In Vitro Effect of Er:YAG Laser on Different Single and Mixed Microorganisms Being Associated with Endodontic Infections.

Objective: The purpose of this in vitro study was to evaluate the antimicrobial effect of activated irrigation with different modes of erbium-doped yttrium aluminum garnet (Er:YAG) laser application on microorganisms related to secondary endodontic infection. Background: Er:YAG laser has been recommended as an adjuvant tool for root canal disinfection during endodontic treatment. Materials and methods: Laser-activated irrigation (LAI) with 300 or 600 µm tips were tested with or without intermittent irrigation with 0.9% sodium chloride (NaCl) solution against different microorganisms (five single strains and dual species (Streptococcus gordonii combined with Actinomyces oris or Fusobacterium nucleatum) in root canals after 3 days of incubation. In a 21-day infection model, LAI was used together with intermittent rinsing with sodium hypochlorite (NaOCl) against the dual-species mixtures; here the incidence of microbial regrowth after up to 7 days was monitored. Results: In the 3-day root infection model, LAI protocols did not show any significant reduction of the microbial load when compared with manual irrigation with saline solution. In the 21-day infection, S. gordonii combined with A. oris were not detectable anymore after applying the LAI protocol with a 600 µm tip (30 mJ/10 pps) up to 7 days after treatment. Conclusions: Application of LAI with a 600 µm tip by using an Er:YAG laser might be advantageous in treatment of endodontic infections.

Effect of ultraviolet treatment on bacterial attachment and osteogenic activity to alkali-treated titanium with nanonetwork structures.

PURPOSE: Alkali-treated titanium with nanonetwork structures (TNS) possesses good osteogenic activity; however, the resistance of this material to bacterial contamination remains inadequate. As such, TNS implants are prone to postoperative infection. In this work, we attempted to alter the biological properties of TNS by treatment with short-duration high-intensity ultraviolet (UV) irradiation. METHODS: TNS discs were treated with UV light (wavelength =254 nm, strength =100 mW/cm2) for 15 minutes using a UV-irradiation machine. We carried out a surface characterization and evaluated the discs for bacterial film formation, protein adsorption, and osteogenic features. RESULTS: The superhydrophilicity and surface hydrocarbon elimination exhibited by the treated material (UV-treated titanium with a nanonetwork structure [UV-TNS]) revealed that this treatment effectively changed the surface characteristics of TNS. Notably, UV-TNS also showed reduced colonization by Actinomyces oris during an initial attachment period and inhibition of biofilm formation for up to 6 hours. Moreover, compared to conventional TNS, UV-TNS showed superior osteogenic activity as indicated by increased levels of adhesion, proliferation, alkaline phosphatase activity, osteogenic factor production, and osteogenesis-related gene expression by rat bone marrow mesenchymal stem cells (rBMMSCs). This inverse relationship between bacterial attachment and cell adhesion could be due to the presence of electron-hole pairs induced by high-intensity UV treatment. CONCLUSION: We suggest that simple UV treatment has great clinical potential for TNS implants, as it promotes the osseointegration of the TNS while reducing bacterial contamination, and can be conducted chair-side immediately prior to implantation.

Characterization of oral bacterial diversity of irradiated patients by high-throughput sequencing.

The objective of this study was to investigate the compositional profiles and microbial shifts of oral microbiota during head-and-neck radiotherapy. Bioinformatic analysis based on 16S rRNA gene pyrosequencing was performed to assess the diversity and variation of oral microbiota of irradiated patients. Eight patients with head and neck cancers were involved in this study. For each patient, supragingival plaque samples were collected at seven time points before and during radiotherapy. A total of 147,232 qualified sequences were obtained through pyrosequencing and bioinformatic analysis, representing 3,460 species level operational taxonomic units (OTUs) and 140 genus level taxa. Temporal variations were observed across different time points and supported by cluster analysis based on weighted UniFrac metrics. Moreover, the low evenness of oral microbial communities in relative abundance was revealed by Lorenz curves. This study contributed to a better understanding of the detailed characterization of oral bacterial diversity of irradiated patients.

In vitro bactericidal effect of Nd:YAG laser on Actinomyces israelii.

A bactericidal effect has been reported by the use of near-infrared laser light on both Gram-positive and Gram-negative bacteria. The aim of this study was to evaluate the effect of Nd:YAG laser on Actinomyces israelii, filamentous bacteria causing cervicofacial actinomycosis. Experiments were realized on bacterial cells in saline suspension or streaked on Mueller-Hinton (MH) agar plates with or without India ink. Laser application was performed in Eppendorf tubes with different powers and frequencies for 40 s; bacterial suspensions were then streaked on agar plates and incubated at 35 °C in proper conditions for 5 days before colony enumeration. A reduction of colony number variable from 60.13 to 100 % for powers of 2, 4, and 6 W at 25-50 Hz of frequency was observed in comparison with growth control. For agar plates, laser application was performed with different powers at 50 Hz for 60 s. A growth inhibition was observed after 5 days of incubation on MH plates with powers of 6 W and on MH-ink plates with all applied powers. This preliminary study showed a bactericidal effect caused by Nd:YAG laser application worthy to be evaluated in further experiments in vivo.

Potential of shock waves to remove calculus and biofilm.

Effective calculus and biofilm removal is essential to treat periodontitis. Sonic and ultrasonic technologies are used in several scaler applications. This was the first feasibility study to assess the potential of a shock wave device to remove calculus and biofilms and to kill bacteria. Ten extracted teeth with visible subgingival calculus were treated with either shock waves for 1 min at an energy output of 0.4 mJ/mm(2) at 3 Hz or a magnetostrictive ultrasonic scaler at medium power setting for 1 min, which served as a control. Calculus was determined before and after treatment planimetrically using a custom-made software using a grey scale threshold. In a second experiment, multispecies biofilms were formed on saliva-preconditioned bovine enamel discs during 64.5 h. They were subsequently treated with shock waves or the ultrasonic scaler (N = 6/group) using identical settings. Biofilm detachment and bactericidal effects were then assessed. Limited efficiency of the shock wave therapy in terms of calculus removal was observed: only 5% of the calculus was removed as compared to 100% when ultrasound was used (P ≤ 0.0001). However, shock waves were able to significantly reduce adherent bacteria by three orders of magnitude (P ≤ 0.0001). The extent of biofilm removal by the ultrasonic device was statistically similar. Only limited bactericidal effects were observed using both methods. Within the limitations of this preliminary study, the shock wave device was not able to reliably remove calculus but had the potential to remove biofilms by three log steps. To increase the efficacy, technical improvements are still required. This novel noninvasive intervention, however, merits further investigation.

Effects of low-energy shock waves on oral bacteria.

We have recently demonstrated that extracorporeal shock-wave therapy (ESWT) is effective in promoting the healing of dermal wounds and in regenerating alveolar bone lost through periodontal disease. The objective of the present study was to determine any antibacterial effect of ESWT on oral bacteria. Monoculture suspensions of 6 bacterial species were treated with 100 to 500 pulses of ESWT at energy flux densities (EFD) of 0.12 mJ/mm(2), 0.22 mJ/mm(2), and 0.3 mJ/mm(2). Following treatment, aliquots were plated for viability determination and compared with untreated controls. ESWT showed a significant microbicidal effect for Streptococcus mutans and an unencapsulated strain of Porphyromonas gingivalis following as few as 100 pulses at 0.3 mJ/mm(2) (p 0.05). These findings suggest that low-energy ESWT may be bactericidal for selected oral bacteria.

Wound healing of periodontal pockets using the diode laser: an interview.

Laser technology appears to have a promising future in dentistry While these devices are still in their infancy it is important to understand the interaction of laser energy on the intraoral structures of the patient, and to evaluate its effect on bacteria responsible for periodontal disease. The following is an interview that relates the details of one important investigation into tissue response as conducted by the Department of Periodontics and Endodontics, SUNY at Buffalo School of Dental Medicine.

Bactericidal effect of Nd:YAG laser irradiation on some endodontic pathogens ex vivo.

AIM: To define the role of neodymium:yttrium-aluminum-garnet (Nd:YAG) lasers in root canal disinfection along with a minimally invasive treatment concept. METHODOLOGY: The hypothesis was tested ex vivo that Nd:YAG laser irradiation has a bactericidal effect on endodontic pathogens inoculated in root canals. Resultant colony-forming unit counts were associated with observations of bacterial cell structural changes using conventional scanning electron microscopy (CSEM) and environmental scanning electron microscopy (ESEM) on inoculated dentine surfaces, following indirect and direct Nd:YAG laser irradiation, respectively. RESULTS: The Nd:YAG laser irradiation (1.5 W, 15 Hz, four times for 5 s) of Enterococcus faecalis inoculated canals resulted in a significant reduction (P < 0.05, Wilcoxon signed rank test) of the bacterial load, meaning a 99.7% kill, but no sterilization. The CSEM procedure verified that the extent of radiation damage was in line with the total amount of laser energy applied. After 2 h of incubation and three cycles of indirect laser treatment (i.e. through a 1-mm-thick dentine disc), no morphologically intact bacteria of Actinomyces naeslundii or Streptococcus anginosus were discernible. However, when micro-colonies of S. anginosus and specially biofilms of E. faecalis were present after 2 days, the in situ experiment using ESEM and direct laser treatment showed that bacterial eradication was reduced in deep layers. CONCLUSIONS: The Nd:YAG laser irradiation is not an alternative but a possible supplement to existing protocols for canal disinfection as the properties of laser light may allow a bactericidal effect beyond 1 mm of dentine. Endodontic pathogens that grow as biofilms, however, are difficult to eradicate even upon direct laser exposure.

Microbiological study and scanning electron microscopic analysis of root canal wall dentin following pumped Diodium Nd:YAG laser irradiation.

The capability of Nd:YAG laser in sterilizing root canals and the alterations of dentinal walls induced by laser treatment were investigated. Thirty root canals were infected by P. aeruginosa ATCC 27853 and thirty canals by A. naeslundii CH-12. Within each infection, 4 groups were selected on the basis of the treatment. Among them, 2 test groups (TGs) were treated by Nd:YAG laser at 15 Hz for 15 s, using 2 different settings: 1 Watt/70 Joule and 1.5 Watt/100 Joule, respectively (n = 10 each). The other 2 groups, used as controls (CGs), were: untreated (positive control, n = 5) and sterilized by 5.25% NaClO group (negative control, n = 5). Observations under scanning electron microscope (SEM) and quantitative bacterial counts were performed. These analyses were performed once per group after infections and treatments. Laser treatments significantly reduced the number of both bacteria. SEM investigation showed melting and crystallization of canal dentin over 1.5 W/100 J. Laser irradiation has a bactericidal effect but it does not completely sterilize the root canal as NaClO 5.25% solution does if the goal of treatment is also to avoid alterations of dentinal walls.

Photodestruction of human dental plaque bacteria: enhancement of the photodynamic effect by photomechanical waves in an oral biofilm model.

BACKGROUND AND OBJECTIVES: Periodontal disease results from the accumulation of subgingival bacterial biofilms on tooth surfaces. There is reduced susceptibility of these biofilms to antimicrobials for reasons that are not known. The goals of this study were to investigate the photodynamic effects of a conjugate between the photosensitizer (PS) chlorin(e6) (c(e6)) and a poly-L-lysine (pL) with five lysine residues on human dental plaque bacteria as well as on biofilms of the oral species Actinomyces naeslundii after their exposure to photomechanical waves (PW) generated by a laser in the presence of the conjugate. STUDY DESIGN/MATERIALS AND METHODS: Subgingival plaque samples from 12 patients with chronic destructive periodontitis were divided in 3 groups that were incubated for 5 minutes with 5 microM c(e6) equivalent from the pL-c(e6) conjugate in the presence of fresh medium (group I), PBS (group II), and 80% PBS/20% ethylenediaminetetra-acetic acid (EDTA) (group III) and were exposed to red light. Also, biofilms of A. naeslundii (formed on bovine enamel surfaces) were exposed to PW in the presence of 5 microM c(e6) equivalent from the pL-c(e6) conjugate and were then irradiated with red light. The penetration depth of the conjugate was measured by confocal scanning laser microscopy (CSLM). In both cases, after illumination serial dilutions were prepared and aliquots were spread over the surfaces of blood agar plates. Survival fractions were calculated by counting bacterial colonies. RESULTS: The PS/light combination achieved almost 90% killing of human dental plaque species. In biofilms of A. naeslundii, CSLM revealed that PW were sufficient to induce a 50% increase in the penetration depth of the pL-c(e6) conjugate into the biofilm. This enabled its destruction (99% killing) after photodynamic therapy (PDT). CONCLUSIONS: PW-assisted photodestruction of dental plaque may be a potentially powerful tool for treatment of chronic destructive periodontal disease.

Bacteriologic evaluation of the effect of Nd:YAG laser irradiation in experimental infected root canals.

The aim of this study was to evaluate the efficacy of the Pumped Diodium-Nd:YAG laser in sterilizing contaminated root canals. After hand instrumentation, 30 teeth were inoculated with Actinomyces naeslundii CH-12 and 30 teeth with Pseudomonas aeruginosa ATCC 27853 and incubated for 24 h. The teeth were divided into three subgroups: subgroup A received no treatment; subgroup B was irradiated with laser (5 Hz for 15 s or 10 Hz for 15 s); and subgroup C was irrigated with 5.25% NaOCl. The number of viable bacteria in each group was evaluated by using the surface-spread plate technique. The results indicated an average of 34.0% decrease in colony-forming units for A. naeslundii CH-12 and 15.7% for P. aeruginosa ATCC 27853 with the 5 Hz/15 s laser treatment, and for the 10 Hz laser frequency, a decrease of the 77.4% for A. naeslundii CH-12 and 85.8% for P. aeruginosa ATCC 27853. No bacteria were detected in the canals treated with 5.25% NaOCl. The results show an antibacterial effect of the Pumped Diodium Nd:YAG laser, depending on the radiation frequency. However, 5.25% NaOCl was more effective than either laser application.

An evaluation of the CO2 laser for endodontic disinfection.

The goal of this study was to evaluate the bactericidal action of the CO2 laser on animal teeth infected with an endodontic bacterial species. After instrumentation, 24 freshly extracted incisors were inoculated with a known concentration of Actinomyces odontolyticus and incubated anaerobically for 18 h. The incisors were separated into three groups: group 1--untreated control teeth; group 2--teeth treated with 3% NaOCl; and group 3--teeth lased with a CO2 laser at 5 W using three successive 9.9 s irradiation periods with 10 s between treatments. For each of the three groups, 60 microliters samples were removed using gel loading capillary pipette tips, and the diluted samples were plated in triplicate on Columbia agar plates. After a 5- to 6-day incubation, the colony-forming units were counted, and the quantitative results were subjected to an analysis of variance. The results of this analysis indicated an average 85% decrease in the colony-forming units in the laser-treated group, compared with the control group. According to Fisher and Scheffé tests, the differences in the averages between the control and laser groups were statistically significant (p < 0.05). The NaOCl treatment was statistically superior to the CO2 laser treatment.

Bacteria in supragingival plaque samples can be killed by low-power laser light in the presence of a photosensitizer.

The purpose of this study was to determine whether bacteria in supragingival plaque samples could be killed by low-power laser light in the presence of a suitable photosensitizer. Plaque samples were obtained from 10 volunteers, treated with either toluidine blue O (TBO) or aluminum disulphonated phthalocyanine (AlPcS2), and then exposed to light from a helium/neon (HeNe) or gallium aluminium arsenide (GaAs) laser respectively. Following irradiation, substantial reductions were achieved in the total anaerobic count as well as in the number of viable streptococci and actinomyces present in the samples. In the absence of laser light, the sensitizers themselves had little effect on the viability of the bacteria in the plaque samples. The HeNe/TBO combination appeared to be more effective than the GaAs/AlPcS2 combination, achieving log10 reductions of 2.95, 5.40 and 3.34 in the total anaerobic count, streptococci and actinomyces respectively with a light energy dose of 1.31 J. If effective in vivo, lethal photosensitization may be useful as a means of eliminating plaque bacteria from a carious lesion prior to its restoration.

[The bactericidal efficiency of ultrasonic in the root canal].

The objective of this paper was to determine the quantitative bactericidal efficiency of ultrasonic in the root canal. Four test organisms found frequently in the root canal were compared. The result showed ultrasonic can kill the test organisms effectively in the root canals. The best bactericidal efficiency occurred at 4 or 5 min. of ultrasonic though the more efficiency as the time longer. Bactericidal irrigation could increase the bactericidal efficiency of ultrasonic and its bactericidal action similar to irrigation bactericidal property.

Comparative bactericidal exposures for selected oral bacteria using carbon dioxide laser radiation.

Although relatively high CO2 laser energies have been shown to sterilize root canals, the response of several bacterial strains to decreasing exposures of CO2 laser energy remains unknown. Freshly grown bacterial cells were irradiated on glass microscope coverslips. A comparison of equivalent energy exposures with differing parameters was made on the bacterial viability. No statistically significant difference was found in the energy required to kill closely related bacterial species. However, the energy density required to kill greater than 99.5% of the bacteria is less than 200 J/cm2, much less than that shown to sterilize in a previous study.

Bactericidal action of carbon dioxide laser radiation in experimental dental root canals.

The ability of a carbon dioxide laser to sterilize the root canal of human teeth has been investigated. Three oral bacteria, Streptococcus sanguis, Streptococcus mutans, and Actinomyces viscosus, and three other bacteria, Bacillus cereus, Staphylococcus aureus, and Pseudomonas aeruginosa were used as experimental organisms. Exposure of cells on glass slides to laser radiation showed there was little difference in the exposure required to kill these six organisms. Complete recovery of bacteria from the root canal was initially a problem and was only achieved when bacterial manipulations and removal were carried out in rapid succession, within 5 min of inoculation. However, the geometry of the instrumented canal and the laser alignment were major factors in achieving consistent cell death of oral bacteria in the root canals. Using sets of 10 teeth, four repeated exposures of 10 W for 1 s was found to sterilize 4 or more of the teeth.

Apollo-Soyuz Test Project on biorhythm of zone-forming fungi: preliminary work.

The purpose of the experiment was to study general and local effects of space flight factors on the rhythm of cellular activity and on the morphological and genetic properties of biological objects. The Pushchino strain, Actinomyces levoris Kras 17-225A-IBFM, isolated at the Institute of Biological Physics, Moscow, was chosen as the main biological object. Under appropriate conditions it gives distinct and continuous rings of spore formation reflecting its intrinsic high degree of synchronism in changing its reproduction forms seen with the unaided eye as transparent rings (vegetative growth) alternate with convex white rings (spore-formation growth). As an additional test object, a film culture of bacteriophage T4Br+ developed at the institute was used. The strains were placed together in one bioblock together with plastic detectors for detecting nuclear particles. The film culture of bacteriophage enabled us to amplify the area of registration of local radiation effects by studying the genetic effects of these: frequency of mutations, induced radiation, their spectrum, subsequent revertability under the action of chemical mutagens with known mechanisms of action on DNA molecules.

[Photochromogeneity of actinomycetes producing heliomycin]. / O fotokhromogennosti aktinomitsetov, obrazyiushchikh geliomitsin

Irradiation with solar rays of actinomycetes producing heliomycin changed the colour and UV-induced luminescence of the growth medium. This was caused by formation of a luminescent pigment, hydroxyquinone. The aerial mycelium reduced in spore formation acquired a pink colour.