Three-dimensional topographic scanning electron microscope and Raman spectroscopic analyses of the irradiation effect on teeth by Nd:YAG, Er: YAG, and CO(2) lasers

A three-dimensional analyzer installed in a scanning electron microscope was used to evaluate the morphology and surface roughness using noncontact profilometry. Observations were carried out on the enamel and dentin surface irradiated by three different lasers: Nd:YAG (wavelength 1.06 microm), Er:YAG (2.94 microm), and CO(2) (10.6 microm). Spectroscopic analysis was done by Raman spectroscopy for nonirradiated and laser-irradiated surfaces. The lasers were applied perpendicularly to vertically sectioned and polished human extracted caries-free molars. The tooth was sectioned at each cavity for cross-section analysis after laser irradiation. Irradiation by Nd:YAG and CO(2) lasers of the enamel surface showed an opaque white color, different from dentin where the surface turned black. The Er:YAG laser induced no changes in color of the dentin. Numerous cracks associated with thermal stress were observed in the CO(2) laser-irradiated dentin. Noncontact surface profile analysis of Er:YAG laser-irradiated enamel and dentin showed the deepest cavities, and direct cross-sectional observations of them showed similar cavity outlines. The CO(2) laser-irradiated dentin had the least surface roughness. Raman spectroscopic analysis showed that fluorescence from the laser-irradiated tooth was generally greater than from nonirradiated teeth. Bands in dentin attributed to organic collagen matrix were lost after Nd:YAG and CO(2) laser irradiation, and a broad peak due to amorphous carbon appeared. The Er:YAG laser-irradiated dentin showed no sign of a carbon band and had more suitable results for dental ablation. Noncontact surface profile analysis was effective to evaluate the structural change in the tooth in the microarea of study after laser irradiation.     

Rapid and conservative ablation and modification of enamel, dentin, and alveolar bone using a high repetition rate transverse excited atmospheric pressure CO2 laser operating at lambda=9.3 micro

Transverse excited atmospheric pressure (TEA) CO(2) lasers tuned to the strong mineral absorption of hydroxyapatite near lambda=9 microm are well suited for the efficient ablation of dental hard tissues if the laser pulse is stretched to greater than 5 to 10 micros to avoid plasma shielding phenomena. Such CO(2) lasers are capable of operating at high repetition rates for the rapid removal of dental hard tissues. The purpose of this study was to test the hypothesis that stretched lambda=9.3-microA CO(2) laser pulses can produce lateral incisions in enamel, dentin, and alveolar bone for dental restorations and implants at repetition rates as high as 400 Hz without peripheral thermal damage. The single pulse ablation rates through enamel, dentin, and bone were determined for incident fluence ranging from (1 to 160 J/m(2)) for laser pulses from 5 to 18 mus in duration. Lateral incisions were produced in hard tissue samples using a computer-controlled scanning stage and water spray, and the crater morphology and chemical composition were measured using optical microscopy and high-resolution synchrotron radiation infrared spectromicroscopy. The residual energy remaining in tooth samples was measured to be 30 to 40% for enamel and 20 to 30% for dentin without water cooling, under optimum irradiation intensities, significantly lower than for longer CO(2) laser pulses. The transmission through 2-m length 300-, 500-, 750-, and 1000-microm silica hollow waveguides was measured and 80% transmission was achieved with 40 mJ per pulse. These results suggest that high repetition rate TEA CO(2) laser systems operating at lambda=9.3 microm with pulse durations of 10 to 20 micros are well suited for dental applications.     

Cryogen spray cooling during laser tissue welding

Cryogen cooling during laser tissue welding was explored as a means of reducing lateral thermal damage near the tissue surface and shortening operative time. Two centimetre long full-thickness incisions were made on the epilated backs of guinea pigs, in vivo. India ink was applied to the incision edges then clamps were used to appose the edges. A 4 mm diameter beam of 16 W, continuous-wave, 1.06 microm, Nd:YAG laser radiation was scanned over the incisions, producing approximately 100 ms pulses. There was a delay of 2 s between scans. The total irradiation time was varied from 1-2 min. Cryogen was delivered to the weld site through a solenoid valve in spurt durations of 20, 60 and 100 ms. The time between spurts was either 2 or 4 s, corresponding to one spurt every one or two laser scans. Histology and tensile strength measurements were used to evaluate laser welds. Total irradiation times were reduced from 10 min without surface cooling to under 1 min with surface cooling. The thermal denaturation profile showed less denaturation in the papillary dermis than in the mid-dermis. Welds created using optimized irradiation and cooling parameters had significantly higher tensile strengths (1.7 +/- 0.4 kg cm(-2)) than measured in the control studies without cryogen cooling (1.0 +/- 0.2 kg cm(-2)) (p < 0.05). Cryogen cooling of the tissue surface during laser welding results in increased weld strengths while reducing thermal damage and operative times. Long-term studies will be necessary to determine weld strengths and the amount of scarring during wound healing.     

The role of mesoscopic modelling in understanding the response of dental enamel to mid-infrared radiation

Human dental enamel has a porous mesostructure at the nanometre to micrometre scales that affects its thermal and mechanical properties relevant to laser treatment. We exploit finite-element models to investigate the response of this mesostructured enamel to mid-infrared lasers (CO(2) at 10.6 microm and Er:YAG at 2.94 microm). Our models might easily be adapted to investigate ablation of other brittle composite materials. The studies clarify the role of pore water in ablation, and lead to an understanding of the different responses of enamel to CO(2) and Er:YAG lasers, even though enamel has very similar average properties at the two wavelengths. We are able to suggest effective operating parameters for dental laser ablation, which should aid the introduction of minimally-invasive laser dentistry. In particular, our results indicate that, if pulses of approximately 10 micros are used, the CO(2) laser can ablate dental enamel without melting, and with minimal damage to the pulp of the tooth. Our results also suggest that pulses with 0.1-1 micros duration can induce high stress transients which may cause unwanted cracking.     

Laser probe temperature control by measuring the returning infra-red radiation

The metal-tipped fibre or ‘laser probe’ developed for angioplasty comprises a metallic probe at the end of an optical fibre. The probe is heated by an argon or Nd: YAG laser and applied against the tissue to be vapourised. The heated probe generates infra-red radiation which is proportional to the temperature of the probe. The paper investigates the feasibility of a feedback control system that measures the temperature of the probe by detecting the infra-red radiation transmitted back through the fibre. The probe was initially heated by physical contact with a hot surface, and then by an argon laser via the optical fibre. The returning IR radiation was sensed by a lead sulphide detector, while probe temperature was simultaneously measured by a thermocouple. Temperatures as low as 200°C were measured through a 5 m long fibre during the laser heating of the probe. The detector signal increased in an exponential fashion as the probe temperature increased. A resolution of 1°C was obtained at a probe temperature of 400°C. It can be concluded that, for the laser probe, it is feasible to use a feedback control system which measures the infra-red radiation transmitted back through the same fibre that carries the heating laser light.     

Adhesion of composite to enamel and dentin surfaces irradiated by IR laser pulses of 0.5-35 micros duration

The characteristics of laser-treated tooth surfaces depend on the laser wavelength, pulse duration, spatial and temporal laser beam quality, incident fluence, surface roughness, and the presence of water during irradiation. Ablated surfaces are most commonly restored with adhesive dental materials and the characteristics of the ablated surfaces influence adhesion of restorative materials. Previous studies suggest that high bond strengths can be achieved using shorter laser pulses that minimize peripheral thermal damage. In this study, Er:YSGG, Er:YAG, and CO(2) lasers were used at irradiation intensities sufficient to simulate efficient clinical caries removal to uniformly irradiate bovine enamel and human dentin surfaces using a motion control system with a microprocessor-controlled water spray. The degree of spatial overlap of adjacent pulses was varied so as to investigate the influence of irradiation uniformity and surface roughness on the bond strength. Composite resin was bonded to the irradiated surfaces and shear bond tests were used to obtain bond strengths in MPa. The highest results were obtained using the Er:YAG pulses with pulse durations less than 35 mus without the necessity for postirradiation acid etching. Some of these groups were not significantly different from nonirradiated, acid-etch-only positive control groups.     

Ultrastructural observations in human oocytes and preimplantation embryos after zona opening using an erbium-yttrium-aluminium-garnet (Er:YAG) laser

For more than 3 years we have performed laser-assisted hatching prior to embryo transfer in patients with recurrent implantation failure using an erbium-yttrium-aluminium-garnet (Er:YAG) laser system that operates in the infrared region of the light spectrum. The laser beam is guided through a quartz fibre and is brought into direct contact with the zona pellucida. This study was undertaken to evaluate the ultrastructural effects of this laser on the zona pellucida and underlying cell membrane of unfertilized human oocytes and pathologically fertilized preimplantation embryos using light and scanning electron microscopy. The Er:YAG laser produces an almost circular zona opening in the shape of a truncated cone tapering off towards the inside, with a mean diameter of 18 mm. The exact diameter of the drilled site depends on the diameter of the fibre tip and the total number of pulses applied. After laser interaction, the zona matrix and the surface of the underlying ooplasm membrane showed no degenerative alterations. We conclude that the Er:YAG laser is an effective microsurgical tool for achieving reproducible, precise zona openings particularly suitable for purposes of assisted hatching because of their characteristic shape.      

Deep reflection-mode photoacoustic imaging of biological tissue

A reflection-mode photoacoustic (PA) imaging system was designed and built to image deep structures in biological tissues. We chose near-infrared laser pulses of 804-nm wavelength for PA excitation to achieve deep penetration. To minimize unwanted surface signals, we adopted dark-field ring-shaped illumination. This imaging system employing a 5-MHz spherically focused ultrasonic transducer provides penetration up to 38 mm in chicken breast tissue. At the 19-mm depth, the axial resolution is 144 microm and the transverse resolution is 560 microm. Internal organs of small animals were imaged clearly.     

Retinal response of Macaca mulatta to picosecond laser pulses of varying energy and spot size

We investigate the relationship between the laser beam at the retina (spot size) and the extent of retinal injury from single ultrashort laser pulses. From previous studies it is believed that the retinal effect of single 3-ps laser pulses should vary in extent and location, depending on the occurrence of laser-induced breakdown (LIB) at the site of laser delivery. Single 3-ps pulses of 580-nm laser energy are delivered over a range of spot sizes to the retina of Macaca mulatta. The retinal response is captured sequentially with optical coherence tomography (OCT). The in vivo OCT images and the extent of pathology on final microscopic sections of the laser site are compared. With delivery of a laser pulse with peak irradiance greater than that required for LIB, OCT and light micrographs demonstrate inner retinal injury with many intraretinal and/or vitreous hemorrhages. In contrast, broad outer retinal injury with minimal to no choriocapillaris effect is seen after delivery of laser pulses to a larger retinal area (60 to 300 microm diam) when peak irradiance is less than that required for LIB. The broader lesions extend into the inner retina when higher energy delivery produces intraretinal injury. Microscopic examination of stained fixed tissues provide better resolution of retinal morphology than OCT. OCT provides less resolution but could be guided over an in vivo, visible retinal lesion for repeated sampling over time during the evolution of the lesion formation. For 3-ps visible wavelength laser pulses, varying the spot size and laser energy directly affects the extent of retinal injury. This again is believed to be partly due to the onset of LIB, as seen in previous studies. Spot-size dependence should be considered when comparing studies of retinal effects or when pursuing a specific retinal effect from ultrashort laser pulses.     

Laser surface modification of hydroxyapatite and glass-reinforced hydroxyapatite

Surface treatment of materials with excimer laser radiation often results in the formation of a rough columnar or cone-shaped surface topography, which leads to a considerable increase in the surface area. As a result, the search for a non-porous bioactive material with adequate mechanical properties and a high surface to volume ratio, similar to porous materials, which could be used for drug delivery in the treatment of periodontitis, justified assessing excimer laser surface treatment to promote controlled roughning of hydroxyapatite (HA) and glass-reinforced hydroxyapatite (GR-HA). A KrF excimer laser with 248 nm radiation wavelength and 30 ns pulse duration was used for surface modification. The laser treatment was carried out in air, using wide ranges of radiation fluence and number of laser pulses. In order to identify the physico-chemical changes induced by the laser treatment and the column formation mechanisms in these materials, the treated surfaces were characterised by laser profilometry, scanning electron microscopy (SEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and Fourier transform infra-red spectroscopy (FTIR). Laser processing induced the formation of a surface topography consisting of cone-shaped features. The constitution of the surface layer was also modified, as revealed by FTIR, XPS and XRD. This work has shown that laser surface modification increases the surface area of HA and GR-HA and is a promising technique to increase the reactivity and drug delivery capability of both materials.     

Dental hard tissue modification and removal using sealed transverse excited atmospheric-pressure lasers operating at lambda=9.6 and 10.6 microm

Pulsed CO(2) lasers have been shown to be effective for both removal and modification of dental hard tissue for the treatment of dental caries. In this study, sealed transverse excited atmospheric pressure (TEA) laser systems optimally tuned to the highly absorbed 9.6 microm wavelength were investigated for application on dental hard tissue. Conventional TEA lasers produce an initial high energy spike at the beginning of the laser pulse of submicrosecond duration followed by a long tail of about 1-4 micros. The pulse duration is well matched to the 1-2 micros thermal relaxation time of the deposited laser energy at 9.6 microm and effectively heats the enamel to the temperatures required for surface modification at absorbed fluences of less than 0.5 J/cm(2). Thus, the heat deposition in the tooth and the corresponding risk of pulpal necrosis from excessive heat accumulation is minimized. At higher fluences, the high peak power of the laser pulse rapidly initiates a plasma that markedly reduces the ablation rate and efficiency, severely limiting applicability for hard tissue ablation. By lengthening the laser pulse to reduce the energy distributed in the initial high energy spike, the plasma threshold can be raised sufficiently to increase the ablation rate by an order of magnitude. This results in a practical and efficient CO(2) laser system for caries ablation and surface modification.     

Improving the penetration depth in multiphoton excitation laser scanning microscopy

More than a threefold increase in multiphoton laser scanning microscopy depth penetration using a passive predispersion compensation system is reported. Using dispersion-controlled pulses to counteract the effects of positive group delay dispersion in the imaging platform, optical sectioning of fluorescent samples to depths in excess of 800 microm was observed, compared with only 240 microm using a noncompensated setup. Experimental results obtained from both the predispersion compensated and noncompensated systems are compared with theoretical values of pulse broadening in a laser scanning microscope. The observed improvement in depth profiling potentially widens the applications and user base of nonlinear microscopy techniques.     

Laser reflectance imaging of human organs and comparison with perfusion images

Based on the principle of backscattering of laser radiation from tissues, a non-invasive PC-AT based reflectance imaging technique is developed. The laser beam from a semiconductor laser operating at 670 nm is guided to the tissue site by an optical fibre. The backscattered radiation is collected by another fibre placed in the same probe, and is detected by a photodiode-amplifier assembly. This probe is moved manually over the organs under observation, and the data after the ADC, interpolation and median filtering are displayed in the form of reflectance image of the organ along with grey scale. By this technique images of the human hands and forearms are obtained, which depend on the variations in their colour, composition and blood flow. A comparison is made with perfusion images, obtained by a Periflux laser Doppler flowmeter. These show that the reflectance images provide greater details of the tissue structure than the perfusion images.     

Cell behavior on laser surface-modified polyethylene terephthalate in vitro

This work has been undertaken to study the cell behavior of L929 fibroblasts on the laser irradiated polyethylene terephthalate (PET) surface. To modify the surface properties of the PET, CO2 pulsed laser at the wavelength of 9.25 microm and KrF excimer laser at 248 nm with various number of pulses were used. Laser irradiation caused some changes in the chemical and physical properties of the laser-treated film surfaces, which were evaluated using different techniques. These changes may affect the cell adhesion and growth on the laser-treated PET. Therefore, cell attachment and spreading were investigated on the laser-treated PET in vitro. The data from in vitro assays showed the fibroblast cells were attached and proliferated extensively on the CO2 and KrF laser-treated films in comparison with the unmodified PET. The results obtained from the cell behavior studies revealed that surface morphology and wettability affected cell adhesion and spreading on the laser-treated PET.     

Selective removal of composite sealants with near-ultraviolet laser pulses of nanosecond duration

It is often necessary to replace pit and fissure sealants and composite restorations. This task is complicated by the necessity for complete removal of the remaining composite to enable suitable adhesion of new composite. Previous studies have shown that 355-nm laser pulses from a frequency-tripled Nd:YAG laser can selectively remove residual composite after orthodontic bracket removal on enamel surfaces. Our objective is to determine if such laser pulses are suitable for selective removal of composite pit and fissure sealants and restorations. Optical coherence tomography is used to acquire optical cross sections of the occlusal topography nondestructively before sealant application, after sealant application, and after sealant removal. Thermocouples are used to monitor the temperature in the pulp chamber during composite removal under clinically relevant ablation rates, i.e., 30 Hz and 30 mJ/pulse. At an irradiation intensity of 1.3 J/cm2, pit and fissure sealants are completely removed without visible damage to the underlying enamel. At intensities above 1.5 J/cm2, incident laser pulses remove the resin layer while at the same time preferentially etching the surface of the enamel. Temperature excursions in the pulp chamber of extracted teeth are limited to less than 5 degrees C if air-cooling is used during the rapid removal (1 to 2 min) of sealants, water-cooling is not necessary. Selective removal of composite restorative materials is possible without damage to the underlying sound tooth structure.     

Absence of large-diameter sensory fibres in a nerve to the cat humerus

A fine branch of the median nerve innervates the periosteum and medullary cavity of the cat humerus. After branching to innervate the periosteum on the medial surface of the humerus, the nerve enters and supplies the medullary cavity via a nutrient foramen, accompanied by a small artery and vein. The composition of the fibres in the nerve was examined using electron microscopy. Myelinated fibres with diameters of 0.8-6.6 microm and unmyelinated fibres with diameters of 0.1-1.4 microm were observed. These diameters indicate that afferent fibres of this nerve are confined within the Group III and IV categories, and may therefore be nociceptive or mechanoreceptive in function. In addition, autonomic efferent fibres may also be present in these fibre groups. As no fibre diameters greater than 7 microm were noted, it appears that Group I and II fibres are absent in this nerve. The fibre distribution suggests that the principal role of this nerve is to relay bone-related nociceptive or mechanoreceptive information to the central nervous system and to provide autonomic regulatory influences on the bone.     

Accelerating protons to therapeutic energies with ultraintense, ultraclean, and ultrashort laser pulses

Proton acceleration by high-intensity laser pulses from ultrathin foils for hadron therapy is discussed. With the improvement of the laser intensity contrast ratio to 10(-1) achieved on the Hercules laser at the University of Michigan, it became possible to attain laser-solid interactions at intensities up to 10(22) W/cm2 that allows an efficient regime of laser-driven ion acceleration from submicron foils. Particle-in-cell (PIC) computer simulations of proton acceleration in the directed Coulomb explosion regime from ultrathin double-layer (heavy ions/light ions) foils of different thicknesses were performed under the anticipated experimental conditions for the Hercules laser with pulse energies from 3 to 15 J, pulse duration of 30 fs at full width half maximum (FWHM), focused to a spot size of 0.8 microm (FWHM). In this regime heavy ions expand predominantly in the direction of laser pulse propagation enhancing the longitudinal charge separation electric field that accelerates light ions. The dependence of the maximum proton energy on the foil thickness has been found and the laser pulse characteristics have been matched with the thickness of the target to ensure the most efficient acceleration. Moreover, the proton spectrum demonstrates a peaked structure at high energies, which is required for radiation therapy. Two-dimensional PIC simulations show that a 150-500 TW laser pulse is able to accelerate protons up to 100-220 MeV energies.     

Fast three-dimensional laser scanning scheme using acousto-optic deflectors

A scheme for fast 3-D laser scanning using acousto-optic deflectors is proposed and demonstrated. By employing counterpropagating acoustic waves that are both chirped and offset in their frequencies, we show that it is possible to simultaneously scan both axially and laterally with frame rates on the order of tens of kilohertz. This scheme was specifically designed for application with multiphoton imaging, particularly of neurons, where it will enable the concurrent monitoring of physiological signals at multiple locations within a microscopic 3-D volume (350 x 350 x 200 microm). When used for this purpose, we demonstrate how this scheme would also inherently compensate for spatial dispersion when ultrafast laser pulses are used in acousto-optic multiphoton microscopy.     

Carbon coated polyethylene: effect of surface energetics and topography on human platelet adhesion

The influence of surface energy and structural properties of carbon coated polyethylene (PE) on the human platelet adhesion was studied. Three types of amorphous carbon coating were obtained by plasma pulse discharge, with the number of pulses grading as 10, 50, 100. Human serum albumin adsorption experiments have been carried out with all samples in vitro. Platelet adhesion analysis by SEM included determination of total quantity of adherent platelets, and respective quantities of platelets at different stages of activation (single, spread, aggregates). Surface topographies ranged from bare PE and such (10 pulses), to globular 0.5 microm in size (50 pulses), and complex fibrillar 3-4 microm structures (100 pulses). Surface free energy varies from 31.7 +/- 0.6 to 40.4 +/- 0.6 mN/m for uncoated PE and 10 pulse coatings, respectively, as determined by contact angle techniques. All studied coatings demonstrate weaker platelet activation properties in comparison with untreated PE. Among all studied coatings, the 50 pulse coated surface seems to be the least suitable for contact with platelets, mainly due to its structural rather than to its energy properties. These data are related to a sharp decrease in the adsorbed protein level for the samples with 50 pulse coatings. The applied analysis of platelet activation enables more accurate characterization of platelet-biomaterial interaction.