Study of the efficacy of 5 ALA-mediated photodynamic therapy on human larynx squamous cell carcinoma (Hep2c) cell line

5-aminolevulanic acid (ALA), a precursor of Protoporphyrin IX, was evaluated as an inducer of photodamage on Hep2c, human larynx squamous cell carcinoma, cell line. Porphyrins are used as active cytotoxic antitumor agents in photodynamic therapy (PDT). The present study evaluates the effects of photodynamic therapy (PDT) with 5-aminolevulinic acid (5-ALA) using human larynx cells as experimental model. Hep2c cell line was irradiated with red light (a diode laser, λ = 635 nm). The influence of different incubation times and concentrations of 5-ALA, different irradiation doses and various combinations of photosensitizer and light doses on the cellular viability of Hep2c cells were studied. The optimal uptake of photosensitizer ALA in Hep-2c cells was investigated by means of spectrometric measurement. Cells viability was determined by means of neutral red assay (NR). It was observed that sensitizer or light doses have no significant effect on cells viability when studied independently. The spectrometric measurements showed that the maximal cellular uptake of 5-ALA occurred after 7 h in vitro incubation. The photocytotoxic assay showed that light dose of 85 J/cm² gives effective PDT outcome for Hep2c cell line incubated with 55 μg/ml of 5-ALA with a conclusion that Hep2c cell line is sensitive to ALA-mediated PDT.     

Efficient treatment of pigmented B16 melanoma using photosensitized long-circulating magnetofullerenosomes

Magnetic targeting was used for melanoma treatment with magnetofullerenosomes. After their intravenous administration, a permanent magnet was attached to the surface of B16 tumors in C57 mice for 24 h, followed by irradiation with an infrared laser pulse and subsequent illumination with a 776 nm diode laser for conventional photodynamic treatment. Tumor response was substantially better than that obtained with either treatment alone.     

Modeling laser irradiation conditions for mucosal tissues in antimicrobial photodynamic therapy

We use computer modeling to analzye empirically selected conditions for antimicrobial photodynamic therapy of mucosal tissues. We calculate the optical and thermal fields for experimental conditions for low-intensity (cold) laser irradiation used in treatment of lesions in mucosal tissues stained by methylene blue: λ = 670 nm, power density 150–300 mW/cm², doses 9–18 J/cm²; λ = 632.8 nm, 15 mW/cm², dose 4.5 J/cm². For numerical estimates, we used the optical characteristics of methylene blue and three layers of mucosal tissues at the laser radiation wavelengths, and also the thermal characteristics of the tissues. The experimental conditions were optimized using the ratio of the tissue penetration depth for the absorbed optical energy and the penetration depth of methylene blue into the lesion, while maintaining safe tissue heating temperatures.     

Photodynamic damage study of HeLa cell line using ALA

The present study evaluates the photodynamic damage with 5-aminolevulinic acid (5-ALA) using HeLa as experimental model. HeLa cell line was irradiated with red light (He-Ne laser, λ = 632.8 CW nm). The influence of different incubation times and concentrations of 5-ALA, different irradiation doses and various combinations of photosensitizer and light doses on the cellular viability of HeLa cells were studied. The optimal uptake of photosensitizer ALA in HeLa cells was investigated by means of PpIX fluorescence intensity by exciting the HeLa cell suspension at 450 nm and a detection wavelength set at 690 nm. Cells viability was determined by means of trypan blue solution. The spectrometric measurements showed that the maximal cellular uptake of 5-ALA occurred after 4 h in vitro incubation. We found that the combination with 5-ALA and laser irradiation leads to time/concentration-dependent increase of cells death and also energy doses-dependent enlarge the cells death. The fluorescence intensity after PDD of carcinoma cells reduce when compared with the control group. The fluorescence emission spectral profiles after PDD of carcinoma cells showed a dip around 425–525 nm when compared with the control group. This may be due to the damage of mitochondria component of cells. The percentage of HeLa cells after PDD shows that the percentage of cells survival rate as function of laser dose (power). Hence it is clear that at 200 μg/ml ALA and 20 mW laser irradiation, more than 70% of HeLa cells were dead after 15 min.     

The application of Raman and anti-stokes Raman spectroscopy for in situ monitoring of structural changes in laser irradiated titanium dioxide materials

The use of Raman and anti-stokes Raman spectroscopy to investigate the effect of exposure to high power laser radiation on the crystalline phases of TiO₂ has been investigated. Measurement of the changes, over several time integrals, in the Raman and anti-stokes Raman of TiO₂ spectra with exposure to laser radiation is reported. Raman and anti-stokes Raman provide detail on both the structure and the kinetic process of changes in crystalline phases in the titania material. The effect of laser exposure resulted in the generation of increasing amounts of the rutile crystalline phase from the anatase crystalline phase during exposure. The Raman spectra displayed bands at 144 cm⁻¹ (A1g), 197 cm⁻¹ (Eg), 398 cm⁻¹ (B1g), 515 cm⁻¹ (A1g), and 640 cm⁻¹ (Eg) assigned to anatase which were replaced by bands at 143 cm⁻¹ (B1g), 235 cm⁻¹ (2 phonon process), 448 cm⁻¹ (Eg) and 612 cm⁻¹ (A1g) which were assigned to rutile. This indicated that laser irradiation of TiO₂ changes the crystalline phase from anatase to rutile. Raman and anti-stokes Raman are highly sensitive to the crystalline forms of TiO₂ and allow characterisation of the effect of laser irradiation upon TiO₂. This technique would also be applicable as an in situ method for monitoring changes during the laser irradiation process.     

Dynamics of radiation scattering by particles of condensed phase in quasicontinuous laser irradiation of metals

Radiation scattering by particles of condensed phase in an ablation plasma plume has been experimentally studied during quasicontinuous laser irradiation (λ = 1.06 μm, q = 0.1–9 MW/cm², τ ∼ 1.5 msec) of duraluminum D16T, aluminum A99, and bismuth. The particle size distribution and the nature of their dispersal during irradiation was studied in scattered light (λ = 0.69 μm) from individual particles that could be visually observed on photographs. It was found that under the pressure developed in the plume, large particles ejected from the irradiated zone can move backward and return to the target (D16T). The plume (Bi) becomes brighter due to ablation of particles in the path of the laser beam. The directional scattering coefficients for scattering from the local zone on the axis of the plume, measured during the laser pulse, were used to study the relationship between the dynamics of entry of condensed phase into the plume, shielding of the target by the particles, and brightening of the plume under the action of the incident laser radiation. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 73, No. 2, pp. 210–219, March–April, 2006.     

High quantum yield of photochemical reactions of protoporphyrin-IX induced by powerful ultrashort laser pulses

The action of powerful pulsed picosecond radiation from a Nd: YAG laser (λ=530 nm, pulse energy: 0.01 J, intensity: 2GW/cm²) and an argon laser (λ=515 nm, power: 50 mW) on protoporphyrin-IX dimethylether in three solvents (trichlormethane, carbon tetrachloride, dioxane) has been studied. Under continuous irradiation the quantum yield and resulting products do not differ materially from the ones produced under mercury lamp irradiation. When irradiation is performed by powerful laser pulses of picosecond duration the quantum yield of photodecomposition of protoporphyrin-IX dimethylether inereases substantially: by 10 in dioxane, by 4 in carbon tetrachloride and by 100 in trichlormethane. It is assumed that a quite different mechanism of multistep excitation is responsible for photodecomposition under powerful picosecond pulses.     

Processing of W/Si and Si/W bilayers and multilayers with single and multiple excimer-laser pulses

Interdiffusion phenomena, thermal damage and ablation of W/Si and Si/W bilayers and multilayers under XeCl-excimer laser (=308 nm) irradiation at fluences of 0.15, 0.3 and 0.6 J/cm² were studied. Samples were prepared by UHV e-beam evaporation onto oxidized Si. The thickness of W and Si layers and the total thickness of the structures were 1–20 nm and 40–100 nm, respectively. 1 to 300 laser pulses were directed to the same irradiation site. At 0.6 J/cm² the samples were damaged even by a single laser pulse. At 0.3 J/cm² WSi₂ silicide formation, surface roughening and ablation were observed. The threshold for significant changes depends on the number of pulses: it was between 3–10 pulses and 10–30 pulses for bilayers with W and Si surfaces, respectively, and more than 100 pulses for multilayers with the same total thickness of tungsten. At 0.15 J/cm² the periodicity of the multilayers was preserved. Temperature profiles in layered structures were obtained by numerical simulations. The observed differences of the resistance of various bilayers and multilayers against UV irradiation are discussed.     

The infrared spectrum of GeO

The high-temperature infrared spectrum of the Δv = 1 sequence of GeO has been investigated from 882 to 955 cm⁻¹ with a tunable diode laser spectrometer. Transitions from ⁷⁰Ge¹⁶O, ⁷²Ge¹⁶O, ⁷³Ge¹⁶O, ⁷⁴Ge¹⁶O, and ⁷⁶Ge¹⁶O were measured for vibrational transitions from v = 1−0 to v = 6−5 and fit to a single set of eight Dunham potential coefficients.     

Characterisation of laser-produced tungsten plasma using optical spectroscopy method

This paper describes results of spectroscopic investigation of laser-produced tungsten plasma. The laser intensity on the target surface reached up to 30 GW/cm² depending on the focusing conditions. Optical spectra emitted from plasma plumes which were formed under vacuum conditions in front of the tungsten target due to the interaction of Nd-YAG laser pulses (1.06 μm, 0.5 J), were characterised by means of an optical spectrometer (λ/Δλ= 900) in the wavelength range from 300 to 1100 nm. The spectra were recorded automatically with the use of a CCD detector with exposition time varied from 100 ns to 50 ms. On the basis of WI and WII lines it was possible to estimate electron temperature and electron density which corresponded to the expansion phase of the plasma. T_e and N_e were measured as 1.1 eV and 8×10¹⁶ cm^-3, respectively. The spectra collected by the ion energy analyser showed that the plasma included tungsten ions up to 6+ ion charge. Signals from the ion collector allowed to estimate the average value of ion energy of tungsten as 4.6 keV. Basing on this value the electron temperature corresponding to the initial stage of the plasma formation was estimated to be about 320 eV. Optical microscope investigation showed that laser irradiation caused structural changes on the surface of the target.     

Cytotoxic and photocytotoxic effect of Photofrin® on human laryngeal carcinoma (Hep2c) cell line

Significant apoptotic effect in Hep2c cell line has been investigated, when diode laser (λ = 635 nm, red) are used as a source of illumination and initiation of photodynamic action. The optimal uptake time of Photofrin® for Hep2c cell line was investigated by means of spectrophotometric measurement. Quantification of the live cell population was determined by means of neutral red assay (NRA). The spectrometry measurements showed that after 46 h incubation, the maximal cellular uptake of Photofrin® was achieved and photocytotoxic assay showed that light dose of 120 J/cm² give effective PDT outcome for Hep2c cell line incubated with 85 μg/ml of Photofrin®. No significant phototoxic and cytotoxic effects on Hep2c cells were observed due to light doses or photosensitize, when studied independently of each other and Photofrin® showed good anti tumor effects.     

Surgical effects on soft tissue produced by a 405-nm violet diode laser in vivo

This study evaluated the surgical performance of a 405-nm diode laser in vivo, using living rat liver tissue. Tissue was incised by irradiation with the laser at low output power ranging from 1 W (722 W/cm²) to 3 W (2165 W/cm²) on a manual control at a rate of 1 mm/s. As a control, incisions using a stainless scalpel were compared. Immediately after operation, the surface of the incisions was macroscopically observed and histopathologically evaluated by microscopy. Laser-ablated liver tissue was smooth with observable signs of remnant carbonization and easily acquired hemostasis. The thickness of the denatured layer increased in proportion to the output power; the coagulation layer did not thicken accordingly. Bleeding could not be stopped for tissues incised with the stainless scalpel. The 405-nm diode laser thus proved to be effective for ablating soft tissue with high hemostatic ability at low power.     

Diode laser irradiation effects on the sealing ability of root canal sealers

This study aimed to test the hypothesis that dentine alterations induced by 810 nm-diode laser may affect the interaction between root canal sealers and the dentin wall. Seventy-two single root human teeth were selected and root canals were enlarged with K-files. Dentine was treated with 0.5% NaOCl and 17% EDTA-T and irradiated (laser group) by diode laser (810 nm/P = 2.5W/I = 1989 W/cm²) or remained non-irradiated (control group). Six samples per group were analyzed by scanning electron microscopy (SEM). The remaining samples of each group were divided into three subgroups (n = 10) and sealed with one of the tested sealers (N-Rickert/AHPlus?/Apexit®). Apical leakage was estimated by evaluating penetration of 0.5% methylene-blue dye. SEM analysis revealed that dentine at the apical third in irradiated samples was melted and fusioned whereas non-irradiated samples exhibited opened dentinal tubules. Despite the morphological changes induced by irradiation, laser did not affect the sealing ability of N-Rickert and AHPlusTM sealers. However, the length of apical leakage in roots filled with Apexit® was lower in irradiated root canals than in non-irradiated samples (p < 0.05). Morphological changes of root canal walls promoted by diode laser irradiation may improve de sealing ability of Apexit®, a calcium hydroxide-based sealer, suggesting that improved sealing promoted by irradiation may represent an additional factor contributing to the endodontic clinical outcome.     

Increased viability of odontoblast-like cells subjected to low-level laser irradiation

Studies have shown that the increase of cell metabolism depends on the low level laser therapy (LLLT) parameters used to irradiate the cells. However, the optimal laser dose to up-regulate pulp cell activity remains unknown. Consequently, the aim of this study was to evaluate the metabolic response of odontoblast-like cells (MDPC-23) exposed to different LLLT doses. Cells at 20000 cells/cm² were seeded in 24-well plates using plain culture medium (DMEM) and were incubated in a humidified incubator with 5% CO₂ at 37°C. After 24 h, the culture medium was replaced by fresh DMEM supplemented with 5% (stress by nutritional deficit) or 10% fetal bovine serum (FBS). The cells were exposed to different laser doses from a near infrared diode laser prototype designed to provide a uniform irradiation of the wells. The experimental groups were: G1: 1.5 J/cm² + 5% FBS; G2: 1.5 J/cm² + 10% FBS; G3: 5 J/cm² + 5% FBS; G4: 5 J/cm² + 10% FBS; G5: 19 J/cm² + 5% FBS; G6: 19 J/cm² + 10% FBS. LLLT was performed in 3 consecutive irradiation cycles with a 24-hour interval. Non-irradiated cells cultured in DMEM supplemented with either 5 or 10% FBS served as control groups. The analysis of the metabolic response was performed by the MTT assay 3 h after the last irradiation. G1 presented an increase in SDH enzyme activity and differed significantly (Mann-Whitney test, p < 0.05) from the other groups. Analysis by scanning electron microscopy showed normal cell morphology in all groups. Under the tested conditions, LLLT stimulated the metabolic activity of MDPC-23 cultured in DMEM supplemented with 5% FBS and exposed to a laser dose of 1.5 J/cm². These findings are relevant for further studies on the action of near infrared lasers on cells with odontoblast phenotype.     

Structure and optical properties of carbon films obtained by multipulse pulsed laser deposition

We have obtained carbon thin films on silicon and glass substrates with multipulse pulsed laser irradiation of graphite under vacuum (p ≈ 2.6 Pa) using a high-frequency series of nanosecond laser pulses (τ = 85 ns, λ = 1060 nm) with pulse repetition frequency f ≈ 10–20 kHz and laser power density q ≈ 15–40 MW/cm². We established the optimal laser power density and laser pulse repetition frequency for obtaining amorphous nanostructured diamond-like films.     

Effects of swift heavy ion irradiation on the electrical characteristics of Au/n-GaAs Schottky diodes

Metal-semiconductor diode of Au/n-GaAs is studied under the irradiation of swift heavy ion (SHI) beam (80 MeV ¹⁶O⁶⁺), using in situ current-voltage characterization technique. The diode parameters like ideality factor, barrier height, and leakage current are observed to vary with irradiation fluence. Significantly, the diode performance improves at a high fluence of 2 × 10¹³ ions cm⁻² with a large decrease of reverse leakage current in comparison to the original as deposited sample. The Schottky barrier height (SBH) also increases with fluence. At a high irradiation fluence of 5 × 10¹³ ions cm⁻² the SBH (0.62 ± 0.01 eV) is much larger than that of the as deposited sample (0.55 ± 0.01 eV). The diode parameters remain stable over a large range of irradiation up to fluence of 8 × 10¹³ ions cm⁻². A prominent annealing effect of the swift ion beam owing to moderate electronic excitation and high ratio of electronic energy loss to the nuclear loss is found to be responsible for the improvement in diode characteristics.     

Effect of <Emphasis Type="Italic">in vivo</Emphasis> irradiation of blood by low-intensity emission from a He–Ne laser on molecular components of blood

We have used the IR absorption spectra of blood, plasma, and packed erythrocytes to study the effect of in vivo exposure of blood to low-intensity emission from a He–Ne laser (λ = 632.8 nm, 4 mW/cm²) on the molecular components of blood. In comparing the IR spectra of samples of blood sample and blood components before and after irradiation, we identified changes in the amide A, amide I, amide II, amide III absorption regions and also in the absorption bands of methyl, methylene, and phosphate groups. We conclude that exposure of blood in vivo to emission from a He–Ne laser leads to a decrease in the contribution of the α-helix conformation in the secondary structure of the blood proteins.     

Antimicrobial photodynamic therapy: The impact of laser radiation on mucous tissue stained with photosensitizer methylene blue

We have calculated the spatial distribution of absorbed light energy within layers of a mucous tissue that contains photosensitizer methylene blue. Under irradiation regimes that are typical of antimicrobial photodynamic therapy (laser radiation wavelength, 670 nm; power density, 150–300 mW/cm²; dose, 9–18 J/cm²), we have analyzed conditions that are necessary for the light penetration and delivery of methylene blue to a nidus of infection on a mucosal surface. We have performed a computer simulation of thermal fields, estimated maximal heating temperatures, and considered the degree at which the heating affects the trans-mission of light through a stained tissue.     

Chemical, morphological and accumulation phenomena in ultrashort-pulse laser ablation of TiN in air

Ultrashort-pulse laser ablation (τ=130 fs, λ=800 nm, repetition rate 2–20 Hz) of titanium nitride was investigated for laser fluences between 0.3 and 4.5 J/cm² using the direct focusing technique in air. The influence of the laser pulse number and the peak fluence was investigated by means of several surface analytical techniques (optical microscopy, dynamic friction atomic force microscopy, scanning Auger electron microscopy and small-spot electron spectroscopy for chemical analysis). The correlation of the results about optical, physical and chemical properties of the irradiated areas allows us to propose a simple oxidation model, which explains different observed phenomena associated with surface damage such as mound formation and crater widening and clarifies the incubation behavior reported earlier for this material. Received: 8 May 2000 / Accepted: 9 May 2000 / Published online: 13 September 2000     

Sum frequency generation at 309 nm using a violet and a near-IR DFB diode laser for detection of OH

We have used a violet diode laser at 404 nm and a distributed feedback diode laser at 1320 nm to produce 0.8 nW of radiation at 309 nm by sum frequency generation in beta-barium borate. The UV radiation was tuned mode-hop-free over 30 GHz and used to detect OH radicals produced in a microwave discharge. By chopping the UV light at 500 Hz, we observed a concentration of 2×10¹² cm^-3 with a signal to noise ratio of 30:1. Received: 16 November 2001 / Revised version: 23 January 2002 / Published online: 14 March 2002