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Journal of Dental Research Jan 2021Osseointegrated dental implants are a revolutionary tool in the armament of reconstructive dentistry, employed to replace missing teeth and restore masticatory,...
Osseointegrated dental implants are a revolutionary tool in the armament of reconstructive dentistry, employed to replace missing teeth and restore masticatory, occlusal, and esthetic functions. Like natural teeth, the orally exposed part of dental implants offers a pristine nonshedding surface for salivary pellicle-mediated microbial adhesion and biofilm formation. In early colonization stages, these bacterial communities closely resemble those of healthy periodontal sites, with lower diversity. Because the peri-implant tissues are more susceptible to endogenous oral infections, understanding of the ecological triggers that underpin the microbial pathogenesis of peri-implantitis is central to developing improved prevention, diagnosis, and therapeutic strategies. The advent of next-generation sequencing (NGS) technologies, notably applied to 16S ribosomal RNA gene amplicons, has enabled the comprehensive taxonomic characterization of peri-implant bacterial communities in health and disease, revealing a differentially abundant microbiota between these 2 states, or with periodontitis. With that, the peri-implant niche is highlighted as a distinct ecosystem that shapes its individual resident microbial community. Shifts from health to disease include an increase in diversity and a gradual depletion of commensals, along with an enrichment of classical and emerging periodontal pathogens. Metatranscriptomic profiling revealed similarities in the virulence characteristics of microbial communities from peri-implantitis and periodontitis, nonetheless with some distinctive pathways and interbacterial networks. Deeper functional assessment of the physiology and virulence of the well-characterized microbial communities of the peri-implant niche will elucidate further the etiopathogenic mechanisms and drivers of the disease.
Topics: Dental Implants; Humans; Microbiota; Peri-Implantitis; Periodontitis; RNA, Ribosomal, 16S
PubMed: 32783779
DOI: 10.1177/0022034520949851 -
Experimental and Therapeutic Medicine Sep 2020Salivary peroxidase and myeloperoxidase are known to display antibacterial activity against oral microbes, and previous indications have pointed to the possibility that...
Salivary peroxidase and myeloperoxidase are known to display antibacterial activity against oral microbes, and previous indications have pointed to the possibility that horseradish peroxidase (HRP) adsorbs onto the membrane of the major oral streptococci, and (). However, the mechanism of interaction between HRP and the bacterial cell wall component is unclear. Dental plaques containing salivary glycoproteins and extracellular microbial products are visualized with 'dental plaque disclosing agent', and are controlled within dental therapy. However, current 'dental plaque disclosing agents' are difficult to evaluate with just dental plaques, since they stain and disclose not only dental plaques but also pellicle formed with salivary glycoproteins on a tooth surface. In this present study, we have demonstrated that HRP interacted with the cell wall component of the major gram-positive bacterial peptidoglycan, but not the major cell wall component of gram-negative bacteria lipopolysaccharide. Furthermore, we observed that the adsorbed HRP labeled with fluorescence was detected on the major oral gram-positive strains and (), but not on a gram-negative strain, (). Furthermore, we have demonstrated that the combination of HRP and chromogenic substrate clearly disclosed the dental plaques and the biofilm developed by , and the major gram-postive bacteria on tooth surfaces, and slightly disclosed the biofilm by . The combination of HRP and chromogenic substrate did not stain either the dental pellicle with the salivary glycoprotein mucin, or naked tooth surfaces. These results have suggested the possibility that the adsorption activity of HRP not only contributes to the evaluation of dental plaque, but that enzymatic activity of HRP may also contribute to improve dental hygiene.
PubMed: 32765778
DOI: 10.3892/etm.2020.9016 -
BMC Oral Health Jun 2020Recent preventive strategies for dental caries focus on targeting the mechanisms underlying biofilm formation, including the inhibition of bacterial adhesion. A...
BACKGROUND
Recent preventive strategies for dental caries focus on targeting the mechanisms underlying biofilm formation, including the inhibition of bacterial adhesion. A promising approach to prevent bacterial adhesion is to modify the composition of acquired salivary pellicle. This in vitro study investigated the effect and possible underlying mechanism of pellicle modification by casein phosphopeptide (CPP) on Streptococcus mutans (S. mutans) initial adhesion, and the impact of fluoride on the efficacy of CPP.
METHODS
The salivary pellicle-coated hydroxyapatite (s-HA) discs were treated with phosphate buffered saline (negative control), heat-inactivated 2.5% CPP (heat-inactivated CPP), 2.5% CPP (CPP) or 2.5% CPP supplemented with 900 ppm fluoride (CPP + F). After cultivation of S. mutans for 30 min and 2 h, the adherent bacteria were visualized by scanning electron microscopy (SEM) and quantitatively evaluated using the plate count method. Confocal laser scanning microscopy (CLSM) was used to evaluate the proportions of total and dead S. mutans. The concentrations of total, free, and bound calcium and fluoride in the CPP and fluoride-doped CPP solutions were determined. The water contact angle and zeta potential of s-HA with and without modification were measured. The data were statistically analyzed using one-way ANOVA followed by a Turkey post hoc multiple comparison test.
RESULTS
Compared to the negative control group, the amount of adherent S. mutans significantly reduced in the CPP and CPP + F groups, and was lowest in the CPP + F group. CLSM analysis showed that there was no statistically significant difference in the proportion of dead S. mutans between the four groups. Water contact angle and zeta potential of s-HA surface significantly decreased in the CPP and CPP + F groups as compared to the negative control group, and both were lowest in the CPP + F group.
CONCLUSIONS
Pellicle modification by CPP inhibited S. mutans initial adhesion to s-HA, possibly by reducing hydrophobicity and negative charge of the s-HA surface, and incorporating fluoride into CPP further enhanced the anti-adhesion effect.
Topics: Bacterial Adhesion; Biofilms; Caseins; Coated Materials, Biocompatible; Dental Caries; Dental Caries Susceptibility; Durapatite; Fluorides; Humans; Phosphopeptides; Saliva; Salivary Proteins and Peptides; Streptococcus mutans; Turkey
PubMed: 32532263
DOI: 10.1186/s12903-020-01158-8 -
Oral Health & Preventive Dentistry Jul 2020During biofilm formation, bacterial species do not attach directly onto the enamel surface, but rather onto the salivary pellicle. Salivary pellicle modification with...
PURPOSE
During biofilm formation, bacterial species do not attach directly onto the enamel surface, but rather onto the salivary pellicle. Salivary pellicle modification with casein and mucin can hinder erosive demineralisation of the enamel, but it should also not promote bacterial adhesion. The aim of our study was to assess whether salivary pellicle modification with casein, or mucin, or a mixture of both proteins (casein and mucin) influence bacterial adhesion, biofilm diversity, metabolism and composition, or enamel demineralisation, after incubation in: (a) a single bacterial model; (b) a five-species biofilm model; or (c) biofilm reformation using the five-species biofilm model after removal of initial biofilm with toothbrushing.
MATERIALS AND METHODS
Enamel specimens were prepared from human molars. Whole-mouth stimulated human saliva was used for pellicle formation. Four pellicle modification groups were established: control (non-modified pellicle); casein - modified with 0.5% casein; mucin - modified with 0.5% mucin; casein and mucin - modified with 0.5% casein and 0.5% mucin. Bacterial adhesion, biofilm diversity, metabolic activity, biofilm mass, and demineralisation (surface hardness) of enamel were assessed after incubation in bacterial broths after 6 h or 24 h.
RESULTS
After 24 h incubation in the five-species biofilm model, the mucin group presented significantly lower biofilm mass than the control (p = 0.028) and the casein and mucin (p = 0.030) groups. No other differences between the groups were observed in any of the other experimental procedures.
CONCLUSION
Pellicle modification with casein and mucin does not promote in vitro bacterial biofilm formation.
Topics: Biofilms; Caseins; Dental Enamel; Dental Pellicle; Humans; Mucins; Saliva
PubMed: 32515418
DOI: 10.3290/j.ohpd.a43351 -
BMC Oral Health Jun 2020Vinegar has been recognized as an effective antimicrobial agent for long. This study intended to elucidate the effect of commercially available vinegar on in situ...
BACKGROUND
Vinegar has been recognized as an effective antimicrobial agent for long. This study intended to elucidate the effect of commercially available vinegar on in situ pellicle formation and existing 24-h biofilms.
METHODS
In situ biofilm formation took place on bovine enamel slabs mounted in individual splints and exposed intraorally over 3 min and 24 h, respectively. After 5 s rinsing with vinegar, all samples were analyzed via fluorescence microscopy (FM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In addition, salivary samples were collected and analyzed via FM. Samples with water rinsing served as controls.
RESULTS
Vinegar caused destruction of the pellicle. Compared to the control group, vinegar rinsing reduced the outer globular layer of the pellicle (p < 0.001), and resulted in formation of subsurface pellicle. Also, vinegar rinsing could reduce bacterial viability and disrupt the 24-h biofilm. Total bacteria amount of saliva samples decreased remarkably (p < 0.001) after vinegar rinsing within 30 min. Reduction of bacterial viability was observed even 120 min after vinegar rinsing in both biofilm and saliva sample (p < 0.001).
CONCLUSION
This in situ study reveals that rinsing with vinegar for only 5 s alters the pellicle layer resulting in subsurface pellicle formation. Furthermore, vinegar rinsing will destruct mature (24-h) biofilms, and significantly reduce the viability of planktonic microbes in saliva, thereby decreasing biofilm formation.
Topics: Acetic Acid; Animals; Biofilms; Cattle; Dental Enamel; Dental Pellicle; Microscopy, Electron, Transmission; Saliva
PubMed: 32503624
DOI: 10.1186/s12903-020-01153-z -
Journal of Dental Research Sep 2020Dental composites are routinely placed as part of tooth restoration procedures. The integrity of the restoration is constantly challenged by the metabolic activities of...
Dental composites are routinely placed as part of tooth restoration procedures. The integrity of the restoration is constantly challenged by the metabolic activities of the oral microbiome. This activity directly contributes to a less-than-desirable half-life for the dental composite formulations currently in use. Therefore, many new antimicrobial dental composites are being developed to counteract the microbial challenge. To ensure that these materials will resist microbiome-derived degradation, the model systems used for testing antimicrobial activities should be relevant to the in vivo environment. Here, we summarize the key steps in oral microbial colonization that should be considered in clinically relevant model systems. Oral microbial colonization is a clearly defined developmental process that starts with the formation of the acquired salivary pellicle on the tooth surface, a conditioned film that provides the critical attachment sites for the initial colonizers. Further development includes the integration of additional species and the formation of a diverse, polymicrobial mature biofilm. Biofilm development is discussed in the context of dental composites, and recent research is highlighted regarding the effect of antimicrobial composites on the composition of the oral microbiome. Future challenges are addressed, including the potential of antimicrobial resistance development and how this could be counteracted by detailed studies of microbiome composition and gene expression on dental composites. Ultimately, progress in this area will require interdisciplinary approaches to effectively mitigate the inevitable challenges that arise as new experimental bioactive composites are evaluated for potential clinical efficacy. Success in this area could have the added benefit of inspiring other fields in medically relevant materials research, since microbial colonization of medical implants and devices is a ubiquitous problem in the field.
Topics: Anti-Infective Agents; Biofilms; Composite Resins; Dental Pellicle; Humans; Microbiota; Mouth; Streptococcus mutans
PubMed: 32479134
DOI: 10.1177/0022034520927690 -
Scanning 2020Hydroxyapatite nanoparticles (nano-HAP) are receiving considerable attention for dental applications, and their adhesion to enamel is well established. However, there...
Hydroxyapatite nanoparticles (nano-HAP) are receiving considerable attention for dental applications, and their adhesion to enamel is well established. However, there are no reports concerning the effects of HAP on other dental materials, and most of the studies in this field are based on designs, neglecting the salivary pellicle-apatite interactions. Thus, this pilot study aims to evaluate the effects of three hydroxyapatite-based solutions and their interactions with different dental material surfaces under oral conditions. Hence, two volunteers carried intraoral splints with mounted samples from enamel and from three dental materials: titanium, ceramics, and polymethyl-methacrylate (PMMA). Three HAP watery solutions (5%) were prepared with different shapes and sizes of nano-HAP (HAP I, HAP II, HAP III). After 3 min of pellicle formation, 10 ml rinse was performed during 30 sec. Rinsing with water served as control. Samples were accessed immediately after rinsing, 30 min and 2 h after rinsing. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to characterize the particles, and SEM evaluated the pellicle-HAP interactions. SEM and TEM results showed a high variation in the size range of the particles applied. A heterogeneous HAP layer was present after 2 h on enamel, titanium, ceramics, and PMMA surfaces under oral conditions. Bridge-like structures were visible between the nano-HAP and the pellicle formed on enamel, titanium, and PMMA surfaces. In conclusion, nano-HAP can adhere not only to enamel but also to artificial dental surfaces under oral conditions. The experiment showed that the acquired pellicle act as a bridge between the nano-HAP and the materials' surface.
Topics: Adult; Bacterial Adhesion; Ceramics; Dental Materials; Durapatite; Female; Humans; Male; Microscopy, Electron, Scanning; Microscopy, Electron, Transmission; Nanoparticles; Pilot Projects; Surface Properties; Titanium; Tooth Erosion
PubMed: 32454927
DOI: 10.1155/2020/6065739 -
International Journal of Molecular... Apr 2020Immune responses triggered by implant abutment surfaces contributed by surface-adsorbed proteins are critical in clinical implant integration. How material...
Immune responses triggered by implant abutment surfaces contributed by surface-adsorbed proteins are critical in clinical implant integration. How material surface-adsorbed proteins relate to host immune responses remain unclear. This study aimed to profile and address the immunological roles of surface-adsorbed salivary proteins on conventional implant abutment materials. Standardized polished bocks (5 × 5 × 1 mm) were prepared from titanium and feldspathic ceramic. Salivary acquired pellicle formed was examined by liquid chromatography-tandem mass spectrometry and gene ontology (GO) analysis to identify and characterize the adsorbed proteins. Out of 759 proteins identified from pooled saliva samples, 396 were found to be attached to the two materials tested-369 on titanium and 298 on ceramic, with 281 common to both. GO annotation of immune processes was undertaken to form a protein-protein interaction network, and 14 hub proteins (≥6 interaction partners) (coding genes: , , , , , , , , , , , , , and ) were identified as the key proteins connecting multiple (6-9) immune processes. The results offered putative immunological prospects of implant abutment material surface-adsorbed salivary proteins, which could potentially underpin the dynamic nature of implant-mucosal/implant-microbial interactions.
Topics: Ceramics; Immunomodulation; Microscopy, Atomic Force; Proteome; Proteomics; Salivary Proteins and Peptides; Surface Properties; Titanium
PubMed: 32349305
DOI: 10.3390/ijms21093083 -
Journal of Dental Research Jun 2020The oral microbiome is one of the most stable ecosystems in the body and yet the reasons for this are still unclear. As well as being stable, it is also highly diverse... (Review)
Review
The oral microbiome is one of the most stable ecosystems in the body and yet the reasons for this are still unclear. As well as being stable, it is also highly diverse which can be ascribed to the variety of niches available in the mouth. Previous studies have focused on the microflora in disease-either caries or periodontitis-and only recently have they considered factors that maintain the normal microflora. This has led to the perception that the microflora proliferate in nutrient-rich periods during oral processing of foods and drinks and starves in between times. In this review, evidence is presented which shows that the normal flora are maintained on a diet of salivary factors including urea, lactate, and salivary protein degradation. These factors are actively secreted by salivary glands which suggests these factors are important in maintaining normal commensals in the mouth. In addition, the immobilization of SIgA in the mucosal pellicle indicates a mechanism to retain certain bacteria that does not rely on the bacterial-centric mechanisms such as adhesins. By examining the salivary metabolome, it is clear that protein degradation is a key nutrient and the availability of free amino acids increases resistance to environmental stresses.
Topics: Dental Pellicle; Immunoglobulin A, Secretory; Microbiota; Mouth; Saliva; Salivary Proteins and Peptides
PubMed: 32283990
DOI: 10.1177/0022034520915486 -
Journal of Applied Oral Science :... 2020The acquired pellicle formation is the first step in dental biofilm formation. It distinguishes dental biofilms from other biofilm types.
INTRODUCTION
The acquired pellicle formation is the first step in dental biofilm formation. It distinguishes dental biofilms from other biofilm types.
OBJECTIVE
To explore the influence of salivary pellicle formation before biofilm formation on enamel demineralization.
METHODOLOGY
Saliva collection was approved by Indiana University IRB. Three donors provided wax-stimulated saliva as the microcosm bacterial inoculum source. Acquired pellicle was formed on bovine enamel samples. Two groups (0.5% and 1% sucrose-supplemented growth media) with three subgroups (surface conditioning using filtered/pasteurized saliva; filtered saliva; and deionized water (DIW)) were included (n=9/subgroup). Biofilm was then allowed to grow for 48 h using Brain Heart Infusion media supplemented with 5 g/l yeast extract, 1 mM CaCl2.2H2O, 5% vitamin K and hemin (v/v), and sucrose. Enamel samples were analyzed for Vickers surface microhardness change (VHNchange), and transverse microradiography measuring lesion depth (L) and mineral loss (∆Z). Data were analyzed using two-way ANOVA.
RESULTS
The two-way interaction of sucrose concentration × surface conditioning was not significant for VHNchange (p=0.872), ∆Z (p=0.662) or L (p=0.436). Surface conditioning affected VHNchange (p=0.0079), while sucrose concentration impacted ∆Z (p<0.0001) and L (p<0.0001). Surface conditioning with filtered/pasteurized saliva resulted in the lowest VHNchange values for both sucrose concentrations. The differences between filtered/pasteurized subgroups and the two other surface conditionings were significant (filtered saliva p=0.006; DIW p=0.0075). Growing the biofilm in 1% sucrose resulted in lesions with higher ∆Z and L values when compared with 0.5% sucrose. The differences in ∆Z and L between sucrose concentration subgroups was significant, regardless of surface conditioning (both p<0.0001).
CONCLUSION
Within the study limitations, surface conditioning using human saliva does not influence biofilm-mediated enamel caries lesion formation as measured by transverse microradiography, while differences were observed using surface microhardness, indicating a complex interaction between pellicle proteins and biofilm-mediated demineralization of the enamel surface.
Topics: Animals; Biofilms; Cattle; Dental Enamel; Dental Pellicle; Hardness; Microradiography; Pasteurization; Reference Values; Saliva; Sucrose; Surface Properties; Tooth Demineralization
PubMed: 32236356
DOI: 10.1590/1678-7757-2019-0501