Sterile Paper Points Research Articles

Association of subgingival Epstein–Barr virus and periodontitis

Background: The Epstein–Barr virus (EBV) is gaining interest as a possible agent in the etiology of periodontitis. Previous studies have shown controversy on whether EBV DNA in the subgingival periodontal pockets is associated with periodontitis. The aim of the present study was to seek the potential relationship between EBV and periodontitis. Methods: Data on socio-demographics, oral health, and periodontal health were recorded, and samples were collected from gingival crevicular fluid, using sterile paper point. This case–control study of 118 participants included 59 subjects with severe periodontitis and 59 control subjects with mild periodontitis. The EBV load was determined by quantitative real-time PCR. Results: EBV DNA was detected in 37.3% of the case samples and in 18.6% of the control samples. There was no significant difference in the load of EBV DNA between severe and mild periodontitis (p>0.05). The observed load of EBV DNA was up to 4.55x105 copies/mL. The detected EBV DNA was significantly associated with the plaque index and the oral hygiene index (all p<0.05). Conclusions: A significant association was not found, but EBV might contribute to periodontitis. Gingival crevicular fluid is useful for monitoring the EBV load by the real-time PCR technique.

COMPARATIVE STUDY OF ANTIBACTERIAL ACTIVITY OF PHOTOACTIVATED DISINFECTION WITH FOTOSAN AND STANDARD ENDODONTIC TREATMENT

Introduction: The purpose of endodontic treatment is to eliminate the bacterial infection in the root canal system and allow healing of apical periodontitis. Sometimes the anatomical complexity of the root canal system makes complete removal of bacteria almost impossible even if the conventional methods of chemo-mechanical debridement are performed strictly according to the protocol. So additional methods of root canal disinfection can be applied such as photoactivated disinfection (PAD). Purpose: The aim of our study was to compare the antibacterial activity of PAD with FotoSan, and conventional endodontic therapy in the treatment of infected root canals. Materials and methods: The study involved 36 teeth of patients who are diagnosed with pulp necrosis or with the periapical chronic periodontitis and require endodontic treatment and divided into two groups of 18 teeth each. The first microbiological sample was taken by placing a sterile paper point in the root canal, after endodontic access cavity preparation. All root canals are prepared by Protaper Universal rotary instruments (Maillefer Instruments SA, Ballaigues, Switzerland). In the first group, the disinfection of the root canals was made with photoactivated disinfection with FotoSan. In the second group was performed the endodontic treatment with the standard protocol of irrigation - 2.5% sodium hypochlorite solution and 17% EDTA. After that, the root canals are dried with sterile points, and a second microbiological sample is taken. Results: In all compared pairs, there was no significantly different effect regarding the number of microorganisms. Conclusions; The disinfection with NaOCl has the strongest antimicrobial effect in this study. Second place is occupied by PAD with FotoSan. We believe that the established antimicrobial effect of PAD make the method appropriate as complementary to routine endodontic treatment.

Microbiological Profiles of Dental Implants in Metabolic Syndrome Patients: A Case-Control Study.

There is a lack of knowledge on the possible influence of systemic conditions on peri-implantitis. The aim of this case-control study is to evaluate the difference in terms of oral pathogens’ concentrations in the peri-implant sulcus of a group of patients affected by metabolic syndrome (Mets) compared to healthy subjects. For each patient, peri-implant sulcular biofilm samples were obtained by inserting two sterile endodontic paper points in the deepest aspect of the peri-implant sulcus for 30 s. The quantitative real-time polymerase chain reaction was performed to evaluate total bacterial counts of six pathogens. Patients were screened for peri-implant diseases and clinical and radiographic parameters were recorded. A total of 50 patients was enrolled in the study, 25 affected by Mets and 25 healthy. Significantly higher bacterial counts were discovered for Aggregatibacter actinomycetemcomitans (p = 0.0008), Prevotella intermedia (p = 0.0477) and Staphylococcus aureus (p = 0.034) in MetS patients compared to healthy subjects. Performing a sub-group analysis, considering peri-implant status and dividing patients by MetS diagnosis, no statistically significant (p < 0.05) differences were found. For the first time, a correlation between MetS presence and a greater prevalence of some bacterial species in the peri-implant sulcus was reported, irrespectively from peri-implant status (health vs. disease).

CLINICAL FEATURES, AGE AND SEX DISTRIBUTIONS, RISK FACTORS AND THE TYPE OF BACTERIA ISOLATED IN PERIODONTITIS PATIENTS IN SANA'A, YEMEN

Background: Periodontitis is an inflammation caused by plaque in the surrounding dental structures. It is a major factor in adult tooth loss. There is lack of information on associated clinical features, risk factors and microbial etiology of periodontitis in Sana’a, Yemen.&#x0D; Aim: The study focused on associated clinical features, risk factors and the separation and classification of bacteria in periodontitis and associated risk factors amongst patients attending dental clinics is Sana’a city. &#x0D; Methods: First, 296 patients were admitted to the dental clinic at the Republican University Hospital and private dental clinics in Sana'a during a period of nearly one year, which began in December 2019 AD and ended in November 2020 AD, when they were diagnosed with dental diseases, then 49 of them were selected who are suffering from periodontitis, of whom 22 are males and 27 are females. Sterile paper points were used for the sample collection. Standard culture and biochemical techniques were used for the isolation and identification. Structured questionnaires were used to record clinical features, demographic variables and other risk factors of periodontitis.&#x0D; Results: A total of 130 microorganisms were isolated from 49 patients with periodontitis. Male patients accounted for 14.4% and females 18.9% of the all 296 patients who attended our clinics. There was no important association between sex and periodontitis occurrence while there was significant association the younger age groups (45.4% in &lt;26 years of age). The most common signs and symptoms were swollen or puffy gums (91.9%), bleed easily gums (96%), halitosis (96%), painful chewing (87.8%), pus between teeth and gums (71.4%), loose teeth or loss of teeth (44.9%), gingival recession (83.7%), spitting out blood when brushing or flossing teeth (79.6%), and tender gums (93.9%).&#x0D; Conclusion: This study is new in Sana'a city. The clinical features of preiodentitis in Yemen and the risk factors are similar to those reported in the literature elsewhere, but the isolated bacteria differ in frequency from those reported elsewhere, as some upper respiratory tract pathogens such as Streptococcus pyogenes are commonly isolated in this study. Knowledge of the clinical features, bacterial causes of gum disease, and risk factors is the key to successful periodontal therapy.&#x0D; &#x0D; Peer Review History: &#x0D; Received 8 January 2021; Revised 5 February; Accepted 25 February, Available online 15 March 2021&#x0D; UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. &#x0D; Received file: Reviewer's Comments:&#x0D; Average Peer review marks at initial stage: 6.0/10&#x0D; Average Peer review marks at publication stage: 7.5/10&#x0D; Reviewer(s) detail:&#x0D; Dr. Tamer Elhabibi,&#x0D; Suez Canal University, Egypt, tamer_hassan@pharm.suez.edu.eg&#x0D; Dr. Rawaa Souhil Al-Kayali, &#x0D; Aleppo University, Syria, rawah67@hotmail.com&#x0D; Rola Jadallah, &#x0D; Arab American University, Palestine, rola@aauj.edu&#x0D; Prof. Dr. Gorkem Dulger,&#x0D; Duzce University, Turkey, gorkemdulger@yandex.com&#x0D; Similar Articles:&#x0D; RISK FACTORS OF PERIODONTAL DISEASES AMONG YEMENI YOUNG DENTAL PATIENTS&#x0D; KNOWLEDGE AND PERCEPTION OF MOLAR INCISOR HYPOMINERALIZATION AMONG DENTAL PRACTITIONERS IN SANA’A CITY- YEMEN&#x0D; PREVALENCE OF STAPHYLOCOCCUS AUREUS IN DENTAL INFECTIONS AND THE OCCURRENCE OF MRSA IN ISOLATES&#x0D; ASSOCIATION BETWEEN THE STREPTOCOCCUS MUTANS BIOFILM FORMATION AND DENTAL CARIES EXPERIENCE AND ANTIBIOTICS RESISTANCE IN ADULT FEMALES

Evaluation of a unique subgingival irrigation with 10% povidone-iodine after scaling and root planing: A randomized clinical trial.

The beneficial effects of antiseptics in the treatment of periodontitis are still controversial. The goal of this randomized split-mouth study was to evaluate the clinical and microbiologic results of a unique subgingival irrigation of 10% povidone-iodine in addition to a full-mouth scaling and root planing for the treatment of chronic periodontitis. Twenty patients with chronic periodontitis took part in this investigation. In each patient, four initially untreated pockets ≥ 4 mm were randomly selected for one subgingival irrigation. After a one-session full-mouth treatment with scaling and root planing (SRP), two pockets of a split mouth received one unique irrigation with 10 mL 0.9% NaCl solution (control group, CG). Two pockets on the other side received 10 mL of povidone-iodine 10% (test group, PIG). Full-mouth plaque score (FMPS), full-mouth bleeding score (FMBS), Plaque Index (PI), Gingival Index (GI), probing pocket depth (PPD), and clinical attachment level (CAL) parameters were assessed at baseline and after 1, 3, and 6 months (M). Bacterial samplings with sterile paper points were taken at M0 and after M1, M3, and M6. These samplings were separately cultured under aerobic and anaerobic conditions before CFU evaluation. Pl, GI, PPD, and CAL were significantly improved in both groups after 6 months. The major difference was seen between M0 and M3 (P < .001). No significant differences were seen between the groups, except for deep pockets (> 6 mm) where a statistical difference was observed in favor of PIG after 6 months; in these subgroups the mean probing reduction was 2.68 ± 0.37 mm for CG versus 3.93 ± 0.23 mm in PIG (P < .001). No significant differences were seen between CG and PIG regarding the microbiologic results after 6 months. One single irrigation of 10% povidone-iodine associated with full-mouth SRP would bring a small additional benefit in deep pockets. More studies are needed to yield additional results.

Essential Oils as Alternatives for Root-Canal Treatment and Infection Control against Enterococcus faecalis—A Preliminary Study

Since natural alternatives are needed in dentistry for the treatment of root canal, where the standard irrigant is NaOCl with significant toxicity, the aim of the study was to assess the antibacterial properties of non-chemical root-canal irrigants (aqueous extracts of oregano, thyme, lemongrass, melaleuca and clove essential oils) against Enterococcus faecalis. For this, aqueous extracts of each essential oil (AqEO) were prepared. A solution of sodium hypochlorite (NaOCl) was used as a positive standard against which the antimicrobial effects of AqEO could be reported. The root canals of seven teeth were inoculated with 20 µL of Enterococcus faecalis ATCC29212 inoculum and incubated overnight at 37 °C. All the teeth canals were instrumented and were irrigated with the corresponding AqEO, NaOCl and saline solution, then rinsed with saline. Bacteriological samples for each canal post-instrumentation were collected with sterile paper points which were inoculated on culture media. A second processing followed the same methodology but involved only irrigation and no instrumentation. Using instrumentation, thyme and clove completely inhibited Enterococcus faecalis growth. Without instrumentation, clove and oregano AqEOs completely reduced the bacterial load as seen in direct inoculation, but bacterial growth was observed in all the samples after enrichment, except for NaOCl. Nevertheless, the turbidity of the enrichment media was lower for the samples irrigated with AqEOs than for control. In conclusion, AqEOs of thyme, oregano and clove showed a promising antibacterial effect, especially when teeth instrumentation was performed.

Antibacterial Activity of Cobalt Ferrite (CoFe&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;4&lt;/sub&gt;) Nanoparticles against Oral Enterococci

This study aimed to show the enhanced effect of nanoparticles cobalt ferrite CoFe2O4 with chlorhexidine on Enterococcus faecium isolated from failure root canal treatment cases. Sol-gel technique was used to prepare the spinel cobalt ferrite CoFe2O4 at 200 °C then sintering at 400 °C &amp; 600 °C, whereas the antimicrobial susceptibility test of nanoparticles was carried out according to the Kirby-Bauer technique compared with chlorhexidine. Fifteen isolates of Enterococcus faecium obtained by inserting sterile paper points in root canals, from patients who consulted the Teaching Hospital of Dentistry College at the University of Mosul, Iraq. For the first time, results showed that the nanoparticle cobalt ferrite CoFe2O4 at 400 °C and 600 °C, when mixed with chlorhexidine, give the mean inhibition zone 8.5334 mm and 8.0667mm respectively, while the pure chlorhexidine was providing a mean inhibition zone of 3.1667 mm, and also the nanoparticles cobalt ferrite in both cases of sintering without mixing, there was no antimicrobial effect. Statistical analysis using Duncan showed significant differences among groups (p &lt; 0.05).

Comparative evaluation of photoactivated disinfection and sonic irrigation as an adjunct to conventional irrigation on Enterococcus faecalis in root canals: An in vitro study

Objective: The aim of the in vitro study was to compare the antibacterial efficacy of photoactivated disinfection (PAD), sonic irrigation as an adjunct to conventional irrigation against Enterococcus faecalis in vitro.Materials and Methods: A total of 75 extracted teeth were selected and prepared followed by inoculation with strains of E. faecalis, and a preirrigation sample was collected using sterile paper points. These teeth were then divided randomly into three groups for irrigation: Group I (Conventional irrigation), Group II (PAD), and Group III (EndoActivator) followed by postirrigation sample collection using the sterile paper points. The samples were swabbed on blood agar plates and incubated followed by the calculation of colony-forming units (CFU's).Results: The results were statistically analyzed using the SPSS software version 18.0. On comparing the mean values among the groups, the reduction in the number of CFU's after the treatment protocol was highly significant for all groups (P < 0.001). With the preirrigation sample, there is a statistically significant difference in the values of Group I and Group II (P = 0.047). However, in case of postirrigation samples, there is nonsignificant difference between Group II and Group III.Conclusion: PAD using 940 nm diode laser and methylene blue and endoactivator were more effective than sodium hypochlorite (NaOCl) in reducing E. faecalis counts.

Effect of calcium hydroxide alone or in combination with ibuprofen and ciprofloxacin on postoperative pain and periapical prostaglandin E2 level: A randomized clinical study

AimThe aim of the present study was to compare pure Ca(OH)2, Ca(OH)2 + ibuprofen and Ca(OH)2 + ciprofloxacin in terms of postoperative pain and prostaglandin E2 (PGE2) level in previously treated teeth with periapical lesions. Materials and methodsSixty-six patients were randomly assigned into 3 groups according to the intracanal medication (Ca(OH)2, Ca(OH)2 + ibuprofen and Ca(OH)2 + ciprofloxacin). After removing gutta-percha from the root canals, the PGE2 sample collection was obtained by introducing three sterile paper points into the root canals through the root apex (2 mm). Selected intracanal medicament was placed into the root canal and the participants were told to record postoperative pain levels at 24, 48, and 72 h and on 1 week after treatment using visual analog scale (VAS). At the second appointment, the medicaments were removed and second sampling was performed using the same method. The PGE2 levels measured by enzyme-linked immunosorbent assay kits, and the data were statistically analyzed. ResultsAll the tested Ca(OH)2 pastes were found to be significantly effective in lowering the preoperative PGE2 levels. However, intergroup analyses revealed that the Ca(OH)2 + ciprofloxacin group had the highest effectiveness in lowering PGE2 with a significant difference when compared with the pure Ca(OH)2 group. There was no statistically significant difference among the groups in terms of pre- and post-operative pain levels. ConclusionThe Ca(OH)2 + ciprofloxacin medication is more effective than the pure Ca(OH)2 medication in lowering periapical PGE2 level. However, addition of ibuprofen or ciprofloxacin to the Ca(OH)2 paste does not provide extra benefit in terms of post-operative pain relief.

Characterization of Root Canal Microbiota in Teeth Diagnosed with Irreversible Pulpitis

IntroductionPrevious studies have shown that in teeth presenting with symptoms of irreversible pulpitis (IP), bacteria and their by-products driving inflammation are confined mainly within the coronal pulpal tissue. The present study aimed to determine the presence and identity of bacteria within pulps presenting with clinical symptoms of IP using molecular methods. MethodsSamples were obtained from 30 adult patients presenting to the dental emergency department with signs and symptoms of IP. After meticulous surface decontamination, the pulp space was accessed, and clinical samples were collected from inflamed pulp tissue using sterile paper points. Genomic DNA was extracted from the clinical samples, and quantification of bacteria was performed using quantitative polymerase chain reaction targeting the conserved 16S ribosomal RNA (rRNA) gene. To characterize the microbial composition, the V3–V5 hypervariable regions of the 16S rRNA gene were amplified and subjected to next-generation sequencing on the MiSeq platform (Illumina, San Diego, CA). ResultsOf the 30 teeth that presented with IP, half of the intracanal samples had a substantial bacterial load (16S rRNA copies) within the IP vital pulp as determined by quantitative polymerase chain reaction. Next-generation sequencing microbial identification was successful in 7 intracanal samples and yielded 187 bacterial operational taxonomic units within the IP samples. The most abundant genera observed among the vital cases were Veillonella (16%), Streptococcus (13%), Corynebacterium (10%), Cutibacterium (9.3%), and Porphyromonas (5.7%). ConclusionsThe current study highlighted the evidence of vital teeth diagnosed as IP harboring considerable bacterial loads and composed of genera reflective of established endodontic pathology and thus may offer insights into the initial events preceding pulpal necrosis.

Evaluation of Antibacterial Activity of Pine Tar on Periodontal Pathogenic Bacteria: An In Vitro Study.

Periodontal pathogens play an important role in etiology and pathogenesis of periodontitis. Microbiological examination of sub gingival plaque is used at the present time in etiological research as well as in clinical treatment of periodontitis to select the appropriate antibiotic agent if indicated. Pine tar has been used for the treatment of various skin diseases. So the study was done to evaluate the effect of Pine Tar oil on bacteria isolated from periodontitis patients. Plaque samples from volunteer patients were collected using sterile paper points. Robertson's Cooked Meat (RCM) medium was used for the transportation and cultivation of aerobic, microaerophilic, and anaerobic microorganisms. The result suggests the use of Pine tar oil for topical application in periodontal diseases. Disc diffusion analysis was sufficient enough to illustrate that 75 µl tar oil solution produced growth inhibition of microbial strains. Pine tar oil has become one of the important areas of research both in pharmaceutical and periodontal research, hence in vivo studies has to be carried out with various form of pine tar.

Efficacy of antimicrobial photodynamic therapy administered using methylene blue, toluidine blue and tetra 2-mercaptopyridine substituted zinc phthalocyanine in root canals contaminated with Enterococcusaecalis.

Traditional chemomechanical treatment procedures are an indispensable part of endodontic treatment, however, additional treatment approaches such as antimicrobial photodynamic therapy (aPDT) may also be recommended for the elimination of residual microorganisms. In this study, the disinfection efficiency of aPDT performed using methylene blue (MB), toluidine blue (TB), and tetra 2-mercaptopyridine substituted zinc phthalocyanine (TM-ZnPc) was compared in the roots contaminated with Enterococcus faecalis (E. faecalis). Forty-nine teeth with a single root and canal were included in this study. The roots were sterilized, and inoculated with E. faecalis. The roots were kept in an incubator for 30 days to form the biofilm. Forty-five teeth were prepared up to the F3 file of the ProTaperNext system under 2.5 % NaOCL irrigation. The samples were divided into three groups according to the type of used photosensitizer (PS) (n = 15); MB (313 μM), TB (327 μM), and TM-ZnPc (6μM). All PSs were irradiation with a light-emitting diode (LED) lamp (630 nm, 2-4 mW/cm²) for the 60 s. Two microbiological samples of the intracanal content were taken (one before and one immediately after additional aPDT in all groups) using sterile paper points. The colony-forming units per milliliter (CFU/mL) were calculated after 24 h of incubation at 37 °C. After all aPDT protocols, intracanal bacterial load decreased significantly compared to the amount after chemomechanical preparation (P < 0.05). No significant difference in the reduction in intracanal bacterial load was found between the PSs (P < 0.05). In the current study, the aPDT protocol performed with TM-ZnPc provided similar antimicrobial efficacy, although it was used at a lower concentration compared to MB and TB. Therefore, the use of TM-ZnPc in intra-canal disinfection in endodontics seems promising.

COMPARATIVE EVALUATION OF ANTIBACTERIAL EFFICACY OF CHLOROQUICKONE STEP IRRIGATING SOLUTION AND CHLORHEXIDINE GLUCONATE SOLUTION ON E. FAECALIS IN ROOT CANAL SYSTEM OF PRIMARY MOLARS: A RANDOMISED EX VIVO STUDY.

Background: Microorganisms and their by-products are considered to be primary etiologic agents in endodontic diseases. Mechanical preparation is the principal mode of reducing the bacterial population in the root canal system, smear layer is still left on the root canal wall. Hence, irrigation of root canal allows for cleaning beyond what might be achieved by root canal instrumentation allows. Aim: To evaluate and compare antibacterial efficacy of Chloroquick- one step irrigating solution and 2% Chlorhexidine Gluconate on E. faecalis in primary molars by Quantitative Reverse Transcriptase Polymerase Chain Reaction (qRT-PCR) study. Methodology: 40 children between age 4-7 years were included in this ex- vivo study based on selection criteria. In Group I (n=20) irrigation was done with chloroquick and normal saline and in Group II (n=20) irrigation was done with 2% chlorhexidine gluconate and normal saline. Pre- and post-irrigation samples were collected using sterile paper points and transferred into transporting medium in Eppendorf tube and sent for qRT-PCR analysis. Results: On intra-group comparison, in Group I a statistically significant reduction in E. Faecalis (colony forming units/ml) count post-irrigation was found (p=0.0130). While for Group II, there was no statistically significant reduction in E. Faecalis (colony forming units/ml) count post-irrigation (p=0.8180). On inter-group comparison between, Group I showed statistically significant reduction in E. Faecalis (colony forming units/ml) count as compared to Group II (p=0.0110) Conclusion: In the present study, continuous soft chelating irrigation protocol has shown promising results for reduction of microbial load from the root canal system of primary teeth

Investigation of microbial profile, levels of endotoxin and lipoteichoic acid in teeth with symptomatic irreversible pulpitis: a clinical study.

To investigate the microbial profile, and levels of endotoxin (LPS) and lipoteichoic acid (LTA), in infected dentine (ID) and root canals (RC) at different phases of root canal treatment in teeth with symptomatic irreversible pulpitis. Ten volunteers were included, and samples were collected from infected dentine (ID) and the root canal lumen (RC) using sterile excavators and paper points, respectively. RC samples were taken before (S1) and after (S2) chemo-mechanical canal preparation (CMP), and after intracanal medication (ICM; S3). Checkerboard DNA-DNA hybridization was used for microbial analysis. The levels of LPS and LTA were evaluated using the limulus amebocyte lysate assay and ELISA, respectively. Shapiro-Wilk's test was used to verify data normality. Friedman's test was used to evaluate statistical differences using checkerboard DNA-DNA hybridization in the ID and RC at the different phases of the RC treatment. Post hoc Dunn's multiple comparison test was used to verify significant differences recorded at the different time-points. The levels of LPS and LTA were analysed statistically by using repeated measures anova and Tukey's post hoc test to evaluate differences in both sites. The significance level was set at 5% (P<0.05). A total of 40 DNA probes were used for microbial investigation of ID and RC samples using checkerboard DNA-DNA hybridization. The levels and complexity of bacteria were similar in the ID and initial RC samples. The levels of LPS and LTA in ID were significantly higher than the initial RC samples (S1; P<0.05). Canal preparation was effective in significantly decreasing the levels of bacteria, LPS and LTA (P<0.05). ICM did not provide additional reduction in the levels of bacteria and LPS (P>0.05). However, a significant reduction in the levels of LTA was observed after ICM (P<0.05). The microbial profile of infected dentine and root canals of teeth with irreversible pulpitis was complex, harbouring different species including Gram-positive and Gram-negative, cocci and bacilli, and facultative and strict anaerobes. Root canal preparation was effective in reducing the levels of bacteria, LPS and LTA from the root canals of teeth with pulpitis.

Microbiological analysis of endodontically treated teeth with apical periodontitis before and after endodontic retreatment

To characterize the microbiota of teeth with endodontic treatment failure by 16S ribosomal RNA genetic sequencing (GS) and PCR at the different phases of the endodontic retreatment and to associate the presence of specific bacteria with clinical and radiographic features in teeth with apical periodontitis. Twenty infected root canals of single-rooted teeth were selected. Samples were collected with sterile paper points before chemo-mechanical preparation (CMP) (S1), after CMP (S2) and after 30 days of intracanal medication (ICM) (S3). Microbial identification was performed using GS and PCR. Tukey-Kramer post hoc test and post hoc ANOVA were used for intergroup analysis. Paired t test and repeated-measures ANOVA were applied for intragroup analysis, at a significance level of 5%. A total of 89 strains were identified using GS. Sixty-five strains were recovered in S1 and 15 strains in S2, and 9 strains remained in S3. Enterococcus faecalis was the most predominant bacteria. Gram-positive cocci bacteria predominated. Gram-negative species were also detected. Using species-specific PCR primers to detect seven species, the most prevalent ones at all the phases of the endodontic retreatment were E. faecalis and Porphyromonas gingivalis. However, Parvimonas micra and P. gingivalis were associated with previous pain, P. gingivalis was associated with tenderness to percussion and E. faecalis, Fusobacterium nucleatum and P. gingivalis were associated with periapical lesion > 3 mm. In conclusion, the microbiota of persistent infection is polymicrobial with predominance of E. faecalis and P. gingivalis in all phases of the endodontic retreatment, regardless of the method used for microbial identification. Associations were found between specific bacteria and clinical/radiographic features. The characterization of the bacteria present at all phases of the endodontic retreatment is important for the monitoring of the effectiveness of the techniques used and to better understand the susceptibility of these species to the disinfection agent used during the procedures.

Improving the Efficiency of Photodynamic Chemotherapy in Root Canals against Enterococcus faecalis In Vitro.

The aim of this study was to evaluate the effect of photoactivated chemotherapy (PACT) on Enterococcus faecalis (E. faecalis) biofilms in root canals using an 90% isopropanol (IPA)-based photosensitizer and removing excess photosensitizer before light incubation. Three hundred and seven extracted human teeth with one root canal were infected with E. faecalis for 72 h and treated in groups: IPA irrigation; PACT; PACT and final rinse with IPA; PACT with photosensitizer removal using either 0.9% NaCl solution or sterile paper points or both; PACT using IPA-based photosensitizer with and without a final rinse of IPA. Root canals were sampled using sterile paper points and dentin chips collected from the root canal walls. Additionally, SEM (Scanning Electron Microscopy) images of the specimens were taken to evaluate the root canal walls for residue bacterial contamination. In all antimicrobial treatment groups treatments E. faecalis counts were significantly reduced in the root canals. Using IPA-based photosensitizer the antimicrobial effect of PACT was significantly enhanced. Irrigation with IPA alone or after PACT significantly increased the antimicrobial effect compared to PACT alone. The collected dentin chips revealed the highest amount of culture negative root canals (10%) after PACT using IPA-based photosensitizer. In the other groups, the culture negative samples ranged from only 0–2 specimens of 30 specimens. REM images show remaining E. faecalis cells on the root canal wall and inside dentin tubules. Using IPA-based photosensitizer significantly enhanced the antimicrobial effect of PACT against E. faecalis in the root canals.

Comparison of endodontic disinfection of primary teeth root canals using rotary and reciprocating system: An in vitro study

Aim: This experimental study in vitro aimed to compare the disinfection of the primary teeth root canals system using the rotary and reciprocating file system. Methodology: Forty-eight root canals from twenty-four primary molar teeth were contaminated with the standard strain of Enterococcus faecalis and randomically splitted into 4 groups (n=12). The Group PN comprised the root canals prepared with the ProTaperTM Next rotary system. The Group WOG was defined by the usage of WaveOneTM Gold reciprocating system. The files from both systems were thermo mechanical processed. The Group PU used the ProTaperTM Universal system, with no thermic treatment, and the Group C (negative control) received no treatment. Samples from the root canals were collected with sterile paper points before and after its preparation, diluted and distributed in plates with Brain Heart Infusion (BHI) agar. The bacterial colonies were counted and the results were statistic analyzed by Wilcoxon and Kruskal-Wallis (Dunn) tests. Results: There was significant bacterial reduction using the systems (p&gt;0,01). However, there was no significant difference among them (p&lt;0,05) (PN=3.38; WOG=3.85; PU=3.26). Conclusion: Both systems (rotary and reciprocating) provide disinfectionon the primary molar teeth root canals system.

Detection of HPV16 E6 DNA in periodontal pockets of middle‐aged and older people

AbstractBackgroundThis study was conducted to investigate the association between oral human papillomavirus type 16 (HPV16) DNA prevalence and periodontitis.Materials and MethodsWe analyzed 89 patients (mean age 71.9 years, range 48‐90 years) who visited the Department of Oral Health of Hiroshima University Hospital. Samples were collected by inserting sterilized paper points into each patient's deepest periodontal pocket to obtain gingival crevicular fluid. Real‐time PCR was performed using type‐specific primers to detect HPV16 DNA. PCR‐targeted bacterial 16S rRNA genes were used to detect periodontal disease‐related bacteria.ResultsOf the 89 participants, four women (4.5%) were HPV16 DNA‐positive. Three of the four HPV16 DNA‐positive participants (75.0%) exhibited bleeding after the paper point was inserted, indicating periodontal tissue inflammation. However, no significant association was found between bleeding and the HPV16 DNA‐positive rate. Importantly, HPV16 DNA positivity was significantly associated with Treponema denticola and Fusobacterium nucleatum (P = 0.01 and P = 0.02, respectively).ConclusionsHuman papilloma virus localizes in periodontal pockets. Inflamed periodontal pockets may indicate oral HPV infection. Periodontitis‐related bacteria may be involved in HPV infection of the periodontal tissues.

Investigation of Filifactor alocis in primary and in secondary endodontic infections: A molecular study

ObjectiveIdentification of specific bacteria in root canals (RCs) in distinct clinical conditions can support the comprehension of pathological processes. Thus, the objective of this clinical study was to investigate the presence of F. alocis in RCs of teeth with primary endodontic infection (PEI) and with persistent/secondary endodontic infection (SEI) by using molecular techniques. It was also aimed to associate its presence with the clinical features. In addition, the levels of F. alocis as well as the total bacterial cells in the samples were also quantitated. DesignOne hundred teeth (50 PEI and 50 SEI) were included. Microbial samples were performed using sterile paper points and assessed by using nested PCR and quantitative Real Time PCR (qPCR). The prevalence of F.alocis in RCs from PEI and SEI were compared by chi-square analysis. Fisher´s exact test or Pearson Chi-square, when appropriate, was used to test associations between clinical and radiographic features and the presence of F. alocis. Significance level was set at 5%. ResultsF. alocis was detected in 23 and 28 (PEI) and 12 and 11 (SEI) RCs using Nested PCR and qPCR, respectively. Statistically significant associations were found between the presence of F. alocis and PEI, pain, wet canals, swelling, abscess and purulent exudate (P < 0.05). Total bacterial count was similar in both conditions (P > 0.05). ConclusionsPEI harbour a significantly higher number of F. alocis than those with SEI. Filifactor alocis was significantly associated with clinical features in primary endodontic infections. Total bacterial count was similar in both clinical conditions.

Clinical Investigation of Microbial Profile and Levels of Endotoxins and Lipoteichoic Acid at Different Phases of the Endodontic Treatment in Teeth with Vital Pulp and Associated Periodontal Disease

IntroductionThe aim of this in vivo study was to investigate the microbial profile as well as the levels of lipopolysaccharide (LPS) and lipoteichoic acid (LTA) at different phases of endodontic treatment in teeth with vital pulp and associated periodontal disease. MethodsTen patients were selected for this clinical study. Samples were taken from periodontal pockets (PPs) and root canals (RCs) using sterile paper points before and after chemomechanical preparation and after intracanal medication. For microbiological analysis, nested polymerase chain reaction and checkerboard DNA-DNA hybridization were used. Levels of LPS and LTA were assessed using limulus amebocyte lysate and enzyme-linked immunosorbent assays, respectively. Data were statistically analyzed at a significance level of 5%. ResultsBacterial DNA from 17 of the 17 species investigated was detected in samples of PPs, whereas 6 of the 17 species were not present in the initial samples of RCs using nested polymerase chain reaction. In the initial samples, 38 of 40 probes were detected in PPs, whereas 12 of 40 probes were detected in RCs using checkerboard DNA-DNA hybridization. Overall, endodontic procedures were efficient in modifying the microbiota of PPs and RCs. Levels of LPS and LTA were reduced after the endodontic procedures, although higher concentrations of both had been found in PPs compared with RCs. ConclusionsThe microbiota of PPs and RCs in teeth with vital pulp and associated periodontal disease is polymicrobial, with the presence of gram-negative, gram-positive, facultative, and strict anaerobes. Chemomechanical preparation and calcium hydroxide–based intracanal medication allowed the reduction of infectious content in both sites.