Bactericidal effects of different laser wavelengths on periodontopathic germs in photodynamic therapy

 This study was an attempt to clarify whether the bactericidal effects of photodynamic therapy (PDT) are wavelength or dose-dependent. We also attempted to create an optimised protocol for a light-based bactericidal modality to eliminate periodontal pathogens. Cultures of Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Porphromonas gingivalis, Prevotella intermedia, and Streptococcus sanguis, were exposed to a He-Ne laser (632.8 nm) with a 30 mW power output, a 100 mW diode laser at 665 nm, or a 100 mW diode laser at 830 nm, in the presence or absence of methylene blue (MB) as a photosensitiser. A control group was also used with exposure to MB alone without laser exposure. The cultures were analysed by viable counts. The results indicated that exposure to the 100 mW laser light could eliminate up to 40% of bacteria on average. In particular, the most effective killing occurred with exposure to laser light in combination with the MB photosensitiser. The results of kinetic studies indicated that the best PDT response rate was achieved with a 60 s (energy density 21.2 J/cm²) exposure to the 665 nm wavelength diode laser in the presence photosensitiser. In this condition, approximately 95% of A. actinomycetemcomitans and F. nucleatum, and 99–100% of the black-pigmented bacteria (P. gingivalis and P. intermedia) and S. sanguis were eliminated. These results showed that both wavelength and energy density are important factors, and that a low power laser of optimal wavelength and dosage, in combination with an appropriate photosensitiser, is a practical bactericidal modality. We concluded that using a diode laser of proper power and wavelength to deliver 60 s of irradiation could be a useful adjunct with mechanical debridement in the prevention of the re-colonisation of subgingival lesions by pathogenic microorganisms. Received: 29 November 2001 / Accepted: 18 June 2002     

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.     

Oral bacteria in multi-species biofilms can be killed by red light in the presence of toluidine blue

BACKGROUND AND OBJECTIVES: Oral bacteria can be killed by light in the presence of a suitable photosensitizer, and this could be used in the treatment of oral infections. In these diseases, however, bacteria are present as biofilms, which are refractive to antimicrobial agents. The purpose of this study was to determine whether oral bacterial biofilms were susceptible to lethal photosensitization. STUDY DESIGN/MATERIALS AND METHODS: Multi-species biofilms of oral bacteria were irradiated with light from a helium/neon laser in the presence of toluidine blue O (TBO) and the survivors enumerated. Controls examining the effects of light and TBO alone were also included. The biofilms were also examined by confocal scanning laser microscopy (CSLM). RESULTS: CSLM revealed that the biofilms had structures similar to those of dental plaque. Although, the biofilms consisted of extremely large numbers of bacteria ( approximately 9 x 10(9)), 97.4% were killed following irradiation with 31.5 J of laser light in the presence of 25 microg/ml TBO. CONCLUSIONS: Substantial numbers of oral bacteria in multi-species biofilms can be killed by light in the presence of TBO. This may be useful in the treatment of dental plaque-related diseases.     

Effect of blood upon the selective ablation of atherosclerotic plaque with a pulsed dye laser

Laser angioplasty systems with laser energy preferentially absorbed by atherosclerotic plaque may offer a safe method of plaque removal. This study evaluated the effect of blood upon selective energy absorption using a pulsed dye laser at 480 nm. Intra-arterial laser irradiation of normal rabbit femoral arteries demonstrated a perforation threshold energy with blood perfusion of 13.1 mJ per pulse compared to 87.9 mJ with saline (P less than .0001), indicating a deleterious effect in the presence of blood. An adverse effect upon arterial healing at 3 days was noted in sheep following intra-arterial irradiation during blood but not saline perfusion. Normal and atherosclerotic human aorta ablation thresholds differed significantly (P less than .0002) under saline (plaque: 20 mJ and normal: 120 mJ) but the difference under blood (plaque: 5 mJ and normal: 20 mJ) was not significant. We conclude that absorption of laser energy by blood can reduce the effect of differential absorption by endogenous chromophores in normal and pathologic vascular tissues and, therefore, removal of blood may be a prerequisite for selective ablation of atherosclerotic plaques.     

Multiple bacteria in aortic aneurysms

OBJECTIVE: The purpose of the present study was to reexamine the possibility that bacteria, particularly anaerobes, are present in aortic aneurysms. METHODS: From December 2000 to November 2001, 53 samples from aneurysm walls were collected from 49 patients during reconstructive surgery. The tissue specimens were sectioned and cultured under anaerobic conditions. Twenty-eight specimens were also subjected to scanning or transmission electron microscopy. RESULTS: Anaerobic cultivation yielded bacteria in 14 of the 53 samples (26.4%). All bacteria were gram-positive cocci or rods from nine genera and 12 species. Five cultures (35%) were mixed, containing two bacterial species. Mixed aerobic and anaerobic species were found in four samples (28.5%). Anaerobic bacteria were recovered from 10 of 14 positive cultures (71%). Among anaerobes found were Propionibacterium acnes, Propionibacterium granulosum, Actinomyces viscosus, Actinomyces naeslundii, and Eggerthella lenta. Coaggregating bacteria of different sizes and structure were found on the aneurysm walls and inside the intravascular plaque at electron microscopy. Bacteria were found in 20 of the 28 samples (71%) examined with scanning or transmission electron microscopy. CONCLUSION: Multiple bacteria, many of which did not belong to the indigenous skin microflora, colonize aortic aneurysms. It is not clear whether the bacteria contribute to weakening of the aortic wall by eliciting inflammation or whether they are secondary colonizers of aneurysms.     

Effects of low-level laser irradiation on human blood lymphocytes in vitro

Low-level laser irradiation (LLLI) has various effects on cultured human lymphocytes in vitro, but little is known about such effects in whole blood. This study investigated whether LLLI affected lymphocyte count in human whole blood in vitro. A total number of 130 blood samples were collected from apparently healthy adult patients through venipuncture into tubes containing EDTA. Each sample was divided into two equal aliquots to be used as a non-irradiated control sample and an irradiated sample. The irradiated aliquot was subjected to laser wavelengths of 405, 589, and 780 nm with different fluences of 36, 54, 72, and 90 J/cm², at a fixed irradiance of 30 mW/cm². A paired student t test was used to compare between non-irradiated and irradiated samples. The lymphocyte counts were measured using a computerized hematology analyzer and showed a significant (P?<?0.02) maximum increase (1.6%) at a fluence of 72 J/cm² when compared with non-irradiated samples. This increase in lymphocyte count upon irradiation was confirmed by flow cytometry. At a wavelength of 589 nm and fluence of 72 J/cm², irradiation of whole blood samples showed a significant increase in CD45 lymphocytes and natural killer (NK) (CD16, CD56) cells, but no significant changes in CD3 T lymphocytes, T-suppressor (CD3, CD8) cells, T-helper (CD3, CD4) cells, and CD19 B lymphocytes when compared with their non-irradiated counterparts. Our results clearly demonstrate that NK cell count is altered by irradiation, which ultimately affects the whole lymphocyte count significantly.     

Thermal damage of blood vessels in a rat skin-flap window chamber using indocyanine green and a pulsed alexandrite laser: A feasibility study

The design criteria and feasibility of specifically targeting blood vessels for thermal damage by using a pulsed alexandrite infra-red laser to heat an intravascularly injected infra-redabsorbing dye, namely indocyanine green (ICG), is demonstrated. Theoretical calculations map the distribution of light and heat in and around the subcutaneous blood vessels in a rat skin-flap window chamber as functions of dye concentration, vessel size, and vessel depth. Theoretical calculations showed that an injected dose of 6–24 mgkg⁻¹ of ICG and a 120-μs, 1-J cm⁻² alexandrite laser pulse at a wavelength of 785 nm would be sufficient to achieve selective vascular damage to a depth of at least 0.15 cm. Feasibility experiments were performed which illustrated that an irradiation of 1.27 J cm⁻² of skin flaps in uninjected control rats showed no evidence of vascular damage while vascular damage was seen in skin flaps using an experimental protocol of 12 mg kg⁻¹ i.v. of ICG and an energy fluence of 0.76 J cm⁻². This procedure could conceivably prove useful in the treatment of vascular lesions or cancer.     

Stimulatory effect of low-level laser therapy on root development of rat molars: a preliminary study

Several studies suggest a biomodulatory influence of low-level laser irradiation in the inflammatory and reparative processes of dental tissues. The aim of this study was to investigate the stimulatory effect of 808-nm laser irradiation on root development of rat molars and also to evaluate the histological reaction of pulp and periapical tissues. Twenty-four 30-day-old Wistar male rats were randomly assigned to three-time and five-time laser therapy groups. After initial x-ray, using mammography equipment, laser energy was applied at a wavelength of 808 nm (2 J/cm², 100 mW, 20 s) to the midroot area of the lower molars of one side of mouth at repeated intervals of the 48 h. The animals were killed 1 day after the final treatment, and root length development of the experimental samples was compared to contra-lateral non-irradiated molars using mammography. The histological reaction of the pulp and periapical tissue was evaluated under light microscopy. Root development was more advanced in irradiated groups than in the non-irradiated controls (p < 0.001). No significant differences, however, could be found between the root development changes in the three-time and five-time laser therapy groups (p > 0.05). Histological findings showed that the occurrence of secondary cement formation was significantly higher in the irradiation groups compared to the controls (p = 0.003). However, there were no statistically significant differences for the frequencies of pulp hyperemia, periodontal ligament fiber organization, or lamina dura remodeling between the groups (p > 0.05). Under the conditions used in this study, 808-nm low-level laser accelerates the rat molar root development in the presence of favorable histological reactions.     

Comparison of the photodynamic fungicidal efficacy of methylene blue, toluidine blue, malachite green and low-power laser irradiation alone against Candida albicans

This study was to evaluate specific effects of photodynamic therapy (energy density 15.8 J/cm², 26.3 J/cm² and 39.5 J/cm²) using methylene blue, toluidine blue and malachite green as photosensitizers and low-power laser irradiation on the viability of Candida albicans. Suspensions of C. albicans containing 10⁶ cells/ml were standardized in a spectrophotometer. For each dye, 120 assays, divided into four groups according to the following experimental conditions, were carried out: laser irradiation in the presence of the photosensitizer; laser irradiation only; treatment with the photosensitizer only; no exposure to laser light or photosensitizer. Next, serial dilutions were prepared and seeded onto Sabouraud dextrose agar for the determination of the number of colony-forming units per milliliter (CFU/ml). The results were subjected to analysis of variance and the Tukey test (P < 0.05). Photodynamic therapy using the photosensitizers tested was effective in reducing the number of C. albicans.. The number of CFU/ml was reduced by between 0.54 log₁₀ and 3.07 log₁₀ and depended on the laser energy density used. Toluidine blue, methylene blue and malachite green were effective photosensitizers in antimicrobial photodynamic therapy against C. albicans, as was low-power laser irradiation alone.     

Determining the optimal dose of 1940-nm thulium fiber laser for assisting the endodontic treatment

Insufficient cleaning, the complex anatomy of the root canal system, inaccessible accessory canals, and inadequate penetration of irrigants through dentinal tubules minimizes the success of the conventional endodontic treatment. Laser-assisted endodontic treatment enhances the quality of conventional treatment, but each laser wavelength has its own its own limitations. The optimal parameters for the antibacterial efficiency of a new wavelength, 1940-nm Thulium Fiber Laser, were firstly investigated in this study. This paper comprises of two preliminary analyses and one main experimental study, presents data about thermal effects of 1940-nm laser application on root canal tissue, effective sterilization parameters for bacteria, Enterococcus faecalis, and finally the antibacterial effectiveness of this 1940-nm Thulium Fiber Laser irradiation in single root canal. Based on these results, the optimal parameter range for safe laser-assisted root canal treatment was investigated in the main experiments. Comparing the antibacterial effects of four laser powers on an E. faecalis bacteria culture in vitro in 96-well plates showed that the most effective group was the one irradiated with 1 W of laser power (antibacterial effect corresponding to a log kill of 3). After the optimal laser power was determined, varying irradiation durations (15, 30, and 60 s) were compared in disinfecting E. faecalis. Laser application caused significant reduction in colony-forming unit values (CFU) compared with control samples in the 17% ethylenediaminetetraacetic acid (EDTA) group. The results of bacteria counts showed that 1 W with 30 s of irradiation with a 1940-nm thulium fiber laser was the optimal dose for safely achieving maximal bactericidal effect.     

Sensitization ofStreptococcus sanguis to killing by light from a helium/neon laser

Solutions of various photosensitizers were added to suspensions of the dental plaqueforming organism,Streptococcus sanguis. These were then exposed to light from a 7.3 mW Helium/Neon (He/Ne) laser for various time periods and the numbers of surviving cells determined by viable counting. Toluidine blue O, haematoporphyrin ester, thionin and methylene blue enabled substantial kills of the organism after irradiation times as short as 10 s which corresponds to an energy dose of 73 mJ. However, haematoporphyrin ester and, to a lesser extent, methylene blue were toxic to the organism in the absence of He/Ne light. Aluminium disulphonated phthalocyanine and crystal violet were ineffective as photosensitizers under the experimental conditions employed. The killing of sensitizedS. sanguis by light from a low-power laser suggests the possibility of a new approach to the control of dental plaque bacteria.     

The 800-nm diode laser irradiation induces skin collagen synthesis by stimulating TGF-β/Smad signaling pathway

The 800-nm diode laser is used clinically for hair removal and leg vein clearance. However, the effects of the laser on skin collagen synthesis have not been established. This study aims to research whether the 800-nm laser can be used for non-ablative rejuvenation and its possible mechanism by using an animal model. Eight 2-month-old rats were irradiated with the 800-nm diode laser at 20, 40, and 60 J/cm², respectively. Skin samples were taken for histological study and dermal thickness measurement at day 30 after laser irradiation. The expression of procollagen type I, III, IV, transforming growth factor-β (TGF-β), Smad2, 3, 4, and phosphorylated-Smad2, 3 in the rat skin was analyzed 24 h after completing all laser treatments by using RT-PCR and Western blot. Immunohistochemistry was performed to evaluate the content of type I collagen in the skin at day 30 after laser irradiation. The 800-nm diode laser treatments markedly improved the histological structure and increased dermal thickness compared to the non-irradiated controls. Laser irradiation at 40 J/cm² significantly up-regulated the expression of procollagen type I and IV, TGF-β and Smad2, 3, 4. The p-Smad2 and p-Smad3 levels were also enhanced in the laser-irradiated skin. The 800-nm laser is effective in improving skin structure and inducing skin new collagen expression. New collagen synthesis induced by the 800-nm laser was mediated by TGF-β/Smad signaling pathway. Thus, it seemed that the 800-nm laser could be used for non-ablative rejuvenation in the future.     

Monitoring of bactericidal action of laser by in vivo imaging of bioluminescent E. coli in a cutaneous wound infection

The worldwide rise in antibiotic resistance necessitates the development of novel antimicrobial strategies. This study aimed to evaluate the bactericidal action of an 810-nm diode laser in a cutaneous wound infection. An Escherichia coli strain was transformed with a shuttle vector (pRB474) containing firefly luciferase gene from Photinus pyralis resulting in a bioluminescent phenotype. Because firefly luciferase is an enzyme and as such is prone to inactivation at elevated temperature, the first phase has consisted in evaluating in vitro the effect of temperature elevation (30, 40, 50, and 60°C for 2 min) on bacteria bioluminescence. The second phase was performed in vivo. Two full-thickness circular, 14-mm diameter wounds (control and laser-irradiated) were induced on rats. Wound infection was carried out using a suspension (50 μl PBS) containing 5x10⁷ cells of bioluminescent E. coli (10⁹ cells/ml). Thirty minutes later, light irradiation was performed with an 810-nm diode laser (P=10 W, ∅=1.4 cm, fluence: 130, 195, and 260 J/cm²). Temperature was measured within each wound with a noncontact infrared thermometer. Light emission of the bioluminescent bacteria was monitored in vivo by a bioluminescence imaging system before and at 4, 8, 24, and 48 h after laser irradiation. In vitro, bacteria bioluminescence is not affected when temperature is maintained at 50°C for 2 min. In vivo, bioluminescence imaging showed that at 4 h, the viability of E. coli was reduced when compared to the control (CTRL) group (p<0.01). This observation was confirmed at 8 h (p<0.001), at 24 h (p<0.001), and finally at 48 h (p<0.001). Loss of viability of E. coli depends on laser fluence. At 48 h, bioluminescent bacteria were not detected (100% loss of viability) in the wound irradiated at 260 J/cm². For this fluence, the temperature reached 45°C at the end of the irradiation. This study confirms previous observations on the bactericidal effect of diode lasers. Because a progressive desiccation of the superficial dermis is usually observed when using laser irradiation, the hypothesis that laser irradiation dries out the wound making the wound an inhospitable place for bacteria is much more relevant than a direct effect of infrared light on chromophores inside bacteria. This is confirmed by the fact that in this latter case, one would expect an immediate drop in luminescence followed by an increase as the surviving bacteria started to divide and repopulate the wound. However, the exact mechanism deserves further studies. This study points out the advantage of using bioluminescence imaging to evaluate laser for the treatment of acute infections in vivo, nondestructively, and noninvasively.     

In vitro impact of laser irradiation on platelet aggregation

The review of blood optical and rheological parameters plays an important role in many medical routine diagnosis and therapeutic applications and is the best way to understand the mechanism of action of low-level laser irradiation on biological tissues. The aim of this study was to investigate the in vitro effect of laser radiation on platelet count and aggregation. Blood samples were obtained from 30 healthy volunteers; each sample was divided into four aliquots, one of them was considered as a control while the other three were exposed to three different laser doses. A wavelength of 532 nm and a low power of 100 mW were used for irradiation with a 4-mm-diameter beam spot. The irradiation times were 1.8, 3.7, and 6.2 s giving doses of irradiation 1.5, 3, and 5 J/cm², respectively. Microsoft Excel was used for statistical analysis. Low laser irradiation induced significant changes in platelet aggregation in the presence of weak agonists such as adenosine diphosphate (ADP) (P?≤?0.05) and epinephrine (P?≤?0.01). Low-level laser therapy has no influence on platelet count; however, it promotes platelet aggregation in response to weak agonists, specifically ADP and epinephrine.     

Low-level Er:YAG laser irradiation enhances osteoblast proliferation through activation of MAPK/ERK

Although the use of high-level Er:YAG laser irradiation has been increasing in periodontal and peri-implant therapy, the effects of low-level Er:YAG laser on surrounding tissues and cells remain unclear. In the present study, the effects of low-level Er:YAG laser irradiation on osteoblast proliferation were investigated. Cells of the osteoblastic cell line MC3T3-E1 were treated with low-level Er:YAG laser irradiation with various combinations of laser settings (fluence 0.7–17.2 J/cm²) and in the absence or presence of culture medium during irradiation. On day 1 and/or day 3, cell proliferation and death were determined by cell counting and by measurement of lactate dehydrogenase (LDH) levels. Further, the role of mitogen-activated protein kinase (MAPK) pathways in laser-enhanced cell proliferation was investigated by inhibiting the MAPK pathways and then measuring MAPK phosphorylation by Western blotting. Higher proliferation rates were found with various combinations of irradiation parameters on days 1 and 3. Significantly higher proliferation was also observed in laser-irradiated MC3T3-E1 cells at a fluence of approximately 1.0–15.1 J/cm², whereas no increase in LDH activity was observed. Further, low-level Er:YAG irradiation induced the phosphorylation of extracellular signal-regulated protein kinase (MAPK/ERK) 5 to 30 min after irradiation. Although MAPK/ERK 1/2 inhibitor U0126 significantly inhibited laser-enhanced cell proliferation, activation of stress-activated protein kinases/Jun N-terminal kinase (SAPK/JNK) and p38 MAPK was not clearly detected. These results suggest that low-level Er:YAG laser irradiation increases osteoblast proliferation mainly by activation of MAPK/ERK, suggesting that the Er:YAG laser may be able to promote bone healing following periodontal and peri-implant therapy.     

Effect of acid etching duration on tensile bond strength of composite resin bonded to erbium:yttrium–aluminium–garnet laser-prepared dentine. Preliminary study

The purpose of this study was to compare the tensile bond strength of composite resin bonded to erbium:yttrium–aluminium–garnet (Er:YAG) laser-prepared dentine after different durations of acid etching. The occlusal third of 68 human third molars was removed in order to expose the dentine surface. The teeth were randomly divided into five groups: group B (control group), prepared with bur and total etch system with 15 s acid etching [37% orthophosphoric acid (H₃PO₄)]; group L15, laser photo-ablated dentine (200 mJ) (laser irradiation conditions: pulse duration 100 μs, air–water spray, fluence 31.45 J/ cm², 10 Hz, non-contact hand pieces, beam spot size 0.9 mm, irradiation speed 3 mm/s, and total irradiation time 2 x 40 s); group L30, laser prepared, laser conditioned and 30 s acid etching; group L60, laser prepared, laser conditioned and 60 s acid etching; group L90, laser prepared, laser conditioned and 90 s acid etching. A plot of composite resin was bonded onto each exposed dentine and then tested for tensile bond strength. The values obtained were statistically analysed by analysis of variance (ANOVA) coupled with the Tukey–Kramer test at the 95 % level. A 90 s acid etching before bonding showed the best bonding value (P < 0.05) when compared with all the other groups including the control group. There is no significance difference between other groups, nor within each group and the control group. There was a significant increase in tensile bond strength of the samples acid etched for 90 s.     

Transmission of laser energy and assessment of target temperature in an experimental model of laser balloon angioplasty

The potential application of laser balloon angioplasty (LBA) is limited because of difficulties in temperature control and dosimetry during trans-balloon laser irradiation. An experimental model of LBA was designed to help understand the polymer membrane effects of two materials on tissue temperature. The durability of two types of polymer films with different optical properties, polyethylene terephthalate (PET), a low scattering polymer and polyetheretherketone (PEEK), a high scattering polymer, were examined as potential balloon material. The PEEK film (thickness: 50 μm) was melted and perforated by light from a diode laser (wavelength: 808±10 nm, irradiance: 1469 W cm⁻², mean exposure time: 30±7 s). On the other hand, the PET film with the same thickness was completely intact after 1 min exposure at the same irradiance. Diode laser irradiation was applied to indocyanine green stained human aortic media in three different exposure methods; directly, through the free PET and through the pressed PET film. Temperatures of laser-irradiated specimens were measured using an infra-red thermal camera with an 8–12 μm bandwidth, and corrected for the emissivity of the tissue and the PET film. Results demonstrated underestimation of surface temperature because of low transmittance of radiated intensity through the free PET film and a significant (p<0.001) increase of corrected temperature (δT=169±32°C) through the pressed-PET exposure compared with the direct exposure (ΔT=81±7°C), even though the same laser irradiance (18.6W cm⁻²) was applied for 5s. That is, the tightly PET-covered tissue develops a significantly higher temperature during diode laser irradiation, and this increased thermal effect can be advantageous for laser welding with less power in laser balloon angioplasty.     

Antimicrobial photodynamic therapy in the non-surgical treatment of aggressive periodontitis: microbiological profile

The aim of this trial was to investigate changes occurring in the subgingival microbiological composition of subjects with aggressive periodontitis, treated with antimicrobial photodynamic therapy (aPDT), in a single episode, or scaling and root planing (SRP), in a split-mouth design on −7, 0, and +90 days. Ten patients were randomly assigned to either aPDT using a laser source in conjunction with a photosensitizer or SRP with hand instruments. Subgingival plaque samples were collected and the counts of 40 subgingival species were determined using checkerboard DNA-DNA hybridization. The data were analyzed using the method of generalized estimating equations (GEE) to test the associations between treatments, evaluated parameters, and experimental times (α = .05). The results indicated that aPDT and SRP affects different bacterial species, with aPDT being effective in reducing numbers of A. actinomycetemcomitans than SRP. On the other hand, SRP was more efficient than aPDT in reducing the presence of periodontal pathogens of the Red Complex. Additionally, a recolonization in the sites treated by aPDT was observed, especially for T. forsythia and P. gingivalis. Under our experimental conditions, this trial demonstrates that aPDT and SRP affected different groups of bacteria, suggesting that their association may be beneficial for the non-surgical treatment of aggressive periodontitis.     

Non-surgical periodontal therapy assisted by potassium–titanyl–phosphate laser: a pilot study

As the American Academy of Periodontology indicates, the treatment of chronic periodontitis should be achieved in the least invasive manner through non-surgical periodontal therapy. However, complete removal of subgingival plaque and calculus is hindered with increasing probing depth (PD) and furcation involvement using hand, sonic or ultrasonic instruments. Many authors have suggested that the use of laser as an adjunct to scaling and root planing (SRP) might improve the effectiveness of conventional periodontal treatment. The aim of this study was to evaluate potassium–titanyl–phosphate (KTP) laser in non-surgical periodontal therapy. Seven hundred and thirty sites with probing depths of 4–6 mm were involved in the study. The sites were divided into four groups: control (SRP, chlorhexidine gel 0.5%), group A (SRP, chlorhexidine gel 0.5%, three sessions of KTP laser irradiation); group B (SRP, three sessions of KTP laser irradiation) and group C (SRP, irrigation with povidone-iodine 10%, three sessions of KTP laser irradiation). KTP laser was used with the following parameters: output power 0.6 W, time on 10 ms, time off 50 ms, 30 s per irradiation, fluence 19 J/cm². All the sites showed improvement in all clinical parameters. Clinical attachment loss (CAL), pocket probing depths (PPDs) and bleeding on probing (BOP), especially in the lased groups, showed significant results (P < 0.001). Our experience showed KTP laser to be a significant help in SRP; nevertheless, more studies are necessary to confirm our results.     

High-frequency low-level diode laser irradiation promotes proliferation and migration of primary cultured human gingival epithelial cells

In periodontal therapy, the use of low-level diode lasers has recently been considered to improve wound healing of the gingival tissue. However, its effects on human gingival epithelial cells (HGECs) remain unknown. The aim of the present study was to examine whether high-frequency low-level diode laser irradiation stimulates key cell responses in wound healing, proliferation and migration, in primary cultured HGECs in vitro. HGECs were derived from seven independent gingival tissue specimens. Cultured HGECs were exposed to a single session of high-frequency (30 kHz) low-level diode laser irradiation with various irradiation time periods (fluence 5.7–56.7 J/cm²). After 20–24 h, cell proliferation was evaluated by WST-8 assay and [³H]thymidine incorporation assay, and cell migration was monitored by in vitro wound healing assay. Further, phosphorylation of the mitogen-activated protein kinase (MAPK) pathways after irradiation was investigated by Western blotting. The high-frequency low-level irradiation significantly increased cell proliferation and [³H]thymidine incorporation at various irradiation time periods. Migration of the irradiated cells was significantly accelerated compared with the nonirradiated control. Further, the low-level diode laser irradiation induced phosphorylation of MAPK/extracellular signal-regulated protein kinase (ERK) at 5, 15, 60, and 120 min after irradiation. Stress-activated protein kinases/c-Jun N-terminal kinase and p38 MAPK remained un-phosphorylated. The results show that high-frequency low-level diode laser irradiation promotes HGEC proliferation and migration in association with the activation of MAPK/ERK, suggesting that laser irradiation may accelerate gingival wound healing.