Proton Pumping ATPases: Rotational Catalysis, Physiological Roles in Oral Pathogenic Bacteria, and Inhibitors.

Proton pumping ATPases, both F-type and V/A-type ATPases, generate ATP using electrochemical energy or pump protons/sodium ions by hydrolyzing ATP. The enzymatic reaction and proton transport are coupled through subunit rotation, and this unique rotational mechanism (rotational catalysis) has been intensively studied. Single-molecule and thermodynamic analyses have revealed the detailed rotational mechanism, including the catalytically inhibited state and the roles of subunit interactions. In mammals, F- and V-ATPases are involved in ATP synthesis and organelle acidification, respectively. Most bacteria, including anaerobes, have F- and/or A-ATPases in the inner membrane. However, these ATPases are not believed to be essential in anaerobic bacteria since anaerobes generate sufficient ATP without oxidative phosphorylation. Recent studies suggest that F- and A-ATPases perform indispensable functions beyond ATP synthesis in oral pathogenic anaerobes; F-ATPase is involved in acid tolerance in Streptococcus mutans, and A-ATPase mediates nutrient import in Porphyromonas gingivalis. Consistently, inhibitors of oral bacterial F- and A-ATPases, such as phytopolyphenols and bedaquiline, strongly diminish growth and survival. Herein, we discuss rotational catalysis of bacterial F- and A-ATPases, and discuss their physiological roles, focusing on oral bacteria. We also review the effects of ATPase inhibitors on the growth and survival of oral pathogenic bacteria. The features of the catalytic mechanism and unique physiological roles in oral bacteria highlight the potential for proton pumping ATPases to serve as targets for oral antimicrobial agents.

The sialyl-O-acetylesterase NanS of Tannerella forsythia encompasses two catalytic modules with different regiospecificity for O7 and O9 of sialic acid.

The periodontal pathogen Tannerella forsythia utilizes host sialic acids as a nutrient source. To also make O-acetylated sialyl residues susceptible to the action of its sialidase and sialic acid uptake system, Tannerella produces NanS, an O-acetylesterase with two putative catalytic domains. Here, we analyzed NanS by homology modeling, predicted a catalytic serine-histidine-aspartate triad for each catalytic domain and performed individual domain inactivation by single alanine exchanges of the triad nucleophiles S32 and S311. Subsequent functional analyses revealed that both domains possess sialyl-O-acetylesterase activity, but differ in their regioselectivity with respect to position O9 and O7 of sialic acid. The 7-O-acetylesterase activity inherent to the C-terminal domain of NanS is unique among sialyl-O-acetylesterases and fills the current gap in tools targeting 7-O-acetylation. Application of the O7-specific variant NanS-S32A allowed us to evidence the presence of cellular 7,9-di-O-acetylated sialoglycans by monitoring the gain in 9-O-acetylation upon selective removal of acetyl groups from O7. Moreover, we established de-7,9-O-acetylation by wild-type NanS as an easy and efficient method to validate the specific binding of three viral lectins commonly used for the recognition of (7),9-O-acetylated sialoglycans. Their binding critically depends on an acetyl group in O9, yet de-7,9-O-acetylation proved advantageous over de-9-O-acetylation as the additional removal of the 7-O-acetyl group eliminated ligand formation by 7,9-ester migration. Together, our data show that NanS gained dual functionality through recruitment of two esterase modules with complementary activities. This enables Tannerella to scavenge 7,9-di-O-acetylated sialyl residues and provides a novel, O7-specific tool for studying sialic acid O-acetylation.

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.

Interactions between the Trimeric Autotransporter Adhesin EmaA and Collagen Revealed by Three-Dimensional Electron Tomography.

Bacterial adhesion to host tissues is considered the first and critical step of microbial infection. The extracellular matrix protein adhesin A (EmaA) is a collagen-binding adhesin of the periodontal pathogen Aggregatibacter actinomycetemcomitans Three 202-kDa EmaA monomers form antenna-like structures on the bacterial surface with the functional domain located at the apical end. The structure of the 30-nm functional domain has been determined by three-dimensional (3D) electron tomography and subvolume averaging. The region exhibits a complex architecture composed of three subdomains (SI to SIII) and a linker between subdomains SII and SIII. However, the molecular interaction between the adhesin receptor complexes has yet to be revealed. This study provides the first detailed 3D structure of reconstituted EmaA/collagen complexes obtained using 3D electron tomography and image processing techniques. The observed interactions of EmaA with collagen were not to whole, intact fibrils, but rather to individual collagen triple helices dissociated from the fibrils. The majority of the contacts with the EmaA functional domain encompassed subdomains SII and SIII and in some cases the tip of the apical domain, involving SI. These data suggest a multipronged mechanism for the interaction of Gram-negative bacteria with collagen.IMPORTANCE Bacterial adhesion is a crucial step for bacterial colonization and infection. In recent years, the number of antibiotic-resistant strains has dramatically increased; therefore, there is a need to search for novel antimicrobial agents. Thus, great efforts are being devoted to develop a clear understanding of the bacterial adhesion mechanism for preventing infections. In host/pathogen interactions, once repulsive forces are overcome, adhesins recognize and tightly bind to specific receptors on the host cell or tissue components. Here, we present the first 3D structure of the interaction between the collagen-binding adhesin EmaA and collagen, which is critical for the development of endocarditis in humans.

Converging findings from linkage between periodontal pathogen with atopic and allergic immune response.

This study aims to explore a relationship between exposures of whole-cell Porphyromonas gingivalis in various doses with atopic inflammatory responses at experimental mice. A pretest-posttest controlled group design, with 16 Wistar rats (Rattus novergicus) randomized into four groups. Group 1 was the control group. Group 2 was given low-dose (9 × 107 colony-forming unit) of P. gingivalis. Group 3 was given medium-dose (9 × 109 colony-forming unit) of P. gingivalis. Group 4 was given high-dose (9 × 1011 colony-forming unit) of P. gingivalis. Interleukin-4, Interleukin-5, Interleukin-17F, Interleukin-21, Immunoglobulin-E, Immunoglobulin-G4, and γ-Interferon were measured by direct-sandwich ELISA just before the treatments began, day-4, and day-11 after treatments. There is a sudden increase of Interleukin-4 in the group 4 (23.79 ±â€¯0.91 pg/ml to 54.17 ±â€¯0.79 pg/ml; p = 0.01) and slight increase of Interleukin-5 in the group 4 (207.60 ±â€¯11.15 pg/ml to 243.40 ±â€¯9.33 pg/ml; p = 0.03). No change was observed for Interleukin-17F in all groups. Serum concentration of Immunoglobulin-E was decreased in group 2 (-10.44 ±â€¯8.13 pg/ml), but increased in group 4 (+1.03 ±â€¯4.57 pg/ml). Taken together, some cytokines are up-regulated and others are down-regulated after exposure to whole-cell P. gingivalis. Moreover, study of host responses during periodontal infection may offer critical key insight that contribute to the development of atopy. CLINICAL IMPLICATIONS: We introduced and explained the potential role of periodontal pathogen Porphyromonas gingivalis in systemic immune responses, along with its virulence factor inside the oral cavity. Our results consider several changes and differences of cytokines and immunoglobulins following whole-cell Porphyromonas gingivalis exposure. However, results of the study need to be interpreted with caution due to its limitations. CAPSULE SUMMARY: Interleukin (IL)-4 and IL-5 had been found increase after exposure to the periodontal pathogens Porphyromonas gingivalis, whereas no or minimal change had been found in the level of IL-17F, Ig-G4, and IFN-γ. The various cytokines and immunoglobulins shown in this study do not prove a causal relationship, and the precise role of Porphyromonas gingivalis in the regulation of atopic immune response warrants further investigation. Nevertheless, these findings may provide some critical key insight into the host responses following Porphyromonas gingivalis infection.

Identification of a highly active tannase enzyme from the oral pathogen Fusobacterium nucleatum subsp. polymorphum.

BACKGROUND: Tannases are tannin-degrading enzymes that have been described in fungi and bacteria as an adaptative mechanism to overcome the stress conditions associated with the presence of these phenolic compounds. RESULTS: We have identified and expressed in E. coli a tannase from the oral microbiota member Fusobacterium nucleatum subs. polymorphum (TanBFnp). TanBFnp is the first tannase identified in an oral pathogen. Sequence analyses revealed that it is closely related to other bacterial tannases. The enzyme exhibits biochemical properties that make it an interesting target for industrial use. TanBFnp has one of the highest specific activities of all bacterial tannases described to date and shows optimal biochemical properties such as a high thermal stability: the enzyme keeps 100% of its activity after prolonged incubations at different temperatures up to 45 °C. TanBFnp also shows a wide temperature range of activity, maintaining above 80% of its maximum activity between 22 and 55 °C. The use of a panel of 27 esters of phenolic acids demonstrated activity of TanBFnp only against esters of gallic and protocatechuic acid, including tannic acid, gallocatechin gallate and epigallocatechin gallate. Overall, TanBFnp possesses biochemical properties that make the enzyme potentially useful in biotechnological applications. CONCLUSIONS: We have identified and characterized a metabolic enzyme from the oral pathogen Fusobacterium nucleatum subsp. polymorphum. The biochemical properties of TanBFnp suggest that it has a major role in the breakdown of complex food tannins during oral processing. Our results also provide some clues regarding its possible participation on bacterial survival in the oral cavity. Furthermore, the characteristics of this enzyme make it of potential interest for industrial use.

Effectiveness of Mentha piperita Leaf Extracts against Oral Pathogens: An in vitro Study.

AIM: The study aims to assess the Mentha piperita leaf extract's effectiveness against oral pathogens. MATERIALS AND METHODS: The leaf extract of M. piperita was prepared using cold water method. The three microbial strains, i.e., Streptococcus mutans, Aggregatibacter actinomycetem-comitans, and Candida albicans were used as microbiological materials. Chlorhexidine 0.2% was used as positive control. The digital caliper was used to measure the zone of inhibition to know the antimicrobial activity at 24 and 48 hours. To compare the activity within and between the different microbial strains, one-way analysis of variance (ANOVA) was used. To analyze the data, Statistical Package for the Social Sciences (SPSS) software version of 21.0 was used. The p-value ≤0.05 was considered as statistically significant. RESULTS: Maximum inhibition zone was seen in both M. piperita extracts and 0.2% chlorhexidine with S. mutans at 24 and 48 hours, followed by A. actinomycetemcomitans, and C. albi-cans respectively. The statistical analysis ANOVA reveals the statistically significant association of M. piperita extracts with p-value <0.001. The comparison with 0.2% chlorhexidine at 24 hours showed a p-value of <0.04 and at 48 hours, it showed a p-value <0.001, which was statistically significant. CONCLUSION: The present study concluded that M. piperita showed antimicrobial activity against the oral microorganisms which are causing major less or more severe oral diseases and it can be administered as an alternative medicine for the conventional treatment. CLINICAL SIGNIFICANCE: The study results serve as a guide in selecting and providing information about the efficacy of M. piperita extracts to the dental professionals. The discovery of a potential herbal medication would be a great development in the field of antimicrobial therapies.

Interactions between Lactobacillus rhamnosus GG and oral micro-organisms in an in vitro biofilm model.

BACKGROUND: Probiotics have shown favourable properties in maintaining oral health. By interacting with oral microbial communities, these species could contribute to healthier microbial equilibrium. This study aimed to investigate in vitro the ability of probiotic Lactobacillus rhamnosus GG (L.GG) to integrate in oral biofilm and affect its species composition. Five oral strains, Streptococcus mutans, Streptococcus sanguinis, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum and Candida albicans were involved. The group setup included 6 mono-species groups, 3 dual-species groups (L.GG + S. mutans/S. sanguinis/C. albicans), and 4 multi-species groups (4/5 species and 4/5 species + L.GG, 4 species were all the tested strains except S. mutans). Cell suspensions of six strains were pooled according to the group setup. Biofilms were grown on saliva-coated hydroxyapatite (HA) discs at 37 °C in anaerobic conditions for 64.5 h. Biofilm medium was added and refreshed at 0, 16.5, and 40.5 h. The pH of spent media was measured. Viable cells of the 16.5 h and 64.5 h biofilms were counted. 64.5 h biofilms were stained and scanned with confocal laser scanning microscopy. RESULTS: Our results showed that L.GG and S. mutans demonstrated stronger adhesion ability than the other strains to saliva-coated HA discs. L.GG, C. albicans, S. mutans and F. nucleatum, with poor ability to grow in mono-species biofilms demonstrated better abilities of adhesion and reproduction in dual- and/or multi-species biofilms. L.GG slightly suppressed the growth of C. albicans in all groups, markedly weakened the growth of S. sanguinis and F. nucleatum in 4sp + L.GG group, and slightly reduced the adhesion of S. mutans in L.GG+ S. mutans group. CONCLUSIONS: To conclude, in this in vitro model L.GG successfully integrated in all oral biofilms, and reduced the counts of S. sanguinis and C. albicans and lowered the biofilm-forming ability of F. nucleatum, but only slightly reduced the adhesion of S. mutans. C. albicans significantly promoted the growth of L.GG.

Serum antibody to Porphyromonas gingivalis in metabolic syndrome among an older Japanese population.

BACKGROUND AND OBJECTIVE: Potentially significant associations between metabolic syndrome (MetS) and periodontal disease have been reported in recent studies; however, there is a dearth of literature regarding the relationship of MetS with serum antibody levels to periodontal pathogens. The aim of this cross-sectional study was to investigate the association between MetS and serum antibody to the periodontal pathogen Porphyromonas gingivalis (P. gingivalis) in 216 Japanese individuals aged 79 years. MATERIALS AND METHODS: Serum antibody levels to P. gingivalis were measured by enzyme-linked immunosorbent assay. An elevated serum antibody response was defined as the upper quartile and was considered as the outcome variable. A multivariable logistic regression model was used to evaluate the association of MetS defined by the modified National Cholesterol Education Program Adult Treatment Panel III criteria with an elevated antibody status. Adjustments for gender, income, education, smoking status, dental-care utilisation patterns and brushing frequency were considered. RESULTS: The prevalence of MetS was 22.2% (n = 48). Study participants with MetS were 2.9 times more likely to have an elevated serum antibody to P. gingivalis (adjusted odds ratio = 2.91, 95% confidence interval = 1.24-6.85) after simultaneous adjustment for other covariates. CONCLUSION: Our findings suggest an independent relationship between MetS and serum antibody levels to P. gingivalis in the Japanese elderly. Additional longitudinal epidemiologic studies with larger, more diversified samples and more complete information are needed to substantiate our findings.

Structure and mechanism of cysteine peptidase gingipain K (Kgp), a major virulence factor of Porphyromonas gingivalis in periodontitis.

Cysteine peptidases are key proteolytic virulence factors of the periodontopathogen Porphyromonas gingivalis, which causes chronic periodontitis, the most prevalent dysbiosis-driven disease in humans. Two peptidases, gingipain K (Kgp) and R (RgpA and RgpB), which differ in their selectivity after lysines and arginines, respectively, collectively account for 85% of the extracellular proteolytic activity of P. gingivalis at the site of infection. Therefore, they are promising targets for the design of specific inhibitors. Although the structure of the catalytic domain of RgpB is known, little is known about Kgp, which shares only 27% sequence identity. We report the high resolution crystal structure of a competent fragment of Kgp encompassing the catalytic cysteine peptidase domain and a downstream immunoglobulin superfamily-like domain, which is required for folding and secretion of Kgp in vivo. The structure, which strikingly resembles a tooth, was serendipitously trapped with a fragment of a covalent inhibitor targeting the catalytic cysteine. This provided accurate insight into the active site and suggested that catalysis may require a catalytic triad, Cys(477)-His(444)-Asp(388), rather than the cysteine-histidine dyad normally found in cysteine peptidases. In addition, a 20-Å-long solvent-filled interior channel traverses the molecule and links the bottom of the specificity pocket with the molecular surface opposite the active site cleft. This channel, absent in RgpB, may enhance the plasticity of the enzyme, which would explain the much lower activity in vitro toward comparable specific synthetic substrates. Overall, the present results report the architecture and molecular determinants of the working mechanism of Kgp, including interaction with its substrates.

Anti-Infection of Oral Microorganisms from Herbal Medicine of Piper crocatum Ruiz & Pav.

The WHO Global Status Report on Oral Health 2022 reveals that oral diseases caused by infection with oral pathogenic microorganisms affect nearly 3.5 billion people worldwide. Oral health problems are caused by the presence of S. mutans, S. sanguinis, E. faecalis and C. albicans in the oral cavity. Synthetic anti-infective drugs have been widely used to treat oral infections, but have been reported to cause side effects and resistance. Various strategies have been implemented to overcome this problem. Synthetic anti-infective drugs have been widely used to treat oral infections, but they have been reported to cause side effects and resistance. Therefore, it is important to look for safe anti-infective alternatives. Ethnobotanical and ethnopharmacological studies suggest that Red Betel leaf (Piper crocatum Ruiz & Pav) could be a potential source of oral anti-infectives. This review aims to discuss the pathogenesis mechanism of several microorganisms that play an important role in causing health problems, the mechanism of action of synthetic oral anti-infective drugs in inhibiting microbial growth in the oral cavity, and the potential of red betel leaf (Piper crocatum Ruiz & Pav) as an herbal oral anti-infective drug. This study emphasises the importance of researching natural components as an alternative treatment for oral infections that is more effective and can meet global needs.

Rapid multiplex real-time PCR assay using a portable device for the detection of oral pathogens.

Colonization by several oral pathogens and the onset of oral diseases, such as dental caries and periodontal diseases, are closely related. Therefore, the analysis of pathogens in oral specimens would be helpful for the risk assessment of oral diseases. We developed a rapid multiplex real-time polymerase chain reaction (PCR) method using a portable device and newly designed probe/primer sets to detect the oral pathogens Streptococcus mutans, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia. The theoretical minimum detectable cell numbers of S. mutans, P. gingivalis, T. denticola, and T. forsythia were 1, 1, 4, and 3, respectively. The multiplex real-time PCR system simultaneously detected the colonization of S. mutans and P. gingivalis in human saliva. These results suggest that the multiplex real-time PCR system may be useful for the risk assessment of oral diseases.

Filifactor alocis enhances survival of Porphyromonas gingivalis W83 in response to H2 O2 -induced stress.

A dysbiotic microbial community whose members have specific/synergistic functions that are modulated by environmental conditions, can disturb homeostasis in the subgingival space leading to destructive inflammation, plays a role in the progression of periodontitis. Filifactor alocis, a gram-positive, anaerobic bacterium, is a newly recognized microbe that shows a strong correlation with periodontal disease. Our previous observations suggested F. alocis to be more resistant to oxidative stress compared to Porphyromonas gingivalis. The objective of this study is to further determine if F. alocis, because of its increased resistance to oxidative stress, can affect the survival of other 'established' periodontal pathogens under environmental stress conditions typical of the periodontal pocket. Here, we have shown that via their interaction, F. alocis protects P. gingivalis W83 under H2 O2 -induced oxidative stress conditions. Transcriptional profiling of the interaction of F. alocis and P. gingivalis in the presence of H2 O2 -induced stress revealed the modulation of several genes, including those with ABC transporter and other cellular functions. The ABC transporter operon (PG0682-PG0685) of P. gingivalis was not significant to its enhanced survival when cocultured with F. alocis under H2 O2 -induced oxidative stress. In F. alocis, one of the most highly up-regulated operons (FA0894-FA0897) is predicted to encode a putative manganese ABC transporter, which in other bacteria can play an essential role in oxidative stress protection. Collectively, the results may indicate that F. alocis could likely stabilize the microbial community in the inflammatory microenvironment of the periodontal pocket by reducing the oxidative environment. This strategy could be vital to the survival of other pathogens, such as P. gingivalis, and its ability to adapt and persist in the periodontal pocket.

Variation in adhesion of Streptococcus mutans and Porphyromonas gingivalis in saliva-derived biofilms on raw materials of orthodontic brackets.

Objective: To evaluate differences in the adhesion levels of the most common oral pathogens, Streptococcus mutans and Porphyromonas gingivalis , in human saliva-derived microcosm biofilms with respect to time and raw materials of orthodontic brackets. Methods: The samples were classified into three groups of bracket materials: 1) monocrystalline alumina ceramic (CR), 2) stainless steel metal (SS), and 3) polycarbonate plastic (PL), and a hydroxyapatite (HA) group was used to mimic the enamel surface. Saliva was collected from a healthy donor, and saliva-derived biofilms were grown on each sample. A real-time polymerase chain reaction was performed to quantitatively evaluate differences in the attachment levels of total bacteria, S. mutans and P. gingivalis at days 1 and 4. Results: Adhesion of S. mutans and P. gingivalis to CR and HA was higher than the other bracket materials (SS = PL < CR = HA). Total bacteria demonstrated higher adhesion to HA than to bracket materials, but no significant differences in adhesion were observed among the bracket materials (CR = SS = PL < HA). From days 1 to 4, the adhesion of P. gingivalis decreased, while that of S. mutans and total bacteria increased, regardless of material type. Conclusions: The higher adhesion of oral pathogens, such as S. mutans and P. gingivalis to CR suggests that the use of CR brackets possibly facilitates gingival inflammation and enamel decalcification during orthodontic treatment.

Does Ultraviolet Radiation Exhibit Antimicrobial Effect against Oral Pathogens Attached on Various Dental Implant Surfaces? A Systematic Review.

Background: Dental implant therapy is currently identified as the most effective treatment for edentulous patient. However, peri-implant inflammations were found to be one of the most common complications that leads to the loss and failure of dental implantation. Ultraviolet (UV) radiation has been proposed to enhance bone integration and reduce bacterial attachment. In this study, we aimed to systematically review the current evidence regarding the antimicrobial effect of UV on different dental implant surfaces. Methods: Five databases including PubMed, Scopus, Web of science, VHL, and Cochran Library were searched to retrieve relevant articles. All original reports that examined the effect of the application of UV radiation on dental implants were included in our study. Results: A total of 16 in vitro studies were included in this systematic review. Polymethyl methacrylate UV radiation has induced a significant decrease in bacterial survival in PMMA materials, with an increased effect by modification with 2.5% and 5% TiO2 nanotubes. UV-C showed a superior effect to UV-A in reducing bacterial attachment and accumulation. UV wavelength of 265 and 285 nm showed powerful bactericidal effects. UV of 365 nm for 24 h had the highest inhibition of bacterial growth in ZnO coated magnesium alloys. In UV-irradiated commercially pure titanium surfaces treated with plasma electrolytic oxidation, silver ion application, heat or alkali had shown significant higher bactericidal effect vs non-irradiated treated surfaces than the treatment with any of them alone. UVC and gamma-ray irradiation increased the hydrophilicity of zirconia surface, compared to the dry heat. Conclusion: UV radiation on Ti surfaces exhibited significant antibacterial effects demonstrated through the reduction in bacterial attachment and biofilm formation with suppression of bacterial cells growth. Combination of UV and treated surfaces with alkali, plasma electrolytic oxidation, silver ion application or heat enhance the overall photocatalytic antimicrobial effect.

Copper-containing nanoparticles: Mechanism of antimicrobial effect and application in dentistry-a narrative review.

Copper has been used as an antimicrobial agent long time ago. Nowadays, copper-containing nanoparticles (NPs) with antimicrobial properties have been widely used in all aspects of our daily life. Copper-containing NPs may also be incorporated or coated on the surface of dental materials to inhibit oral pathogenic microorganisms. This review aims to detail copper-containing NPs' antimicrobial mechanism, cytotoxic effect and their application in dentistry.

Ultrashort Peptide Hydrogels Display Antimicrobial Activity and Enhance Angiogenic Growth Factor Release by Dental Pulp Stem/Stromal Cells.

Recent studies on peptide hydrogels have shown that ultrashort peptides (<8 amino acids) can self-assemble into hydrogels. Ultrashort peptides can be designed to incorporate antimicrobial motifs, such as positively charged lysine residues, so that the peptides have inherent antimicrobial characteristics. Antimicrobial hydrogels represent a step change in tissue engineering and merit further investigation, particularly in applications where microbial infection could compromise healing. Herein, we studied the biocompatibility of dental pulp stem/stromal cells (DPSCs) with an ultrashort peptide hydrogel, (naphthalene-2-ly)-acetyl-diphenylalanine-dilysine-OH (NapFFεKεK-OH), where the epsilon (ε) amino group forms part of the peptide bond rather than the standard amino grouping. We tested the antimicrobial properties of NapFFεKεK-OH in both solution and hydrogel form against Staphylococcus aureus, Enterococcus faecalis and Fusobacterium nucleatum and investigated the DPSC secretome in hydrogel culture. Our results showed NapFFεKεK-OH hydrogels were biocompatible with DPSCs. Peptides in solution form were efficacious against biofilms of S. aureus and E. faecalis, whereas hydrogels demonstrated antimicrobial activity against E. faecalis and F. nucleatum. Using an angiogenic array we showed that DPSCs encapsulated within NapFFεKεK-OH hydrogels produced an angiogenic secretome. These results suggest that NapFFεKεK-OH hydrogels have potential to serve as novel hydrogels in tissue engineering for cell-based pulp regeneration.

Periodontal Pathogens' strategies disarm neutrophils to promote dysregulated inflammation.

Periodontitis is an irreversible, chronic inflammatory disease where inflammophilic pathogenic microbial communities accumulate in the gingival crevice. Neutrophils are a major component of the innate host response against bacterial challenge, and under homeostatic conditions, their microbicidal functions typically protect the host against periodontitis. However, a number of periodontal pathogens developed survival strategies to evade neutrophil microbicidal functions while promoting inflammation, which provides a source of nutrients for bacterial growth. Research on periodontal pathogens has largely focused on a few established species: Tannerella forsythia, Treponema denticola, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, and Porphyromonas gingivalis. However, advances in culture-independent techniques have facilitated the identification of new bacterial species in periodontal lesions, such as the two Gram-positive anaerobes, Filifactor alocis and Peptoanaerobacter stomatis, whose characterization of pathogenic potential has not been fully described. Additionally, there is not a full understanding of the pathogenic mechanisms used against neutrophils by organisms that are abundant in periodontal lesions. This presents a substantial barrier to the development of new approaches to prevent or ameliorate the disease. In this review, we first summarize the neutrophil functions affected by the established periodontal pathogens listed above, denoting unknown areas that still merit a closer look. Then, we review the literature on neutrophil functions and the emerging periodontal pathogens, F. alocis and P. stomatis, comparing the effects of the emerging microbes to that of established pathogens, and speculate on the contribution of these putative pathogens to the progression of periodontal disease.

Multicomponent Peptide Hydrogels as an Innovative Platform for Cell-Based Tissue Engineering in the Dental Pulp.

In light of the increasing levels of antibiotic resistance, nanomaterials and novel biologics are urgently required to manage bacterial infections. To date, commercially available self-assembling peptide hydrogels have not been studied extensively for their ability to inhibit micro-organisms relevant to tissue engineering sites such as dental root canals. In this work, we assess the biocompatibility of dental pulp stem/stromal cells with commercially available multicomponent peptide hydrogels. We also determine the effects of dental pulp stem/stromal cell (DPSC) culture in hydrogels on growth factor/cytokine expression. Furthermore, to investigate novel aspects of self-assembling peptide hydrogels, we determine their antimicrobial activity against the oral pathogens Staphylococcus aureus, Enterococcus faecalis, and Fusobacterium nucleatum. We show that self-assembling peptide hydrogels and hydrogels functionalized with the adhesion motif Arg-Gly-Asp (RGD) are biocompatible with DPSCs, and that cells grown in 3D hydrogel cultures produce a discrete secretome compared with 2D-cultured cells. Furthermore, we show that soluble peptides and assembled hydrogels have antimicrobial effects against oral pathogens. Given their antibacterial activity against oral pathogens, biocompatibility with dental pulp stem/stromal cells and enhancement of an angiogenic secretome, multicomponent peptide hydrogels hold promise for translational use.

Chemical composition and biological effects of three different types (tear, paste, and mass) of bitter Ferula assa-foetida Linn. gum.

Ferula assa-foetida gum oleoresin (FAGO) is traditionally used for treatment of different ailments. The subject of this article was to evaluate the chemical composition and antibacterial effects of the different types of bitter FAGO essential oils in the natural habitat of Southwest Iran. α-pinene (38.2%), and (Z)-propenyl sec-butyl disulfide (12.4%) were the main components of F. assa-foetida tear type essential oil, with high antibacterial, anti-biofilm and cytotoxic activities, followed by paste and mass types, respectively. α-pinene (27.8%), and (Z)-propenyl-sec-butyl disulfide (24.2%) were identified as main components of paste type. α-pinene (33.4%), ß-pinene (16.3%), and (Z)-propenyl sec-butyl disulfide (16.1%) were the main components of asafoetida mass type gum oleoresin essential oil. FAGO essential oil specially tear one could serve as suitable candidate for further studies.