Selective photoantisepsis.

BACKGROUND AND OBJECTIVE: Selective killing of pathogens by laser is possible due to the difference in absorption of photon energy by pathogens and host tissues. The optical properties of pathogenic microorganisms are used along with the known optical properties of soft tissues in calculations of the laser-induced thermal response of pathogen colonies embedded in a tissue model. The objective is to define the laser parameters that optimize pathogen destruction and depth of the bactericidal effect. MATERIALS AND METHODS: The virtual periodontium is a computational model of the optical and time-dependent thermal properties of infected periodontal tissues. The model simulates the periodontal procedure: Laser Sulcular Debridement.1 Virtual pathogen colonies are placed at different depths in the virtual periodontium to determine the depth for effective bactericidal effects given various laser parameters (wavelength, peak power, pulse duration, scan rate, fluence rate) and differences in pathogen sensitivities. RESULTS: Accumulated background heat from multiple passes increases the depth of the bactericidal effect. In visible and near-IR wavelengths the large difference in absorption between normal soft tissue and Porphyromonas gingivalis (Pg) and Prevotella intermedia (Pi) results in selective destruction. Diode laser (810 nm) efficacy and depth of the bactericidal effect are variable and dependent on hemin availability. Both pulsed-Nd:YAG and the 810 nm diode lasers achieve a 2-3 mm deep damage zone for pigmented Pg and Pi in soft tissue without surface damage (selective photoantisepsis). The model predicts no selectivity for the Er:YAG laser (2,940 nm). Depth of the bactericidal effect is highly dependent on pathogen absorption coefficient. Highly sensitive pathogens may be destroyed as deep as 5-6 mm in soft tissue. Short pulse durations enable confinement of the thermal event to the target. Temporal selectivity is achieved by adjusting pulse duration based on target size. CONCLUSION: The scatter-limited phototherapy model of the infected periodontium is applied to develop a proper dosimetry for selective photoantisepsis. Dosimetry planning is essential to the development of a new treatment modality. Lasers Surg. Med. 48:763-773, 2016. © 2016 Wiley Periodicals, Inc.

The Black Bug Myth: Selective photodestruction of pigmented pathogens.

BACKGROUND AND OBJECTIVE: It is commonly believed that pigmented pathogens are selectively targeted by dental lasers. To test this notion optical diffuse reflection spectroscopy (DRS) was used to obtain absorption spectra for the periodontal pathogens, Porphyromonas gingivalis (Pg) and Prevotella intermedia (Pi). MATERIALS AND METHODS: Spectra from 400 to 1,100 nm wavelengths of Pg colonies cultured with different concentrations of hemin were obtained to test the hypothesis that "visual pigmentation" predicts absorption of near-infrared (IR) dental laser energy. Ablation threshold at 1,064 nm [1] was measured for the pathogenic fungus, Candida albicans (Ca). RESULTS: The hypothesis was demonstrated to be true at 810 nm, it was false at 1,064 nm. Diode laser (810 nm) efficacy and "depth of kill" is dependent on hemin availability from 400 to about 900 nm. Pg and Pi absorption at 1,064 nm (µa = 7.7 ± 2.6 cm(-1) ) is independent of hemin availability but is determined by another unknown chromophore. Ca is non-pigmented but very sensitive to 1,064 nm irradiation. CONCLUSIONS: The amount of visual pigmentation does not necessarily predict sensitivity to dental laser irradiation. Spectra in visible and near-IR wavelengths demonstrate a large difference in absorption between soft tissue and Pg or Pi. This difference represents a host/pathogen differential sensitivity to laser irradiation, the basis for selective photoantisepsis. Lasers Surg. Med. 48:706-714, 2016. © 2016 Wiley Periodicals, Inc.

Phototargeting human periodontal pathogens in vivo.

The effects of blue light at 455 nm were investigated on the bacterial composition of human dental plaque in vivo. Eleven subjects who refrained from brushing for 3 days before and during phototherapy participated in the study. Light with a power density of 70 mW/cm(2) was applied to the buccal surfaces of premolar and molar teeth on one side of the mouth twice daily for 2 min over a period of 4 days. Dental plaque was harvested at baseline and again at the end of 4 days from eight posterior teeth on both the exposed side and unexposed sides of the mouth. Microbiological changes were monitored by checkerboard DNA probe analysis of 40 periodontal bacteria. The proportions of black-pigmented species Porphyromonas gingivalis and Prevotella intermedia were significantly reduced on the exposed side from their original proportions by 25 and 56 %, respectively, while no change was observed to the unexposed side. Five other species showed the greatest proportional reduction of the light-exposed side relative to the unexposed side. These species were Streptococcus intermedius, Fusobacterium nucleatum ss. vincentii, Fusobacterium nucleatum ss. polymorphum, Fusobacterium periodonticum, and Capnocytophaga sputigena. At the same time, the percentage of gingival areas scored as being red decreased on the side exposed to light from 48 to 42 %, whereas the percentage scored as red increased on the unexposed side from 53 to 56 %. No adverse effects were found or reported in this study. The present study proposes a new method to modify the ecosystem in dental plaque by phototherapy and introduces a new avenue of prophylactic treatment for periodontal diseases.

Photobleaching of red fluorescence in oral biofilms.

BACKGROUND AND OBJECTIVE: Many species of oral bacteria can be induced to fluoresce due to the presence of endogenous porphyrins, a phenomenon that can be utilized to visualize and quantify dental plaque in the laboratory or clinical setting. However, an inevitable consequence of fluorescence is photobleaching, and the effects of this on longitudinal, quantitative analysis of dental plaque have yet to be ascertained. MATERIAL AND METHODS: Filter membrane biofilms were grown from salivary inocula or single species (Prevotella nigrescens and Prevotella intermedia). The mature biofilms were then examined in a custom-made lighting rig comprising 405 nm light-emitting diodes capable of delivering 220 W/m(2) at the sample, an appropriate filter and a digital camera; a set-up analogous to quantitative light-induced fluorescence digital. Longitudinal sets of images were captured and processed to assess the degradation in red fluorescence over time. RESULTS: Photobleaching was observed in all instances. The highest rates of photobleaching were observed immediately after initiation of illumination, specifically during the first minute. Relative rates of photobleaching during the first minute of exposure were 19.17, 13.72 and 3.43 arbitrary units/min for P. nigrescens biofilms, microcosm biofilm and P. intermedia biofilms, respectively. CONCLUSION: Photobleaching could be problematic when making quantitative measurements of porphyrin fluorescence in situ. Reducing both light levels and exposure time, in combination with increased camera sensitivity, should be the default approach when undertaking analyses by quantitative light-induced fluorescence digital.

Bactericidal effect of Er,Cr:YSGG laser irradiation on endodontic biofilm: An ex vivo study.

AIM: This ex vivo study aimed to evaluate the of Er,Cr:YSGG laser effectiveness in the decontamination of an endodontic biofilm. MATERIALS AND METHODS: Seventy-three single rooted human teeth, freshly were chosen. Each tooth was exposed to four associated species in an endodontic biofilm (Enterococcus faecalis, Streptococcus salivarius, Porphyromonas gingivalis, and Prevotella intermedia) and randomly allocated to one of the seven experimental groups. The group 1 (7 teeth) was used to finalize the reliable biofilm-forming technique. The groups 2 and 3 (15 teeth each group) were irradiated with two different Er;Cr:YSGG laser settings (0,75 W - 40 Hz and 4 W - 40 Hz, respectively). The groups 4 and 5 (15 teeth each group) were irrigated with two different solutions and laser irradiated with the same settings (1,5 W - 15 Hz). The group 6 (6 teeth) was the control group treated only with 4 ml 2,5% NaOCl irrigation during 60 s. RESULTS: The observations of group 2 and 3 specimens showed the ripeness of the biofilm with the presence of Enterococcus faecalis and Streptococcus salivarius in chains but in group 3 thermal edge effects produced by the optic fiber in the canal walls were present. The group 4 specimens observation showed an average cleaning of the root canal walls while on the canal walls of group 5 samples the apical third presented several debris and smear layer and in the centre cracks and melting dentin of the radicular wall were observed. CONCLUSION: In those experimental conditions, this study, demonstrated that Er,Cr:YSGG laser has a canals decontamination ability when associated to NaOCl irrigation.

Photodynamic therapy in periodontal therapy: microbiological observations from a private practice.

In recent years, the combination of laser light and photosensitizer known as photodynamic therapy (PDT) has been used in periodontal therapy. However, there are not enough clinical studies to fully evaluate the effects of PDT on the periodontal tissues. This microbiological study examined the effects of PDT on the periodontal bacteria in combination with scaling and root planing (SRP) in the same group of patients by randomly selecting PDT or SRP for use in different quadrants of the mouth. For the present study, PDT was compared with a diode laser (980 nm) and an Nd:YA G laser (1,064 nm). Microbiological samples were examined and evaluated over a period of three months. Significant bacterial reduction has been observed in all cases. The diode laser with SRP presented long-term positive results, while PDT showed a significant bacteria reduction during the entire observation period.

The bactericidal effect of a Genius Nd:YAG laser.

PURPOSE: To evaluate the 'in vitro' bactericidal effect of the Nd:YAG laser (Genius, MØlsgaard Dental, Copenhagen, Denmark) on six periodontal pathogens. METHODS: Suspensions of six different periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Fusobacterium nucleatum and Parvimonas micra) were prepared in small Eppendorff tubes, and exposed to a Nd:YAG laser for five different periods of time. Laser settings used: Power 6 Watt (on a scale of 1-12 W), Frequency 50 Hz, Pulse duration 250 mus. After exposure to the laser, aliquots of the suspensions were spread on blood agar plates for bacterial counting. RESULTS: After 5 s of laser exposure, there was a decrease in total colony forming units for all six selected microorganisms. After 15, 30 and 45 s, no viable bacterial cells could be retrieved. CONCLUSION: In this 'in vitro' model, 15 s of Nd:YAG laser use was found to be effective for total killing of the six tested periodontal pathogens.

The Effects of Ultraviolet Light-Emitting Diodes with Different Wavelengths on Periodontopathic Bacteria In Vitro.

Objective: The aim of this study was to examine effects of recently developed ultraviolet light-emitting diodes (UV LEDs) wavelengths on in vitro growth and gene expression of cultural periodontopathic bacteria, and on viability of experimental gingival fibroblasts. Materials and methods: Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, and Streptococcus oralis were irradiated by UV LEDs (265, 285, 310, 365, and 448 nm) at 600 mJ/cm2 and grown anaerobically in vitro. The colony forming units were counted after 1 week. Cell morphology was observed using a scanning electron microscope (SEM). Quantitative real-time polymerase chain reaction was performed to investigate gene expression changes by 310 nm irradiation. Viability of the irradiated human gingival fibroblasts was evaluated using WST-8 assay. Results: Both 265 and 285 nm resulted in the complete death of bacteria and fibroblasts, whereas 310 nm caused partial killing and suppression of bacterial growth and much less damage to the fibroblasts in vitro. Both 365 and 448 nm resulted in no significant change. SEM showed that P. gingivalis cells gradually degraded from day 2 or 3 and were severely destructed on day 5 for 265, 285, and 310 nm. The 310 nm irradiation transiently suppressed the transcripts of SOS response- and cell division-relative genes. Conclusions: Both 265 and 285 nm may induce powerful bactericidal effects and severe fibroblast phototoxicity, and 310 nm may induce partial killing or growth suppression of bacterial cells with much less fibroblast phototoxicity. UV lights may have potential for bacterial suppression, with situations dependent on wavelength, in periodontal and peri-implant therapy.

Chair-side detection of Prevotella Intermedia in mature dental plaque by its fluorescence.

BACKGROUND: Prevotella intermedia/nigrescens is one of the well-known pathogens causing periodontal diseases, and the red florescence excited by the visible blue light caused by the protoporphyrin IX in the bacterial cells could be useful for the chair-side detection. The aim of this study was to evaluated levels of periodontal pathogen, especially P. intermedia in clinical samples of red fluorescent dental plaque. METHODS: Thirty two supra gingival plaque samples from six individuals were measured its fluorescence at 640nm wavelength excited by 409nm. Periodontopathic bacteria were counted by the Invader PLUS PCR assay. Co-relations the fluorescence intensity and bacterial counts were analyzed by Person's correlation coefficient and simple and multiple regression analysis. Positive and negative predictive values of the fluorescence intensities for with or without P. intermedia in supragingival plaque was calculated. RESULTS: When relative fluorescence unit (RFU) were logarithmic transformed, statistically significant linear relations between RFU and bacterial counts were obtained for P. intermedia, Porphyromonas gingivalis and Tannerella forsythia. By the multiple regression analysis, only P. intermedia had statistically significant co-relation with fluorescence intensities. All of the fluorescent dental plaque contained P. intermedia m. In contrast, 28% of non-fluorescent plaques contained P. intermedia. CONCLUSION: To check the fluorescence dental plaque in the oral cavity could be the simple chair-side screening of the mature dental plaque before examining the periodontal pathogens especially P. intermedia by the PCR method.

Comparison of Riboflavin and Toluidine Blue O as Photosensitizers for Photoactivated Disinfection on Endodontic and Periodontal Pathogens In Vitro.

Photoactivated disinfection has a strong local antimicrobial effect. In the field of dentistry it is an emerging adjunct to mechanical debridement during endodontic and periodontal treatment. In the present study, we investigate the effect of photoactivated disinfection using riboflavin as a photosensitizer and blue LED light for activation, and compare it to photoactivated disinfection with the widely used combination of toluidine blue O and red light. Riboflavin is highly biocompatible and can be activated with LED lamps at hand in the dental office. To date, no reports are available on the antimicrobial effect of photoactivated disinfection using riboflavin/blue light on oral microorganisms. Planktonic cultures of eight organisms frequently isolated from periodontal and/or endodontic lesions (Aggregatibacter actinomycetemcomitans, Candida albicans, Enterococcus faecalis, Escherischia coli, Lactobacillus paracasei, Porphyromonas gingivalis, Prevotella intermedia and Propionibacterium acnes) were subjected to photoactivated disinfection with riboflavin/blue light and toluidine blue O/red light, and survival rates were determined by CFU counts. Within the limited irradiation time of one minute, photoactivated disinfection with riboflavin/blue light only resulted in minor reductions in CFU counts, whereas full kills were achieved for all organisms when using toluidine blue O/red light. The black pigmented anaerobes P. gingivalis and P. intermedia were eradicated completely by riboflavin/blue light, but also by blue light treatment alone, suggesting that endogenous chromophores acted as photosensitizers in these bacteria. On the basis of our results, riboflavin cannot be recommended as a photosensitizer used for photoactivated disinfection of periodontal or endodontic infections.

Comparative evaluation of antibacterial effects of Nd:YAG laser irradiation in root canals and dentinal tubules.

The antibacterial effects of the Nd:YAG laser on contaminated root canals and dentinal tubules were observed as the aim of this study. The samples were inoculated with Streptococcus sanguis (NCTC 7853) and Prevotella intermedia (NCTC 93336), and the effects of Nd:YAG laser were tested on these teeth. The specimens were lased with 1.8 W and 2.4 W Nd:YAG laser for 30 s, and the presence of bacteria in tubules was observed under light microscopy. The 1.8 W laser sterilized the tubules in 86.3% of sections inoculated with S. sanguis, whereas 2.4 W laser sterilized in 98.5% of the sections. Both laser powers sterilized all samples inoculated with P. intermedia. The scanning electron microscopic observations supported the light microscopic findings.

Clinical and microbiologic follow-up evaluations after non-surgical periodontal treatment with erbium:YAG laser and scaling and root planing.

BACKGROUND: This study compared erbium-doped: yttrium, aluminum, and garnet (Er:YAG) laser irradiation (100 mJ/pulse; 10 Hz; 12.9 J/cm(2)) with or without conventional scaling and root planing (SRP) to SRP only for treatment of periodontal pockets. METHODS: Nineteen patients with pockets from 5 to 9 mm were included. In a split-mouth design, each site was allocated to a treatment group: 1) SRPL, SRP and laser; 2) L, laser; 3) SRP, SRP only; and 4) C, no treatment. Clinical parameters of probing depth (PD), gingival recession, and clinical attachment level (CAL) were evaluated at baseline and 1, 3, 6, and 12 months after treatment. Visible plaque index, gingival bleeding index (GI), bleeding on probing (BOP), and subgingival plaque samples were also measured 12 days postoperatively, in addition to the above mentioned months. Intergroup and intragroup statistical analyses were performed (P <0.05). RESULTS: GI decreased for SRPL and increased for L, SRP, and C (P <0.05) 12 days postoperatively and decreased for SRPL and SRP (P <0.05) 3, 6, and 12 months after baseline; BOP and PD decreased for all treated groups (P <0.01) 3, 6, and 12 months after treatment. CAL gain was significant for SRPL, L, and SRP (P <0.05) 3, 6, and 12 months postoperatively. SRPL and L presented a significant reduction in the percentage of sites with bacteria 6 and 12 months after treatment (P <0.05). CONCLUSION: Non-surgical periodontal treatment with Er:YAG laser may be an alternative treatment for reduction and control of the proliferation of microorganisms in persistent periodontitis.

Phototargeting oral black-pigmented bacteria.

We have found that broadband light (380 to 520 nm) rapidly and selectively kills oral black-pigmented bacteria (BPB) in pure cultures and in dental plaque samples obtained from human subjects with chronic periodontitis. We hypothesize that this killing effect is a result of light excitation of their endogenous porphyrins. Cultures of Prevotella intermedia and P. nigrescens were killed by 4.2 J/cm2, whereas P. melaninogenica required 21 J/cm2. Exposure to light with a fluence of 42 J/cm2 produced 99% killing of P. gingivalis. High-performance liquid chromatography demonstrated the presence of various amounts of different porphyrin molecules in BPB. The amounts of endogenous porphyrin in BPB were 267 (P. intermedia), 47 (P. nigrescens), 41 (P. melaninogenica), and 2.2 (P. gingivalis) ng/mg. Analysis of bacteria in dental plaque samples by DNA-DNA hybridization for 40 taxa before and after phototherapy showed that the growth of the four BPB was decreased by 2 and 3 times after irradiation at energy fluences of 4.2 and 21 J/cm2, respectively, whereas the growth of the remaining 36 microorganisms was decreased by 1.5 times at both energy fluences. The present study suggests that intraoral light exposure may be used to control BPB growth and possibly benefit patients with periodontal disease.

Bacterial reduction in periodontal pockets through irradiation with a diode laser: a pilot study.

This study examines the application of a diode laser with a wavelength of 805 nm for periodontal treatment. While the use of the diode laser in this field has not been investigated so far, several authors have reported on the use of neodymium:yttrium-aluminum-garnet (Nd:YAG) laser for such applications. The aim of this study was to examine the immediate effect of the diode laser in reducing the bacterial concentration in periodontal pockets. Important periodontal indices (PBI, CPITN) were assessed in 50 patients to obtain initial values for a planned long-term study and to select appropriate periodontal pockets for this study. The periodontal pockets were required to have a minimum depth of 4 mm. Only proximal pockets were included in this study. The patients were subdivided into two groups. After microbiological samples had been collected with sterile paper tips, the group selected for laser treatment was subjected to scaling. One week after scaling, the patients underwent laser treatment. One week later, a second series of microbiological samples were obtained and the patients were subjected again to scaling; this time, however, they did not undergo laser treatment after 1 week. Two weeks after scaling, another series of microbiological samples was collected. The microbiological samples were evaluated to verify bacterial elimination from the periodontal pockets. A comparison between the initial and the final bacterial counts revealed that irradiation with the diode laser facilitates considerable bacterial elimination, especially of Actinobacillus actinomycetemcomitans, from periodontal pockets.

Treatment of periodontal pockets with a diode laser.

BACKGROUND AND OBJECTIVE: The aim of this study is to examine the long-term effect of diode laser therapy on periodontal pockets with regard to its bactericidal abilities and the improvement of periodontal condition. STUDY DESIGN/MATERIALS AND METHODS: Fifty patients were randomly subdivided into two groups (laser-group and control-group) and microbiologic samples were collected. There have been six appointments for 6 months following an exact treatment scheme. After evaluating periodontal indices (bleeding on probing, Quigley-Hein) including pocket depths and instruction of patients in oral hygiene and scaling therapy of all patients, the deepest pockets of each quadrant of the laser-group's patients were microbiologically examined. Afterwards, all teeth were treated with the diode laser. The control-group received the same treatment but instead of laser therapy were rinsed with H2O2. Each appointment also included a hygienic check-up. After 6 months the final values of the periodontal indices and further microbiologic samples were measured. The total bacterial count as well as specific bacteria, such as Actinobacillus actinomycetemcomitans, Prevotella intermedia, and Porphyromonas gingivalis, were assessed semiquantitatively. RESULTS: The bacterial reduction with diode laser therapy was significantly better than in the control group. The index of bleeding on probing improved in 96.9% in the laser-group, whereas only 66.7% in the control group. Pocket depths could be more reduced in the laser group than in the control group. CONCLUSION: The diode laser reveals a bactericidal effect and helps to reduce inflammation in the periodontal pockets in addition to scaling. The diode laser therapy, in combination with scaling, supports healing of the periodontal pockets through eliminating bacteria.

Elimination of bacteria on different implant surfaces through photosensitization and soft laser. An in vitro study.

Microbiologic examinations of implants have shown that certain microorganisms described as periodontal pathogens may have an influence on the development and the progression of peri-implant disease. This experimental study aimed to examine the bactericidal effect of irradiation with a soft laser on bacteria associated with peri-implantitis following exposure to a photosensitizing substance. Platelets made of commercially pure titanium, either with a machined surface or with a hydroxyapatite or plasma-flame-sprayed surface or with a corundum-blasted and etched surface, were incubated with a pure suspension of Actinobacillus actinomycetemcomitans or Porphyromonas gingivalis or Prevotella intermedia. The surfaces were then treated with a toluidine blue solution and irradiated with a diode soft laser with a wave length of 905 nm for 1 min. None of the smears obtained from the thus treated surfaces showed bacterial growth, whereas the smears obtained from surfaces that had been subjected to only one type of treatment showed unchanged growth of every target organism tested (P < 0.0006). Electron microscopic inspection of the thus treated platelets revealed that combined dye/laser treatment resulted in the destruction of bacterial cells. The present in vitro results indicate that lethal photosensitization may be of use for treatment of peri-implantitis.