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Objectives: Gingivitis is the initial stage of periodontitis, one of the most common oral diseases and the primary cause of tooth loss. This study aims to evaluate the effect of toothpaste containing 2% zinc citrate on gingival health and the abundance of three bacteria related to gingivitis and periodontitis., Methods and Materials: Eleven volunteers with the same oral health status were randomly assigned to the treatment (n = 5) and control (n = 6) groups. The control group used fluoride toothpaste, while the treatment group used fluoride toothpaste supplemented with 2% zinc citrate for 3 months. The plaque index, gingival index, and bleeding index were measured at baseline (0 day), 3 weeks, and 3 months. Dental plaque from four areas of the mouth (FDI criteria) was collected at the same timepoints. A total of 132 dental plaque samples were analyzed using quantitative PCR (qPCR) to monitor the abundance of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythia., Results: Toothpaste containing 2% zinc citrate significantly lowered the gingival index and reduced gum bleeding but did not affect the plaque index. It also reduced the total abundance of the three bacteria related to gingivitis and periodontitis in dental plaque over a long-term period., Conclusions: Toothpaste with 2% zinc citrate persistently improves gingival health and reduces the presence of gingivitis-associated bacteria in dental plaque., Trial Registration: Chinese Clinical Trial Registry (Clinical trial registration no.: ChiCTR1900020592) (09/01/2019)., (© 2024 The Author(s). Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)

Background: Considering that antimicrobial resistance among oral pathogens is a significant concern in dental practice, with broader implications for overall health due to the oral microbiota serving as a reservoir for antibiotic resistance genes (ARGs), research into natural products is crucial for addressing this issue., Objective: This study aimed to evaluate tea tree oil (TTO) and chitosan (CH) performance against oral pathogens, including mixed-species biofilm, and its effects on bacteria growth, in addition to chemical characterization and cytotoxicity of TTO., Methods: Tea Tree Oil and low molecular weight chitosan were used in this study. The chemical composition of TTO was analyzed using gas chromatography coupled with mass spectrometry (GC-MS). To evaluate TTO's antimicrobial properties, time-kill and cell viability assays were conducted. Additionally, minimum inhibitory concentration (MIC), minimum microbiocidal concentration (MMC), checkerboard, and biofilm assays were performed using TTO and CH alone and in combination., Results: TTO chromatography peaks found consistent with the standard ISO4730:2017 and literature. TTO and CH exhibited inhibitory activity against all tested microorganisms. The predominantly microbiostatic activity of TTO is probably related to terpinen-4-ol associated with terpinene. The oil at MIC value was able to delay the log phase of Aggregatibacter actinomycetemcomitans growth. Fibroblasts (L929) viability remained above 70 % during 24 h for TTO concentrations ranging from 0.5 to 0.0625 mg/ml. TTO-CH combination showed a synergistic activity (FIC = 0.5) against A. actinomycetemcomitans and Streptococcus sanguinis, at a concentration of 0,25MIC for both species. The compounds at MIC concentration inhibited both monospecies and mixed-species biofilms studied bacteria to the same extent as the azithromycin control., Conclusion: TTO and CH demonstrated efficacy in combating oral pathogens and TTO-CH combination offers a promising approach to confront microbial resistance in the oral environment., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ana Carolina Morais Apolonio reports financial support was provided by Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG- APQ-01165-22). If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Aim: To retrospectively compare two approaches for the adjunctive use of systemic antibiotics in non-surgical periodontal therapy: one based on the detection of Aggregatibacter actinomycetemcomitans (Aa) and the other on age and severity of periodontitis (Age & PPD). We also assessed the additional benefit of antibiotics in reducing the need for further surgical therapy in each group., Materials and Methods: Patients of the Department of Periodontology, Goethe University Frankfurt, Germany, were screened for microbiological testing between 2008 and 2018. Patients were categorized by their microbiological result (Aa+/-) and demographic/clinical data (Age & PPD+/-). Agreement on antibiotic indication was tested. The clinical evaluation focussed on teeth with probing pocket depths (PPDs) ≥ 6 mm., Results: Analysis of 425 patients revealed 30% categorized as Age & PPD+ and 34% as Aa+. Sixty-three percent had consistent antibiotic recommendations (phi coefficient 0.14, p = .004). Patients in the Age & PPD+ group receiving antibiotics showed the most substantial reduction in the number of teeth with PPD ≥ 6 mm after non-surgical periodontal therapy., Conclusions: Both strategies resulted in a significant clinical improvement compared with those without antibiotic treatment and restricted antibiotic use similarly, but targeted different patient groups. Younger individuals with severe periodontitis benefited most from antibiotics, reducing the need for additional surgeries. The study was registered in an international trial register (German Clinical Trial Register number DRKS00028768, registration date 27 April 2022, https://drks.de/search/en/trial/DRKS00028768)., (© 2024 The Author(s). Journal of Clinical Periodontology published by John Wiley & Sons Ltd.)

Dental Implants are expected to possess both excellent osteointegration and antibacterial activity because poor osseointegration and infection are two major causes of titanium implant failure. In this study, we constructed layer-by-layer self-assembly films consisting of anionic casein phosphopeptides-amorphous calcium phosphate (CPP-ACP) and cationic poly (L-lysine) (PLL) on sandblasted and acid etched (SLA) titanium surfaces and evaluated their osseointegration and antibacterial performance in vitro and in vivo. The surface properties were examined, including microstructure, elemental composition, wettability, and Ca 2+ ion release. The impact the surfaces had on the adhesion, proliferation and differentiation abilities of MC3T3-E1 cells were investigated, as well as the material's antibacterial performance after exposure to the oral microorganisms such as Porphyromonas gingivalis (P. g) and Actinobacillus actinomycetemcomitans (A. a). For the in vivo studies, SLA and Ti (PLL/CA-3.0) 10 implants were inserted into the extraction socket immediately after extracting the rabbit mandibular anterior teeth with or without exposure to mixed bacteria solution (P. g & A. a). Three rabbits in each group were sacrificed to collect samples at 2, 4, and 6 weeks of post-implantation, respectively. Radiographic and histomorphometry examinations were performed to evaluate the implant osseointegration. The modified titanium surfaces were successfully prepared and appeared as a compact nano-structure with high hydrophilicity. In particular, the Ti (PLL/CA-3.0) 10 surface was able to continuously release Ca 2+ ions. From the in vitro and in vivo studies, the modified titanium surfaces expressed enhanced osteogenic and antibacterial properties. Hence, the PLL/CPP-ACP multilayer coating on titanium surfaces was constructed via a layer-by-layer self-assembly technology, possibly improving the biofunctionalization of Ti-based dental implants., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objectives: The present study aimed to assess and compare the effect of Morus alba and chlorhexidine gel as an adjunct to scaling and root planing (SRP) in treating stage II periodontitis., Methods: A single-blind, randomized controlled trial was conducted on 180 patients with stage II periodontitis who received full-mouth SRP. They were randomly assigned to receive chlorhexidine digluconate (CHX) gel, Morus alba (MA) and placebo gel for Groups A, B and C, respectively, at the baseline, 15 days and 30 days. Plaque index (PI), Gingival index (GI), periodontal pocket depth (PPD) and quantitative analysis (culture) of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia were assessed at baseline and 45 days. Analysis of variance was used to compare the significant difference in PI, GI, PPD and microbiological parameters between the three groups after the intervention, followed by post hoc Mann-Whitney U and Tukey's HSD test for clinical and microbiological parameters, respectively., Results: Intergroup comparison of the PI, GI and microbiological parameters between the MA and CHX groups at the end of 45 days did not show a statistically significant difference (p > 0.05), whereas a statistically significant difference was observed for PPD between MA and CHX groups with the mean difference of 0.18 mm (p = 0.002)., Conclusion: Morus alba gel was found to be effective in decreasing PPD. However, there was no difference between Morus alba and chlorhexidine gel as an adjunct to SRP in treating stage II periodontitis., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Titanium implants are subject to bacterial adhesion and peri-implantitis induction, and biosurfactants bring a new alternative to the fight against infections. This work aimed to produce and characterize the biosurfactant from Bacillus subtilis ATCC 19,659, its anti-adhesion and antimicrobial activity, and cell viability. Anti-adhesion studies were carried out against Streptococcus sanguinis, Staphylococcus aureus, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Proteus mirabilis as the minimum inhibitory concentration and the minimum bactericidal concentration. Cell viability was measured against osteoblast and fibroblast cells. The biosurfactant was classified as lipopeptide, with critical micelle concentration at 40 µg mL - 1 , and made the titanium surface less hydrophobic. The anti-adhesion effect was observed for Staphylococcus aureus and Streptococcus sanguinis with 54% growth inhibition and presented a minimum inhibitory concentration of 15.7 µg mL - 1 for Streptococcus sanguinis and Aggregatibacter actinomycetemcomitans. The lipopeptide had no cytotoxic effect and demonstrated high potential application against bacterial biofilms., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: There is insufficient clinical and microbiological evidence to support the use of diode laser and air-polishing with erythritol as supplements to scaling and root planning(SRP). The aim of the current study is to evaluate the clinical and microbiologic efficacy of erythritol subgingival air polishing and diode laser in treatment of periodontitis., Methods: The study encompassed twenty-four individuals seeking periodontal therapy and diagnosed with stage I and stage II periodontitis. Eight patients simply underwent SRP. Eight more patients had SRP followed by erythritol subgingival air polishing, and eight patients had SRP followed by diode laser application. At baseline and six weeks, clinical periodontal parameters were measured, including Plaque Index (PI), Gingival Index (GI), periodontal Probing Depth (PPD), and Clinical Attachment Level (CAL). The bacterial count of Aggregatibacter actinomycetemcomitans(A.A), Porphyromonas gingivalis (P.G) was evaluated at different points of time., Results: The microbiological assessment revealed significant differences in the count of A.A. between the laser and erythritol groups immediately after treatment, indicating a potential impact on microbial levels. However, the microbial levels showed fluctuations over the subsequent weeks, without statistically significant differences. Plaque indices significantly decreased post-treatment in all groups, with no significant inter-group differences. Gingival indices decreased, and the laser group showed lower values than erythritol and control groups. PPD and CAL decreased significantly across all groups, with the laser group exhibiting the lowest values., Conclusion: The supplementary use of diode laser and erythritol air polishing, alongside SRP, represents an expedited periodontal treatment modality. This approach leads to a reduction in bacteria and improvement in periodontal health., Trial Registration: This clinical trial was registered on Clinical Trials.gov (Registration ID: NCT06209554) and released on 08/01/2024., (© 2024. The Author(s).)

Porphyromonas gingivalis has been associated with progression of periodontitis, characterized by inflammation and destruction of periodontal tissues. Here, we report that matcha, a product of Camellia sinensis , hampers the adherence and survival of P. gingivalis through multiple tactics. Matcha extract (ME) inhibited the growth not only of P. gingivalis but also of Prevotella nigrescen s and Fusobacterium nucleatum , while it did not inhibit growth of nine species of oral streptococci and Aggregatibacter actinomycetemcomitans . ME-mediated P. gingivalis growth inhibition was characterized by both morphological and physiological changes at the bacterial envelope, which were accompanied by nano-particle formation and decreased membrane fluidity/permeability without loss of membrane integrity. ME also triggered autoaggregation of P. gingivalis in a major fimbriae (FimA)-dependent manner. In addition, adherence of P. gingivalis was dramatically inhibited by ME, irrespective of fimbriae. Furthermore, a structure-activity relationship study tested a series of catechins isolated from ME and identified the pyrogallol-type B-ring of catechins as essential for P. gingivalis growth inhibition. In a clinical study to assess the microbiological and therapeutic effects of matcha mouthwash in patients with periodontitis, the P. gingivalis number in saliva was significantly reduced by matcha mouthwash compared to the pre-intervention level. A tendency toward improvement in probing pocket depth was observed in the matcha group, although the difference was not statistically significant. Taken together, we present a proof of concept, based on the multimodal inhibitory effect of matcha against P. gingivalis , and that matcha may have clinical applicability for prevention and treatment of periodontitis., Importance: Periodontitis, a multifactorial inflammatory disease of the oral cavity, results in alveolar bone destruction, and is a major cause of tooth loss of humans. In addition, emerging evidence has demonstrated associations between periodontitis and a wide range of other chronic inflammation-driven disorders, including diabetes mellitus, preterm birth, cardiovascular disease, aspiration pneumonia, rheumatoid arthritis, cognitive disorder, and cancer. In the present study, we report that matcha, a product of Camellia sinensis , hampers Porphyromonas gingivalis , a major periodontal pathobiont, in not only a series of in vitro experiments but also a pilot intervention clinical trial of patients with periodontitis, in which matcha mouthwash statistically significantly reduced the P. gingivalis number in saliva, as compared to the pre-intervention level. Taken together, we suggest that matcha may have clinical applicability for prevention and treatment of periodontitis., Competing Interests: The authors declare no conflict of interest.

Antibiotic resistance has become an urgent threat to health care in recent years. The use of drug delivery systems provides advantages over conventional administration of antibiotics and can slow the development of antibiotic resistance. In the current study, we developed a toxin-triggered liposomal antibiotic delivery system, in which the drug release is enabled by the leukotoxin (LtxA) produced by the Gram-negative pathogen, Aggregatibacter actinomycetemcomitans. LtxA has previously been shown to mediate membrane disruption by promoting a lipid phase change in nonlamellar lipids, such as 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-methyl (N-methyl-DOPE). In addition, LtxA has been observed to bind strongly and nearly irreversibly to membranes containing large amounts of cholesterol. Here, we designed a liposomal delivery system composed of N-methyl-DOPE and cholesterol to take advantage of these interactions. Specifically, we hypothesized that liposomes composed of N-methyl-DOPE and cholesterol, encapsulating antibiotics, would be sensitive to LtxA, enabling controlled antibiotic release. We observed that liposomes composed of N-methyl-DOPE were sensitive to the presence of low concentrations of LtxA, and cholesterol increased the extent and kinetics of content release. The liposomes were stable under various storage conditions for at least 7 days. Finally, we showed that antibiotic release occurs selectively in the presence of an LtxA-producing strain of A. actinomycetemcomitans but not in the presence of a non-LtxA-expressing strain. Together, these results demonstrate that the designed liposomal vehicle enables toxin-triggered delivery of antibiotics to LtxA-producing strains of A. actinomycetemcomitans., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Angela C. Brown reports financial support was provided by National Institutes of Health. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Introduction: Quorum-quenching enzyme Est816 hydrolyzes the lactone rings of N -acyl homoserine lactones, effectively blocking the biofilm formation and development of Gram-negative bacteria. However, its applications in the oral field is limited. This study aimed to evaluate the efficacy of enzyme Est816 in combination with antibiotics against periodontitis induced by Aggregatibacter actinomycetemcomitans in vitro and in vivo ., Methods: The antimicrobial efficacy of enzyme Est816 in combination with minocycline, metronidazole, and amoxicillin was determined using the minimum inhibitory concentration test. The anti-biofilm effect of enzyme Est816 was assessed using scanning electron microscopy, live/dead bacterial staining, crystal violet staining, and real-time quantitative PCR. Biocompatibility of enzyme Est816 was assessed in human gingival fibroblasts (HGF) by staining. A rat model of periodontitis was established to evaluate the effect of enzyme Est816 combined with minocycline using micro-computed tomography and histological staining., Results: Compared to minocycline, metronidazole, and amoxicillin treatment alone, simultaneous treatment with enzyme Est816 increased the sensitivity of biofilm bacteria to antibiotics. Enzyme Est816 with minocycline exhibited the highest rate of biofilm clearance and high biocompatibility. Moreover, the combination of enzyme Est816 with antibiotics improved the antibiofilm effects of the antibiotics synergistically, reducing the expression of the virulence factor leukotoxin gene ( ltxA ) and fimbria-associated gene ( rcpA ). Likewise, the combination of enzyme Est816 with minocycline exhibited a remarkable inhibitory effect on bone resorption and inflammation damage in a rat model of periodontitis., Discussion: The combination of enzyme Est816 with antibiotics represents a prospective anti-biofilm strategy with the potential to treat periodontitis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Wang, Ju, Guo, Shan, Wu, Zhang and Zhang.)

Objective: To evaluate the antibacterial effectiveness of a combination of ε-poly-L-lysine (ε-PL), funme peptide (FP) as well as domiphen against oral pathogens, and assess the efficacy of a BOP® mouthwash supplemented with this combination in reducing halitosis and supragingival plaque in a clinical trial., Materials and Methods: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the compound against Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans, and Aggregatibacter actinomycetemcomitans were determined by the gradient dilution method. Subsequently, the CCK-8 assay was used to detect the toxicity of mouthwash on human gingival fibroblastst, and the effectiveness in reducing halitosis and supragingival plaque of the mouthwash supplemented with the combination was analyzed by a randomized, double-blind, parallel-controlled clinical trial., Results: The combination exhibited significant inhibitory effects on tested oral pathogens with the MIC < 1.56% (v/v) and the MBC < 3.13% (v/v), and the mouthwash containing this combination did not inhibit the viability of human gingival fibroblasts at the test concentrations. The clinical trial showed that the test group displayed notably lower volatile sulfur compounds (VSCs) at 0, 10, 24 h, and 7 d post-mouthwash (P < 0.05), compared with the baseline. After 7 days, the VSC levels of the and control groups were reduced by 50.27% and 32.12%, respectively, and notably cutting severe halitosis by 57.03% in the test group. Additionally, the Plaque Index (PLI) of the test and control group decreased by 54.55% and 8.38%, respectively, and there was a significant difference in PLI between the two groups after 7 days (P < 0.01)., Conclusions: The combination of ε-PL, FP and domiphen demonstrated potent inhibitory and bactericidal effects against the tested oral pathogens, and the newly formulated mouthwash added with the combination exhibited anti-dental plaque and anti-halitosis properties in a clinical trial and was safe., Trial Registration: The randomized controlled clinical trial was registered on Chinese Clinical Trial Registry (No. ChiCTR2300073816, Date: 21/07/2023)., (© 2024. The Author(s).)

Purpose: To investigate the microbiological outcomes obtained with either subgingival debridement (SD) in conjunction with a gel containing sodium hypochlorite and amino acids followed by subsequent application of a cross-linked hyaluronic acid gel (xHyA) gel, or with SD alone., Materials and Methods: Forty-eight patients diagnosed with stages II-III (grades A/B) generalised periodontitis were randomly treated with either SD (control) or SD plus adjunctive sodium hypochlorite/amino acids and xHyA gel (test). Subgingival plaque samples were collected from the deepest site per quadrant in each patient at baseline and after 3 and 6 months. Pooled sample analysis was performed using a multiplex polymerase chain reaction (PCR)-based method for the identification of detection frequencies and changes in numbers of the following bacteria: Aggregatibacter actinomycetemcomitans (A.a), Porphyromonas gingivalis (P.g), Tannerella forsythia (T.f), Treponema denticola (T.d), and Prevotella intermedia (P.i)., Results: In terms of detection frequency, in the test group, statistically significant reductions were found for P.g, T.f, T.d and P.i (p < 0.05) after 6 months. In the control group, the detection frequencies of all investigated bacterial species at 6 months were comparable to the baseline values (p > 0.05). The comparison of the test and control groups revealed statistically significant differences in detection frequency for P.g (p = 0.034), T.d (p < 0.01) and P.i (p = 0.02) after 6 months, favouring the test group. Regarding reduction in detection frequency scores, at 6 months, statistically significant differences in favour of the test group were observed for all investigated bacterial species: A.a (p = 0.028), P.g (p = 0.028), T.f (p = 0.004), T.d (p <0.001), and P.i (p = 0.003)., Conclusions: The present microbiological results, which are related to short-term outcomes up to 6 months post-treatment, support the adjunctive subgingival application of sodium hypochlorite/amino acids and xHyA to subgingival debridement in the treatment of periodontitis.

Background: The aim of this study was to investigate an in vitro dynamic bioreactor model by evaluating the antimicrobial effect of clinically relevant amoxicillin doses on polymicrobial microcosm biofilms derived from subgingival plaque., Methods: Biofilms from pooled subgingival plaque were grown for 108  hours in control and experimental dynamic biofilm reactors. Amoxicillin was subsequently infused into the experimental reactor to simulate the pharmacokinetic profile of a standard 500 mg thrice-daily dosing regimen over 5 days and biofilms were assessed by live/dead staining, scanning electron microscopy, and quantitative polymerase chain reaction., Results: Following establishment of the oral microcosm biofilms, confocal imaging analysis showed a significant increase in dead bacteria at 8 hours (p = 0.0095), 48 hours (p = 0.0070), 96 hours (p = 0.0140), and 120 hours (p < 0.0001) in the amoxicillin-treated biofilms compared to the control biofilms. Nevertheless, viable bacteria remained in the center of the biofilm at all timepoints. Significant reductions/elimination in Campylobacter rectus, Tannerella forsythia, Aggregatibacter actinomycetemcomitans, and Peptostreptococcus anaerobius was observed among the amoxicillin-treated biofilms at the 96 and 120 hour timepoints., Conclusion: A novel in vitro dynamic model of oral microcosm biofilms was effective in modeling the antimicrobial effect of a pharmacokinetically simulated clinically relevant dose of amoxicillin., (© 2023 The Authors. Journal of Periodontology published by Wiley Periodicals LLC on behalf of American Academy of Periodontology.)

Introduction: Development of bacterial resistance and antimicrobial side-effect has shifted the focus of research toward Ethnopharmacology. A biologically active compound derived from the plants may increase the effectiveness of antibiotic when used in combination. The present study aims to determine the synergistic antibacterial effect of ethanolic extracts of Punica granatum (pericarp), Commiphora molmol, Azadirachta indica (bark) in combination with amoxicillin, metronidazole, tetracycline, and azithromycin on periodontopathic bacteria: Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola and Aggregatibacter actinomycetemcomitans., Methodology: Periodontopathic bacterial strains were isolated from the plaque sample that was collected from periodontitis patients and grown under favorable conditions. Susceptibility of bacteria to the antibiotics and extracts was determined by disc diffusion method by measuring the diameter of the inhibition zones. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of plant extracts were evaluated against each bacterium. Synergistic effect of plant extract in combination with antibiotics was tested against each bacterium by measuring the diameter of zone of inhibition (ZOI)., Results: Findings revealed that all plant extracts exhibited an inhibitory effects on the proliferation and growth of periodontopathic bacteria. The maximum antibacterial effect was exhibited by C. molmol on P. gingivalis (ZOI = 20 ± 0.55 mm, MIC = 0.53 ± 0.24 mg/mL and MBC = 5.21 ± 1.81 mg/mL) (p < 0.05), meanwhile, no antibacterial activity was exhibited by P. granatum on T. forsythia. Synergistic antibacterial effect was recorded when plant extracts were used in combination with antibiotics. The best synergism was exhibited by P. granatum with amoxicillin against A. actinomycetemcomitans (24 ± 1.00 mm) (p < 0.05)., Conclusions: The synergistic test showed significant antibacterial activity when plant extracts were combined with antibiotics against all the experimented bacteria., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2021 Shahabe Abullais Saquib, Nabeeh Abdullah AlQahtani, Irfan Ahmad, Suraj Arora, Shaik Mohammed Asif, Mukhatar Ahmed Javali, Nazima Nisar.)

Cranberries are widely recognized as a functional food that can promote oral health. However, the high concentration of organic acids in cranberry juice can cause tooth enamel erosion. Electrodialysis with bipolar membrane (EDBM) is a process used for the deacidification of cranberry juice. The present study investigated whether the removal of organic acids (0%, 19%, 42%, 60%, and 79%) from cranberry juice by EDBM affects its antibacterial activity against major periodontopathogens as well as its anti-inflammatory properties in an oral epithelial cell model. A deacidification rate ≥60% attenuated the bactericidal effect against planktonic and biofilm-embedded Aggregatibacter actinomycetemcomitans but had no impact on Porphyromonas gingivalis and Fusobacterium nucleatum . Cranberry juice increased the adherence of A. actinomycetemcomitans and P. gingivalis to oral epithelial cells, but reduced the adherence of F. nucleatum by half regardless of the deacidification rate. F. nucleatum produced more hydrogen sulfide when it was exposed to deacidified cranberry juice with a deacidification rate ≥42% compared to the raw beverage. Interestingly, the removal of organic acids from cranberry juice lowered the cytotoxicity of the beverage for oral epithelial cells. Deacidification attenuated the anti-inflammatory effect of cranberry juice in an in vitro oral epithelial cell model. The secretion of IL-6 by lipopolysaccharide (LPS)-stimulated oral epithelial cells exposed to cranberry juice increased proportionally with the deacidification rate. No such effect was observed with respect to the production of IL-8. This study provided evidence that organic acids, just like phenolic compounds, might contribute to the health benefits of cranberry juice against periodontitis.

The prevalence of periodontal disease poses a significant global health burden. Treatments for these diseases, primarily focused on removal and eradication of dental plaque biofilms, are challenging due to limited access to periodontal pockets where these oral pathogens reside. Herein, we report on the development and characterization of nitric oxide (NO)-releasing carboxymethylcellulose (CMC) derivatives and evaluate their in vitro bactericidal efficacy against planktonic Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans, two prominent periodontopathogens. Bactericidal exposure assays revealed that three of the synthesized NO-releasing polymers were capable of reducing bacterial viability of both species by 99.9% in 2 hr at concentrations of 4 mg ml -1 or lower, reflecting NO's potent and rapid bactericidal action. The NO-releasing CMCs elicited minimal toxicity to human gingival fibroblasts at their bactericidal concentrations following 24-hr exposure., (© 2020 Wiley Periodicals LLC.)

The present investigation reports an in-vitro study using combination of laccase and an enhancer capable of inhibiting the growth of pathogenic microorganisms, preventing biofilm formation, and whitening teeth. Laccase-cinnamic acid system remarkably inhibited the growth of Aggregatibacter actinomycetemcomitans, Candida albicans, S. aureus, and Streptococcus mutans whilst showed no significant effects on Gram-negative bacteria. Data presented that cinnamic acid (10 mM) with laccase (0.125 U ml -1 ) led to a maximum decrease of about 90%, in S. mutans biofilm formation. The confocal laser scanning microscopy showed considerable detachment of S. mutans cells from glass substratum. The combined laccase-cinnamic acid system could remove teeth discoloration caused by coffee. SEM of the teeth surface exhibited no damages such as surface cracking or fracture. Liquid chromatography-tandem mass spectrometry (LC-MS) and cyclic voltammetry (CV) studies showed that laccase can catalyze the one-electron oxidation of cinnamic acid to the respective radical. This radical can then undergo several fates, including recombination with another radical to form a dimeric species, dismutation of the radical back to cinnamic acid or decarboxylation to give various reduced oxygen species. Therefore, the redox potential values of phenolic monomers/oligomers are related with their biological activities., (Copyright © 2021. Published by Elsevier B.V.)

Objectives: We and others have previously shown that epigallocatechin gallate (EGCg) inhibits the activity of an important virulence factor, leukotoxin (LtxA), produced by the oral bacterium Aggregatibacter actinomycetemcomitans, suggesting the potential use of this molecule as an anti-virulence strategy to treat periodontal infections. Here, we sought to better understand the effects of EGCg on toxin secretion and A. actinomycetemcomitans pathogenicity in a co-culture model., Methods: We used a quantitative immunoblot assay to determine the concentrations of LtxA in the bacterial supernatant and on the bacterial cell surface. Using a co-culture model, consisting of A. actinomycetemcomitans and THP-1 cells, we studied the impact of EGCg-mediated changes in LtxA secretion on the toxicity of A. actinomycetemcomitans., Key Findings: EGCg increased production of LtxA and changed the localization of secreted LtxA from the supernatant to the surface of the bacterial cells. In the co-culture model, a single low dose of EGCg did not protect host THP-1 cells from A. actinomycetemcomitans-mediated cytotoxicity, but a multiple dosing strategy had improved effects., Conclusions: Together, these results demonstrate that EGCg has important, but complicated, effects on toxin secretion and activity; new dosing strategies and comprehensive model systems may be required to properly develop these anti-virulence activities., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Royal Pharmaceutical Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objective: To determine the mechanism of growth inhibition of Aggregatibacter actinomycetemcomitans by Maillard reaction products (MRP)., Design: Growth and cell viabilities in the presence or absence of MRP were measured for both the rough and smooth variants of the bacteria. Effects of addition of ferrous and ferric ions on the inhibition of the bacteria by MRP were determined., Results: MRPs decreased the extent of complex formation of Chrome Azurol S with iron suggesting that MRPs can chelate iron effectively. The chelation causes growth inhibition of both the rough and smooth strains. At low concentrations of the inhibitor, lag time was extended by approximately 12 h while at high concentrations, cells were killed, decreasing cell viability by up to 8 orders of magnitude. Growth of both the rough and smooth strains could be restored to original level by addition of iron. For the rough strain, both ferrous and ferric ions could relieve the inhibition by MRP while for the smooth strain only ferrous ion was effective., Conclusion: MRPs inhibit the growth of A. actinomycetemcomitans by chelating iron and the inhibition can be relieved by addition of iron., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Dental implants are the most innovative and superior treatment modality for tooth replacement. However, titanium implants still suffer from insufficient antibacterial capability and peri-implant diseases remain one of the most common and intractable complications. To prevent peri-implant diseases, a composite coating containing a new antibacterial agent, (Z-)-4-bromo-5-(bromomethylene)-2(5H)-furanone (BBF) was fabricated on titanium. This study was designed to investigate the antibacterial activity of the composite coating against two common peri-implant pathogens ( Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans ). The morphology of the composite coating showed that BBF-loaded poly(L-lactic acid) nanospheres were well-distributed in the pores of the microarc oxidation coating, and cross-linked with each other and the wall pores by gelatin. A release study indicated that the antibacterial coating could sustain the release of BBF for 60 d, with a slight initial burst release occurring during the first 4 h. The antibacterial rate of the composite coating for adhering bacteria was the highest (over 97%) after 1 d and over 90% throughout a 30-day incubation period. The total fluorescence intensity of the composite coating was the lowest, and the vast majority of the fluorescence was red (dead bacteria). Moreover, real-time polymerase chain reaction analysis confirmed that the relative gene expression of the adherent bacteria on the composite coating was down-regulated. It was therefore concluded that the composite coating fabricated on titanium, which showed excellent and relatively long-term antibacterial activity against Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans , is a potential and promising strategy to be applied on dental implants for the prevention of peri-implant diseases.

Periodontitis is a chronic inflammatory disease caused by various periodontal pathogens. Weissella cibaria CMU (oraCMU) is a probiotic that promotes oral health. However, its anti‑inflammatory effects against periodontal pathogens have not yet been investigated. The present study evaluated the anti‑inflammatory effects of live oraCMU against stimulation with the formalin‑inactivated periodontal pathogen Aggregatibacter actinomycetemcomitans in RAW 264.7 macrophages. Cell viability was analyzed by the MTS assay in a dose‑dependent manner (at multiplicities of infection of 0.1, 1, 10, 100 and 1,000). Nitric oxide (NO) was monitored using the Griess test. The mRNA expression of proinflammatory cytokines such as interleukin (IL)1β and IL6 was assessed by reverse transcription‑quantitative PCR. Western blotting was used to examine the effects of oraCMU on the phosphorylation of NF‑κB inhibitor α (IκBα) and IκBα kinase (IKK), the nuclear translocation of the NF‑κB subunit p65 and the expression of inducible NO synthase (iNOS). Live oraCMU had no cytotoxic effects on RAW 264.7 macrophages. In A. actinomycetemcomitans‑stimulated RAW 264.7 macrophages, oraCMU reduced NO production by suppressing iNOS expression and downregulating the mRNA expression of proinflammatory cytokines in a dose‑dependent manner. IKK phosphorylation and IκBα degradation were dose‑dependently inhibited by oraCMU and the nuclear translocation of p65 via the canonical NF‑κB pathway was simultaneously reduced. The results indicated that oraCMU possessed anti‑inflammatory activity associated with the inhibition of NF‑κB signal activation in response to periodontal pathogens. This suggests that oraCMU is a beneficial anti‑inflammatory probiotic that can aid in the maintenance of oral health.

Bacteria easily adhere, colonize, and form biofilm on oral implants subsequently causing periimplantation periarthritis and mechanical loosening. Previous studies show that a high potential surface on polymeric implants can achieve surface bacteriostasis without side effects. In this study, a high surface potential is introduced to zirconia ceramics to mitigate bacterial infection. Carbon and nitrogen plasma immersion ion implantation (C-PIII and N-PIII) are conducted on zirconia ceramic samples sequentially to elevate the surface potential. The surface with a high potential but without ion leaching exhibits excellent antibacterial effects against oral bacteria and little bacterial resistance is triggered. The surface also has high strength and excellent biocompatibility. The nitrogen-containing inorganic structure with high potential can actualize bacteriostasis and biocompatibility on zirconia ceramics simultaneously and this new strategy can enhance the antibacterial ability of oral implants., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Periodontal diseases are mainly the results of infections and inflammation of the gum and bone that surround and support the teeth. In this study, the alveolar bone destruction in periodontitis is hypothesized to be treated with novel Mg-Cu alloy grafts due to their antimicrobial and osteopromotive properties. In order to study this new strategy using Mg-Cu alloy grafts as a periodontal bone substitute, the in vitro degradation and antibacterial performance were examined. The pH variation and Mg 2+ and Cu 2+ release of Mg-Cu alloy extracts were measured. Porphyromonas gingivalis ( P. gingivalis ) and Aggregatibacter actinomycetemcomitans ( A. actinomycetemcomitans ), two common bacteria associated with periodontal disease, were cultured in Mg-Cu alloy extracts, and bacterial survival rate was evaluated. The changes of bacterial biofilm and its structure were revealed by scanning electron microscopy (SEM) and transmission electronic microscopy (TEM), respectively. The results showed that the Mg-Cu alloy could significantly decrease the survival rates of both P. gingivalis and A. actinomycetemcomitans . Furthermore, the bacterial biofilms were completely destroyed in Mg-Cu alloy extracts, and the bacterial cell membranes were damaged, finally leading to bacterial apoptosis. These results indicate that the Mg-Cu alloy can effectively eliminate periodontal pathogens, and the use of Mg-Cu in periodontal bone grafts has a great potential to prevent infections after periodontal surgery., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2020 Xue Zhao et al.)

This study investigated the bactericidal effect, the underlying mechanisms of treatment, and recovery of biocompatibility of the infected titanium surface using a combination treatment of silver ion application and ultraviolet-A (UV-A) light irradiation. Streptococcus mutans and Aggregatibacter actinomycetemcomitans were used in suspension and as a biofilm on a titanium surface to test for the bactericidal effect. The bactericidal effect of the combination treatment was significantly higher than that of silver ion application or UV-A light irradiation alone. The bactericidal effect of the combination treatment was attributable to hydroxyl radicals, which generated from the bacterial cell wall and whose yield increased with the silver concentration. To assess the biocompatibility, proliferation and calcification of MC3T3E1 cells were evaluated on the treated titanium surface. The treated titanium screws were implanted into rat tibias and the removal torques were measured 28 days post-surgery. The titanium surface that underwent the combination treatment exhibited recovery of biocompatibility by allowing cellular proliferation or calcification at levels observed in the non-infected titanium surfaces. The removal torque 28 days after surgery was also comparable to the control values. This approach is a novel treatment option for peri-implantitis.

For dental implants the accumulation of anaerobic bacteria is a main reason for peri-implant inflammation, which untreated can lead to implant loss. Oxygen releasing substances may act as antibacterial agents. In this study glucose-1-phosphate (Glc-1P) biofunctionalized zinc peroxide (ZnO₂) nanoparticles of four different synthesis ratio (1-10:1) and sizes (4-5 nm) were tested against the anaerobes Fusobacterium nucleatum, Porphyromonas gingivalis, and Prevotella intermedia, as well as against Aggregatibacter actinomycetemcomitans, Enterococcus faecalis, Staphylococcus aureus, Lactobacillus paracasei, and the yeast Candida albicans. Nanoparticles stabilized with o-phosphorylethanolamine, bis[2-(methacryloyloxy)ethyl] phosphate, or dioctyl sulfosuccinate instead of glucose were used as controls. For every combination of test strain and nanoparticle both, the minimal inhibitory (MIC) and minimal microbicidal concentration (MBC or MFC) were determined under different pH conditions in microtiter plates. Furthermore, transmission electron (TEM) and fluorescence microscopy after live-dead-staining was performed on selected combinations of pathogen and nanoparticle in order to visualize the interactions. The ZnO 2 /Glc-1P nanoparticles had an inhibitory effect on gram-negative anaerobes and on A. actinomycetemcomitans with a pH-dependent MIC ≥25 μg/ml and MBC ≥50 μg/ml, while the gram-positive species tested and C. albicans were not inhibited. In TEM images, attachment of nanoparticle-chains to the bacterial outer membrane and subsequent penetration was found together with an intracellular oxygen release. For nanoparticles with other stabilizers than glucose an invasion was only seen in elongated, dividing cells, possibly because of the more porous cell wall in the parting layer. Decorating ZnO 2 by glucose-1-phosphate is a Trojan horse approach to permit their uptake in gram-negative oxygen-sensitive bacterial cells., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Probiotics can stabilize gut flora, regulate intestinal immunity and protect the host from enteric diseases; however, their roles in oral health have received little attention compared to their roles in gut health. Nowadays, the prevalence of sugar-sweetened foods and abuse of antibiotics contribute towards dysbiosis of oral microbiota and drug resistance development in oral pathogens, resulting in various intractable oral diseases. We screened the antibacterial activities of viable and heat-killed probiotic strains against the oral pathogens Streptococcus mutans, Porphyromonas gingivalis, Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans. The probiotic strains Lactobacillus salivarius subsp. salicinius AP-32, L. rhamnosus CT-53, L. paracasei ET-66 and Bifidobacterium animalis subsp. lactis CP-9 displayed strong antipathogenic activities, whereas heat-killed AP-32, CT-53 and ET-66 displayed high levels of pathogen inhibition. The antibacterial activities of these probiotics were not associated with their H 2 O 2 production; L. acidophilus TYCA02 produced high levels of H 2 O 2 but merely exhibited moderate antibacterial activities. Oral tablets containing probiotics showed positive inhibitory effects against oral pathogens, particularly those containing viable probiotics. Our results indicate that probiotics prevent the growth of oral pathogens and improve oral health, providing insights into the antipathogenic efficacy of different probiotic species and their potential role in functional foods that improve oral health. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study provides insights into the antipathogenic efficacy of different probiotic species and their potential roles in developing functional foods to improve oral health. We showed that the probiotic strains Lactobacillus salivarius subsp. salicinius AP-32, L. rhamnosus CT-53, L. paracasei ET-66 and Bifidobacterium animalis subsp. lactis CP-9 have great potential for use in the development of functional foods to improve oral health. Since active probiotics may provide strong and long-term protection, the development of functional food products should favour the use of viable bacteria., (© 2020 The Society for Applied Microbiology.)

Purpose: Electrospun PLA fiber devices were investigated in the form of fiber mats and disks. Metronidazole was used as an active agent; its concentration was 12.2 and 25.7 wt% in the devices., Methods: The structure was studied by X-ray diffraction and scanning electron microscopy, drug release by dissolution measurements, while the antimicrobial efficiency was tested on five bacterial strains., Results: The XRD study showed that the polymer was partially crystalline in both devices, but a part of metronidazole precipitated and was in the form of crystals among and within the fibers. Liquid penetration and dissolution were different in the two devices, they were faster in disks and slower in fiber mats, due to the morphology of the device and the action of capillary forces. Disks released the drug much faster than fiber mats. Although the release study indicated fast drug dissolution, the concentration achieved a plateau value in 24 hrs for the disks; the inhibition effect lasted much longer, 13 days for bacteria sensitive to metronidazole. The longer inhibition period could be explained by the slower diffusion of metronidazole located inside the fibers of the device., Conclusion: The results suggest that the devices may be effective in the treatment of periodontitis., Competing Interests: The authors report no conflicts of interest in this work., (© 2020 Budai-Szűcs et al.)

Background: Recently, antimicrobial photodynamic therapy (aPDT) as an alternative treatment modality has been used adjunctively in the treatment of periodontitis and peri-implantitis. Photosensitizing agents in the form of nanoparticles have been designed for improving the efficiency of aPTD. Graphene quantum dots are a special type of nanocrystals that can promote aPDT when coupled with curcumin (Cur). The main objective of the present study was to investigate the effects of photoexcited GQD-Cur on the metabolic activity of perio-pathogen mixed biofilms., Materials and Methods: GQD-Cur was synthesized and characterized by scanning electron microscopy (SEM), dynamic light scattering (DLS), fourier transform infrared (FTIR) spectroscopy, ultraviolet-visible spectrometry (UV-Vis), and X-ray diffraction (XRD). The cell cytotoxicity effect of GQD-Cur was evaluated on primary human gingival fibroblast (HuGu) cells. Perio-pathogen mixed biofilms including Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia photosensitized with GQD doped with Cur were irradiated with a blue LED at a wavelength of 435 ± 20 nm for 1 min, and then bacterial viability measurements were performed. The antimicrobial susceptibility profile, biofilm formation ability, amount of reactive oxygen species (ROS) released, and variations of gene expressions involved in biofilm formation were assessed., Results: The SEM, DLS, FTIR, UV-Vis spectrometry, and XRD pattern confirmed that GQD-Cur was synthesized successfully. According to the results, GQD-Cur exhibited no cytotoxicity against HuGu cells. Photoexcited GQD-Cur resulted in a significant reduction in cell viability (93%) and biofilm formation capacity (76%) of peri-pathogens compared to the control group (P < 0.05). According to the results, a significant concentration-dependent increase in the ROS generation was observed in perio-pathogens mixed cells treated with different doses of GQD-Cur-aPDT. Moreover, rcpA, fimA, and inpA gene expression profiles were downregulated by 8.1-, 9.6-, and 11.8-folds, respectively., Conclusions: Based on the results, photoexcited GQD-Cur have a high potency of perio-pathogens suppression in planktonic and biofilm forms and downregulation of the biofilm genes expression pattern was exploited as a nanoscale-based platform for periodontitis., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A 45-year-old- man presented with left chest wall pain, swelling and cough. Over a 2-month period he developed abscesses in the right foot, right anterior thigh, left buttock and left chest. Incision and drainage of the soft tissue abscesses and video-assisted thoracoscopic surgery to drain the loculated empyema contiguous with the chest wall abscess were performed as surgical management. Gram stain showed beaded Gram-positive rods and the culture initially grew Aggregatibacter actinomycetemcomitans and Eikenella corrodens Pathological evaluation of the pleura showed sulfur granules and organisms consistent with Actinomyces spp. on Gomori methenamine silver stain; Actinomyces israelii was recovered in culture with extended incubation. The patient was treated for 3 weeks with ceftriaxone and oral metronidazole, followed by oral amoxicillin. Culture of A. actinomycetemcomitans with other findings consistent with actinomycosis warrants 6-12 months of antibiotic therapy., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2019. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Antimicrobial Photodynamic therapy (aPDT) is a novel modality suggested for treatment of peri-implantitis. This study aimed to assess the effect of aPDT with toluidine blue (TBO) and indocyanine green (ICG) and 635 nm and 808 nm diode laser on Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) biofilm formed on Laser-Lok titanium discs., Materials and Methods: Eighty sterile Laser-Lok titanium discs were inoculated with A. actinomycetemcomitans to form biofilm and were randomly divided into 8 groups (n = 10) of control, chlorhexidine (CHX), TBO, ICG, 635 nm diode laser with 220 mW power, 808 nm diode laser with 250 mW power, 100 μg/mL TBO+635 nm diode laser and ICG+808 nm diode laser. Number of colony forming units (CFUs) on the surface of each disc was counted after the intervention. Data were analyzed using the Kruskal-Wallis test., Results: Significant differences were noted in colony count among the eight groups after the intervention (P = 0.001). Pairwise comparisons with adjusted P value test showed that aPDT with TBO+635 nm laser and ICG+808 nm laser caused significant reduction of bacterial biofilm compared to the control group (P = 0.0001). TBO alone caused significant reduction of biofilm compared to the control group (P = 0.004). No other significant differences were noted (P > 0.05)., Conclusion: Within the limitations of this study, the results showed that aPDT is a potential modality for decontamination of implant surface and reduction of A. actinomycetemcomitans biofilm in vitro. In this study, aPDT with TBO+635 nm diode laser and ICG+808 nm diode laser decreased the bacterial load on titanium discs., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The concentration of methylene blue (MB) photosensitizer could affect the eradication efficacy of antimicrobial photodynamic therapy (aPDT) in the treatment of contaminated implants, which is linked to the osseointegration of the implant. We evaluated osteoblast functions on the contaminated SLA (sandblasting, large-grit and acid-etching) Ti alloy surfaces after the concentration-dependent use of MB-aPDT. Totally 1164 SLA discs were randomly distributed for the analyses of antibacterial efficacy and osteoblast functions. Gram-negative (Aggregatibacter actinomycetemcomitans; A. actinomycetemcomitans) or Gram-positive (Streptococcus mutans; S. mutans) adhered on disc samples was subjected to aPDT with different MB concentrations (200, 250, 300, 350, and 400 μg/mL) using 660 nm diode laser with maximum output 80 mW for 1 min irradiation (4.8 J/cm 2 ). Bactericidal effect was examined by viability, morphology, and lipopolysaccharide (LPS) assays. The disinfected disc surfaces by MB-aPDT to support osteoblast-like MG63 attachment, proliferation, differentiation, and mineralization were assessed for the predetermined culture time intervals. The statistical differences between the means were performed using a one-way analysis of variance (ANOVA) with a post hoc Scheffe test. The results of the morphology observation and bacterial survival examination consistently indicated a remarkably lower quantity of bacterial colonies on biofilm-contaminated surfaces after the aPDT treatment with higher MB concentration. Similarly, the higher MB concentration in aPDT resulted in the lower LPS amounts remaining on the A. actinomycetemcomitans-contaminated surfaces. Intriguingly, the expression of osteoblast cultured on disinfected surfaces using aPDT with higher MB concentration was comparable to the control without contamination. Within the limits of this in vitro model, this formulation of 400 μg/mL MB used in aPDT may be not only the lethal concentration against the 2 bacteria-contaminated implants, but it could also enhance the osteoblast functions on the contaminated implants. Nevertheless, the efficacy in the clinical practice for peri-implantitis therapy remains to be studied., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Antimicrobial photodynamic therapy (aPDT) is an effective adjunctive therapeutic modality for the treatment of local infections, including periodontitis and peri-implantitis. After receiving aPDT, microbial cells in the biofilm structure may produce and/ or release soluble biofilm-derived effectors (BDEs), which may affect the biology of the host cells in the community context of their surrounding microenvironment. Given the fact that no study has investigated the role of BDEs following aPDT in the pathogenesis of infectious diseases, the aim of the current study was to determine the effect of BDEs of Aggregatibacter actinomycetemcomitans following exposure to sub-lethal doses of indocyanine green (ICG)-aPDT on human gingival fibroblasts (HGFs) in terms of cytokines produced., Materials and Methods: In this study, we evaluated the effect of biofilm-conditioned medium (BCM) resulting from the treatment of A. actinomycetemcomitans biofilm with a sub-lethal dose of aPDT on cytokines production, including IL-6, IL-8, CXCL10, TGF-β, and bFGF of HGFs using enzyme-linked immunoassays (ELISA). The sensitivity of cytokines to BDEs was determined by micro-titer plates., Results: The maximal sub-lethal dose of ICG-PDT was 20.15 μM/mL ICG at a fluence of 31.2 J/cm 2 . The BCM of ICG-PDT-treated viable A. actinomycetemcomitans significantly reduced IL-6, IL-8, and CXCL10 levels compared to the BCM of untreated viable A. actinomycetemcomitans (78-, 93-, and 61.6-fold reduction, respectively; all P < 0.01). TGF-β and bFGF were strongly induced by BCM of ICG-PDT treated viable A. actinomycetemcomitans (by 57.7 and 36.1 folds, respectively; both P < 0.05). The BCM of untreated viable A. actinomycetemcomitans degraded most of the CxCL10, TGF-β and bFGF (58.8, 61.5, and 71.6%, respectively) in 24 h, while it degraded 9.3% of IL-6 and 15.1% of IL-8 after 24 h., Conclusion: The results of the current study revealed that a sub-lethal dose of ICG-aPDT through the effect of BCM on HGFs could not only significantly reduce the production of pro-inflammatory cytokines but also promoted their role in periodontal regeneration due to increasing the bFGF level. Altogether, ICG-aPDT, with it's antimicrobial effects reduces inflammation and induces of tissue regeneration resulting from BCM, can be considered an efficient adjunctive therapeutic method for the treatment of local infections., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Previous research findings support an antimicrobial effect of polyphenols against a variety of pathogens, but there is no evidence of this effect against periodontal pathogens in complex biofilms. The purpose of this study was to evaluate the antimicrobial activity of red wine and oenological extracts, rich in polyphenols, against the periodontal pathogens Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum and total bacteria growing in an in vitro oral biofilm static model., Methods: A previously validated biofilm model, including Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, F. nucleatum, P. gingivalis and A. actinomycetemcomitans was developed on sterile hydroxyapatite discs. Red wine (and dealcoholized wine), and two polyphenols-rich extracts (from wine and grape seeds) were applied to 72 h biofilms by dipping the discs during 1 and 5 min in the wine solutions and during 30 s and 1 min in the oenological extracts. Resulting biofilms were analyzed by confocal laser scanning microscopy and viable bacteria (colony forming units/mL) were measured by quantitative polymerase chain reaction combined with propidium monoazide. A generalized linear model was constructed to determine the effect of the tested products on the viable bacterial counts of A. actinomycetemcomitans, P. gingivalis and F. nucleatum, as well on the total number of viable bacteria., Results: The results showed that red wine and dealcoholized red wine caused reduction in viability of total bacteria within the biofilm, with statistically significant reductions in the number of viable P. gingivalis after 1 min (p = 0.008) and in A. actinomycetemcomitans after 5 min of exposure (p = 0.011) with red wine. No evidence of relevant antibacterial effect was observed with the oenological extracts, with statistically significant reductions of F. nucleatum after 30 s of exposure to both oenological extracts (p = 0.001)., Conclusions: Although moderate, the antimicrobial impact observed in the total bacterial counts and counts of A. actinomycetemcomitans, P. gingivalis and F. nucleatum, encourage further investigations on the potential use of these natural products in the prevention and treatment of periodontal diseases.

There is evidence that pathogenic bacteria can adapt to antiseptics upon repeated exposure. More alarming is the concomitant increase in antibiotic resistance that has been described for some pathogens. Unfortunately, effects of adaptation and cross-adaptation are hardly known for oral pathogens, which are very frequently exposed to antiseptics. Therefore, this study aimed to determine the in vitro increase in minimum inhibitory concentrations (MICs) in oral pathogens after repeated exposure to chlorhexidine or cetylpyridinium chloride, to examine if (cross-)adaptation to antiseptics/antibiotics occurs, if (cross-)adaptation is reversible and what the potential underlying mechanisms are. When the pathogens were exposed to antiseptics, their MICs significantly increased. This increase was in general at least partially conserved after regrowth without antiseptics. Some of the adapted species also showed cross-adaptation, as shown by increased MICs of antibiotics and the other antiseptic. In most antiseptic-adapted bacteria, cell-surface hydrophobicity was increased and mass-spectrometry analysis revealed changes in expression of proteins involved in a wide range of functional domains. These in vitro data shows the adaptation and cross-adaptation of oral pathogens to antiseptics and antibiotics. This was related to changes in cell surface hydrophobicity and in expression of proteins involved in membrane transport, virulence, oxidative stress protection and metabolism.

Background: Antimicrobial photodynamic therapy (aPDT), is a promising approach proposed as an adjunct for the decontamination of dental implant surfaces. This study aimed to investigate the effect of aPDT with laser or light emitting diode (LED) compared with conventional chlorhexidine treatment on the titanium-attached biofilms of Aggregatibacter actinomycetemcomitans., Methods and Materials: Thirty-six acid-etched and sandblasted (SLA) titanium discs were allocated to six groups and incubated with the titanium-adherent biofilms of A. actinomycetemcomitans as follows. Negative control (no treatment applied), positive control (0.2% chlorhexidine solution), 0.1 mg/mL Toluidine Blue [TBO] group, aPDT-treated groups subjected either to diode laser with a wavelength of 635 nm wavelength or LED with the peak wavelength of 630 nm with TBO as photosensitizer and sterile control (not contaminated). Following sonication and transferring the specimens to the microplate, the number of colony-forming units (CFUs) per disc was calculated. Data were analyzed using one-way analysis of variance (ANOVA) and additional post hoc tests with the level of significance set at P < 0.05., Results: aPDT using TBO + LED was significantly more effective (0.93 ± 0.24 × 10 4 ) in the suppression of A. actinomycetemcomitans compared with TBO + Laser (2.65 ± 0.7 × 10 4 ). However, the lowest mean of CFU count was found in sterile, and chlorhexidine groups, respectively (P < 0.0001) and the highest bacterial count was observed in the negative control group (P < 0.0001)., Conclusions: LEDs and diode lasers have a lower ability to suppress A. actinomycetemcomitans biofilms compared to 0.2% chlorhexidine in vitro. However, the aPDT with the use of LED as a light source and TBO as a photosensitive agent could be an appropriate alternative to conventional chlorhexidine treatment., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Hinokitiol, a component of the essential oil isolated from Cupressaceae, possesses antibacterial and antifungal activities and has been used in oral care products. In this study, the antibacterial activities of hinokitiol toward various oral, nasal and nasopharyngeal pathogenic bacteria, including Streptococcus mutans, Streptococcus sobrinus, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Fusobacterium nucleatum, methicillin-resistant and -susceptible Staphylococcus aureus, antibiotic-resistant and -susceptible Streptococcus pneumoniae, and Streptococcus pyogenes were examined. Growth of all these bacterial strains was significantly inhibited by hinokitiol, minimal inhibitory concentrations of hinokitiol against S. mutans, S. sobrinus, P. gingivalis, P. intermedia, A. actinomycetemcomitans, F. nucleatum, methicillin-resistant S. aureus, methicillin-susceptible S. aureus, antibiotic-resistant S. pneumoniae isolates, antibiotic-susceptible S. pneumoniae, and S. pyogenes being 0.3, 1.0, 1.0, 30, 0.5, 50, 50, 30, 0.3-1.0, 0.5, and 0.3 μg/mL, respectively. Additionally, with the exception of P. gingivalis, hinokitiol exerted bactericidal effects against all bacterial strains 1 hr after exposure. Hinokitiol did not display any significant cytotoxicity toward the human gingival epithelial cell line Ca9-22, pharyngeal epithelial cell line Detroit 562, human umbilical vein endothelial cells, or human gingival fibroblasts, with the exception of treatment with 500 μg/mL hinokitiol, which decreased numbers of viable Ca9-22 cells and gingival fibroblasts by 13% and 12%, respectively. These results suggest that hinokitiol exhibits antibacterial activity against a broad spectrum of pathogenic bacteria and has low cytotoxicity towards human epithelial cells., (© 2019 The Societies and John Wiley & Sons Australia, Ltd.)

Background: The bystander effects, whereby naive (bystander) microbial cells near microbial cells directly exposed to certain treatment show responses that would not have happened in the absence of the directly targeted microbial cells, is recently documented in the field of microbiology. In this article, we discuss that substantial bystander responses are also observed after antimicrobial photodynamic therapy (aPDT) using curcumin (Cur)., Materials and Methods: Bystander effects induced by whole bacterial cell suspension (WBCS T ), cell-free supernatants fluid (CFSF T ), and bacterial cell pellet (BCP T ) obtained from A. actinomycetemcomitans culture treated with Cur-aPDT on cell survival, quorum sensing (QS) ability, metabolic activity and efflux capacity of A. actinomycetemcomitans were determined using microbial viability assay, Escherichia coli-based bioassay, XTT reduction method, and ethidium bromide (EtBr) accumulation assay, respectively., Results: A. actinomycetemcomitans cell survival reduced by 82.7% (P = 0.001) and 76.2% (P = 0.01) after exposure to WBCS T and CFSF T , respectively. The A. actinomycetemcomitans population increased by 5.5% (P = 0.7) after exposure to BCP T . Bacterial metabolic activity decreased by 42.6% (P = 0.02), 35.3% (P = 0.03), and 9.4% (P = 0.5) after exposure to WBCS T , CFSF T , and BCP T , respectively. A. actinomycetemcomitans exposed to WBCS T , CFSF T , and BCP T showed a reduction of 83.2% (P = 0.001), 77.2% (P = 0.01) and 21.9% (P = 0.09) in the QS mediator compared to the WBCS U , CFSF U , and BCP U of untreated A. actinomycetemcomitans, respectively. No significant change of the EtBr accumulation was observed in the three preparations of the Cur-aPDT-treated culture (i.e. WBCS T , CFSF T , and BCP T ) compared to their respective controls., Conclusions: The results of the current study revealed that Cur-aPDT could significantly reduce microbial cell survival, cell metabolic activity, efflux capacity, and QS ability through the bystander effects. As a result, the bystander effects of Cur-aPDT along with the direct effect of Cur-aPDT can enhance the efficiency of aPDT as an adjunct therapeutic strategy for treatment of local infections., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The main treatment of periodontal disease is the mechanical removal of supra and subgingival biofilm. Adjuvant therapies as antimicrobial photodynamic therapy (aPDT) may offer improved clinical and microbiological results. The aim of this in vitro study was to evaluate the effect of toluidine and methylene blue dyes, associated with red laser and LED, on elimination of a suspension of Aggregatibacter actinomycetemcomitans (A.a). Experimental groups (n = 29) consisted of positive (broth) and negative (gentamicin) controls, three different dyes concentrations (0.05; 0.1; 10 mg/ml) alone or associated with laser (660 nm) at two power settings (70 and 100 mW) and LED (627 ± 10 nm). Bacterial suspension received all treatments, and after serial dilutions they were cultured for 24 h in petri dishes for colony forming unit counts. Data were analyzed by ANOVA complemented by Tukey's test (p < 0.05). The results showed that both dyes, at a concentration of 10 mg/ml, alone or associated with laser and LED, caused 100% of death similar to the negative control (p > 0.05). It can be concluded that blue dyes for aPDT, at high concentration (10 mg/ml), are capable of eliminating A.a without adjuvant use of light sources., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Streptococcus oralis subspecies dentisani is explored as an anti-cariogenic probiotic. Here, subjecting freshly stimulated saliva samples of 35 healthy volunteers, six epidemiologically unrelated and two related strains were isolated (prevalence around 20%) applying a newly developed three-step procedure. Furthermore, the probiotic strain S. dentisani 7746 (AB-Dentisanium®) was tested under a variety of environmental conditions for its inhibitory effect on six S. mutans , two S. sobrinus , 15 other oral or intestinal streptococci, 15 S. dentisani strains, and six representatives of other species including periodontopathogens. All except one of the S. mutans strains were inhibited by 7746 colonies or culture supernatant concentrate but only if either the test cell number was low or the producer or its bacteriocin concentration, respectively, was high. S. sanguinis OMI 332, S. salivarius OMI 315, S. parasanguinis OMI 335, S. vestibularis OMI 238, and the intestinal S. dysgalactiae OMI 339 were not inhibited, while the other 10 streptococcal strains (especially S. oralis OMI 334 and intestinal S. gallolyticus OMI 326) showed a certain degree of inhibition. From the panel of other bacterial species only Aggregatibacter actinomycetemcomitans was slightly inhibited. With the exception of OMI 285 and OMI 291 that possessed a 7746 bacteriocin-like gene cluster, all S. dentisani strains and especially type strain 7747 T were strongly inhibited by 7746. In conclusion, probiotic strain 7746 might antagonize the initiation and progression of dental caries by reducing S. mutans if not too abundant. S. dentisani strains inhibit each other, but strains with similar bacteriocin-related gene clusters, including immunity genes, are able to co-exist due to cross-resistance. In addition, development of resistance and adaptation to 7746-bacteriocins was observed during our study and needs attention. Hence, mechanisms underlying such processes need to be further investigated using omics-approaches. On the manufacturing level, probiotic strains should be continuously tested for function. Further clinical studies investigating inhibition of S. mutans by AB-Dentisanium® are required that should also monitor the impact on the oral microbiome composition including resident S. dentisani strains.

The objectives of present research were to develop and characterize thermosensitive and mucoadhesive polymer-based sustained release moxifloxacin in situ gels for the treatment of periodontal diseases. Poloxamer- and chitosan-based in situ gels are in liquid form at room temperature and transform into gel once administered into periodontal pocket due to raise in temperature to 37 °C. Besides solution-to-gel characteristic of polymers, their mucoadhesive nature aids the gel to adhere to mucosa in periodontal pocket for prolonged time and releases the drug in sustained manner. These formulations were prepared using cold method and evaluated for pH, solution-gel temperature, syringeability and viscosity. In vitro drug release studies were conducted using dialysis membrane at 37 °C and 50 rpm. Antimicrobial studies carried out against Aggregatibacter actinomycetemcomitans (A.A.) and Streptococcus mutans (S. Mutans) using agar cup-plate method. The prepared formulations were clear and pH was at 7.01-7.40. The viscosity of formulations was found to be satisfactory. Among the all, formulations comprising of 21% poloxamer 407 and 2% poloxamer 188 (P5) and in combination with 0.5% HPMC (P6) as well as 2% chitosan and 70% β-glycerophosphate (C6) demonstrated an ideal gelation temperature (33-37 °C) and sustained the drug release for 8 h. Formulations P6 and C6 showed promising antimicrobial efficacy with zone of inhibition of 27 mm for A.A. and 55 mm for S. Mutans. The developed sustained release in situ gel formulations could enhance patient's compliance by reducing the dosing frequency and also act as an alternative treatment to curb periodontitis.

Bacterial cellulose (BC) has shown high potential as innovative wound dressing and drug delivery system. Bringing both together, drug-loaded BC was investigated for applications in dental therapies such as dental extraction or mucosal transplantation. Both applications would benefit from a material which degrades under physiological conditions, and from an antibiotic environment. Consequently, periodate-oxidation of BC was investigated to facilitate modified degradation behaviour. A periodate concentration of 0.14 mol/L at ϑ = 25 °C and t = 8 h resulted in a material loss of <10%, but at the same time a sufficient degree of degradation. Additionally, native and oxidised BC loaded with doxycycline was tested for prophylaxis against infection. An in vitro-toxicity test (MTT assay) provided a first confirmation of biocompatibility, whereas agar diffusion tests proved antibiotic efficiency against pathogenic oral bacteria. Release studies of the drug from native and oxidised BC confirmed a comparative biphasic release behaviour., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: The aim of this study was to evaluate the effect of multiple sessions of antimicrobial photodynamic therapy (aPDT) on the treatment of experimental periodontitis (EP) in rats treated with chemotherapy., Methods: Chemotherapy using 5-fluorouracil (5-FU) consisted of intraperitoneal administration of 60 and 40 mg/kg of 5-FU. 120 rats were subjected to chemotherapy with 5-FU and divided into groups: PT (periodontal treatment); PT+1aPDT (PT and single aPDT session); PT+4aPDT(PT and 4 sessions of aPDT); 1aPDT (single aPDT session); 4aPDT(4 sessions of aPDT). EP was induced in the mandibular molars via ligature placement. The alveolar bone loss (ABL) area in the furcation region was analysed histometrically. Proliferating cell nuclear antigen (PCNA), tartrate-resistant acid phosphatase (TRAP), receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG) and cleaved caspase-3 (CC3) were analysed by immunohistochemistry. Prostaglandin E2 was quantified using an ELISA, tumor necrosis factor (TNF)-α and interleukin (IL)-6 were assessed using a multiplex method. The prevalence of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella nigrescens, Prevotella intermedia and Fusobacterium nucleatum was assessed using PCR. The data were statistically analysed (P < 0.05)., Results: The PT+4aPDT group showed lower ABL than the PT or 1aPDT groups on day 7. Rats treated with aPDT showed a higher number of PCNA-positive cells with reduced immunolabeling of RANKL. Significant reductions in Prevotella nigrescens were observed in the PT+4aPDT group and in Aggregatibacter actinomycetemcomitans for the 1aPDT and 4aPDT groups., Conclusion: Repeated sessions of aPDT as an adjunct or alternative therapy were effective at reducing ABL, regulating bone metabolism, and reducing Prevotella nigrescens and Aggregatibacter actinomycetemcomitans., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Peptide nucleic acids (PNA) are DNA/RNA analogs in which the sugar-phosphate backbone is replaced by N-2-aminoethylglycine. PNA are widely used for experimental antisense therapy due to their strong affinity to mRNA. By targeting specific genes, protein synthesis and the growth of bacteria or cancer cells can be inhibited by PNA. Here, we report the design and evaluation of antisense PNA for selective growth inhibition of Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans, potent pathogens associated with periodontitis. Antisense PNA against groEL and acpP were prepared with carrier peptide (KFFKFFKFFK). Anti-groEL PNA for P. gingivalis specifically inhibited growth in a dose-dependent manner, and growth was inhibited for 5 h at a concentration of 3 μM. Anti-groEL PNA for A. actinomycetemcomitans inhibited growth for 2 h at a concentration of 3 μM with reduced GroEL protein expression. Anti-acpP PNA did not show a marked growth inhibitory effect on either species. Although further studies are needed to develop more effective antisense PNA for both species, anti-groEL PNA may be potentially useful species-specific antibacterial tools against oral pathogens., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

It is important to address the periodontitis-associated bacteria in the residual subgingival plaque after scaling and root planing to successfully treat periodontitis. In this study, we explored the possibility of exploiting the ion pairing/complexation of minocycline, Ca 2+ , and sulfate/sulfonate-bearing biopolymers to develop an intrapocket delivery system of minocycline as an adjunct to scaling and root planing. Minocycline-calcium-dextran sulfate complex microparticles were synthesized from minocycline, CaCl 2 , and dextran sulfate. They were characterized using Fourier-transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. An in vitro release study was conducted to evaluate the release kinetics of minocycline from these microparticles. Agar disk diffusion assays and biofilm-grown bacteria assays were used to assess antibacterial capability. High loading efficiency (96.98% ± 0.12%) and high loading content (44.69% ± 0.03%) for minocycline were observed for these complex microparticles. Mino-Ca-DS microparticles achieved sustained release of minocycline for at least 9 days at pH 7.4 and 18 days at pH 6.4 in phosphate-buffered saline, respectively. They also demonstrated potent antimicrobial effects against Streptococcus mutans and Aggregatibacter actinomycetemcomitans in agar disk diffusion and biofilm assays. These results suggested that the ion pairing/complexation of minocycline, Ca 2+ , and sulfonate/sulfate-bearing biopolymers can be exploited to develop complex microparticles as local delivery systems for periodontitis treatment., (Copyright © 2018 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

The conventional treatment of periodontal disease does not solve the high incidence of recolonization of periodontal pockets by pathogens. Here, we introduce an innovative concept of incorporating autochthonous bacteria as potential probiotics into nanofibers for local treatment. We selected and isolated the strain 25.2.M from the oral microbiota of healthy volunteers. It was identified as Bacillus sp. based on 16S rRNA sequence analyses. The strain is nonpathogenic, produces antimicrobial substances, and can grow over the periodontal pathogen Aggregatibacter actinomycetemcomitans in vitro, making it a promising probiotic candidate. The strain 25.2.M was successfully incorporated into the nanofibers in the form of spores (10 7 CFU/mg), the viabilities of which were exceptional (max. change of 1 log unit) both during electrospinning and after 12 months of storage. The release of the bacteria was delayed from chitosan/poly(ethylene oxide) compared to poly(ethylene oxide) nanofibers, and the antimicrobial activity against A. actinomycetemcomitans was confirmed. The developed nanodelivery system for administration into periodontal pockets thus offers a promising approach for the inhibition of periodontal pathogens and restoration of healthy oral microbiota.

This study describes the successful synthesis of nitric oxide (NO)-releasing compounds with biodegradable and injectable properties and demonstrates that the kinetics of NO release vary according to the type of NO donor. The antimicrobial activity of NO-releasing compounds against three common periodontal pathogens, i.e., Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Actinomyces israelii, was investigated using a susceptibility assay. Human gingival fibroblasts were treated with NO-releasing compounds at the minimum concentrations required for bacterial growth and cytotoxicity was evaluated using the MTT cell proliferation assay. Our results suggest that NO-releasing compounds can be used topically to treat both gram-negative and gram-positive periodontal pathogens. Comparison of the antimicrobial activity and cytotoxicity assay results between the NO-releasing compounds revealed that an NO donor comprising a macromolecule without surface charge, a lower instantaneous NO concentration, and an adequate supply of NO were associated with a strong bactericidal effect and low cytotoxicity. NO-releasing compounds with these properties may be suitable for treatment of periodontitis., Competing Interests: The authors have declared that no competing interests exist.

Aggregatibacter actinomycetemcomitans is well-known as the pathogen of gingivitis or periodontitis, and discitis or vertebral osteomyelitis cases caused by this organism have rarely been reported. Ampicillin or amoxicillin has been used in the previously reported discitis cases; however, no cases have been reported that is treated with levofloxacin. We report the first published case we chose levofloxacin to treat. We failed to perform the susceptibility testing because of the poor growth and fastidious nature of the organism, and the result of susceptibility of amoxicillin was unclear. Levofloxacin, which A. actinomycetemcomitans is usually susceptible to, can be an effective alternative oral antimicrobial agent in such cases., (Copyright © 2018 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

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.

Purpose: The aim of this study was to evaluate the efficacy of photodynamic therapy (PDT) and light-activated disinfection (LAD) against a 3-day-old bacterial suspension prepared from three different bacterial species present on titanium dental implants, and to analyze the possible alterations of the implant surfaces as a result of the PDT and LAD., Materials and Methods: The study was conducted on 72 titanium dental implants contaminated with a bacterial suspension prepared from three bacterial species: Prevotella intermedia, Aggregatibacter actinomycetemcomitans, and Porphyromonas gingivalis. The contaminated implants were incubated under anaerobic conditions for 72 hours and then were randomly divided into four experimental groups and two control groups (n = 12 each), according to the following treatment protocols: group 1 (PDT1): PDT (660 nm, 100 mW, 60 seconds) with toluidine blue; group 2 (PDT2): PDT (660 nm, 100 mW, 60 seconds) with phenothiazine chloride dye; group 3 (LAD): light-emitting diode (LED) with toluidine blue; group 4 (toluidine blue): treatment with only toluidine blue for 60 seconds. In the positive control group, the implants were treated with a 0.2% chlorhexidine-based solution for 60 seconds, and in the negative control group, no treatment was used., Results: The highest bacterial reduction was recorded in the PDT1 (98.3%) and PDT2 (97.8%) groups. The results of this study showed that there was a statistically significant reduction of bacteria in the PDT1 and PDT2 groups compared with the negative control group (P < .05), individually for each bacterial species as well as for all three species together. LAD was less effective than PDT1 and PDT2, and did not show a statistically significant difference compared with the negative control or any other treatment group. Toluidine blue was the least effective treatment in terms of both the total bacterial count and the individual count for each bacterial species., Conclusion: Both PDT1 and PDT2 protocols showed a high efficacy against a 3-day-old bacterial biofilm on dental implants and were more effective compared with LAD.

Background and Objective: Curcumin, one of an established curcuminoid substances extracted from Curcuma longa, has been used as a photosensitizer (PS) in photodynamic therapy (PDT). Curcuminoid substances has been reported to have benefits in treating dental chronic infection and inflammation diseases, such as chronic periodontitis. The purpose of this study was to find the optimum concentration of Curcuma longa (CL) extract, containing all curcuminoid substances, and the power density of blue light (BL) in photodynamic therapy against periodontally pathogenic bacteria, A. actinomycetemcomitans., Methods: Antibacterial activity of various concentrations of CL extract against A. actinomycetemcomitans was determined. Exponentially growing bacteria were combined with 2-fold dilution of CL extract solution ranging from 25 to 0.098 μg/ml. Co-culture bacteria treated with 0.12% chlorhexidine (CHX) served as the positive control. The effect of photostimulation with light emitting diode (LED) 420-480 nm at 16.8 J/cm 2 for 1 min on the selected concentration of CL extract was examined. Bacteria viability was determined by plate counting technique. In addition, production of free radicals was tested by electron spin resonance spectroscope (ESR) with 5,5-dimethyl-1-pyrroline N-oxide (DMPO)., Results: The antibacterial activity of CL extract was dose dependent. Without BL, 25 μg/ml CL extract showed 6.03 ± 0.39 log 10 A. actinomycetemcomitans. Interestingly, the combination of BL and 0.78 μg/ml CL extract solution showed complete absence of A. actinomycetemcomitans. Peak signal intensity of hydroxyl radical production was also detected with the combination of BL and CL., Conclusions: CL extract not only had antimicrobial activity but also could be used as an effective PS when stimulated with BL in PDT. The optimal antibacterial effect of CL extract with BL was equal to the standard oral disinfectant, 0.12% CHX., (Copyright © 2018 Elsevier B.V. All rights reserved.)