Antimicrobial Efficacy of Salvadora persica Extracts on a Monospecies Biofilm on Orthodontic Brackets In Vitro.

PURPOSE: The oral cavity is a rich ecosystem with a plethora of microorganisms, and different components of fixed orthodontic appliances may contribute to a shift in the balance of oral ecology. The purpose of this study was to investigate the antimicrobial potential of hexane and ethanol extracts of Salvadora persica on a monospecies biofilm model established on orthodontic brackets in vitro. MATERIALS AND METHODS: Streptococcus mutans biofilm was formed on mini diamond orthodontic brackets following three days of anaerobic incubation at 37˚C. The bacterial cell viability of this biofilm was measured after their exposure to saline, hexane extract of S. persica, ethanol extract of S. persica and 0.2% chlorhexidine using 3-(4, 5-dimethylthiazol- 2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrazolium (MTS) assay. On half of the brackets, the colony forming units (CFU) were counted. Both experiments were performed in triplicate. RESULTS: The absorbance values obtained from the MTS reduction assay after exposure to the different test agents showed a decline in the bacterial cell viability of the S. mutans biofilm as follows: chlorhexidine (+)0.05). The CFU counts of S. mutans obtained from chlorhexidine exposure were lower than from hexane and ethanol extracts. CONCLUSION: S. persica extracts were found to have antimicrobial effects on S. mutans biofilm established in vitro on orthodontic brackets suggestive of its potential use as an oral antimicrobial agent for orthodontic patients.

Obturation Techniques Allow Microbial Leakage Unless Protected.

PURPOSE: To evaluate the quality of the apical 5-mm seal produced by different filling techniques using a bacterial leakage model. MATERIALS AND METHODS: Sixty-five extracted single-rooted human teeth were decoronated, prepared, and instrumented. Roots were randomly divided into three experimental groups (15 roots each) and control groups (10 roots each). The apical 5 mm was filled with cold lateral condensation (CLC) technique, continuous wave of condensation (CWC), or injectable thermoplasticized gutta-percha (Obtura II) using AH26 Plus as a sealer. Positive controls were filled with gutta-percha without sealer, whereas negative controls were filled with a CLC technique and covered completely with two layers of nail varnish, including the orifice. A dual-chamber leakage model using Enterococcus faecalis as a microbial marker was used for leakage evaluation. Bacterial penetration was monitored over a 60-day period, and leakage was recorded when turbidity was observed in the lower chamber. RESULTS: All positive controls exhibited turbidity in the lower chamber within 24 hours. All negative controls demonstrated no bacterial leakage for the entire 60-day observation period. The estimated mean day for leakage was 32 for CLC, 35 for CWC, and 30 days for Obtura II. Wilcoxon test showed no statistically significant difference (p = 0.98) in the survival time between the experimental groups. CONCLUSION: The three filling techniques produced similar resistance to bacterial leakage when used to fill the apical 5-mm segment of the canal while leaving the rest of the canal unfilled.

Antibiofilm effect of different concentrations of silver nanoparticles combined with calcium hydroxide against Enterococcus faecalis biofilm: An ex vivo study.

This study aimed to evaluate the antibiofilm activity of different concentrations of silver nanoparticles (AgNPs) in combination with calcium hydroxide [Ca(OH)2] against Enterococcus faecalis biofilm. On an E. faecalis biofilm on dentin discs, the following medicaments were applied for 7 days (n = 13/group): 0.005% AgNPs+Ca(OH)2, 0.01% AgNPs+Ca(OH)2, 0.02% AgNPs + Ca(OH)2, Ca(OH)2 and saline/control. Specimens were stained with LIVE/DEAD® BacLight™ dye and analysed with confocal laser scanning microscopy. Proportion of dead bacteria was calculated and analysed. There was a significant reduction in E. faecalis biofilm in all medicament groups (43.5%, 49.1%, 69.1%, 48.7%) respectively, compared with control group (2.54%) (p < 0.001). The 0.02% AgNPs + Ca(OH)2 group demonstrated the most significantly superior antibiofilm effect, with no significant difference between remaining groups. In conclusion, combining 0.02% AgNPs enhanced the antibiofilm effect of Ca(OH)2 on E. faecalis biofilm compared with lower AgNPs concentrations.

Biomarkers of non-communicable chronic disease: an update on contemporary methods.

Chronic diseases constitute a major global burden with significant impact on health systems, economies, and quality of life. Chronic diseases include a broad range of diseases that can be communicable or non-communicable. Chronic diseases are often associated with modifications of normal physiological levels of various analytes that are routinely measured in serum and other body fluids, as well as pathological findings, such as chronic inflammation, oxidative stress, and mitochondrial dysfunction. Identification of at-risk populations, early diagnosis, and prediction of prognosis play a major role in preventing or reducing the burden of chronic diseases. Biomarkers are tools that are used by health professionals to aid in the identification and management of chronic diseases. Biomarkers can be diagnostic, predictive, or prognostic. Several individual or grouped biomarkers have been used successfully in the diagnosis and prediction of certain chronic diseases, however, it is generally accepted that a more sophisticated approach to link and interpret various biomarkers involved in chronic disease is necessary to improve our current procedures. In order to ensure a comprehensive and unbiased coverage of the literature, first a primary frame of the manuscript (title, headings and subheadings) was drafted by the authors working on this paper. Second, based on the components drafted in the preliminary skeleton a comprehensive search of the literature was performed using the PubMed and Google Scholar search engines. Multiple keywords related to the topic were used. Out of screened papers, only 190 papers, which are the most relevant, and recent articles were selected to cover the topic in relation to etiological mechanisms of different chronic diseases, the most recently used biomarkers of chronic diseases and finally the advances in the applications of multivariate biomarkers of chronic diseases as statistical and clinically applied tool for the early diagnosis of chronic diseases was discussed. Recently, multivariate biomarkers analysis approach has been employed with promising prospect. A brief discussion of the multivariate approach for the early diagnosis of the most common chronic diseases was highlighted in this review. The use of diagnostic algorithms might show the way for novel criteria and enhanced diagnostic effectiveness inpatients with one or numerous non-communicable chronic diseases. The search for new relevant biomarkers for the better diagnosis of patients with non-communicable chronic diseases according to the risk of progression, sickness, and fatality is ongoing. It is important to determine whether the newly identified biomarkers are purely associations or real biomarkers of underlying pathophysiological processes. Use of multivariate analysis could be of great importance in this regard.

Gut Microbiota Dynamics in Relation to Long-COVID-19 Syndrome: Role of Probiotics to Combat Psychiatric Complications.

Increasing numbers of patients who recover from COVID-19 report lasting symptoms, such as fatigue, muscle weakness, dementia, and insomnia, known collectively as post-acute COVID syndrome or long COVID. These lasting symptoms have been examined in different studies and found to influence multiple organs, sometimes resulting in life-threating conditions. In this review, these symptoms are discussed in connection to the COVID-19 and long-COVID-19 immune changes, highlighting oral and psychiatric health, as this work focuses on the gut microbiota's link to long-COVID-19 manifestations in the liver, heart, kidney, brain, and spleen. A model of this is presented to show the biological and clinical implications of gut microbiota in SARS-CoV-2 infection and how they could possibly affect the therapeutic aspects of the disease. Probiotics can support the body's systems in fighting viral infections. This review focuses on current knowledge about the use of probiotics as adjuvant therapies for COVID-19 patients that might help to prevent long-COVID-19 complications.

Influence of Preclinical Training on Root Canal Treatment Technical Quality and Confidence Level of Undergraduate Dental Students.

OBJECTIVES: The aim of this study was to investigate the influence of exposure to additional preclinical endodontic training on undergraduate students' technical quality of root canal treatment and overall confidence levels in endodontics. METHODS: Technical quality of root canal treatment performed clinically by fifth-year undergraduate students was evaluated and divided into two groups: Group 1, teeth treated by students who had attended both a preclinical endodontic block course and an elective preclinical course. Group 2: teeth treated by students who had not attended the elective preclinical course. All students were also invited to participate in a survey to rate their undergraduate endodontic training and confidence levels performing endodontic treatment. Statistical analysis of data was performed using Person chi-square test, Fisher Freeman Halton exact test, and t-test. A p-value <0.05 was considered statistically significant. RESULTS: There was no significant difference between the two groups in overall obturation quality (p=0.619). However, more teeth treated by attendees were of adequate obturation length (p=0.015) and lacked procedural errors (p=0.004). Significantly more elective course attendees rated their undergraduate endodontic training as adequate (p=0.002), but there was no significant difference in the level of confidence between the attendees and the non-attendees. CONCLUSION: Within the limitations of this study, additional preclinical training showed minimal effect on overall quality of root canal treatment performed clinically by undergraduate students and did not enhance their confidence levels; however, it was associated with more satisfaction with their undergraduate endodontic education.

Comparative analysis of prevalence of apical periodontitis in smokers and non-smokers using cone-beam computed tomography.

OBJECTIVE: The aim of this study was to compare the prevalence and size of periapical lesions among smokers and non-smokers using cone-beam computed tomography (CBCT). MATERIALS AND METHODS: Retrievable CBCT datasets for 46 male patients ≥18 years during a consecutive period from 2008 to 2016 were examined. The medical, smoking history and other clinical findings (signs of previous dental trauma; Decayed Missing Filled Teeth (DMFT) scores; the percentage of root filled teeth; and oral hygiene status) were obtained. Periapical status of all included teeth was assessed by CBCT images. Statistical analysis was conducted using t-test, Pearson correlation and multiple regression. RESULTS: The prevalence of apical periodontitis was 13.93% in smokers and 14.26% in non-smokers with no significant difference (p = 0.936). The mean of the average size of lesions between the two groups were almost comparable, 3.50 mm in smokers and 2.89 mm in non-smokers (p = 0.567). Pearson correlation and multiple regression analysis showed that the percentage of lesion present and the average lesion size were not correlated to any independent variable. CONCLUSIONS: While smoking is considered a risk factor for marginal periodontitis, there was no difference between smokers and non-smokers in terms of apical periodontitis.

Mutations in MSX1, PAX9 and MMP20 genes in Saudi Arabian patients with tooth agenesis.

Tooth agenesis in human being is the most common congenital anomaly associated with dental development. Mutations in many genes such as MSH homeobox 1 (MSX1), paired box gene 9 (PAX9), ectodysplasin A (EDA) and EDA receptor (EDAR) have been associated with familial form of this condition. However, in large majority of patients, genetic cause could not be identified. The primary aim of present study was to identify the causative mutation(s) in these genes in Saudi Arabian families diagnosed with non-syndromic form of disease. Direct sequencing of coding regions, including exon-intron boundaries of these genes was carried out. All identified nucleotide variations were also tested to exclude possibility of being rare polymorphisms. The sequence analysis of exons and exon-intronic regions of these genes revealed five new mutations that include four in MSX1, one in PAX9 and one single nucleotide polymorphism (SNP) in majority of the patients in MMP20. One novel mutation in exon 1 of MSX1 gene (5354C > G; A40G) was found in three patients. In addition, another novel mutation was detected in two patients in exon 3 (PAX9) as g.10672A > T which changes asparagine to isoleucine at position 40. These mutations were not found in any of the control subjects. A single SNP in MMP20 genes (g.5066A > C) that changes lysine to threonine at position 18 was found in 10% controls as well. Our results for the first time demonstrates that mutations in MSX1 gene might play an important role in hypodontia cases involving pre-molars and is a risk factor for this ethnic population mainly of Arabs and is first report linking these mutations with tooth agenesis.

Combined effect of a mixture of tetracycline, acid, and detergent, and nisin against Enterococcus faecalis and Actinomyces viscosus biofilms.

OBJECTIVES: To evaluate the combined effect of a mixture of tetracycline, acid, and detergent (MTAD) and Nisin against Enterococcus faecalis (E. faecalis) and Actinomyces viscosus (A. viscosus) biofilms. METHODS: This study was conducted between June and December 2013 in collaboration with Dental Caries Research Chair, College of Dentistry, King Saud University, Riyadh, Saudi Arabia. Single-species biofilms (n=9/species/observation period) were generated on membrane filter discs and subjected to 5, 10, or 15 minute incubation with MTADN (MTAD with 3% Nisin), 5.25% sodium hypochlorite (NaOCl), or normal saline. The colony forming units were counted using the Dark field colony counter. RESULTS: A 100% bactericidal effect of 5.25% NaOCl was noted during the 3 observation periods; a significant reduction (p=0.000) in mean survival rates of E. faecalis (77.3+13.6) and A. viscosus (39.6+12.6) was noted after 5 minutes exposure to MTADN compared with normal saline (78000000+5291503) declining to almost no growth after 10 and 15 minutes. The survival rates of the E. faecalis and A. viscosus biofilm were no different after treatment with MTADN and 5.25% NaOCl at the 3 observation periods (p=1.000). CONCLUSION: A combination of MTAD and Nisin was as effective as NaOCl against E. faecalis and A. viscosus biofilms.

Attachment and morphological behavior of human periodontal ligament fibroblasts to mineral trioxide aggregate: a scanning electron microscope study.

The attachment and morphology of human periodontal ligament fibroblasts to mineral trioxide aggregate (MTA) was evaluated using a scanning electron microscope. The material was placed at an apical cavity of 30 single-rooted slices of extracted human teeth. The specimens were divided into two groups of 15 root slices each (freshly mixed and set state). For each experimental group, five root slices were used per observation period (4, 8, and 24 h). A set of two glass slides was used per observation period for the control group. The experiments were performed in tissue-culture cluster 96-well plates in which 1 ml of human periodontal ligament fibroblast cell suspension was placed over the MTA filling and the control glass slides. For the positive-control group, 0.5 ml of methyl methacrylate 2% (vol/vol) was added to the cell suspensions before being dispensed into the wells. Results showed the normal cell morphology in the negative controls. Few round cells with less smooth surfaces and many rough blebs were seen in the positive control, and most of these cells did not show any attachment to the substratum. Similar observations were seen with the freshly prepared-MTA group. In the set-MTA group, cells were round and flattened, displayed smooth surfaces, and appeared to be tightly attached to MTA. It was concluded that the quality and quantity of cell attachment to the retrofilling material could be used as a criterion to evaluate material's toxicity. This research (FN#1077) is registered with the College of Dentistry research center, King Saud University, Riyadh, Saudi Arabia. The author thanks the administration of the King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia, and in particular Dr. M. N. Al-Ahdal for providing the use of the Molecular Virology and Infectious Disease Laboratory, Mr. Yunus Siddiqui for his support, and Dr. Saad AL-Nazhan for his assistance in preparing the manuscript.

Cytotoxic effect of Salvadora persica extracts on human gingival fibroblast cells.

OBJECTIVES: To assess the cytotoxic potential of Salvadora persica (S. persica) extracts on human gingival fibroblast (HGF) cells. METHODS: This study was conducted between January and May 2012 in collaboration with Dental Caries Research Chair, College of Dentistry, King Saud University, Riyadh, Saudi Arabia. Extracts of S. persica using hexane, ethylacetate, and ethanol as solvents at concentrations of 0.5 mg/ml and 1 mg/ml were evaluated for their cytotoxic activity against HGFs using the 3 cytotoxic assays: (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole) (MTS), lactic dehydrogenase (LDH), and crystal violet (CV). International standards for the evaluation of medical materials recommended cut-off value of cell survival >70% was used for interpretation of the results. RESULTS: Ethanol extract of S. persica at 0.5 mg/ml and 1 mg/ml and hexane extract of S. persica at 0.5 mg/ml were completely devoid of cytotoxic activity, hexane extract at 1 mg/ml in comparison with controls  demonstrated some cytotoxicity with cell survival of 88% (p=0.045) in MTS, 86% (p=0.01) in LDH, and 88% (p=0.002) in CV assays. Similarly, ethyl acetate extract of S. persica at 0.5 mg/ml maintained cell viability of 91% in MTS, 81% in LDH, and 80% in CV assays. Maximum cytotoxicity against HGFs was observed with ethyl acetate extract of S. persica at 1 mg/ml with cell survival of 60% in MTS, 40% in LDH, and 66% CV assays (p=0.0001).   CONCLUSION: The acceptable level of cytotoxicity associated with S. persica ethanol and hexane extracts requires further evaluation to be used as irrigation solutions in endodontic treatment.