Comparison of the effectiveness of Morus alba and chlorhexidine gels as an adjunct to scaling and root planing on stage II periodontitis - A randomized controlled clinical trial.

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.

Synergistic antibacterial activity of herbal extracts with antibiotics on bacteria responsible for periodontitis.

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.

Effect of cranberry juice deacidification on its antibacterial activity against periodontal pathogens and its anti-inflammatory properties in an oral epithelial cell model.

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.

Essential Oil from Psidium cattleianum Sabine (Myrtaceae) Fresh Leaves: Chemical Characterization and in vitro Antibacterial Activity Against Endodontic Pathogens

Abstract Endodontic infections result from oral pathogenic bacteria which reach and infect dental pulp, as well as surrounding tissues, through cracks, unrepaired caries and failed caries restorations. This study aims to determine the chemical composition of essential oil from Psidium cattleianum leaves (PC-EO) and to assess its antibacterial activity against endodontic bacteria. Antibacterial activity of PC-EO was evaluated in terms of its minimum inhibitory concentration (MIC) values by the broth microdilution method on 96-well microplates. Bacteria Porphyromonas gingivalis (MIC = 20 µg/mL), Prevotella nigrescens (MIC = 62.5 µg/mL), Fusobacterium nucleatum (MIC = 12.5 µg/mL), Actinomyces naeslundii (MIC = 50 µg/mL), Bacteroides fragilis (MIC = 12.5 µg/mL), Aggregatibacter actinomycetemcomitans (MIC = 6.25 µg/mL) and Peptostreptococcus anaerobius (MIC = 62.5 µg/mL) were evaluated and compared to chlorhexidine dihydrochloride (CDH), the positive control. PC-EO was obtained by hydrodistillation with the use of a Clevenger-type apparatus whereas its chemical composition was analyzed by gas chromatography-flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Viridiflorol (17.9%), β-caryophyllene (11.8%), 1,8-cineole (10.8%) and β-selinene (8.6%) were the major constituents found in PC-EO, which exhibited high antibacterial activity against all endodontic pathogens under investigation. Therefore, PC-EO, a promising source of bioactive compounds, may provide therapeutic solutions for the field of endodontics.

Antimicrobial activity of red wine and oenological extracts against periodontal pathogens in a validated oral biofilm model.

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.

Changes of microbial cell survival, metabolic activity, efflux capacity, and quorum sensing ability of Aggregatibacter actinomycetemcomitans due to antimicrobial photodynamic therapy-induced bystander effects.

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 (WBCST), cell-free supernatants fluid (CFSFT), and bacterial cell pellet (BCPT) 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 WBCST and CFSFT, respectively. The A. actinomycetemcomitans population increased by 5.5% (P = 0.7) after exposure to BCPT. Bacterial metabolic activity decreased by 42.6% (P = 0.02), 35.3% (P = 0.03), and 9.4% (P = 0.5) after exposure to WBCST, CFSFT, and BCPT, respectively. A. actinomycetemcomitans exposed to WBCST, CFSFT, and BCPT 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 WBCSU, CFSFU, and BCPU 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. WBCST, CFSFT, and BCPT) 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.

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.

Antibacterial Activity of Homeopathic Medications Lycopodium clavatum and Arsenicum album Against Periodontal Bacteria / Evaluación antibacteriana de los medicamentos homeopáticos sobre bacterias periodontales

Abstract There are several controversies regarding the efficacy of homeopathic substances; however, these remedies are used in many countries for the treatment of various pathological conditions. The purpose of this study was to evaluate the in vitro antibacterial activity of two homeopathic tinctures Arsenicum album (mineral extract) and Lycopodium clavatum (plant extract) on the periodontal bacteria Actinomyces israelii, Streptococcus sanguinis, Prevotella intermedia, Aggregatibacter actinomycetemcomitans and Phorphyromonas gingivalis (P. gingivalis). Materials and methods: Equal numbers of bacteria were seeded on agar plates containing enriched media with the homeopathic solutions at 1dH and 1cH dilutions. After 7 days of incubation under anaerobic conditions, colony forming units (CFUs) were counted. The antibacterial effect was calculated based on the total number of CFUs observed on non-tincture containing agar, and on the tincture containing plates. Results: No visible growth of any of the strains was observed on the plates containing Arsenicum album at any of the dilutions tested. In contrast, when Lycopodium clavatum at 1cH dilution was tested, only P. gingivalis was susceptible to this compound. Conclusions: The results suggest that the mineral extract tincture had a greater antibacterial activity than the plant extract tincture, also Lycopodium clavatum preparation could be an effective inhibitor of periodontal pathogens bacteria such as P. gingivalis.

Resumen Se necesita un mayor número de estudios in vitro e in vivo para validar estos resultados.

Evaluation of the Efficacy of Guava Extract as an Antimicrobial Agent on Periodontal Pathogens.

AIM: The present study was undertaken to assess the inhibitory effect of guava extracts on Porphyromonas gingivalis and Aggregatibacteractinomycetemcomitans, to assess the time-kill curve of P. gingivalis and A. actinomycetemcomitans, and to determine the antiproteolytic activity of guava on P. gingivalis. MATERIALS AND METHODS: Kanamycin blood agar was used to isolate P. gingivalis and A. actinomycetemcomitans. Ethanolic guava extract (EGE) and aqueous guava extract (AGE) were prepared and the inhibitory effects of these extracts for two periodontal pathogens were tested by minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) procedures. Antibacterial activity of guava extracts was determined by well diffusion method. Antiproteolytic activity of guava on protease of P. gingivalis was determined by gelatin liquefaction test. RESULTS: The MIC determined for AGE and EGE was at 75 µL/mL concentration for P. gingivalis, whereas EGE exhibited the activity at 75 µL/mL on P. gingivalis. The MIC determined for AGE was at 50 µL/mL for A. actinomycetemcomitans, whereas MIC determined for EGE was at 3.12 µL/mL for A. actinomycetemcomitans. Porphyromonas gingivalis was susceptible to EGE compared with AGE. Aggregatibacter actinomycetemcomitans was more susceptible to guava extracts compared with P. gingivalis. CONCLUSION: Guava extract may be a potential therapeutic agent for periodontitis as it shows significant activity against both P. gingivalis and A. actinomycetemcomitans. CLINICAL SIGNIFICANCE: Guava leaves extract can be used as economical and suitable adjuvant to synthetic drugs and can be a potential therapeutic agent for periodontitis.

In vitro Antimicrobial Activity of Ocimum sanctum (Tulsi) Extract on Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis.

Aim: The present study was conducted with an aim to assess the antimicrobial activity of Ocimum sanctum (tulsi) extract on Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis.Materials and methods: Tulsi extract with ethanol was prepared using cold extraction method in the present in vitro study. Various concentrations (2, 4, 6, and 8%) were then obtained by dilution with dimethylformamide. A 0.2% chlorhexidine served as the positive control, whereas the negative control was dimeth-ylformamide. Zones of inhibition were measured, each for A. actinomycetemcomitans and P. gingivalis. For comparison within the group and between the study groups, one-way analysis of variance (ANOVA) and Tukey's post hoc tests were used. A statistical significance level of p < 0.05 was established. Results: The 8% concentration of the tulsi extract showed maximum zone of inhibition against A. actinomycetemcomitans and P. gingivalis (40.10 ± 0.90, 33.79 ± 1.82 mm), followed by the 6, 4, and 2% concentrations. The 0.2% chlorhexidine, which was the positive control, had 39.80 ± 1.24 and 32.28 ± 1.28 mm zones of inhibition; dimethylformamide showed 13.55 ± 1.92 and 10.21 ± 2.16 mm zones of inhibition against both the microorganisms. The ANOVA showed highly statistically significant (p < 0.0001) results between and within the groups. The antimicrobial activity of tulsi extract at 6 and 8% concentrations, and 0.2% chlorhexidine against A. actinomycetemcomitans showed statistically significant differences between the groups. The concentration of tulsi extract at 8 and 0.2% chlorhexidine on P. gingivalis showed statistically significant differences between the groups. Conclusion: It was concluded that 8% concentration of O. sanctum (tulsi) extract showed the maximum antimicrobial activity against A. actinomycetemcomitans and P. gingivalis. It is thus recommended that this may be useful as an adjunc-tive to mechanical therapy in the prevention and treatment of periodontal diseases. Clinical significance:O. sanctum (tulsi) is a herb that is abundantly available, easily accessible, economically feasible, and culturally acceptable. Therefore, it is very useful in the management of oral diseases and also for overcoming many barriers that exist for the utilization of dental services, such as affordability, accessibility, availability, and acceptability. Keywords:Aggregatibacter actinomycetemcomitans, Antimicrobial, Ocimum sanctum, Porphyromonas gingivalis.

Photodynamic therapy of Curcuma longa extract stimulated with blue light against Aggregatibacter actinomycetemcomitans.

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/cm2 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 ±â€¯0.39 log10A. 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.

Dual action of highbush blueberry proanthocyanidins on Aggregatibacter actinomycetemcomitans and the host inflammatory response.

BACKGROUND: The highbush blueberry (Vaccinium corymbosum) has a beneficial effect on several aspects of human health. The present study investigated the effects of highbush blueberry proanthocyanidins (PACs) on the virulence properties of Aggregatibacter actinomycetemcomitans and macrophage-associated inflammatory responses. METHODS: PACs were isolated from frozen highbush blueberries using solid-phase chromatography. A microplate dilution assay was performed to determine the effect of highbush blueberry PACs on A. actinomycetemcomitans growth as well as biofilm formation stained with crystal violet. Tight junction integrity of oral keratinocytes was assessed by measuring the transepithelial electrical resistance (TER), while macrophage viability was determined with a colorimetric MTT assay. Pro-inflammatory cytokine and MMP secretion by A. actinomycetemcomitans-stimulated macrophages was quantified by ELISA. The U937-3xκB-LUC monocyte cell line transfected with a luciferase reporter gene was used to monitor NF-κB activation. RESULTS: Highbush blueberry PACs reduced the growth of A. actinomycetemcomitans and prevented biofilm formation at sub-inhibitory concentrations. The treatment of pre-formed biofilms with the PACs resulted in a loss of bacterial viability. The antibacterial activity of the PACs appeared to involve damage to the bacterial cell membrane. The PACs protected the oral keratinocytes barrier integrity from damage caused by A. actinomycetemcomitans. The PACs also protected macrophages from the deleterious effect of leukotoxin Ltx-A and dose-dependently inhibited the secretion of pro-inflammatory cytokines (IL-1ß, IL-6, CXCL8, TNF-α), matrix metalloproteinases (MMP-3, MMP-9), and sTREM-1 by A. actinomycetemcomitans-treated macrophages. The PACs also inhibited the activation of the NF-κB signaling pathway. CONCLUSION: The antibacterial and anti-inflammatory properties of highbush blueberry PACs as well as their ability to protect the oral keratinocyte barrier and neutralize leukotoxin activity suggest that they may be promising candidates as novel therapeutic agents.

Microbiologic Response to Periodontal Therapy and Multivariable Prediction of Clinical Outcome.

BACKGROUND: This study assesses the microbiologic effects of a two-phase antimicrobial periodontal therapy and tested microbiologic, clinical, and biologic markers as prognostic indicators for clinical success. METHODS: Eighty patients with chronic or aggressive periodontitis received periodontal treatment supplemented with 375 mg amoxicillin plus 500 mg metronidazole, three times daily for 7 days. In group A, antibiotics were given during the first non-surgical phase (T1); in group B, antibiotics were given during the second surgical phase (T2). Six microorganisms, group assignment, demographic and clinical variables, peak values of 15 cytokines, and nine acute-phase proteins in serum were evaluated as potential predictors of at least one site with probing depth (PD) >4 mm and bleeding on probing (BOP) at 12 months post-therapy. RESULTS: T1 decreased the counts of Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia (Pi), and Treponema denticola significantly more in group A than group B. Aggregatibacter actinomycetemcomitans and Parvimonas micra (Pm) showed a significant decrease only if the treatment was supplemented with antibiotics, i.e., T1 in group A, or T2 in group B. After T2, differences between groups were no longer significant. A multivariable model including four parameters revealed a predictive value of Pm (odds ratio [OR] = 4.38, P = 0.02) and Pi (OR = 3.44, P = 0.049) and yielded moderate accuracy for predicting the treatment outcome (area under the curve = 0.72). Host-derived factors and treatment sequence were not significantly associated with the outcome. CONCLUSIONS: Long-term microbiologic outcomes of periodontal therapy with adjunctive antibiotics either in T1 or T2 were similar. Detection of Pm before therapy was a predictor for persistence of sites with PD >4 mm and BOP at 12 months post-treatment.

Photo-activated elimination of Aggregatibacter actinomycetemcomitans in planktonic culture: Comparison of photodynamic therapy versus photothermal therapy method.

BACKGROUND: Periodontal pathogens are the main factors responsible for periodontal diseases and considering the limitations of conventional mechanical debridement, new treatment approaches are under investigation. This study was designed to evaluate and compare the antibacterial effects of two different systems of photodynamic and photothermal therapy on Aggregatibacter actinomycetemcomitans as the main pathogen involved in aggressive Periodontitis. METHODS: Cultures of Aggregatibacter actinomycetemcomitans were exposed to 662nm laser in presence of Radachlorin® photosensitizer (photodynamic group) or 810nm laser in presence of EmunDo® photosensitizer (photothermal group), then bacterial suspension of each well in the study groups were diluted and subcultured on the surface of Muller-Hinton agar plates. subsequently the number of colony forming units per milliliter of the wells were determined and checked by analysis of variance and Tukey test (p<0.05). RESULTS: Aggregatibacter actinomycetemcomitans suspensions showed significant reduction in both groups of photodynamic and photothermal therapy with no priority. CONCLUSION: Based on the results of this study, photodynamic and photothermal therapy can be proposed as a new promising approaches for bacterial elimination in periodontal diseases.

Monitoring gene expression of rcpA from Aggregatibacter actinomycetemcomitans versus antimicrobial photodynamic therapy by relative quantitative real-time PCR.

BACKGROUND: Aggregatibacter actinomycetemcomitans is an important pathogen that is frequently found in various infections, particularly aggressive periodontitis. In this study, we described the outcome of the expression level of A. actinomycetemcomitans virulence factor following treatment by antimicrobial photodynamic therapy (aPDT) with indocyanine green (ICG) as a photosensitizing agent. MATERIALS AND METHODS: To determine the aPDT effect on the cell-surviving assay and expression ratio of the rcpA gene in A. actinomycetemcomitans by a colony-forming unit and relative quantitative (q) real-time PCR (qRT-PCR) assays, respectively, the proper dosing of sub-lethal aPDT was specified. RESULTS: The results of the current study showed that ICG-mediated aPDT, using 250-1000µg/mL, showed a significant reduction in A. actinomycetemcomitans growth when compared to the control group (P<0.05). Also, a sub-lethal dose of aPDT against A. actinomycetemcomitans was 125µg/mL ICG, with a 30s diode laser irradiation time at fluency of 15.6J/cm2 that could reduce the expression of rcpA gene approximately 6-fold. DISCUSSION: aPDT with ICG could reduce the cell survival and the virulence agent of A. actinomycetemcomitans. Thus, use of the appropriate aPDT dosage can be used for the successful treatment of periodontitis in vivo.

Application of diverse natural polymers in the design of oral gels for the treatment of periodontal diseases.

The objective of this study was to prepare periodontal gels using natural polymers such as badam gum, karaya gum and chitosan. These gels were tested for their physical and biochemical properties and assessed for their antibacterial activity against Aggregatibacter actinomycetemcomitans and Streptococcus mutans, two pathogens associated with periodontal disease. Badam gum, karaya gum and chitosan were used to prepare gels of varying concentrations. Moxifloxacin hydrochloride, a known antimicrobial drug was choosen in the present study and it was added to the above gels. The gels were then run through a battery of tests in order to determine their physical properties such as pH and viscosity. Diffusion studies were carried out on the gels containing the drug. Antimicrobial testing of the gels against various bacteria was then carried out to determine the effectiveness of the gels against these pathogens. The results showed that natural polymers can be used to produce gels. These gels do not have inherent antimicrobial properties against A. actinomycetemcomitans and S. mutans. However, they can be used as a transport vehicle to carry and release antimicrobial drugs.

Antimicrobial efficiency of mouthrinses versus and in combination with different photodynamic therapies on periodontal pathogens in an experimental study.

BACKGROUND AND OBJECTIVE: In the therapy of destructive periodontal disease, chemical antimicrobial agents and increasingly photodynamic therapy (PDT) play an important adjunctive role to standard mechanical anti-infective treatment procedures. However, both antiseptic methods have their shortcomings in terms of eliminating periodontal pathogens. The aim of the study was to compare the antibacterial efficacy of different antiseptic mouthrinses, of a conventional and a new, modified PDTplus as well as of the different antiseptic mouthrinses combined with either the conventional or the modified PDTplus against periopathogens. MATERIAL AND METHODS: Six representative periodontitis-associated bacterial strains were grown for 24 h under anaerobic conditions. After mixing the individual cell pellets they were exposed to 10 different antiseptic mouthrinse formulations: chlorhexidine (0.2%, 0.06%, CHX); CHX + cetylpyridinium chloride (each 0.05%); sodium hypochlorite (0.05%); polyhexanide (0.04%, PHMB1; 0.1%, PHMB2); octenidine dihydrochloride (0.1%); fluoride (250 ppm); essential oils; povidone iodine (10%); and saline (0.9%, NaCl) as control. Furthermore, the bacteria were treated with conventional PDT based on light-emitting diodes and a new modified photodisinfection combining photosensitizer with hydrogen peroxide to PDTplus also based on light-emitting diodes. In addition to the single treatments, a combined application of antiseptic exposure followed by use of PDT or PDTplus was performed. The microbial viability was characterized by analyzing colony growth and fluorescence-based vitality proportions. RESULTS: Nearly all mouthrinses caused a statistically significant growth inhibition. The most effective antiseptics, CHX (0.2%), CHX/cetylpyridinium chloride and octenidine dihydrochloride, inhibited bacterial growth completely. Conventional PDT resulted in moderate reduction of colony growth. The modified PDTplus achieved maximum antimicrobial effect. The combination of antiseptic exposure and PDT against periopathogens predominantly increased antibacterial efficacy compared to the single applications. The mouthrinse containing essential oil seemed to interfere with PDT. CONCLUSION: A combination therapy of preceding chemotherapeutical exposure and subsequent photodisinfection may be a more effective and promising antibacterial treatment than single applications of the antiseptic methods. The modified PDTplus using oxygen-enriched toluidine showed a superior antibacterial effect on periodontal pathogens to conventional PDT and to the majority of the investigated mouthrinses.

Antimicrobial effects of citrus sinensis peel extracts against periodontopathic bacteria: an in vitro study.

BACKGROUND: Use of plant extracts and phytochemicals with known antimicrobial properties may have great significance in therapeutic treatments. OBJECTIVE: To assess the in vitro antimicrobial potential and also determine the minimum inhibitory concentration (MIC) of Citrus sinensis peel extracts with a view of searching a novel extract as a remedy for periodontal pathogens. MATERIALS AND METHODS: Aqueous and ethanol (cold and hot) extracts prepared from peel of Citrus sinensis were screened for in vitro antimicrobial activity against Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia, using agar well diffusion method. The lowest concentration of every extract considered as the minimal inhibitory concentration (MIC) values were determined for both test organisms. Confidence level and level of significance were set at 95% and 5% respectively. RESULTS: Prevotella intermedia and Porphyromonas gingivalis were resistant to aqueous extracts while Aggregatibacter actinomycetemcomitans was inhibited at very high cncentrations. Hot ethanolic extracts showed significantly higher zone of inhibition than cold ethanolic extract. Minimum inhibitory concentration of hot and cold ethanolic extracts of Citrus sinensis peel ranged between 12-15 mg/ml against all three periodontal pathogens. CONCLUSIONS: Both extracts were found sensitive and contain compounds with therapeutic potential. Nevertheless, clinical trials on the effect of these plants are essential before advocating large-scale therapy.

Essential Oil from Berries of Lebanese Juniperus excelsa M. Bieb Displays Similar Antibacterial Activity to Chlorhexidine but Higher Cytocompatibility with Human Oral Primary Cells.

Chlorhexidine (CHX), one of the most effective drugs administered for periodontal treatment, presents collateral effects including toxicity when used for prolonged periods; here, we have evaluated the bactericidal potency and the cytocompatibility of Juniperus excelsa M. Bieb essential oil (EO) in comparison with 0.05% CHX. The EO was extracted from berries by hydrodistillation and components identified by gas chromatography and mass spectrometry. Bacterial inhibition halo analysis, quantitative cell viability 2,3-bis(2-methoxy-4-nitro-5-sulphophenyl)-5-[(phenyl amino) carbonyl]-2H-tetrazolium hydroxide assay (XTT), and colony forming unit (CFU) count were evaluated against the two biofilm formers Aggregatibacter actinomycetemcomitans and Streptococcus mutans. Finally, cytocompatibility was assessed with human primary gingival fibroblasts (HGF) and mucosal keratinocytes (HK). The resulting EO was mainly composed of monoterpene hydrocarbons and oxygenated monoterpenes. An inhibition halo test demonstrated that both bacteria were sensitive to the EO; XTT analysis and CFU counts confirmed that 10-fold-diluted EO determined a statistically significant (p < 0.05) reduction in bacteria count and viability towards both biofilm and planktonic forms in a comparable manner to those obtained with CHX. Moreover, EO displayed higher cytocompatibility than CHX (p < 0.05). In conclusion, EO exhibited bactericidal activity similar to CHX, but a superior cytocompatibility, making it a promising antiseptic alternative to CHX.

Antimicrobial photodynamic effect to treat residual pockets in periodontal patients: a randomized controlled clinical trial.

AIM: A randomized controlled clinical trial was designed to evaluate the efficacy of the photodynamic therapy (PDT) in the treatment of residual pockets of chronic periodontitis patients. MATERIAL AND METHODS: Thirty-four patients with at least four residual periodontal pockets undergoing maintenance care were included and randomly assigned to test group (PDT, n = 18) or control group (sham procedure, n = 16). The intervention was performed at baseline, 3, 6 and 12 months. Clinical parameters such as pocket probing depth (PPD), clinical attachment level (CAL), bleeding on probing (BoP) and plaque index (PI) were measured before intervention and after 3, 6 and 12 months. Subgingival samples were obtained at baseline, and after 7 days, 3, 6 and 12 months to quantify Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia by real-time polimerase chain reaction (PCR). RESULTS: All clinical variables showed significant improvement during the study, but there was no significant difference between test and control groups. The microbiological analyses showed no differences between groups at any time during the study. CONCLUSION: Within the limits of this clinical trial and considering the laser and photosensitizer protocol used, PDT failed to demonstrate additional clinical and bacteriological benefits in residual pockets treatment.