Ultrasonic agitation reduces the time of calcium hydroxide antimicrobial effect and enhances its penetrability.

OBJECTIVES: The objective of the present work was to evaluate the ultrasonic agitation, time and vehicle (propylene glycol or distilled water) on the antimicrobial potential and penetrability of calcium hydroxide pastes on infected dentin by means of Confocal Laser Scanning Microscopy (CLSM) and microbiological culture (MC). MATERIALS AND METHODS: Dentin specimens were infected with Enterococcus faecalis using a new contamination protocol of 5 days. The specimens were divided into eight groups and dressed with the pastes for 7 or 15 days: G1) calcium hydroxide (CH) + propylene glycol (prop)/7 days (d), G2) CH + prop/7d + ultrasonic agitation (U), G3) CH + distilled water (dw)/7d, G4) CH + dw/7d + U, G5) CH + prop/15d, G6) CH + prop/15d + U, G7) CH + dw/15d, G8) CH + dw/15d + U. The ultrasonic activation was made for 1 min in both directions with a plain point insert. After medications removal, the images obtained by CLSM showed the viable (green) and dead (red) bacteria with Live and Dead dye. By the MC, the dentinal wall debris obtained by burs were collected for colony counts. For the penetration test, the Rodamine B dye was added to the CH pastes and analyzed by CLSM. RESULTS: The 7 and 15-days CH + prop+U pastes performed better antimicrobial efficacy, followed by the CH + dw+U/15d paste. CONCLUSIONS: All pastes demonstrated better penetration and antimicrobial activity against E. faecalis when agitated with ultrasound, even in periods of up to seven days. The propylene glycol vehicle showed better results. CLINICAL RELEVANCE: Agitation of the dressing that remains for less time inside the root canal can optimize the decontamination of endodontic treatment.

Study of microbiocenosis of canine dental biofilms.

Dental biofilm is a complex microbial community influenced by many exogenous and endogenous factors. Despite long-term studies, its bacterial composition is still not clearly understood. While most of the research on dental biofilms was conducted in humans, much less information is available from companion animals. In this study, we analyzed the composition of canine dental biofilms using both standard cultivation on solid media and amplicon sequencing, and compared the two approaches. The 16S rRNA gene sequences were used to define the bacterial community of canine dental biofilm with both, culture-dependent and culture-independent methods. After DNA extraction from each sample, the V3-V4 region of the 16S rRNA gene was amplified and sequenced via Illumina MiSeq platform. Isolated bacteria were identified using universal primers and Sanger sequencing. Representatives of 18 bacterial genera belonging to 5 phyla were isolated from solid media. Amplicon sequencing largely expanded this information identifying in total 284 operational taxonomic units belonging to 10 bacterial phyla. Amplicon sequencing revealed much higher diversity of bacteria in the canine dental biofilms, when compared to standard cultivation approach. In contrast, cultured representatives of several bacterial families were not identified by amplicon sequencing.

The facultative human oral pathogen Prevotella histicola in equine cheek tooth apical/ periapical infection: a case report.

BACKGROUND: Prevotella histicola is a facultative oral pathogen that under certain conditions causes pathologies such as caries and periodontitis in humans. Prevotella spp. also colonize the oral cavity of horses and can cause disease, but P. histicola has not yet been identified. CASE PRESENTATION: A 12-year-old Tinker mare was referred to the clinic for persistent, malodorous purulent nasal discharge and quidding. Conservative antibiotic (penicillin), antiphlogistic (meloxicam), and mucolytic (dembrexine-hydrochloride) treatment prior to referral was unsuccessful and symptoms worsened. Oral examination, radiography, sino-/ rhinoscopy, and standing computed tomography revealed severe apical/ periapical infection of the upper cheek tooth 209 with accompanying unilateral sinonasal inflammation and conchal necrosis. The tooth exhibited extensive subocclusal mesial infundibular cemental hypoplasia and caries, and an occlusal fissure fracture. After mechanical debridement and thermoplastic resin filling of the spacious subocclusal carious infundibular lesion, the tooth was extracted intraorally. The sinusitis and conchal necrosis were treated transendoscopically. Selective bacteriological swab cultures of affected tooth roots and subsequent matrix-assisted laser desorption ionization-time of flight mass spectrometry showed an infection with the obligate anaerobic, Gram-negative bacterium P. histicola. Surgical intervention and adapted antibiotic therapy led to normal healing without complications. CONCLUSIONS: This study provides the first documented case of dental infection in a horse caused by P. histicola at once indicating necessity of more sufficient microbiological diagnostics and targeted antibiotic treatment in equine dental practice. This finding is also conducive to understand species-specific Prevotella diversity and cross-species distribution.

Effects of Cold-Light Bleaching on Enamel Surface and Adhesion of Streptococcus mutans.

Tooth bleaching is becoming increasingly popular among patients with tooth staining, but the safety of bleaching agents on tooth structure has been questioned. Primarily thriving on the biofilm formation on enamel surface, Streptococcus mutans has been recognized as a major cariogenic bacterial species. The present study is aimed at investigating how cold-light bleaching would change enamel roughness and adhesion of Streptococcus mutans. Human premolars were divided into 72 enamel slices and allocated into 3 groups: (1) control, (2) cold-light bleaching with 35% hydrogen peroxide (Beyond™), and (3) 35% hydrogen peroxide (Beyond™) alone. Biofilms of Streptococcus mutans were cultivated on enamel slices in 5% CO2 (v/v) at 37°C for 1 day or 3 days. Enamel surfaces and biofilms were observed using scanning electron microscope (SEM). Atomic force microscopy (AFM) was applied to quantify the roughness of enamel surface, and the amounts of biofilms were measured by optical density of scattered biofilm and confocal laser scanning microscopy (CLSM). Cold-light bleaching significantly increased (p < 0.05) surface roughness of enamel compared to controls, but significantly inhibited (p < 0.05) adhesion of Streptococcus mutans on enamel in the bacterial cultures of both 1 day and 3 days. In conclusion, cold-light bleaching could roughen enamel surface but inhibit Streptococcus mutans adhesion at the preliminary stage after the bleaching treatment.

Bacteriological Evaluation of Gingival Crevicular Fluid in Teeth Restored Using Fixed Dental Prostheses: An In Vivo Study.

The present in vivo study determined the microbiological counts of the gingival crevicular fluid (GCF) among patients with fixed dental prostheses fabricated using three different techniques. A total of 129 subjects were divided into three study groups: first, cobalt-chrome-based, metal-ceramic prostheses fabricated by the conventional method (MC, n = 35); the second group consisted of cobalt-chrome-based, metal-ceramic prostheses fabricated by the computer-aided design and computer-aided manufacturing (CAD/CAM) technique (CC-MC, n = 35); the third group comprised zirconia-based ceramic prostheses fabricated using the CAD/CAM technique (CC-Zr, n = 35). The control consisted of 24 patients using prostheses fabricated with either MC, CC-MC, or CC-Zr. The GCF was obtained from the subjects before treatment, and 6 and 12 months after the prosthetic treatment. Bacteriological and bacterioscopic analysis of the GCF was performed to analyze the patients' GCF. The data were analyzed using SPSS V20 (IBM Company, Chicago, IL, USA). The number of microorganisms of the gingival crevicular fluid in all groups at 12 months of prosthetic treatment reduced dramatically compared with the data obtained before prosthetic treatment. Inflammatory processes in the periodontium occurred slowly in the case of zirconium oxide-based ceramic constructions due to their biocompatibility with the mucous membranes and tissues of the oral cavity as well as a reduced risk of dental biofilm formation. This should be considered by dentists and prosthodontists when choosing restoration materials for subjects with periodontal pathology.

Exodontia associated bacteremia in horses characterized by next generation sequencing.

Bacteremia resulting from dental surgery is increasingly recognized as a health risk, especially in older and immunocompromised patients. Dentistry-associated bacteremia can lead to remote infections, as exemplified by valvular endocarditis. Emerging evidence points to a novel role played by oral cavity commensals in the pathogenesis of diabetes, respiratory disease, cardiovascular disease, and adverse pregnancy outcomes. Whether dental extraction, a commonly undertaken procedure in old horses, causes bacteremia has not been reported extensively. In a prospective clinical study using next generation sequencing (based on bacterial 16S rRNA), the circulating blood microbiome was characterized before and at 1 h following extraction of incisor, canine or cheek teeth from 29 adult horses with dental disease. 16S rRNA gene sequencing results from the blood microbiome were compared with those from gingival swab samples obtained prior to extraction at the location of the diseased tooth. Bacteremia associated with translocated gingival commensals was demonstrated in horses undergoing exodontia and was, in some cases, still evident one hour post-operatively.

Understanding the Basis of METH Mouth Using a Rodent Model of Methamphetamine Injection, Sugar Consumption, and Streptococcus mutans Infection.

"METH mouth" is a common consequence of chronic methamphetamine (METH) use, resulting in tooth decay and painful oral tissue inflammation that can progress to complete tooth loss. METH reduces the amount of saliva in the mouth, promoting bacterial growth, tooth decay, and oral tissue damage. This oral condition is worsened by METH users' compulsive behavior, including high rates of consumption of sugary drinks, recurrent tooth grinding, and a lack of frequent oral hygiene. Streptococcus mutans is a Gram-positive bacterium found in the oral cavity and associated with caries in humans. Hence, we developed a murine model of METH administration, sugar intake, and S. mutans infection to mimic METH mouth in humans and to investigate the impact of this drug on tooth colonization. We demonstrated that the combination of METH and sucrose stimulates S. mutans tooth adhesion, growth, and biofilm formation in vivo METH and sucrose increased the expression of S. mutans glycosyltransferases and lactic acid production. Moreover, METH contributes to the low environmental pH and S. mutans sucrose metabolism, providing a plausible mechanism for bacterium-mediated tooth decay. Daily oral rinse treatment with chlorhexidine significantly reduces tooth colonization in METH- and sucrose-treated mice. Furthermore, human saliva inhibits S. mutans colonization and biofilm formation after exposure to either sucrose or the combination of METH and sucrose. These findings suggest that METH might increase the risk of microbial dental disease in users, information that may help in the development of effective public health strategies to deal with this scourge in our society.IMPORTANCE "METH mouth" is characterized by severe tooth decay and gum disease, which often causes teeth to break or fall out. METH users are also prone to colonization by cariogenic bacteria such as Streptococcus mutans In addition, this oral condition is aggravated by METH users' compulsive behavior, including the consumption of beverages with high sugar content, recurrent tooth grinding, and a lack of frequent oral hygiene. We investigated the effects of METH and sugar consumption on S. mutans biofilm formation and tooth colonization. Using a murine model of METH administration, sucrose ingestion, and oral infection, we found that the combination of METH and sucrose increases S. mutans adhesion and biofilm formation on the teeth of C57BL/6 mice. However, daily chlorhexidine-based oral rinse treatment reduces S. mutans tooth colonization. Similarly, METH has been associated with dry mouth or hyposalivation in users. Hence, we assessed the impact of human saliva on biofilm formation and demonstrated that surface preconditioning with saliva substantially reduces S. mutans biofilm formation. Our results are significant because to our knowledge, this is the first basic science study focused on elucidating the fundamentals of METH mouth using a rodent model of prolonged drug injection and S. mutans oral infection. Our findings may have important translational implications for the development of treatments for the management of METH mouth and more effective preventive public health strategies that can be applied to provide effective dental care for METH users in prisons, drug treatment centers, and health clinics.

Fossil microbial shark tooth decay documents in situ metabolism of enameloid proteins as nutrition source in deep water environments.

Alteration of organic remains during the transition from the bio- to lithosphere is affected strongly by biotic processes of microbes influencing the potential of dead matter to become fossilized or vanish ultimately. If fossilized, bones, cartilage, and tooth dentine often display traces of bioerosion caused by destructive microbes. The causal agents, however, usually remain ambiguous. Here we present a new type of tissue alteration in fossil deep-sea shark teeth with in situ preservation of the responsible organisms embedded in a delicate filmy substance identified as extrapolymeric matter. The invading microorganisms are arranged in nest- or chain-like patterns between fluorapatite bundles of the superficial enameloid. Chemical analysis of the bacteriomorph structures indicates replacement by a phyllosilicate, which enabled in situ preservation. Our results imply that bacteria invaded the hypermineralized tissue for harvesting intra-crystalline bound organic matter, which provided nutrient supply in a nutrient depleted deep-marine environment they inhabited. We document here for the first time in situ bacteria preservation in tooth enameloid, one of the hardest mineralized tissues developed by animals. This unambiguously verifies that microbes also colonize highly mineralized dental capping tissues with only minor organic content when nutrients are scarce as in deep-marine environments.

Five millennia of Bartonella quintana bacteraemia.

During the two World Wars, Bartonella quintana was responsible for trench fever and is now recognised as an agent of re-emerging infection. Many reports have indicated widespread B. quintana exposure since the 1990s. In order to evaluate its prevalence in ancient populations, we used real-time PCR to detect B. quintana DNA in 400 teeth collected from 145 individuals dating from the 1st to 19th centuries in nine archaeological sites, with the presence of negative controls. Fisher's exact test was used to compare the prevalence of B. quintana in civil and military populations. B. quintana DNA was confirmed in a total of 28/145 (19.3%) individuals, comprising 78 citizens and 67 soldiers, 20.1% and 17.9% of which were positive for B. quintana bacteraemia, respectively. This study analysed previous studies on these ancient samples and showed that the presence of B. quintana infection followed the course of time in human history; a total of 14/15 sites from five European countries had a positive prevalence. The positive rate in soldiers was higher than those of civilians, with 20% and 18.8%, respectively, in the 18th and 19th centuries, but the difference in frequency was not significant. These results confirmed the role of dental pulp in diagnosing B. quintana bacteraemia in ancient populations and showed the incidence of B. quintana in both civilians and soldiers.

Anti-Biofouling Coatings on the Tooth Surface and Hydroxyapatite.

Dental plaque is one type of biofouling on the tooth surface that consists of a diverse population of microorganisms and extracellular matrix and causes oral diseases and even systematic diseases. Numerous studies have focused on preventing bacteria and proteins on tooth surfaces, especially with anti-biofouling coatings. Anti-biofouling coatings can be stable and sustainable over the long term on the tooth surface in the complex oral environment. In this review, numerous anti-biofouling coatings on the tooth surface and hydroxyapatite (as the main component of dental hard tissue) were summarized based on their mechanisms, which include three major strategies: antiprotein and antibacterial adhesion through chemical modification, contact killing through the modification of antimicrobial agents, and antibacterial agent release. The first strategy of coatings can resist the adsorption of proteins and bacteria. However, these coatings use passive strategies and cannot kill bacteria. The second strategy can interact with the cell membrane of bacteria to cause bacterial death. Due to the possibility of delivering a high antibacterial agent concentration locally, the third strategy is recommended and will be the trend of local drug use in dentistry in the future.

Microbiomes of colored dental biofilms in children with or without severe caries experience.

BACKGROUND: Biofilm coloration can compromise maturation and increase the risk of oral disease in adulthood, though children with colored biofilm do not always demonstrate a poor oral health status. AIM: The microbial compositions of colored and white biofilms in children were compared. DESIGN: Thirty-two dental biofilm samples from 16 children (age < 13 years) were analyzed using 16S rRNA pyrosequencing, then the subjects were divided into severe caries and healthy (caries-free) groups. Correlations between microbiomes and oral health status were also examined. RESULTS: Phylogenetic analysis revealed no distinctly different patterns between colored and white biofilms. In the severe caries group, genus Actinomyces, Cardiobacterium, Kingella, Lautropia, and Veillonella, and family Neisseriaceae were detected, though abundance was significantly different between colored and white biofilm specimens, in contrast to the healthy group. In addition, five colored biofilm samples from the severe caries group contained greater than 15% Actinomyces, which led us to consider that genus to be possibly associated with formation of colored biofilm in children. CONCLUSIONS: Our findings indicate that differences in bacterial composition between colored and white biofilms are higher in individuals with severe caries. Additional research may reveal the significance of colored dental biofilm in children.

Characterizing the postmortem human bone microbiome from surface-decomposed remains.

Microbial colonization of bone is an important mechanism of postmortem skeletal degradation. However, the types and distributions of bone and tooth colonizing microbes are not well characterized. It is unknown if microbial communities vary in abundance or composition between bone element types, which could help explain differences in human DNA preservation. The goals of the present study were to (1) identify the types of microbes capable of colonizing different human bone types and (2) relate microbial abundances, diversity, and community composition to bone type and human DNA preservation. DNA extracts from 165 bone and tooth samples from three skeletonized individuals were assessed for bacterial loading and microbial community composition and structure. Random forest models were applied to predict operational taxonomic units (OTUs) associated with human DNA concentration. Dominant bacterial bone colonizers were from the phyla Proteobacteria, Actinobacteria, Firmicutes, Bacteroidetes, and Planctomycetes. Eukaryotic bone colonizers were from Ascomycota, Apicomplexa, Annelida, Basidiomycota, and Ciliophora. Bacterial loading was not a significant predictor of human DNA concentration in two out of three individuals. Random forest models were minimally successful in identifying microbes related to human DNA concentration, which were complicated by high variability in community structure between individuals and body regions. This work expands on our understanding of the types of microbes capable of colonizing the postmortem human skeleton and potentially contributing to human skeletal DNA degradation.

Analysis of oral microbiome from fossil human remains revealed the significant differences in virulence factors of modern and ancient Tannerella forsythia.

BACKGROUND: Recent advances in the next-generation sequencing (NGS) allowed the metagenomic analyses of DNA from many different environments and sources, including thousands of years old skeletal remains. It has been shown that most of the DNA extracted from ancient samples is microbial. There are several reports demonstrating that the considerable fraction of extracted DNA belonged to the bacteria accompanying the studied individuals before their death. RESULTS: In this study we scanned 344 microbiomes from 1000- and 2000- year-old human teeth. The datasets originated from our previous studies on human ancient DNA (aDNA) and on microbial DNA accompanying human remains. We previously noticed that in many samples infection-related species have been identified, among them Tannerella forsythia, one of the most prevalent oral human pathogens. Samples containing sufficient amount of T. forsythia aDNA for a complete genome assembly were selected for thorough analyses. We confirmed that the T. forsythia-containing samples have higher amounts of the periodontitis-associated species than the control samples. Despites, other pathogens-derived aDNA was found in the tested samples it was too fragmented and damaged to allow any reasonable reconstruction of these bacteria genomes. The anthropological examination of ancient skulls from which the T. forsythia-containing samples were obtained revealed the pathogenic alveolar bone loss in tooth areas characteristic for advanced periodontitis. Finally, we analyzed the genetic material of ancient T. forsythia strains. As a result, we assembled four ancient T. forsythia genomes - one 2000- and three 1000- year-old. Their comparison with contemporary T. forsythia genomes revealed a lower genetic diversity within the four ancient strains than within contemporary strains. We also investigated the genes of T. forsythia virulence factors and found that several of them (KLIKK protease and bspA genes) differ significantly between ancient and modern bacteria. CONCLUSIONS: In summary, we showed that NGS screening of the ancient human microbiome is a valid approach for the identification of disease-associated microbes. Following this protocol, we provided a new set of information on the emergence, evolution and virulence factors of T. forsythia, the member of the oral dysbiotic microbiome.

Equine saliva components during mastication, and in vivo pH changes in the oral biofilm of sound and carious tooth surfaces after sucrose exposure.

BACKGROUND: The role of saliva composition and dietary sugar in development of infundibular caries in equine cheek teeth is not fully understood. This study analysed electrolyte and urea concentrations in saliva in relation to different forage and measured pH changes after sucrose application in vivo in sound and carious cheek teeth. RESULTS: Forage type had no effect on the equine saliva electrolyte concentrations, which varied considerably both intra- and inter-individually. Chewing resulted in increased values for all electrolytes except bicarbonate. Compared with stimulated human saliva, horse saliva after mastication, contained higher amounts of potassium, calcium and bicarbonate, and less phosphate. The in vivo pH measurements showed a lower resting pH and a more pronounced pH drop after sucrose application in carious teeth compared to sound teeth. CONCLUSIONS: No large differences were found between the composition of equine saliva and human saliva. A more pronounced acidogenicity was found for the carious than sound teeth. Thus, the caries process in equine cheek teeth seems to follow the same pattern as in human teeth, caused by acid production by oral microorganisms after sugar consumption.

Quaternary ammonium silane, calcium and phosphorus-loaded PLGA submicron particles against Enterococcus faecalis infection of teeth: An in vitro and in vivo study.

Refractory root canal infection of human teeth is the primary cause of dental treatment failure. Enterococcus faecalis is the major cause of refractory root canal infection. In the present study, poly(D,L-lactic-co-glycolide) (PLGA) submicron particles were used as carriers to deliver an antimicrobial quaternary ammonium silane (code-named K21) as well as calcium and phosphorus elements. The release profiles, antibacterial ability against E. faecalis, extent of infiltration into dentinal tubules, biocompatibility and in vitro mineralization potential of the particles were investigated. In addition, the antimicrobial effects of the particles against E. faecalis infection were evaluated in vivo in the teeth of beagle dogs. The encapsulated components were released from the PLGA particles in a sustained-release manner. The particles also displayed good biocompatibility, in vitro mineralization ability and antibacterial activity against E. faecalis. The particles could be driven into dentinal tubules of dentin slices by ultrasonic activation and inhibited E. faecalis colonization. In the root canals of beagle dogs, PLGA submicron particles loaded with K21, calcium and phosphorus demonstrated strong preventive effects against E. faecalis infection. The system may be developed into a new intracanal disinfectant for root canal treatment.

How does ultrasonic cavitation remove dental bacterial biofilm?

Bacterial biofilm accumulation is problematic in many areas, leading to biofouling in the marine environment and the food industry, and infections in healthcare. Physical disruption of biofilms has become an important area of research. In dentistry, biofilm removal is essential to maintain health. The aim of this study is to observe biofilm disruption due to cavitation generated by a dental ultrasonic scaler (P5XS, Acteon) using a high speed camera and determine how this is achieved. Streptococcus sanguinis biofilm was grown on Thermanox™ coverslips (Nunc, USA) for 4 days. After fixing and staining with crystal violet, biofilm removal was imaged using a high speed camera (AX200, Photron). An ultrasonic scaler tip (tip 10P) was held 2 mm away from the biofilm and operated for 2 s. Bubble oscillations were observed from high speed image sequences and image analysis was used to track bubble motion and calculate changes in bubble radius and velocity on the surface. The results demonstrate that most of the biofilm disruption occurs through cavitation bubbles contacting the surface within 2 s, whether individually or in cavitation clouds. Cleaning occurs through shape oscillating microbubbles on the surface as well as through fluid flow.

Inhibitory potential of EGCG on Streptococcus mutans biofilm: A new approach to prevent Cariogenesis.

Dental caries is a common cause for tooth loss and Streptococcus mutans is identified as the etiologic pathogen. This study evaluates the inhibitory potential of Epigallocatechin gallate (EGCG) on S.mutans glucansucrase enzyme and its biofilm. Glucansucrase binding and the inhibitory potential of EGCG was validated using AutoDock tool and enzyme inhibitory assay. Biofilm inhibitory potential was also confirmed using Scanning Electron Microscopic (SEM) analysis in human tooth samples. Molecular docking revealed that EGCG interacted with GLU 515 and TRP 517 amino acids and binds to glucansucrase. SEM analysis revealed inhibition of S.mutans biofilm by various concentrations of EGCG on surfaces of tooth samples. Bioinformatics and biological assays confirmed that EGCG potentially binds to the S. mutans glucansucrase and inhibits its enzymatic activity. Enzymatic inhibition of glucansucrase attenuated biofilm formation potential of S. mutans on tooth surface. Thus, we conclude that EGCG inhibitory potential of S. mutans biofilm on the tooth surface is a novel approach in prevention of dental caries.

A 3D printed microfluidic flow-cell for microscopy analysis of in situ-grown biofilms.

Biofilm phenomena ranging from metabolic processes to attachment, detachment and quorum sensing are influenced by the fluid flow across the biofilm. A number of commercially available flow-cells allow for microscopy analysis of laboratory biofilms under flow, but there is a lack of shear controlled microfluidic devices that accommodate biofilms grown in situ on carriers or tissue samples. Therefore, we developed a flow-cell with adjustable geometry for microscopy analysis of in situ-grown biofilm samples under shear-controlled flow. The flow-cells were designed as one-piece disposable models, 3D-printed in resin and sealed with a coverslip after insertion of the biofilm sample. As a proof of concept, we studied the impact of stimulated saliva flow on pH developments in in situ-grown dental biofilms exposed to sucrose. Under static conditions, pH dropped in the biofilms, with pronounced differences between individual biofilms, but also between different microscopic fields of view within one biofilm. pH in the top layer of the biofilms tended to be lower than pH in the bottom layer. Under conditions of stimulated saliva flow (5 mm/min), pH rose to neutral or slightly alkaline values in all biofilms, and the vertical gradients were reversed, with the biofilm bottom becoming more acidic than the top. Hence, the present work demonstrates the importance of flow for the study of pH in dental biofilms.

Validation of an adherence assay to detect group mutans streptococci in saliva samples.

The aim of the present study was to validate and establish a cut off point and the predictive value of an adhesion test (AA-MSMG), as a microbiological method for evaluating cariogenic risk. The study is based on a variant (20% sucrose) of a selective medium descripted by Gold et al. (MSMG). This method differentiates mutans group streptococci (MGS) by exacerbating the production of insoluble extracellular polysaccharide which gives adhesion to surfaces such as glass, plastic and dental enamel. Caries assessment according to ICDAS was conducted in 154 patients (aged >21 years) who were attended at Preventive and Community Dentistry Department, School of Dentistry, University of Buenos Aires, Argentina, between August 2017 to August 2018. The study population was assigned to groups according to the presence/ absence of caries lesions: Group A: ICDAS lesion code = 0 (L=0) on all dental surfaces (n=23); and Group B: L>1 (n=131). After mouth-rinsing with distilled water, saliva samples were collected with fasting and hygiene protocol, and sent immediately to the Microbiological Diagnosis Laboratory, Microbiology Department, School of Dentistry, University of Buenos Aires. Samples were homogenized and serially diluted to the tenth. 100 pl of the dilutions were cultured in 25 cm2 sterile plastic flasks containing 9.9 ml of modified selective medium described by Gold (MSMG-selective and differential medium). Cultures were incubated in an anaerobic atmosphere at 36 ± 1°C for 48 hours. The supernatants were eluted and the samples washed with sterile distilled water. Colony forming unit counts were performed by calibrated researchers (Kappa >0.75) using a stereoscopic microscope at 50X. Mutans group streptococci (MGS) counts ranged from 1x104 to 1x105 CFU/ml in group A, and were higher than 1x106 CFU/ml in Group B. Statically analysis of results (ROC) showed that the AAMSMG has a satisfactory predictive value (91%) and established a cutoff point in 1.68x105 UFC / ml. This would indicate that individuals whose MGS saliva counts are higher than the cutoff value would be 5 times more likely to develop dental caries. Adherence assay could be a useful microbiological predictor of caries risk.

El objetivo del presente estudio fue validar, establecer el punto de corte y valor predictivo de una técnica microbiológica para evaluar el nivel de estreptococos del grupo mutans en saliva. La técnica consiste en un test de adherencia que emplea un medio selectivo modificado (20% sacarosa) descripto por Gold et al. (TA-MSMG). Este método permite diferenciar a los estreptococos del grupo mutans (SGM) exacerbando la producción del polisacárido extracelular insoluble que le confiere adhesión a superficies como vidrio, plástico y esmalte dental. De acuerdo con los criterios de ICDAS se sembraron 154 salivas de pacientes mayores de edad, que asistieron al Servicio de Odontología Preventiva y Comunitaria de la Facultad de Odontología de la Universidad de Buenos Aires entre los meses de agosto de los años 2017 y 2018. La población estudiada fue asignada a dos grupos según la presencia / ausencia de lesiones de caries: Grupo A: código de lesión ICDAS = 0 (L = 0) en todas las superficies dentales (n = 23); y Grupo B: L> 1 (n = 131). Después de realizar un enjuague bucal con agua destilada, las muestras de saliva se recogieron según protocolo (ayuno de 4 horas y suspensión de higiene dental de 12 hs). Las muestras se remitieron de inmediato al Laboratorio de Diagnóstico Microbiológico, Departamento de Microbiología de la Facultad de Odontología, Universidad de Buenos Aires. Para su procesamiento, las muestras fueron homogeneizadas y diluidas al décimo. Se cultivaron 100 pl de las diluciones en botellas de plástico estériles de 25 cm2 que contenían 9,9 ml de medio de Gold modificado (MSMG-20% sacarosa). Los cultivos se incubaron en atmósfera anaeróbica a 36 ± 1°C durante 48 horas. El sobrenadante se eluyó y las muestras se lavaron con agua destilada estéril. Los recuentos de unidades formadoras de colonias SGMfueron realizados por investigadores calibrados (Kappa >0.75) utilizando un microscopio estereoscópico a 50X. Los recuentos de SGM presentaron una variación entre 1x104y 1x105 UFC/ml en el grupo A, mientras que en el Grupo B fueron superiores a 1x106 UFC/ml. El análisis estadístico de los resultados determinó una curva ROC que establece para el TA-MSMG un valor predictivo del 91% y un punto de corte en 1.68x105 UFC SGM / ml. Esto indicaría que los individuos cuyos recuentos en saliva de SGM sean superiores al valor de corte, tendrían 5 veces más posibilidades de desarrollar caries (5:1). Este método podría ser un instrumento útil al momento de evaluar (indicador microbiológico) el riesgo cariogénico del paciente.

Computer simulations of food oral processing to engineer teeth cleaning.

Oral biofilm accumulation in pets is a growing concern. It is desirable to address this problem via non-invasive teeth cleaning techniques, such as through friction between teeth and food during chewing. Therefore, pet food design tools are needed towards optimising cleaning efficacy. Developing such tools is challenging, as several parameters affecting teeth cleaning should be considered: the food's complex mechanical response, the contacting surfaces topology as well as the wide range of masticatory and anatomical characteristics amongst breeds. We show that Finite Element (FE) models can efficiently account for all these parameters, through the simulation of food deformation and fracture during the first bite. This reduces the need for time consuming and costly in-vivo or in-vitro trials. Our in-silico model is validated through in-vitro tests, demonstrating that the initial oral processing stage can be engineered through computers with high fidelity.