BACKGROUND: Aggregatibacter actinomycetemcomitan (A.a) and Actinomyces naeslundii (A.n) are two gram-negative chromogenic bacteria involved in the formation of dental black stainings. Our study aimed to investigate the antibacterial effect of photodynamic therapy (aPDT) using two photosensitizers, Methylene Blue (MB) and Indocyanine Green (ICG).
METHODS: In this in-vitro study, two isolates of each selected bacterium were cultured and treated as follows; Negative control with no treatment; CHX as a positive control; ICG; MB; ICG with 808 nm laser activation; and MB with 660 nm laser activation. The number of colonies (CFU/mL) was determined to compare the groups. The qualitative evaluation of biofilm formation was done by scanning electron microscopy of treated enamel pieces. The logarithmic values of the colony counts were compared using One-way ANOVA and the Welch test Tukey HSD and Games-Howell tests were used for multiple comparisons. P-values of less than 0.05 were considered statistically significant.
RESULTS: The use of ICG alone or along with laser irradiation at the wavelength of 808 nm significantly reduced the number of colonies of A.a and A.n bacteria. Comparing the colony counts in the MB group with the positive control showed no significant decrease in bacterial load. On the contrary, activation of MB with 660 nm radiation of diode laser showed a significant antibacterial effect. The density of bacterial biofilm was significantly lower in the groups treated with MB and ICG without laser activation than in the control group; however, the reduction in bacteria biofilm density was more robust using photodynamic therapy with ICG.
CONCLUSIONS: aPDT using MB with 660 nm laser and ICG with 808 nm laser significantly reduced the number of chromogenic A.a and A.n bacteria, and photodynamic therapy with ICG was proven to be significantly more effective than MB with or without laser radiation.

Antimicrobial Photodynamic Therapy (aPDT) based on the action of visible light and photosensitizers has emerged as a promising microbial reduction and alternative to antibiotics resistance to cariogenic pathogens. The present research aims to evaluate the antimicrobial effect of aPDT mediated by a new photosensitizer (amino acid porphyrin conjugate 4i) on Streptococcus mutans (S. mutans) biofilm. Qualitative morphologic characteristics of S. mutans biofilms are shown by scanning electron microscopy (SEM). The colony plate counting method is used to measure the dark toxicity and the phototoxicity of different concentrations of 4i-aPDT to S. mutans biofilms. MTT assay is conducted to investigate the effect of 4i mediated aPDT on the metabolic activity of S. mutans biofilm. Changes in structure morphology, bacterial density and extracellular matrix of S. mutans biofilm are observed by SEM. The distribution of living and dead bacteria in biofilm is detected using Confocal laser microscopy (CLSM). The results indicate that single laser irradiation has no antibacterial effect on S. mutans biofilms. With the increase of 4i concentration or the prolongation of laser irradiation time, the antibacterial effect of 4i-mediated aPDT on S. mutans biofilm is more statistically significant compared to the control. When the concentration of 62.5 µmol/L 4i is continuously illuminated for 10 min, the logarithm of the colonies in the biofilm shows a reduction of 3.4 log10. MTT assay detected absorbance values of biofilm by 4i-mediated aPDT are the lowest, indicating a significant decrease in biofilm metabolic activity. SEM analysis shows that 4i mediated aPDT reduced the quantity and density of S. mutans. A dense red fluorescence image of the 4i-aPDT treated biofilm is observed under CLSM, indicating that the dead bacteria are widely distributed.

Until now, in clinical dentistry, antibacterial photodynamic therapy (aPDT) has been restricted to in-office treatments, which hampers repeated applications. This pilot study tested the benefit of a commercially available Lumoral® device designed for regular periodontal dual-light aPDT treatment at home. Seven patients with peri-implant disease applied dual-light aPDT daily in addition to their normal dental hygiene for four weeks. A single Lumoral® treatment includes an indocyanine green mouth rinse followed by 40 J/cm2 radiant exposure to a combination of 810 nm and 405 nm light. A point-of-care analysis of active-matrix metalloproteinase (aMMP-8), visible plaque index (VPI), bleeding on probing (BOP), and peri-implant pocket depth (PPD) measurements was performed on day 0, day 15, and day 30. Reductions in aMMP-8 (p = 0.047), VPI (p = 0.03), and BOP (p = 0.03) were observed, and PPD was measured as being 1 mm lower in the implant (p = ns). These results suggest a benefit of regular application of dual-light aPDT in peri-implantitis. Frequently repeated application can be a promising approach to diminishing the microbial burden and to lowering the tissue destructive proteolytic and inflammatory load around dental implants. Further studies in larger populations are warranted to show the long-term benefits.

OBJECTIVE: The aim of this pilot study was to evaluate the clinical and microbiological outcomes of light-activated disinfection (LAD) alone or combined with probiotics as an adjunct to non-surgical periodontal treatment.
METHODS: In this single-blinded, randomized, controlled clinical pilot study, 48 patients (28 females and 20 males) with untreated periodontitis (stages II and III, grade B) were included. Using a parallel-group design, patients were randomly assigned into 3 groups to receive subgingival debridement (SD) alone (group 1, n = 16), SD with LAD (group 2, n = 16), or SD with LAD plus probiotic treatment (group 3, n = 16). Probing pocket depth (PPD), clinical attachment level (CAL), bleeding on probing (BOP), gingiva-index simplified (GIs), plaque-control record (PCR), and subgingival microbiological samples were analyzed at baseline, 3 months, and 6 months of follow-up.
RESULTS: All treatment modalities demonstrated clinical improvements in PPD and CAL at 6 months compared to baseline but without a statistical significant difference between the groups. The combination of SD + LAD + probiotic treatment (group 3) demonstrated significantly greater reductions in BOP, GIs, and red complex bacteria P. gingivalis and T. forsythia compared with other groups at 6 months (p < 0.05).
CONCLUSIONS: A single application of LAD as an adjunct to SD provided no additional clinical and microbiological benefits compared to SD alone. The combination of SD + LAD + probiotic treatment in group 3 led to further improvements of the inflammatory parameters.
CONCLUSIONS: The additional use of probiotics in periodontal treatment can be a useful approach to support inflammation and infection control of periodontal tissues. Further studies are necessary to determine the extent of added benefit for this treatment approach.

We investigated the influence of femtosecond laser irradiation on the growth of the two most common infectious bacterial pathogens in wounds; Staphylococcus aureus and Pseudomonas aeruginosa as an attempt to validate optimum parameters for a laser-based bactericidal modality to be used clinically. Bacterial cultures were exposed to femtosecond laser irradiation at different wavelengths, exposure times, and laser powers. The source of femtosecond laser was INSPIRE HF100 laser system, Spectra-Physics, which is pumped by a mode-locked femtosecond Ti: sapphire laser MAI TAI HP, Spectra-Physics. After irradiation, bacterial cells\' survival was monitored by observing the clear zones of inhibition in cultured agar plates. Results for all strains indicated that the exposure to femtosecond laser irradiation with a wavelength ranging from ultraviolet (λ > 350 nm) to blue laser light (λ < 480 nm), for a period above 20 min and with a power density of ≈ 0.063 W/cm2, was enough to inhibit both bacterial pathogens with the results maintained for 1 week following irradiation.

BACKGROUND: At present, very limited data are available on the clinical and microbiological outcomes obtained following repeated application of aPDT following one single mechanical debridement.
OBJECTIVE: To evaluate clinically and microbiologically the outcomes following one single session of subgingival mechanical debridement (scaling and root planing; e.g. SRP) followed by 1x immediate application of aPDT and 2 x subsequent use of aPDT without SRP.
METHODS: Forty patients diagnosed with generalized chronic periodontitis that were enrolled in periodontal maintenance (supportive periodontal therapy) program, were randomly assigned to one of the two treatments: 1. SRP by means of ultrasonic and hand instruments followed by one single session of SRP followed by 1x immediate application of aPDT and 2 x subsequent applications of aPDT without SRP (test) or 2. SRP alone (control). The following clinical parameters were recorded at baseline, at 3 and 6 months: Full-Mouth Plaque Scores (FMPS), Full-Mouth Bleeding Scores (BOP), Probing Pocket Depth (PPD), Clinical Attachment Level (CAL) and Gingival Recession (RC). Additionally, microbiological samples were evaluated at baseline and six months after treatment. The primary outcome variable was BOP.
RESULTS: Both treatments improved statistically significantly (p < 0.05) the FMPS, PPD and CAL values, while no statistically significant changes occurred in terms of RC. In the test group, BOP decreased statistically significantly (p < 0.05) after 3 and 6 months, while in the control group the respective values decreased statistically significantly only at 3 months. Both treatments reduced statistically significantly the total bacteria counts (TBC) after 6 months (p < 0.05). At 6 months, the use of SRP and aPDT resulted in a statistically significant decrease in the number of all tested bacteria except A. actinomycetemcomitans while the use of SRP alone resulted only in a statistically significant decrease in the numbers of P. gingivalis, T. denticola and T. forsythia.
CONCLUSIONS: In periodontal patients enrolled in a maintenance program one single session of SRP followed by 3x application of aPDT, enhanced the clinical and microbiological outcomes compared to SRP alone.