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Histatin-5 (Hst5) is a member of the histatin superfamily of cationic, His-rich, Zn(II)-binding peptides in human saliva. Hst5 displays antimicrobial activity against fungal and bacterial pathogens, often in a Zn(II)-dependent manner. In contrast, here we showed that under in vitro conditions that are characteristic of human saliva, Hst5 does not kill seven streptococcal species that normally colonize the human oral cavity and oropharynx. We further showed that Zn(II) does not influence this outcome. We then hypothesized that Hst5 exerts more subtle effects on streptococci by modulating Zn(II) availability. We initially proposed that Hst5 contributes to nutritional immunity by limiting nutrient Zn(II) availability and promoting bacterial Zn(II) starvation. By examining the interactions between Hst5 and Streptococcus pyogenes as a model Streptococcus species, we showed that Hst5 does not influence the expression of Zn(II) uptake genes. In addition, Hst5 did not suppress growth of a ΔadcAI mutant strain that is impaired in Zn(II) uptake. These observations establish that Hst5 does not promote Zn(II) starvation. Biochemical examination of purified peptides further confirmed that Hst5 binds Zn(II) with high micromolar affinities and does not compete with the AdcAI high-affinity Zn(II) uptake protein for binding nutrient Zn(II). Instead, we showed that Hst5 weakly limits the availability of excess Zn(II) and suppresses Zn(II) toxicity to a ΔczcD mutant strain that is impaired in Zn(II) efflux. Altogether, our findings led us to reconsider the function of Hst5 as a salivary antimicrobial agent and the role of Zn(II) in Hst5 function.

Dental procedures produce aerosols which may remain suspended and travel significant distances from the source. Dental aerosols and droplets contain oral microbes and there is potential for infectious disease transmission and major disruption to dental services during infectious disease outbreaks. One method to control hazardous aerosols often used in industry is Local Exhaust Ventilation (LEV). The aim of this study was to investigate the effect of LEV on aerosols and droplets produced during dental procedures. Experiments were conducted on dental mannequins in an 825.4 m3 open plan clinic, and a 49.3 m3 single surgery. 10-minute crown preparations were performed with an air-turbine handpiece in the open plan clinic, and 10-minute full mouth ultrasonic scaling in the single surgery. Fluorescein was added to instrument irrigation reservoirs as a tracer. In both settings, Optical Particle Counters (OPCs) were used to measure aerosol particles between 0.3 – 10.0 μm and liquid cyclone air samplers were used to capture aerosolised fluorescein tracer. Additionally, in the open plan setting fluorescein tracer was captured by passive settling onto filter papers in the environment. Tracer was quantified fluorometrically. An LEV device with High Efficiency Particulate Air (HEPA) filtration and a flow rate of 5,000 L/min was used. LEV reduced aerosol production from the air-turbine handpiece by 90% within 0.5 m, and this was 99% for the ultrasonic scaler. OPC particle counts were substantially reduced for both procedures, and air-turbine settled droplet detection reduced by 95% within 0.5 m. The effect of LEV was substantially greater than suction alone for the air-turbine and was similar to the effect of suction for the ultrasonic scaler. LEV reduces aerosol and droplet contamination from dental procedures by at least 90% in the breathing zone of the operator and it is therefore a valuable tool to reduce the dispersion of dental aerosols.

Extracellular DNA (eDNA) is an important component of biofilm matrix that serves to maintain biofilm structural integrity, promotes genetic exchange within the biofilm, and provides protection against antimicrobial compounds. Advances in microscopy techniques have provided evidence of the cobweb- or lattice-like structures of eDNA within biofilms from a range of environmental niches. However, methods to reliably assess the abundance and architecture of eDNA remain lacking. This study aimed to address this gap by development of a novel, high-throughput image acquisition and analysis platform for assessment of eDNA networks in situ within biofilms. Utilizing Streptococcus gordonii as the model, the capacity for this imaging system to reliably detect eDNA networks and monitor changes in abundance and architecture (e.g., strand length and branch number) was verified. Evidence was provided of a synergy between glucans and eDNA matrices, while it was revealed that surface-bound nuclease SsnA could modify these eDNA structures under conditions permissive for enzymatic activity. Moreover, cross talk between the competence and hexaheptapeptide permease systems was shown to regulate eDNA release by S. gordonii. This novel imaging system can be applied across the wider field of biofilm research, with potential to significantly advance interrogation of the mechanisms by which the eDNA network architecture develops, how it can influence biofilm properties, and how it may be targeted for therapeutic benefit. IMPORTANCE Extracellular DNA (eDNA) is critical for maintaining the structural integrity of many microbial biofilms, making it an attractive target for the management of biofilms. However, our knowledge and targeting of eDNA are currently hindered by a lack of tools for the quantitative assessment of eDNA networks within biofilms. Here, we demonstrate use of a novel image acquisition and analysis platform with the capacity to reliably monitor the abundance and architecture of eDNA networks. Application of this tool to Streptococcus gordonii biofilms has provided new insights into how eDNA networks are stabilized within the biofilm and the pathways that can regulate eDNA release. This highlights how exploitation of this novel imaging system across the wider field of biofilm research has potential to significantly advance interrogation of the mechanisms by which the eDNA network architecture develops, how it can influence biofilm properties, and how it may be targeted for therapeutic benefit.

Infection control is critical for the safe delivery of dental care. Infection control practices must be responsive to emerging and re-emerging infectious diseases and outbreaks, as was clearly seen during the peak of the COVID-19 pandemic. An emerging global outbreak of the monkeypox virus has again raised potential challenges for infection control in dentistry. Monkeypox is an infectious disease, characterised by a rash affecting the skin and soft tissues, including the oral cavity. Previously, cases were mostly seen following contact with infected animals in Central and West Africa, with limited human-to-human transmission within and outside of these areas. However, since May 2022, sustained human-to-human transmission has occurred globally. Monkeypox can be transmitted via close contact with an infected person, contaminated objects and surfaces, or by droplets and possibly aerosols, which is therefore of potential importance to dental settings. This article discusses the relevance of monkeypox to dental professionals, the typical presentation of the disease, its potential impact on infection prevention and control practices and the delivery of dental services. The current monkeypox outbreak highlights the need for a more sustained programme of research into dental infection control that can provide a solid evidence base to underpin preparedness planning for future outbreaks and pandemics.

Background Dental procedures often produce aerosol and splatter which have the potential to transmit pathogens such as SARS‐CoV‐2. The existing literature is limited. Objective(s) To develop a robust, reliable and valid methodology to evaluate distribution and persistence of dental aerosol and splatter, including the evaluation of clinical procedures. Methods Fluorescein was introduced into the irrigation reservoirs of a high‐speed air‐turbine, ultrasonic scaler and 3‐in‐1 spray, and procedures were performed on a mannequin in triplicate. Filter papers were placed in the immediate environment. The impact of dental suction and assistant presence were also evaluated. Samples were analysed using photographic image analysis, and spectrofluorometric analysis. Descriptive statistics were calculated and Pearson’s correlation for comparison of analytic methods. Results All procedures were aerosol and splatter generating. Contamination was highest closest to the source, remaining high to 1–1.5 m. Contamination was detectable at the maximum distance measured (4 m) for high‐speed air‐turbine with maximum relative fluorescence units (RFU) being: 46,091 at 0.5 m, 3,541 at 1.0 m, and 1,695 at 4 m. There was uneven spatial distribution with highest levels of contamination opposite the operator. Very low levels of contamination (≤0.1% of original) were detected at 30 and 60 minutes post procedure. Suction reduced contamination by 67–75% at 0.5–1.5 m. Mannequin and operator were heavily contaminated. The two analytic methods showed good correlation (r=0.930, n=244, p

In the past decade, there has been a tremendous increase in studies on the link between oral microbiome and systemic diseases. However, variations in study design and confounding variables across studies often lead to inconsistent observations. In this narrative review, we have discussed the potential influence of study design and confounding variables on the current sequencing-based oral microbiome-systemic disease link studies. The current limitations of oral microbiome-systemic link studies on type 2 diabetes mellitus, rheumatoid arthritis, pregnancy, atherosclerosis, and pancreatic cancer are discussed in this review, followed by our perspective on how artificial intelligence (AI), particularly machine learning and deep learning approaches, can be employed for predicting systemic disease and host metadata from the oral microbiome. The application of AI for predicting systemic disease as well as host metadata requires the establishment of a global database repository with microbiome sequences and annotated host metadata. However, this task requires collective efforts from researchers working in the field of oral microbiome to establish more comprehensive datasets with appropriate host metadata. Development of AI-based models by incorporating consistent host metadata will allow prediction of systemic diseases with higher accuracies, bringing considerable clinical benefits.

Extracellular DNA (eDNA) is a key component of many microbial biofilms including dental plaque. However, the roles of extracellular deoxyribonuclease (DNase) enzymes within biofilms are poorly understood. Streptococcus gordonii is a pioneer colonizer of dental plaque. Here, we identified and characterised SsnA, a cell wall-associated protein responsible for extracellular DNase activity of S. gordonii. The SsnA-mediated extracellular DNase activity of S. gordonii was suppressed following growth in sugars. SsnA was purified as a recombinant protein and shown to be inactive below pH 6.5. SsnA inhibited biofilm formation by Streptococcus mutans in a pH-dependent manner. Further, SsnA inhibited the growth of oral microcosm biofilms in human saliva. However, inhibition was ameliorated by the addition of sucrose. Together, these data indicate that S. gordonii SsnA plays a key role in interspecies competition within oral biofilms. Acidification of the medium through sugar catabolism could be a strategy for cariogenic species such as S. mutans to prevent SsnA-mediated exclusion from biofilms.

Normative approaches have been developed with the aim of providing high-quality methods and strict criteria that, when applied correctly, lead to reliable results. Standards, specifications, and guidelines are needed to facilitate exchange of goods or information and secure comparability of data derived from different laboratories and sources. They are available along the whole flow from study development to test selection, study conduct, and reporting and are widely used for the evaluation of medical devices, market approval, and harmonization of terms and devices. Standards are developed by specific national and international organizations or by dedicated interest groups, mainly scientists in their respective fields. ISO (International Organization for Standardization) standards are developed following stringent regulations, and groups of experts formulate such standards. They should come from different areas (multistakeholder approach) to have as much and as broad input as possible and to avoid single-interest dominance. However, the presence of academia in such groups has been comparatively low. There is a clear need and responsibility of the oral health community to participate in the development of normative documents to provide methodological knowledge and experience, balance the interests of other stakeholders, and finally improve oral health. This will help to ensure that rapidly advancing fields of research, such as the oral health impacts of COVID-19 or the application of artificial intelligence in dentistry, benefit from standardization of approaches and reporting.

Cell-cell adhesion between oral bacteria plays a key role in the development of polymicrobial communities such as dental plaque. Oral streptococci such as Streptococcus gordonii and Streptococcus oralis are important early colonizers of dental plaque and bind to a wide range of different oral microorganisms, forming multispecies clumps or “coaggregates.” S. gordonii actively responds to coaggregation by regulating gene expression. To further understand these responses, we assessed gene regulation in S. gordonii and S. oralis following coaggregation in 25% human saliva. Coaggregates were formed by mixing, and after 30 min, RNA was extracted for dual transcriptome sequencing (RNA-Seq) analysis. In S. oralis, 18 genes (6 upregulated and 12 downregulated) were regulated by coaggregation. Significantly downregulated genes encoded functions such as amino acid and antibiotic biosynthesis, ribosome, and central carbon metabolism. In total, 28 genes were differentially regulated in Streptococcus gordonii (25 upregulated and 3 downregulated). Many genes associated with transporters and a two-component (NisK/SpaK) regulatory system were upregulated following coaggregation. Our comparative analyses of S. gordonii-S. oralis with different previously published S. gordonii pairings (S. gordonii-Fusobacterium nucleatum and S. gordonii-Veillonella parvula) suggest that the gene regulation is specific to each pairing, and responses do not appear to be conserved. This ability to distinguish between neighboring bacteria may be important for S. gordonii to adapt appropriately during the development of complex biofilms such as dental plaque. IMPORTANCE Dental plaque is responsible for two of the most prevalent diseases in humans, dental caries and periodontitis. Controlling the formation of dental plaque and preventing the transition from oral health to disease requires a detailed understanding of microbial colonization and biofilm development. Streptococci are among the most common colonizers of dental plaque. This study identifies key genes that are regulated when oral streptococci bind to one another, as they do in the early stages of dental plaque formation. We show that specific genes are regulated in two different oral streptococci following the formation of mixed-species aggregates. The specific responses of S. gordonii to coaggregation with S. oralis are different from those to coaggregation with other oral bacteria. Targeting the key genes that are upregulated during interspecies interactions may be a powerful approach to control the development of biofilm and maintain oral health.

Since its inception in 1919, the Journal of Dental Research has continually published high-quality articles that span the breadth of research topics relevant to dentistry, oral surgery, and medicine. As part of the journal’s centennial celebration, we conducted an electronic search on Scopus to identify and analyze the top 100 most cited articles from 1919 to 2018. Since Scopus does not capture older citations, we conducted an additional analysis by Google Scholar to identify key articles published in the first 50 y of the journal. Based on Scopus, the articles were ranked in descending order per their citation counts. The citation counts of the 100 most cited articles varied from 262 to 1,503. The year in which the largest number of top 100 articles were published was 2004 ( n = 6). Within the top 100, the majority of articles originated from the United States ( n = 52). Research Reports–Biomaterials & Bioengineering was the most frequent category of cited articles ( n = 35). There was no significant association between total citation count and time since publication (correlation coefficient = –0.051, P = 0.656). However, there was a significant negative association of citation density (correlation coefficient = –0.610, P < 0.01) with time since publication. Our analyses demonstrate the broad reach of the journal and the dynamics in citation patterns and research agenda over its 100-y history. There is considerable evidence of the high variance in research output, when measured via citations, across the globe. Moreover, it remains unclear how patients’ priorities and dental health care needs are aligned with the perceived influence of single research pieces identified by our search. Our findings may help to inspire future research in tackling these inequalities and highlight the need for conceptualizing research priorities.

Desensitising agents are added to dentifrices to occlude exposed dentine tubules and reduce pain associated with dentine hypersensitivity. In occluding the tubules these agents may alter the surface layer of the dentine and consequently affect bacterial biofilm formation. This research sought to examine the effects of desensitising agents on dentinal biofilms using an in vitro model. A constant depth film fermenter (CDFF) was selected to mimic the oral environment and human dentine with exposed tubules was analysed. Calcium sodium phosphosilicate (CSPS) was selected as a model desensitising agent. Dentine discs were treated with pumice or CSPS-containing dentifrices with or without fluoride, or left untreated (control). Dual-species biofilms of Streptococcus mutans and Streptococcus sobrinus were grown in artificial saliva and analysed by viable counts, polymerase chain reaction (PCR) and scanning electron microscopy (SEM). SEM images confirmed the presence of occluded tubules after CSPS application and demonstrated the formation of biofilms containing extracellular matrix material. Analysis of PCR and viable count data using a one-way ANOVA showed no significant differences for bacterial composition for any of the four treatments. There were, however, trends towards increased numbers of bacteria for the pumice and CSPS treated samples which was reversed by the addition of fluoride to CSPS. In conclusion, CSPS was not found to have a significant effect on biofilms and an in vitro model for testing desensitising agents has been developed, however, further work is required to improve the reproducibility of the biofilms formed and to explore the trends seen.

Objectives High-speed dental instruments produce aerosol and droplets. The objective of this study was to evaluate aerosol and droplet production from a novel electric micromotor handpiece (without compressed air coolant) in real world clinical settings. Methods 10-minute upper incisor crown preparations were performed in triplicate in an open-plan clinic with mechanical ventilation providing 3.45 air changes per hour. A 1:5 ratio electric micromotor handpiece which allows water coolant without compressed air (Ti-Max Z95L, NSK) was used at three speeds: 60,000 (60 K), 120,000 (120 K), and 200,000 (200 K) revolutions per minute. Coolant solutions contained fluorescein sodium as a tracer (2.65 mmol L − 1). High-speed air-turbine positive control, and negative control conditions were conducted. Aerosol production was evaluated at 3 locations (0.5 m, 1.5 m, and 1.7 m) using: (1) an optical particle counter (OPC; 3016-IAQ, Lighthouse) to detect all aerosol; and (2) a liquid cyclone air sampler (BioSampler, SKC Ltd.) to detect aerosolised fluorescein, which was quantified by spectrofluorometric analysis. Settled droplets were detected by spectrofluorometric analysis of filter papers placed onto a rig across the open-plan clinic. Results Local (within treatment bay) settled droplet contamination was elevated above negative control for all conditions, with no difference between conditions. Settled droplet contamination was not detected above negative controls outside the treatment bay for any condition. Aerosol detection at 1.5 m and 1.7 m, was only increased for the air-turbine positive control condition. At 0.5 m, aerosol levels were highly elevated for the air-turbine, minimally elevated for 200 K and 120 K, and not elevated for 60 K. Conclusions Electric micromotor handpieces which use water-jet coolant alone without compressed air produce localised (within treatment bay) droplet contamination, but are unlikely to produce aerosol contamination beyond the immediate treatment area (1.5 m), allowing them to be used safely in most open-plan clinic settings.

Dental plaque is the key etiological agent in caries formation and the development of the prevalent chronic oral inflammatory disease, periodontitis. The dental plaque biofilm comprises a diverse range of microbial species encased within a rich extracellular matrix, of which extracellular DNA (eDNA) has been identified as an important component. The molecular mechanisms of eDNA release and the structure of eDNA have yet to be fully characterized. Nonetheless, key functions that have been proposed for eDNA include maintaining biofilm structural integrity, initiating adhesion to dental surfaces, acting as a nutrient source, and facilitating horizontal gene transfer. Thus, eDNA is a potential therapeutic target for the management of oral disease–associated biofilm. This review aims to summarize advances in the understanding of the mechanisms of eDNA release from oral microorganisms and in the methods of eDNA detection and quantification within oral biofilms.

The extracellular matrix is a critical component of microbial biofilms, such as dental plaque, maintaining the spatial arrangement of cells and coordinating cellular functions throughout the structure. The extracellular polymeric substances that comprise the matrix include carbohydrates, nucleic acids, proteins, and lipids, which are frequently organized into macromolecular complexes and/or are associated with the surfaces of microbial cells within the biofilm. Cariogenic dental plaque is rich in glucan and fructan polysaccharides derived from extracellular microbial metabolism of dietary sucrose. By contrast, the matrix of subgingival dental plaque is a complex mixture of macromolecules that is still not well understood. Components of the matrix escape from microbial cells during lysis by active secretion or through the shedding of vesicles and serve to anchor microbial cells to the tooth surface. By maintaining the biofilm in close association with host tissues, the matrix facilitates interactions between microorganisms and the host. The outcome of these interactions may be the maintenance of health or the development of dental disease, such as caries or periodontitis. The matrix affords microbial cells protection against chemical and physical insults and hinders the eradication of pathogenic dental plaque. Therefore, strategies to control the matrix are critical to maintain oral health. This review discusses recent advances in our understanding of the composition, origins, and function of the dental plaque matrix, with a focus on subgingival dental plaque. New strategies to control subgingival dental plaque based on targeting the biofilm matrix are also considered.

Understanding the pathophysiology of the coronavirus disease 2019 (COVID-19) infection remains a significant challenge of our times. The gingival crevicular fluid being representative of systemic status and having a proven track record of detecting viruses and biomarkers forms a logical basis for evaluating the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The study aimed to assess gingival crevicular fluid (GCF) for evidence of SARS-CoV-2 in 33 patients who were deemed to be COVID-19 positive upon nasopharyngeal sampling. An attempt was also made to comparatively evaluate it with saliva in terms of its sensitivity, as a diagnostic fluid for SARS-CoV-2. GCF and saliva samples were collected from 33 COVID-19–confirmed patients. Total RNA was extracted using NucliSENS easyMAG (bioMérieux) and eluted in the elution buffer. Envelope gene (E gene) of SARS-CoV-2 and human RNase P gene as internal control were detected in GCF samples by using the TRUPCR SARS-CoV-2 RT qPCR kit V-2.0 (I) in an Applied Biosystems 7500 real-time machine. A significant majority of both asymptomatic and mildly symptomatic patients exhibited the presence of the novel coronavirus in their GCF samples. Considering the presence of SARS-CoV-2 RNA in the nasopharyngeal swab sampling as gold standard, the sensitivity of GCF and saliva, respectively, was 63.64% (confidence interval [CI], 45.1% to 79.60%) and 64.52% (CI, 45.37% to 80.77%). GCF was found to be comparable to saliva in terms of its sensitivity to detect SARS-CoV-2. Saliva samples tested positive in 3 of the 12 patients whose GCF tested negative, and likewise GCF tested positive for 2 of the 11 patients whose saliva tested negative on real-time reverse transcription polymerase chain reaction. The results establish GCF as a possible mode of transmission of SARS-CoV-2, which is the first such report in the literature, and also provide the first quantifiable evidence pointing toward a link between the COVID-19 infection and oral health.

Introduction Dental procedures produce splatter and aerosol which have potential to spread pathogens such as SARS-CoV-2. Mixed evidence exists on the aerosol-generating potential of orthodontic procedures. The aim of this study was to evaluate splatter and/or settled aerosol contamination during orthodontic debonding. Material and methods Fluorescein dye was introduced into the oral cavity of a mannequin. Orthodontic debonding was undertaken with surrounding samples collected. Composite bonding cement was removed using a speed-increasing handpiece with dental suction. A positive control condition included a water-cooled, high-speed air-turbine crown preparation. Samples were analysed using digital image analysis and spectrofluorometric analysis. Results Contamination across the eight-metre experimental rig was 3% of the positive control on spectrofluorometric analysis and 0% on image analysis. Contamination of the operator, assistant and mannequin was 8%, 25% and 28% of the positive control, respectively. Discussion Splatter and settled aerosol from orthodontic debonding is distributed mainly within the immediate locality of the mannequin. Widespread contamination was not observed. Conclusions Orthodontic debonding is unlikely to produce widespread contamination via splatter and settled aerosol, but localised contamination is likely. This highlights the importance of personal protective equipment for the operator, assistant and patient. Further work is required to examine suspended aerosol. Zusatzmaterial online: Zu diesem Beitrag sind unter 10.1038/s41415-020-2503-9 für autorisierte Leser zusätzliche Dateien abrufbar., Key points Orthodontic debonding, including removal of composite using a slow-speed handpiece with dental suction, appears to pose little risk of widespread distribution of settled contamination.Splatter and settled aerosol was produced during the debonding procedure; however, this was mainly localised to the patient, operator and assistant.Further work is required to examine aerosol which remains suspended in the air. Zusatzmaterial online: Zu diesem Beitrag sind unter 10.1038/s41415-020-2503-9 für autorisierte Leser zusätzliche Dateien abrufbar.

Helicobacter pylori is associated with chronic gastritis, gastric or duodenal ulcers, and gastric cancer. Since the oral cavity is the entry port and the first component of the gastrointestinal system, the oral cavity has been discussed as a potential reservoir of H. pylori. Accordingly, a potential oral-oral transmission route of H. pylori raises the question concerning whether close contact such as kissing or sharing a meal can cause the transmission of H. pylori. Therefore, this topic has been investigated in many studies, applying different techniques for detection of H. pylori from oral samples, i.e. molecular techniques, immunological or biochemical methods and traditional culture techniques. While molecular, immunological or biochemical methods usually yield high detection rates, there is no definitive evidence that H. pylori has ever been isolated from the oral cavity. The specificity of those methods may be limited due to potential cross-reactivity, especially with H. pylori-like microorganisms such as Campylobacter spp. Furthermore, the influence of gastroesophageal reflux has not been investigated so far. This review aims to summarize and critically discuss previous studies investigating the potential colonization of H. pylori in the oral cavity and suggest novel research directions for targeting this critical research question.

The concept of slippery lubricant-infused surfaces has shown promising potential in antifouling for controlling detrimental biofilm growth. In this study, nontoxic silicone oil was either impregnated into porous surface nanostructures, referred to as liquid-infused surfaces (LIS), or diffused into a polydimethylsiloxane (PDMS) matrix, referred to as a swollen PDMS (S-PDMS), making two kinds of slippery surfaces. The slippery lubricant layers have extremely low contact angle hysteresis, and both slippery surfaces showed superior antiwetting performances with droplets bouncing off or rolling transiently after impacting the surfaces. We further demonstrated that water droplets can remove dust from the slippery surfaces, thus showing a "cleaning effect". Moreover, "coffee-ring" effects were inhibited on these slippery surfaces after droplet evaporation, and deposits could be easily removed. The clinically biofilm-forming species P. aeruginosa (as a model system) was used to further evaluate the antifouling potential of the slippery surfaces. The dried biofilm stains could still be easily removed from the slippery surfaces. Additionally, both slippery surfaces prevented around 90% of bacterial biofilm growth after 6 days compared to the unmodified control PDMS surfaces. This investigation also extended across another clinical pathogen, S. epidermidis, and showed similar results. The antiwetting and antifouling analysis in this study will facilitate the development of more efficient slippery platforms for controlling biofouling.

IntroductionDental procedures often produce splatter and aerosol which have potential to spread pathogens such as SARS-CoV-2. Mixed guidance exists on the aerosol generating potential of orthodontic procedures. The aim of this study was to evaluate aerosol and/or splatter contamination during an orthodontic debonding procedure.Material and MethodsFluorescein dye was introduced into the oral cavity of a mannequin. Orthodontic debonding was carried out in triplicate with filter papers placed in the immediate environment. Composite bonding cement was removed using a slow-speed handpiece with dental suction. A positive control condition included a high-speed air-turbine crown preparation. Samples were analysed using digital image analysis and spectrofluorometric analysis.ResultsContamination across the 8-metre experimental rig was 3% of the positive control on spectrofluorometric analysis and 0% on image analysis. There was contamination of the operator, assistant, and mannequin, representing 8%, 25%, and 28% of the positive control spectrofluorometric measurements, respectively.DiscussionOrthodontic debonding produces splatter within the immediate locality of the patient. Widespread aerosol generation was not observed.ConclusionsOrthodontic debonding procedures are low risk for aerosol generation, but localised splatter is likely. This highlights the importance of personal protective equipment for the operator, assistant, and patient.Three ‘In brief’ pointsOrthodontic debonding, including removal of composite using a slow speed handpiece with dental suction, appears to be a low risk procedure for aerosol generation.Splatter was produced during the debonding procedure, however this was mainly localised to the patient, operator and assistant.A single positive reading was identified 3.5 meters away from the patient, highlighting the need for suitable distancing and/or barriers in open clinical environments.

Surface topography designed to achieve spatial segregation has shown promise in delaying bacterial attachment and biofilm growth. However, the underlying mechanisms linking surface topography to the inhibition of microbial attachment and growth still remain unclear. Here, we investigated bacterial attachment, cell alignment and biofilm formation of Pseudomonas aeruginosa on periodic nano-pillar surfaces with different pillar spacing. Using fluorescence and scanning electron microscopy, bacteria were shown to align between the nanopillars. Threadlike structures (“bacterial nanotubes”) protruded from the majority of bacterial cells and appeared to link cells directly with the nanopillars. Using ΔfliM and ΔpilA mutants lacking flagella or pili, respectively, we further demonstrated that cell alignment behavior within nano-pillars is independent of the flagella or pili. The presence of bacteria nanotubes was found in all cases, and is not linked to the expression of flagella or pili. We propose that bacterial nanotubes are produced to aid in cell–surface or cell–cell connections. Nano-pillars with smaller spacing appeared to enhance the extension and elongation of bacterial nanotube networks. Therefore, nano-pillars with narrow spacing can be easily overcome by nanotubes that connect isolated bacterial aggregates. Such nanotube networks may aid cell–cell communication, thereby promoting biofilm development.

BackgroundDental procedures often produce aerosol and splatter which are potentially high risk for spreading pathogens such as SARS-CoV-2. The existing literature is limited.Objective(s)To develop a robust, reliable and valid methodology to evaluate distribution and persistence of dental aerosol and splatter, including the evaluation of clinical procedures.MethodsFluorescein was introduced into the irrigation reservoirs of a high-speed air-turbine, ultrasonic scaler and 3-in-1 spray and procedures performed on a mannequin in triplicate. Filter papers were placed in the immediate environment. The impact of dental suction and assistant presence were also evaluated. Samples were analysed using photographic image analysis, and spectrofluorometric analysis. Descriptive statistics were calculated and Pearson’s correlation for comparison of analytic methods.ResultsAll procedures were aerosol and splatter generating. Contamination was highest closest to the source, remaining high to 1-1.5 m. Contamination was detectable at the maximum distance measured (4 m) for high-speed air-turbine with maximum relative fluorescence units (RFU) being: 46,091 at 0.5 m, 3,541 at 1.0 m, and 1,695 at 4 m. There was uneven spatial distribution with highest levels of contamination opposite the operator. Very low levels of contamination (≤0.1% of original) were detected at 30 and 60 minutes post procedure. Suction reduced contamination by 67-75% at 0.5-1.5 m. Mannequin and operator were heavily contaminated. The two analytic methods showed good correlation (r=0.930, n=244, pConclusionDental procedures have potential to deposit aerosol and splatter at some distance from the source, being effectively cleared by 30 minutes in our setting.

The oral ecosystem is shaped by complex interactions between systemic health disease and the resident oral microbiota. Research in the last two decades has produced datasets describing the genetics and physiology of the host and the oral microbiome in health and disease. There are inter-individual differences in the ability to tolerate oral disease-promoting challenges. Identification of the key factors that drive a healthy and resilient oral ecosystem is urgently needed. So far, progress is being made towards replicating the host-microbiota interplay in vitro. Clinical studies may shed light on the mechanisms of oral health resilience. However, most clinical studies are cross-sectional and are insufficient for understanding resilience or for identifying biomarkers that correlate with the point of transition from oral health to dysbiosis. Mathematical and computational models, including artificial intelligence approaches, offer an opportunity to inform the design of clinical studies by identifying key biomarkers and interaction networks in complex datasets and predicting important parameters. This paper discusses some of the challenges and opportunities for understanding the biological basis of resilience of the oral ecosystem. It discusses the current status and challenges, and proposes a way forward to better understand resilience towards oral diseases.

Background This systematic scoping review aimed to identify changes in biomarkers of microbiological, immunological and biochemical origin during experimental gingivitis (EG) studies that might indicate resistance and resilience. Methods The term ‘experimental gingivitis’ was run in PubMed from inception to April 11th, 2018. From the 411 studies retrieved, 22 studies were included for this review. Results Studies reporting data on biomarker changes during and after full mouth EG trial were included. Two studies reported findings on changes in biomarkers of microbiological, 12 on immunological and eight on biochemical origin. Changes were reported in the induction phase, and occasionally in the resolution phase. The microbiological composition of both supragingival and subgingival dental plaque changed over the course of EG to a more pathogenic direction, but showed a shift back to a more normal composition. This indicates resilience of the oral microbiome. For immunological biomarkers, it was challenging to retrieve a robust pattern of changes across multiple studies. IL-1β and IL-6 in saliva and in gingival crevicular fluid increased during induction phase and returned in the resolution phase below baseline values. The biochemical parameters cystatin-SN, cystatin-S and lactoferrin in saliva were increased at the end of induction phase, however also here no clear pattern emerged based on all available studies. Conclusions More research is needed to investigate which microbiological, immunological, and biochemical biomarkers can be useful for future investigations into the resistance and resilience of the oral cavity to experimental gingivitis.

The ability of opportunistic pathogens such as Group A Streptococcus (GAS) to transition between mucosal colonisation and invasive disease requires complex systems for adapting to markedly different host environments. The battle to acquire essential trace metals such as manganese and iron from the host is central to pathogenesis. Using a molecular genetic approach, Turner et al. [Biochem. J. (2019) 476, 595–611] show that it is not just individual metal concentrations that are important, but the ratio of iron to manganese within cells. Increasing this ratio by knocking out pmtA, encoding the Fe(II) exporter PmtA, or by disrupting mtsA, encoding an MtsABC Mn(II)-import system component, led to reductions in superoxide dismutase (SodA) activity and increased sensitivity to oxidative stress. The authors show that SodA is at least 4-fold more active with Mn bound than with Fe and speculate that high intracellular Fe:Mn ratios reduce superoxide dismutase activity through the mismetalation of SodA. Challenging wild-type GAS with 1 mM H2O2 led to a decrease in Fe:Mn ratio and a 3-fold increase in SodA activity, indicating that modulation of the balance between intracellular Fe and Mn may play an important role in adaptation to oxidative stress. This work unravels some of the key mechanisms for maintaining appropriate Mn and Fe concentrations within bacterial cells and underscores the need for future studies that take an holistic view to metal ion homeostasis in bacteria. Strategies aimed at interfering with the balance of intracellular metal ions represent a promising approach for the control of invasive microbial infections.

Many oral bacteria form macroscopic clumps known as coaggregates when mixed with a different species. It is thought that these cell-cell interactions are critical for the formation of mixed-species biofilms such as dental plaque. Here, we assessed the impact of coaggregation between two key initial colonizers of dental plaque, Streptococcus gordonii and Veillonella parvula, on gene expression in each partner. These species were shown to coaggregate in buffer or human saliva. To monitor gene regulation, coaggregates were formed in human saliva and, after 30 minutes, whole-transcriptomes were extracted for sequencing and Dual RNA-Seq analysis. In total, 272 genes were regulated in V. parvula, including 39 genes in oxidoreductase processes. In S. gordonii, there was a high degree of inter-sample variation. Nevertheless, 69 genes were identified as potentially regulated by coaggregation, including two phosphotransferase system transporters and several other genes involved in carbohydrate metabolism. Overall, these data indicate that responses of V. parvula to coaggregation with S. gordonii are dominated by oxidative stress-related processes, whereas S. gordonii responses are more focussed on carbohydrate metabolism. We hypothesize that these responses may reflect changes in the local microenvironment in biofilms when S. gordonii or V. parvula immigrate into the system.

Novel nicotine products, particularly electronic cigarettes (e-cigarettes), have become increasingly popular over the past decade. E-cigarettes are sometimes regarded as a less harmful alternative to tobacco smoking, and there is some evidence of their potential role as a smoking cessation aid. However, there are concerns about their health consequences, particularly in users who are not tobacco smokers, and also when used long term. Given the mode of delivery of these products, there is potential for oral health consequences. Over the past few years, there have been an increasing number of studies conducted to explore their oral health effects. In vitro studies have reported a range of cellular effects, but these are much less pronounced than those resulting from exposure to tobacco smoke. Microbiological studies have indicated that e-cigarette users have a distinct microbiome, and there is some indication this may be more pathogenic compared to nonusers. Evidence of oral health effects from clinical trials is still limited, and most studies to date have been small in scale and usually cross-sectional in design. Epidemiological studies highlight concerns over oral dryness, irritation, and gingival diseases. Interpreting data from e-cigarette studies is challenging, given the different populations that have been investigated and the continual emergence of new products. Overall, studies reveal potential oral health harms, underscoring the importance of efforts to reduce use in nonsmokers. However, in smokers who are using e-cigarettes as an aid to help them quit, the benefits of quitting tobacco smoking may outweigh any negative oral health impacts of e-cigarette use, particularly in the short term. Future research is needed to understand the clinical significance of some of the biological changes observed by following different cohorts of users longitudinally in carefully designed clinical studies and pragmatic trials supported by high-quality in vitro studies.