingivalis (P. gingivalis) in periapical granulomas predicts the generation of citrullinated proteins in the lesion. Citrullination of proteins may lead to the formation of anti-citrullinated autoantibodies (ACPA-s) initiating the formation of an autoimmune loop which may contribute to the perpetuation of inflammatory reactions and tissue damage in chronic apical periodontitis. The objective of this study was to demonstrate the formation of citrullinated proteins in chronic apical periodontitis and whether they can act as autoantigens. METHODOLOGY: Twenty-five periapical granulomas (n = 25) were investigated in the study. Healthy periodontal tissue samples served as normal control tissue (n = 6). The peptidyl-citrulline level was determined with the dot blot method. ACPA levels were analysed using anti-citrullinated cyclic peptide (anti-CCP) EDIA kit. Differences between periapical granuloma and control samples were assessed using Mann-Whitney U tests. p Values <.05 were considered as statistically significant. RESULTS: Protein concentrations, peptidyl-citrulline levels and anti-CCP ratios were compared between periapical granuloma and healthy control groups. Multiple linear regression analysis revealed significant (p = .042) hypercitrullination in periapical granuloma samples. Moreover, there was a significant difference in the ACPA ratios between periapical granuloma (2.03 ± 0.30) and healthy control (0.63 ± 0.17) groups (p = .01). Seventeen of 25 periapical granuloma samples (17/25; 68%), whereas one out of six control samples (1/6; 17%) were shown to be positive for the presence of ACPA. CONCLUSIONS: This is the first study detecting the presence of citrullinated peptides and APCA in periapical granuloma, suggesting the contribution of autoimmune reactions in the pathogenesis and perpetuation of chronic apical periodontitis.

Heterogeneity of T cells in periapical lesions and in vitro validation of the proangiogenic effect of GZMA on HUVECs.

AIM: T cells are key immunomodulatory cells in periapical lesions. This study aimed to explore the roles of T cells in chronic apical periodontitis (CAP) using single-cell RNA sequencing and to further investigate Granzyme A (GZMA) in angiogenesis regulation. METHODOLOGY: A total of five CAP samples were collected for single-cell RNA sequencing. We performed subcluster and lineage-tracing analyses for T cells. According to differential gene expression, distinct biological functions enriched in T cells of CAP were presented by gene set enrichment analysis (GSEA) and compared with healthy gingiva (data obtained from the GEO database). CellChat was used to explore potential ligand-receptor interactions between T cells and endothelial cells in CAP. The coculture of primary human umbilical vein endothelial cells (HUVECs) and Jurkat T cells, as well as the addition of GZMA recombinant protein, was used to validate the predicted pair of GZMA and coagulation factor II thrombin receptor (F2R) by RT-PCR, angiogenesis and migration assays. RESULTS: A transcriptomic atlas of 44 746 individual cells was constructed from the periapical lesions of five patients with CAP by single-cell RNA-seq, and eight cell types were identified. We identified nine subsets of T cells and deciphered the cellular heterogeneity of T cells in CAP at the functional level by subclustering and GSEA. Lineage tracing revealed a distinct lineage of T cells in CAP and predicted the transition of the T cellular state upon CAP. GSEA revealed multiple biological processes and relevant angiogenesis genes upregulated in CAP T cells. GZMA-F2R pairs were predicted by cell-cell interactions in CAP. High expression of GZMA and F2R was observed in the coculture of HUVECs and Jurkat T cells, and the proangiogenic capacity of the GZMA recombinant protein was emphasized by in vitro experiments. CONCLUSIONS: Our study provides novel insights into the heterogeneity of T cells in periapical lesions and reveals the potential role of GZMA in T cells in regulating angiogenesis in HUVECs.

0.1% Nano-silver mediates PD-1/PD-L1 pathway and alleviates chronic apical periodontitis in rats.

This study was to investigate whether the programmed death-1 (PD-1)/programmed death-ligand 1 (PD-L1) and T-helper 17 (Th17)/regulatory T (Treg) balance are associated with chronic apical periodontitis (CAP) relived by 0.1% nano-silver. CAP rat models were established by opening the first molars of the right and left mandible and exposing the pulp cavity to the oral cavity. CAP model was verified by cone-beam computed tomography, X-ray digital radiovisiography, and hematoxylin-eosin (H and E) staining. The rats were randomly divided into the sham, Ca(OH)2, and 0.1% nano-silver groups (n = 12 in each group) 2 weeks after surgery. The pathological changes in the apical area were detected by H and E staining. PD-1, PD-L1, RORγT, IL-17, and Foxp3 in periapical tissues were detected by qRT-PCR and immunohistochemistry. Th17/Treg and PD-1/PD-L1 were analyzed by flow cytometry. After 7, 14, and 21 days of 0.1% nano-silver treatment, inflammatory cells in the apical region were slightly reduced and inflammatory infiltration was relieved compared with the sham group. RORγT, IL-17, PD-1, and PD-L1 decreased and Foxp3 increased after 7, 14, and 21 days of 0.1% nano-silver treatment compared with the sham group (p < 0.05); however, there were no significant differences with Ca(OH)2 group (p > 0.05). Flow cytometry revealed that 0.1% nano-silver solution decreased Th17/Treg and PD-1/PD-L1 ratio. 0.1% Nano-silver significantly reduced the inflammation of CAP in rats. PD-1/PD-L1 was included in Th17/Treg balance restored by 0.1% nano-silver.

Milk-Derived Small Extracellular Vesicles Promote Osteogenic Differentiation and Inhibit Inflammation via microRNA-21.

Chronic apical periodontitis (CAP) is a disease with characteristics of inflammation and bone loss. In this study, our objective was to examine the function of small extracellular vesicles (sEVs) obtained from milk in encouraging osteogenic differentiation and inhibiting inflammation by miR-21 in CAP. The expression of miR-21 was detected using qRT-PCR in human CAP samples. The impact of miR-21 on the process of osteogenic differentiation was investigated using CCK-8, qRT-PCR, immunofluorescence staining, and Western blot analysis. The evaluation of RAW 264.7 cell polarization and the assessment of inflammatory factor expression were conducted through qRT-PCR. The influence of sEVs on MC3T3-E1 cells and RAW 264.7 cells was examined, with a particular emphasis on the involvement of miR-21. In human CAP samples, a decrease in miR-21 expression was observed. MiR-21 increased the expression of osteogenesis-related genes and M2 polarization genes while decreasing the expression of M1 polarization genes and inflammatory cytokines. Treatment with milk-derived sEVs also promoted osteogenesis and M2 polarization while inhibiting M1 polarization and inflammation. Conversely, the addition of miR-21 inhibitors resulted in opposite effects. Our results indicated that sEVs derived from milk had a positive effect on bone formation and activation of anti-inflammatory (M2) macrophages and simultaneously reduced inflammation by regulating miR-21 in CAP.

Chronic apical periodontitis exacerbates atherosclerosis in apolipoprotein E-deficient mice and leads to changes in the diversity of gut microbiota.

AIM: To investigate the impact of chronic apical periodontitis (CAP) on atherosclerosis and gut microbiota by establishing a Porphyromonas gingivalis (P. gingivalis)-induced CAP in an apolipoprotein E-deficient (apoE-/- ) mice model. METHODOLOGY: Twenty-eight male apoE-/- mice were divided into two groups with 14 in each: CAP group and control group. In the CAP group, sterile cotton wool containing 108 colony-forming units of P. gingivalis was placed into the pulp chamber after pulp exposure followed by coronal resin filling in bilateral maxillary first and second molars. The mice were fed with a chow diet to induce atherosclerosis. Animals were euthanized 16 weeks after the operation, and the periapical lesions of bilateral maxillary first and second molars were assessed by micro-CT. After collection of aortic arches, atherosclerotic lesions were measured by Oil Red O staining. Serum levels of high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), and triglycerides (TG) were measured. Stools were collected to detect alterations in gut microbiota by 16S rRNA gene sequencing. Independent samples t-test was used to calculate the difference between the two groups. RESULTS: CAP was observed in 98.2% of molars. A significant increase in atherosclerotic plaque formation in the aortic arches was found in the CAP groups (CAP: 2.001% ± 0.27%, control: 0.927% ± 0.22%, p = .005). No significant difference was observed between sevum level of HDL-C (CAP: 2.295 ± 0.31 mmol/L, Control: 3.037 ± 0.55 mmol/L, p = .264) or LDL-C (CAP: 17.066 ± 3.95 mmol/L, Control: 10.948 ± 1.69 mmol/L, p = .177) in CAP group and Control group. There were no significant differences in TG (CAP: 1.076 ± 0.08 mmol/L, control: 1.034 ± 0.13 mmol/L, p = .794) or TC (CAP: 6.372 ± 0.98 mmol/L, control: 6.679 ± 0.75 mmol/L, p = .72) levels between the two groups (p > .05). The alpha diversity was elevated in the CAP group. In terms of beta diversity, the CAP and control groups were clearly distinguished by the microbial community. CONCLUSION: In a mouse experimental model, pulp infection with P. gingivalis -induced CAP, thus aggravating the development of atherosclerosis. Meanwhile, CAP increased alpha diversity and altered the beta diversity of the gut microbiota.

Association between maternal chronic apical periodontitis (CAP) and low birth weight preterm birth (LBWPT).

OBJECTIVE: To determine the association between maternal chronic apical periodontitis and low birth weight preterm birth. METHODS: The case-control study was conducted at the Gynaecology Ward of the Civil Hospital, Karachi, from September 2017 to April 2018, and comprised women aged 19-48 years with singleton pregnancy delivering spontaneously. The subjects were examined for the presence of periodontitis. The mothers who delivered low birth weight preterm babies were the cases in group A and those who delivered normal birth weight babies were the controls in group B. On the delivery day, after the subject having been moved to the room, data was collected through a questionnaire to record demographic details, history of pregnancy and information about the newborn. The radiographs were assessed for the presence of chronic apical periodontitis. The association between maternal chronic apical periodontitis and low birth weight preterm birth was subsequently determined. Data was analysed using SPSS 24. RESULTS: Of the 200 subjects, 100(50%) were in group A with a mean age of 27.17±5.11 years, and 100(50%) were in group B with a mean age of 27.08±4.90 years. Low birth weight preterm birth was associated with education level and family size (p<0.05). There was no association between maternal chronic apical periodontitis and low birth weight preterm birth (p>0.05). CONCLUSIONS: There was no association between maternal chronic apical periodontitis and low birth weight preterm birth.

Wnt antagonist secreted frizzled-related protein I (sFRP1) may be involved in the osteogenic differentiation of periodontal ligament cells in chronic apical periodontitis.

AIM: To explore the mechanism of secreted frizzled-related protein I (sFRP1) involvement in the osteogenic differentiation of human periodontal ligament cells (hPDLCs) under inflammatory conditions. METHODOLOGY: hPDLCs were cultured in an osteogenic differentiation-inducing medium (odi) and subjected to the stimulation of Porphyromonas gingivalis lipopolysaccharide (P. gingivalis LPS) with or without the inhibition of sFRP1. Quantitative real-time polymerase chain reaction, Western blot and enzyme-linked immunosorbent assay were carried out to evaluate the expression of osteogenic markers as well as the classic Wnt signalling pathway. Periapical periodontitis was induced in Wistar rats to further confirm the effect of sFRP1 inhibitor on bone loss in vivo. After the Shapiro-Wilk normality test, data were analysed by Student's paired t-test or one-way Anova test with a P value less than 0.05 as the level of statistical significance. RESULTS: Significantly decreased mRNA and protein expression of osteogenic markers were detected in hPDLCs treated with P. gingivalis LPS during osteogenic induction (P < 0.001). Increased expression of sFRP1 was observed (P < 0.01), whilst Wnt/ß-catenin signalling pathway was inhibited by the addition of P. gingivalis LPS (P < 0.01). After the addition of the sFRP1 inhibitor, the decrease of osteogenic markers (P < 0.05) and the inhibition of Wnt/ß-catenin signalling pathway (P < 0.05) were reversed significantly. The animal experiment further confirmed that the sFRP1 inhibitor significantly reduced bone loss of periapical lesions in vivo (P < 0.0001). CONCLUSIONS: Wnt antagonist sFRP1 was involved in the osteogenic differentiation of hPDLCs under inflammation. Modulation of the Wnt/ß-catenin signalling pathway through the inhibition of sFRP1 may offer a new perspective on the treatment of chronic apical periodontitis.

Effects of chronic apical periodontitis on the inflammatory response of the aorta in hyperlipemic rats.

OBJECTIVES: To study the effects of chronic apical periodontitis (CAP) on the inflammatory response and initial lesion of aorta in hyperlipemic rats. MATERIALS AND METHODS: Sprague-Dawley (SD) rats aged 14 weeks were randomly divided into 4 experimental groups (n = 8), namely, normal diet (ND), high-fat diet (HFD), CAP, and HFD + CAP. The rats were raised under controlled conditions and fed with diet specified for each group. All subjects were euthanatized after 14 weeks for histopathological analysis. Serum cytokines were analyzed to assess changes in gene and protein expression of aorta via enzyme-linked immunosorbent assay and real-time polymerase chain reaction. Results were analyzed by one-way ANOVA. RESULTS: Low-density lipoprotein cholesterol levels in rats in HFD + CAP group were significantly higher than those in other groups. By comparison, low-density lipoprotein cholesterol levels in rats in both the HFD and HFD + CAP groups were significantly lower than those in the other groups. No significant difference among all groups was observed in terms of CRP level. However, levels of IL-2, IL-6, and IL-10 increased in the experimental CAP rats compared with the control rats. mRNA expression levels of MCP-1, TLR-4, and NF-κB p65 were markedly elevated in rats in the HFD group compared with those in rats in the ND group. TLR-4 mRNA expression was significantly higher in rats in the HFD + CAP group than that in rats in the HFD group. CONCLUSIONS: CAP mediated the high expression of cytokines and induced the initial inflammatory response in the aorta. Apical periodontitis may affect the level of inflammatory cytokines (IL-2, IL-6, and IL-10) depending on the immune response. CLINICAL RELEVANCE: CAP may trigger a systemic inflammatory response and affect the aorta of patients.

Clinical and microbiological assessment of root canal decontamination in chronic apical periodontitis using the ultrasound.

OBJECTIVE: The aim: To study the effectiveness of chronic apical periodontitis treatment by the combined use of ultrasonic treatment of root canals and multicomponent antimicrobial gel according to the results of clinical and microbiological researches. PATIENTS AND METHODS: Materials and methods: 64 patients with chronic apical periodontitis at the age of 18-56 years were treated. Patients were divided into two groups: the main and control. In the main group the root canals of 36 teeth were sonicated in combination with multicomponent antimicrobial gel, in the control - 35 teeth were treated by a standard method. The content of the root canals for microbiological studies was obtained before endodontic treatment and before permanent obturation. Frequency of content and number of bacteria in the samples were evaluated. RESULTS: Results: All samples before treatment were positive for the presence of variable bacterial flora, among which Staphylococcus epidermidis (43.9%), Enterococcus faecalis (37.9%), Streptococcus spp. (24.8%), Candida albicans (24.4%), Fusibacterium (9.4%) were the most dominant, their number was 7.4-4.8 lg CFU/ml. Repeated research after the proposed and standard method of root canal decontamination has shown a significant decrease in microflora. According to the data of clinical and microbiological examination, the method which was developed by us revealed a positive result in 86% of cases compared with 63% when treated by the standard method. CONCLUSION: Conclusions: The effectiveness of a complex treatment method with combined use of the ultrasonic irrigation and multicomponent antimicrobial gel for root canals decontamination in chronic apical periodontitis is demonstrated. Significant reduction of microflora growth and destruction of microbial associations, good penetration of multicomponent antimicrobial gel into endodontic structures due to ultrasound compared with a standard method were achieved.

[Clinical and radiographic rationale for endodontic treatment of permanent teeth with periapical destruction]. / Kliniko-rentgenologicheskoe obosnovanie éndodonticheskogo lecheniia postoiannykh zubov s periapikal'noi destruktsiei.

In the course of the present study four endodontic treatment schemes were tested and the most effective scheme was determined through the use of calcium hydroxide-based paste being applied three times at an interval of 7-14 days with the subsequent long-term temporary obturation using calcium hydroxide with iodoform, ultrasonic activation of sodium hypochlorite in the root canal, hydrodynamic irrigation and a diode laser. During the entire treatment period, the most effective scheme has revealed a reduction in defect size by 2,57±0,17 (p>0,0001) as well as a 1,84 degree (p>0,0001) decrease in PAI score. Based on optical density data, it was concluded that the application of calcium hydroxide with iodoform in the treatment scheme leads to an 2,2-4,2% improvement in bone tissue regeneration in the periapical zone. The greatest and complete inhibition of microorganisms was determined in patients of the first and third experimental groups in which a diode laser was used. The use of a diode laser in endodontic dental treatment with periapical destruction, enhances antibacterial activity and contributes to the complete inhibition of pathogenic microflora in the root canals.

[Clinical evaluation of the innovative concept of teeth saving]. / Klinicheskaia otsenka innovatsionnoi kontseptsii sokhraneniia zubov.

AIM: To evaluate the effectiveness of the innovative concept of dental medical technologies in patients with resected (amputated) roots of teeth with and without periodontal diseases. MATERIAL AND METHODS: Of the 516 examined patients with periapical destructive foci of infection of various genesis, 4 clinical groups of 24 people were randomly formed: (1) with a diagnosis of apical periodontitis of incisors, canines or premolars with individual milled transdental implants made of zirconium dioxide; (2) patients with a similar diagnosis and concomitant periodontitis of moderate severity with the same implants; (3) patients with periapical destructive process of molars without periodontitis with the same implants. Control group (4) included patients with similar diagnoses with resection of the root apex without implantation. Patients of the study groups were operated in accordance with the developed clinical protocols, including the manufacture and installation of individual transdental implants with fixation in the postoperative period, developed within the framework of tooth-preserving technologies of immobilizing structures. RESULTS: The reinforcement of teeth with resected roots with transdental implants together with immobilization of these teeth in the postoperative period restores the biomechanical characteristics of the tooth 2.7 times more effectively. However, there is a slight change in the mobility of the operated teeth in an earlier period (after 3 months), which is associated with the resorption of the bone-replacing agent used to fill the intraoperative defect and with the defective formation of bone tissue. The values of peritelomeric 6 months after the operation differ from the control by 1.4 units only.

microRNA-146a and Hey2 form a mutual negative feedback loop to regulate the inflammatory response in chronic apical periodontitis.

Chronic apical periodontitis (CAP) is defined as chronic inflammation of the dental pulp and root canal system. Porphyromonas endodontalis lipopolysaccharide ( P. endodontalis LPS) plays an important role in inducing an inflammatory response in CAP. microRNA-146a (miR-146a) is a key regulator of inflammation and is induced by LPS. Hairy and enhancer-of-split related with YRPW motif 2 (Hey2) has been confirmed to be induced by the Notch signaling pathway, which is involved in tooth development, pulp regeneration, and repair after injury. Our study aimed to investigate the functional role of miR-146a via the targeting of Hey2 in CAP as well as the underlying mechanism. Compared with 13 healthy controls, miR-146a and Hey2 expressions were significantly higher in 20 patients with CAP. In addition, miR-146a, Hey2, interleukin (IL)-6, IL-1ß, and tumor necrosis factor (TNF)-α expressions were significantly increased in MC3T3-E1 cells stimulated with different concentrations (0-20 µg/mL) of P. endodontalis LPS for different amounts of time (0-48 hours). Moreover, miR-146a, which acts as an anti-inflammatory mediator, negatively regulated the expression of IL-6, IL-1ß, and TNF-α, and Hey2 was confirmed as a target gene of miR-146a by a luciferase reporter assay. Hey2 also negatively regulated miR-146a, IL-6, IL-1ß, and TNF-α expressions, and P. endodontalis LPS strongly induced Hey2 recruitment to the IL-6 promoter (-400 ~ -200 bp). These findings suggest that miR-146a and Hey2 form a mutual negative feedback regulatory loop, demonstrating a novel mechanism that regulates inflammatory responses in CAP.

Micro-CT analysis of chronic apical periodontitis induced by several specific pathogens.

AIM: To conduct three-dimensional micro-CT analysis of bone destruction, periapical sclerotic changes and inflammatory root resorption (IRR) to compare the differences between Enterococcus faecalis (Ef)- and Porphyromonas gingivalis (Pg)-induced chronic apical periodontitis (CAP). METHODOLOGY: Mono-species bacteria-induced CAP was established in the first and second molars of the right maxilla and mandible of male Sprague-Dawley rats. Fifteen animals were divided into three groups with five rats in each group: control group, Ef-CAP group and Pg-CAP group. The maxilla and mandible were harvested and then scanned by micro-CT. Alveolar bone destruction was evaluated by measuring the volume of alveolar bone resorption, bone volume fraction (BV/TV), trabecular thickness (Tb. Th), trabecular number (Tb. N) and trabecular spacing (Tb. Sp). The results were analysed using one-way analysis of variance and Fisher's least significant difference methods (LSD). The sclerotic changes in the periapical bone were graded and the IRR indexes were scored. The data were analysed using the chi-square test. RESULTS: The alveolar bone resorption volume of the Pg-CAP group was significantly larger than that of the Ef-CAP group (P < 0.01). In the maxilla, both Pg-CAP and Ef-CAP groups had a significant decrease in BV/TV and increase in Tb. Sp (P < 0.01) with the more significant changes of trabecular bone in the Pg-CAP group. A significant reduction of Tb. N was only found in the Pg-CAP group (P < 0.05). There was no significant change in Tb. Th in either group (P > 0.05). In the mandible, except for an increase in Tb. Sp in the Pg-CAP group (P < 0.05), there was no significant change in BV/TV, Tb. Th or Tb. N (P > 0.05). No obvious sclerotic change was observed. IRR was detected in significantly more root surfaces in the Pg-CAP group (240 surfaces, 60%) than those in the Ef-CAP group (178 surfaces, 44.5%) (P < 0.001). The IRR extension was also significantly more advanced in the Pg-CAP group (P < 0.05). CONCLUSIONS: Pg-CAP caused more substantial alveolar bone destruction and IRR than Ef-CAP. The maxilla was more susceptible to CAP in terms of microstructural changes of trabecular bone than the mandible. Tb. Sp was the most sensitive index for evaluating the residual alveolar bone of CAP.

[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.