Flow path system of ultraviolet C irradiation from xenon flash to reduce bacteria survival in platelet products containing a platelet additive solution.

BACKGROUND: Our previous study showed that ultraviolet C (UVC) from xenon (Xe) flash without any photoreactive compounds inactivated bacteria in platelet concentrates (PCs) with less damage to platelets (PLTs) as compared with Xe flash containing ultraviolet A, ultraviolet B, and visible light. Here, we report a UVC irradiation system for PCs under flow conditions consisting of a flow path-irradiation sheet, a peristaltic pump, and a collection bag. STUDY DESIGN AND METHODS: Platelet concentrates containing Ringer's solution (R-PCs) inoculated with bacteria were injected into a flow path sheet using a peristaltic pump, being irradiated with UVC from Xe flash. The quality of the irradiated PCs containing platelet additive solution (PAS-PCs) was assessed based on PC variables, PLT surface markers, and aggregation ability. RESULTS: Streptococcus dysgalactiae (12 tests) and Escherichia coli (11) were all negative on bacterial culture, while Staphylococcus aureus (12) and Klebsiella pneumoniae (14) grew in one and two R-PCs, respectively. Bacillus cereus spores were inactivated in 7 of 12 R-PCs. PC variables became significantly different between irradiated and nonirradiated PAS-PCs. P-selectin, first procaspase-activating compound (PAC-1) binding, and phosphatidylserine increased by irradiation. Aggregability stimulated by adenosine diphosphate, collagen, or thromboxane A2 increased in the irradiated PAS-PCs, while that by thrombin became smaller compared with nonirradiated controls. CONCLUSION: This newly developed system inactivated bacteria including spores in R-PCs. PAS-PCs irradiated by this system retained acceptable in vitro quality and aggregability. Usage of a peristaltic pump instead of agitator during irradiation may enable this system to be directly combined with an apheresis blood cell separator.

Bacterial endotoxins and microorganisms in the oral cavities of patients on cancer therapy.

OBJECTIVES: This study investigated the presence of Streptococci, Staphylococci, aerobic gram negative bacteria (AGNB), Candida and bacterial endotoxins in the oral cavities of patients receiving chemo- and/or radiotherapy for cancer. METHODS: Samples of oral cavity rinse were collected from 100 patients on cancer treatment and 70 healthy individuals. Demographic and clinical data were recorded. Samples were cultured onto various agar plates for qualitative and quantitative analysis and tested for the presence of endotoxin. Results were analysed using the Mann-Whitney and chi-square tests. RESULTS: In cancer patients, S. aureus counts were high and 66.7% of patients on chemo- and radiotherapy carried these bacteria (p=<0.05). The Candida carrier rate was significantly (p < 0.01) high in cancer patients (54%). No significant difference was found in the carrier rate of Streptococci and AGNB between the healthy and cancer group as well as between the cancer patients with chemo and radio- and chemotherapy alone. No significant difference was found in the level of endotoxin between the cancer patients and healthy individuals, and cancer patients with and without AGNB. CONCLUSIONS: No differences in the prevalence of bacteria and bacterial endotoxins were found between the cancer patients and healthy individuals. Oral cavity endotoxins did not correlate with the carriage of AGNB. However, due to the high prevalence in cancer patients, the role of Candida species and S. aureus in the pathology may not be excluded.

Bactericidal effects of 310 nm ultraviolet light-emitting diode irradiation on oral bacteria.

BACKGROUND: Ultraviolet (UV) light is used for phototherapy in dermatology, and UVB light (around 310 nm) is effective for treatment of psoriasis and atopic dermatitis. In addition, it is known that UVC light (around 265 nm) has a bactericidal effect, but little is known about the bactericidal effect of UVB light. In this study, we examined the bactericidal effects of UVB-light emitting diode (LED) irradiation on oral bacteria to explore the possibility of using a 310 nm UVB-LED irradiation device for treatment of oral infectious diseases. METHODS: We prepared a UVB (310 nm) LED device for intraoral use to examine bactericidal effects on Streptococcus mutans, Streptococcus sauguinis, Porphyromonas gingivalis, and Fusobacterium nucleatum and also to examine the cytotoxicity to a human oral epithelial cell line (Ca9-22). We also examined the production of nitric oxide and hydrogen peroxide from Ca9-22 cells after irradiation with UVB-LED light. RESULTS: Irradiation with the 310 nm UVB-LED at 105 mJ/cm2 showed 30-50% bactericidal activity to oral bacteria, though 17.1 mJ/cm2 irradiation with the 265 nm UVC-LED completely killed the bacteria. Ca9-22 cells were strongly injured by irradiation with the 265 nm UVC-LED but were not harmed by irradiation with the 310 nm UVB-LED. Nitric oxide and hydrogen peroxide were produced by Ca9-22 cells with irradiation using the 310 nm UVB-LED. P. gingivalis was killed by applying small amounts of those reactive oxygen species (ROS) in culture, but other bacteria showed low sensitivity to the ROS. CONCLUSIONS: Narrowband UVB-LED irradiation exhibited a weak bactericidal effect on oral bacteria but showed low toxicity to gingival epithelial cells. Its irradiation also induces the production of ROS from oral epithelial cells and may enhance bactericidal activity to specific periodontopathic bacteria. It may be useful as a new adjunctive therapy for periodontitis.

Low-dose irradiation affects the functional behavior of oral microbiota in the context of mucositis.

The role of host-microbe interactions in the pathobiology of oral mucositis is still unclear; therefore, this study aimed to unravel the effect of irradiation on behavioral characteristics of oral microbial species in the context of mucositis. Using various experimental in vitro setups, the effects of irradiation on growth and biofilm formation of two Candida spp., Streptococcus salivarius and Klebsiella oxytoca in different culture conditions were evaluated. Irradiation did not affect growth of planktonic cells, but reduced the number of K. oxytoca cells in newly formed biofilms cultured in static conditions. Biofilm formation of K. oxytoca and Candida glabrata was affected by irradiation and depended on the culturing conditions. In the presence of mucins, these effects were lost, indicating the protective nature of mucins. Furthermore, the Galleria melonella model was used to study effects on microbial virulence. Irradiated K. oxytoca microbes were more virulent in G. melonella larvae compared to the nonirradiated ones. Our data indicate that low-dose irradiation can have an impact on functional characteristics of microbial species. Screening for pathogens like K. oxytoca in the context of mucosits could be useful to allow early detection and immediate intervention.

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.

The effect of radiofrequency ablation on microbiology of the tonsils.

OBJECTIVE: To investigate the effect of tonsil size reduction using temperature controlled radiofrequency on the number of pathogenic bacteria in the tonsil tissue. MATERIALS AND METHODS: This study was performed on 25 patients who had undergone tonsillectomy under general anesthesia at our clinic. Immediately after the cold knife tonsillectomy both tonsils were removed, one was included in the control and the other one was included in the study group. In vitro radiofrequency was applied to the tonsil in the study group at eight distinct points, each lasting 15s. Biopsy materials were taken under sterile conditions from the center of each tonsil for further culturing. RESULTS: The difference in bacterial number was investigated between the two groups. The bacterial number following radiofrequency administration was found to be significantly very lower compared to the control group (p<0.01). Radiofrequency administration significantly reduced growth of all types of bacteria. CONCLUSION: The radiofrequency tonsil ablation technique, which is used safely and effectively in the management of obstructive tonsil hypertrophy, currently has no indication for the treatment of patients with chronic and recurrent tonsillitis. However, when the right conditions are provided, the radiofrequency tonsil ablation technique may be applied to patients with chronic and recurrent tonsillitis and further studies investigating the differences in the frequency of patients' tonsillitis episodes should be undertaken.

Decontamination of rough titanium surfaces with diode lasers: microbiological findings on in vivo grown biofilms.

OBJECTIVES: The bactericidal efficacy of diode lasers has already been demonstrated in vitro. We investigated the reduction of aerobe bacteria - colonizing rough titanium samples in biofilms intraorally grown - by diode lasers of different wave lengths. MATERIAL AND METHODS: Twenty-two volunteers participated in the trial. They were fitted for 10 days with custom-made intraoral plastic splints carrying titanium sleeves. A part of the sleeves was then irradiated with diode lasers in different modes. The other part remained non-irradiated and served as control. Directly after irradiation, the sleeves were swabbed and the gained bacteria were first examined microscopically and then were cultured under aerobic conditions. RESULTS: The bacteria in the controls and in the treated samples were quantified. A comparison with the controls revealed a marked overall reduction of bacterial colonization in all irradiated sleeves. Continuous irradiation for 20 s reduced bacteria counts by 99.67% at 810 nm and 99.58% at 980 nm. Repeating the 20 s exposure five times reduced counts by 99.98% at 810 nm and by 99.39% at 980 nm. A 98.86% reduction was seen after irradiation in pulsed mode. A further analysis in respect of different isolated bacteria revealed that the streptococci group was reduced by 99.29-99.99%, while the staphylococci group was reduced to a lesser extent in the range 94.67-99.99%. CONCLUSION: The results are of clinical relevance. In comparison with the mean bacterial counts of the untreated samples, all irradiation programs studied in this investigation reduced mean bacterial colonization in a biofilm on intraoral rough titanium surfaces by more than 98%. The actual extent of reduction was dependent on the bacteria species as well as on the irradiation mode.

In vivo effect of carbon dioxide laser-skin resurfacing and mechanical abrasion on the skin's microbial flora in an animal model.

BACKGROUND: Although beam-scanning carbon dioxide (CO2) lasers have provided a highly efficient tool for esthetic skin rejuvenation there has been no comprehensive animal studies looking into microbial skin changes following CO2 laser skin resurfacing. OBJECTIVE: To evaluate the in vivo effects of CO2 laser skin resurfacing in an experimental rat model in comparison with mechanical abrasion on the skin microbial flora. METHODS: Four separate cutaneous sections of the right dorsal surface of 10 Wistar rats were treated with a CO2 laser, operating at 18 W and delivering a radiant energy of 5.76 J/cm2, while mechanical abrasions of the skin were created on four sections of the left dorsal surface using a scalpel. Samples for culture and biopsies were obtained from the skin surfaces of the rats on day 1 of application of the CO2 laser or mechanical abrasion, as well as 10, 30, and 90 days after the procedure. The presence of four microorganisms (staphylococci, streptococci, diphtheroids, and yeasts) was evaluated as a microbe index for the skin flora, and colony counts were obtained using standard microbiological methods. RESULTS: Skin biopsy specimens, following CO2 laser treatment, initially showed epidermal and papillary dermal necrosis and later a re-epithelization of the epidermis as well as the generation of new collagen on the upper papillary dermis. The reduction in microbial counts on day 1 of the CO2 laser-inflicted wound was statistically significant for staphylococci and diphtheroids compared with the baseline counts (p=.004 and p<.001, respectively), and for staphylococci, diphtheroids, and yeasts compared with the scalpel-inflicted wound on the same day (p=0.029, p<.001, and p=.030, respectively). CONCLUSIONS: Skin resurfacing using CO2 lasers considerably reduces microbial counts of most microorganisms in comparison with either normal skin flora or a scalpel-inflicted wound. This might contribute to the positive clinical outcome of laser skin resurfacing.

Antimicrobial efficacy of semiconductor laser irradiation on implant surfaces.

PURPOSE: This study was conducted to investigate the antimicrobial effect of an 809-nm semiconductor laser on common dental implant surfaces. MATERIALS AND METHODS: Sandblasted and acid-etched (SA), plasma-sprayed (TPS), and hydroxyapatite-coated (HA) titanium disks were incubated with a suspension of S. sanguinis (ATCC 10556) and subsequently irradiated with a gallium-aluminum-arsenide (GaAlAs) laser using a 600-microm optical fiber with a power output of 0.5 to 2.5 W, corresponding to power densities of 176.9 to 884.6 W/cm2. Bacterial reduction was calculated by counting colony-forming units on blood agar plates. Cell numbers were compared to untreated control samples and to samples treated with chlorhexidine digluconate (CHX). Heat development during irradiation of the implants placed in bone blocks was visualized by means of shortwave thermography. RESULTS: In TPS and SA specimens, laser irradiation led to a significant bacterial reduction at all power settings. In an energy-dependent manner, the number of viable bacteria was reduced by 45.0% to 99.4% in TPS specimens and 57.6% to 99.9% in SA specimens. On HA-coated disks, a significant bacterial kill was achieved at 2.0 W (98.2%) and 2.5 W (99.3%) only (t test, P < .05). For specimens treated with CHX, the bacterial counts were reduced by 99.99% in TPS and HA-coated samples and by 99.89% in SA samples. DISCUSSION: The results of the study indicate that the 809-nm semiconductor laser is capable of decontaminating implant surfaces. Surface characteristics determine the necessary power density to achieve a sufficient bactericidal effect. The bactericidal effect, however, was lower than that achieved by a 1-minute treatment with 0.2% CHX. The rapid heat generation during laser irradiation requires special consideration of thermal damage to adjacent tissues. CONCLUSION: No obvious advantage of semiconductor laser treatment over conventional methods of disinfection could be detected in vitro.

Non-mutans streptococci in patients receiving radiotherapy in the head and neck area.

OBJECTIVE: To study mutans and non-mutans streptococci in patients after radiotherapy of the head and neck. METHODS: Oral rinse samples collected from nasopharyngeal carcinoma patients before and after radiotherapy were diluted and cultured on nonselective and selective media for enumeration of total cultivable plaque flora, mutans and non-mutans streptococci and lactobacilli. Non-mutans streptococci were identified biochemically and by 16S rDNA sequence homology analysis. RESULTS: After irradiation, mutans streptococci were not isolated; the levels of Streptococcus mitis and lactobacilli increased significantly. The level of Streptococcus salivarius increased, but the significance was the borderline. The level of Streptococcus sanguis decreased significantly after irradiation. The abundance of other oral streptococci species showed no significant changes. CONCLUSIONS: S. mitis and S. salivarius are the predominant non-mutans streptococci in the high-caries-risk oral flora following radiotherapy.

Nd:YAG laser irradiation of infected root canals in combination with microbiological examinations.

In this in vivo study, 30 subjects with infected root canals were treated with the neodymium: yttrium-aluminum-garnet, or Nd:YAG, laser using standard laser settings and procedures. In microbiological examinations conducted before irradiation, the authors found streptococci in 30 cases and staphylococci in 15 cases. After the first irradiation, the authors found that 19 root canals showed minimal streptococcal growth and 10 root canals showed minimal staphylococcal growth.

Thermal effects and antibacterial properties of energy levels required to sterilize stained root canals with an Nd:YAG laser.

Thermal effects and antibacterial properties of an Nd:YAG laser were studied to establish clinically safe levels of energy to deliver into the root canal and to determine the energy level needed to sterilize infected root canals. The results indicate that lasing cycles of 3 J-s for 15 s followed by a 15-s recovery interval can be continued for prolonged periods without risk of thermal damage to surrounding tissues. In vitro lasing of root canals inoculated with dark stained bacteria showed that two such lasing cycles sterilized only two out of eight canals, whereas when four cycles were used seven out of eight canals were sterilized. Guidelines for energy levels in endodontic Nd:YAG laser work are discussed, and base data for calculating appropriate energy levels are given.

Lethal photosensitization of bacteria in subgingival plaque from patients with chronic periodontitis.

Subgingival plaque samples from patients with chronic periodontitis were exposed to light from a 7.3 mW Helium/Neon laser for 30 s in the presence and absence of 50 micrograms/ml toluidine blue O as a photosensitizer. Viable counts of various groups and species of bacteria were carried out before and after irradiation. The median numbers of viable bacteria initially present in the 30-microliters aliquots irradiated were 1.13 x 10(5) cfu (aerobes), 4.08 x 10(5) cfu (anaerobes), 4.92 x 10(3) cfu (black-pigmented anaerobes), 4.75 x 10(2) cfu (Porphyromonas gingivalis), 6.15 x 10(3) cfu (Fusobacterium nucleatum) and 1.7 x 10(4) cfu (streptococci). The dye/laser combination achieved significant reductions in the viability of these organisms, the median reductions in the viable counts being 91.1% for aerobes, 96.6% for anaerobes, 100% for black-pigmented anaerobes, P. gingivalis and F. nucleatum and 94.2% for streptococci. Overall, the viability of bacteria in the 20 plaque samples was not significantly decreased by the dye alone. However, in a small minority of samples there were indications of light-independent, dye-induced toxicity. Low-power lasers, in conjunction with appropriate photosensitizers, may be a useful adjunct to mechanical debridement in the treatment of inflammatory periodontal diseases if a similar effectiveness against subgingival plaque bacteria can be achieved in vivo.

Dye-mediated bactericidal effect of He-Ne laser irradiation on oral microorganisms.

Little attention has been given to the bactericidal effect of laser irradiation, particularly using low-power energy lasers. It has been demonstrated that He-Ne laser light has an inhibitory action on dental plaque. The purpose of this study was to investigate the bactericidal effect of He-Ne laser irradiation on cariogenic microorganisms. The bactericidal effect was determined by the formation of a growth-inhibitory zone or by the counting of viable bacterial colonies. Streptococcus sobrinus AHT that is a Gram-positive microorganism was sensitive to He-Ne laser light, but Escherichia coli, a Gram-negative microorganism, was resistant. The effect of several dyes necessary to instigate a bactericidal action was also examined. A growth-inhibitory zone was observed using 10 kinds of blue, purple, or green dyes, which were mainly phenylmethane dyes. The leakage of potassium from S. sobrinus AHT following laser irradiation was determined using an atomic absorption spectrophotometer. The leakage began to increase following irradiation for 2 min, and reached a plateau following irradiation for 30-60 min. Moreover, to examine some changes in the dye itself following laser irradiation in the absence of bacteria, ultraviolet-visible absorption spectra and 1H NMR spectra were recorded. In this study, it was indicated that the bactericidal effect on cariogenic bacteria by He-Ne laser irradiation was efficient only in the presence of specific dyes. It is suggested that this laser may be suitable for clinical applications in preventive dentistry.

[Use of UV radiation for the disinfection of water. V. Microbiological studies of the behavior of bacterial cells from the logarithmic and from the stationary phase in cold and warm drinking water]. / Anwendung von UV-Strahlen zur Desinfektion von Wasser V. Mitteilung: Mikrobiologische Untersuchungen über das Verhalten von Bakterienzellen aus der logarithmischen und aus der stationären Phase in kaltem und in warmem Trinkwasser.

The u.v.-susceptibility of E. cloacae, E. coli and E. faecium was tested with a flow-through u.v. light treatment apparatus. Drinking water of three ranges of temperature (ca. 13 degrees C, ca. 33 degrees C, and ca. 48 degrees C) was used. The u.v.-susceptibility was tested with bacteria harvested in the exponential phase of growth as well as with bacteria harvested in the stationary phase. In all ranges of temperature and both phases of growth reductions of at least 99.9999% of E. cloacae and E. coli were obtained by a dosage of less than or equal to 25 mWs/cm2. For E. faecium harvested in the exponential phase of growth this could be confirmed in cold water of 13 degrees C, while in warm water of 48 degrees C a dosage of ca. 47 mWs/cm2 for a reduction of 99.9999% was necessary. To kill E. faecium-cells, which were harvested in the stationary phase, dosage between 39 and 45 mWs/cm2 were necessary in water of the three ranges of temperature.

Effectiveness of ultrasonic files in the disruption of root canal bacteria.

The physical mechanisms of ultrasound, namely cavitation and acoustic streaming, generated by the Enac-Osada ultrasonic unit were investigated for effectiveness in disrupting Streptococcus mitis. In addition, the bactericidal effect of ultrasound in the presence of 2.5% sodium hypochlorite was examined. Bacterial suspensions were irradiated directly with ultrasound in simulated root canals, and the viability of bacteria was examined after growth on a blood agar medium under anaerobic conditions at 37 degrees C for 5 days. The results indicated that ultrasound per se failed to disrupt bacteria but resulted in increases in the viable counts; the former was considered to be because of the lack of cavitation and the latter because of the dispersal effects of acoustic streaming. The 2.5% sodium hypochlorite solution demonstrated powerful bactericidal activity.

[Use of UV radiation for the disinfection of water. IV. Microbiological studies of the UV sensitivity of different aged cells of E. faecium, E. coli and P. aeruginosa]. / Anwendung von UV-Strahlen zur Desinfektion von Wasser. IV. Mitteilung: Mikrobiologische Untersuchungen zur UV-Sensibilität von Zellen unterschiedlichen Alters von E. faecium, E. coli und P. aeruginosa.

With a flow-through u.v. light treatment apparatus the u.v.-susceptibility of E. faecium, E. coli and P. aeruginosa in drinking water was tested. The investigations were done with cells harvested in the exponential phase of growth. One part of the suspensions was used immediately (= 0 days old cells), two other parts were stored for 5 and 7 days in sterilised drinking water in the dark at room temperature (= 5 and 7 days old cells). In either case--0 or 5 and 7 days old cells--reductions of at least 99.999% were obtained by a dosage of 25 mWs/cm2. No difference concerning the u.v.-susceptibility could be noticed between the different kind of cells of E. faecium, E. coli, and P. aeruginosa.

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.

[Effect of helium-neon-laser radiation on microbes]. / Wirksamkeit eines Helium-Neon-Lasers auf Mikroorganismen.

In a well-defined in-vitro experiment the effects of a He-Ne-laser of 6 mW output on some dominating microbes in the oral cavity were studied by using a scanning electron microscope. No changes on the surface in the irradiated colonies could be seen even after an exposition lasting for 60 minutes (continuous or pulsated wave). Therefore an antibacterial effect of softlaser-irradiation sometimes described in literature in questionable.

[Effect of low-intensity laser radiation on wound microflora]. / K voprosu o vliianii lazernogo izlucheniia nizkoi intensivnosti na mikrofloru ran.

The authors have established that the laser radiation of low intensity fails to exert bactericide influence on the wound microflora. The positive effect of the laser light of low intensity in the complex treatment of purulent wounds and trophic ulcers seems to be due to the total influence on the patient's organism and the local pathological process.