Controlled ablation of microtubules using a picosecond laser.

The use of focused high-intensity light sources for ablative perturbation has been an important technique for cell biological and developmental studies. In targeting subcellular structures many studies have to deal with the inability to target, with certainty, an organelle or large macromolecular complex. Here we demonstrate the ability to selectively target microtubule-based structures with a laser microbeam through the use of enhanced yellow fluorescent protein (EYFP) and enhanced cyan fluorescent protein (ECFP) variants of green fluorescent protein fusions of tubule. Potorous tridactylus (PTK2) cell lines were generated that stably express EYFP and ECFP tagged to the alpha-subunit of tubulin. Using microtubule fluorescence as a guide, cells were irradiated with picosecond laser pulses at discrete microtubule sites in the cytoplasm and the mitotic spindle. Correlative thin-section transmission electron micrographs of cells fixed one second after irradiation demonstrated that the nature of the ultrastructural damage appeared to be different between the EYFP and the ECFP constructs suggesting different photon interaction mechanisms. We conclude that focal disruption of single cytoplasmic and spindle microtubules can be precisely controlled by combining laser microbeam irradiation with different fluorescent fusion constructs. The possible photon interaction mechanisms are discussed in detail.

Er:YAG laser modification of root canal dentine: influence of pulse duration, repetitive irradiation and water spray.

The aim of this study was to determine the effects of varying parameters of Er:YAG laser irradiation with and without water spray cooling on root canal dentine in vitro. After horizontally removing tooth crowns from extracted human teeth, roots were axially sectioned into thin slices, exposing the root canal surface. An Er:YAG laser delivered 10-30 J/cm(2) into a 0.4-mm diameter laser spot on the root canal surface. Single pulses of different lengths (80-280 micro s) were applied with and without water spray cooling/irrigation, and sequences of three pulses at a repetition rate of 30 Hz were applied at selected pulse parameters. The irradiated samples were investigated using both confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). At most irradiation conditions, the root canal dentine surface was ablated. Three-dimensional images from CLSM revealed that the cavity walls were not smooth. Depths of the cavities revealed significant differences between the cavities. No debris was observed at the surface of cavities at any irradiation condition. Strong melting and recrystallisation, or unusually flat surfaces with open dentinal tubules were obtained with sequences of three pulses without water cooling. CLSM is an effective tool for investigation of laser effects on root canal dentine. By varying the irradiation conditions, the Er:YAG laser can induce different modifications of root canal surface, which may be very interesting for root canal preparation.

Simple organ cornea culture model for re-epithelialization after in vitro excimer laser ablation.

BACKGROUND AND OBJECTIVE: Most of the in vitro work to characterize the effects of clinical laser surgery on corneal tissues has concentrated on the effects on stromal keratocytes and endothelium with little attention being paid to corneal epithelium. Our purpose is to describe the epithelial healing rates observed in freshly cultured rabbit corneas treated with phototherapeutic keratectomy (PTK). STUDY DESIGN/MATERIALS AND METHODS: Corneas were placed in a simple organ culture system, with media change every 2 days. A clinical excimer laser was used to perform a 6 mm diameter, 100 microm depth transepithelial PTK on 24 cultured rabbit corneas, 1 day after culture initiation. For each post-treatment day, one experimental and one control cornea were removed from culture and stained with fluorescein, photographed, and fixed for histology. Epithelial defect area was measured with digital imaging software and analyzed statistically to assess the re-epithelialization rate. RESULTS: Control corneas, maintained in culture for 1-4 days, had no epithelial defects. Those corneas treated with PTK exhibited an immediate epithelial defect that slowly healed over 3 days. This was confirmed on histopathological analysis. A significant linear trend in re-epithelialization across the time points studied was found (F = 80.48, P = 0.0029). The slope of the linear regression model showed an estimate rate of re-epithelialization of -6.70 over the 3 days. CONCLUSION: We have described the development of a simple, whole organ, rabbit cornea culture model for re-epithelialization after PTK. Our rates of epithelial healing resemble those found in the literature in live rabbit models. Therefore, this model may possibly be used to monitor epithelial wound healing in different corneal diseases or injuries.

Intravitreal VEGF and bFGF produce florid retinal neovascularization and hemorrhage in the rabbit.

PURPOSE: Vascular endothelial growth factor (VEGF) causes widespread retinal vascular dilation, produces breakdown of the blood-retinal barrier, and is implicated in ocular neovascularization (NV). Basic fibroblast growth factor (bFGF) also has been implicated in the production of ocular NV. This study was performed to investigate the ability of simultaneous sustained intravitreal release of both VEGF and bFGF to induce robust retinal NV in the rabbit. METHODS: Intravitreal implantation of sustained-release Hydron polymeric pellets containing both 20 microg of VEGF and 20 microg of bFGF was performed on adult male Dutch belted rabbits. In other animals either 20 microg or 50 microg bFGF-containing pellets was implanted intravitreally; also, either 20 microg VEGF or 50 microg VEGF-containing pellets was implanted. Control rabbits received either blank polymeric pellets or a pellet containing 30 microg bovine serum albumin. Eyes were examined by indirect ophthalmoscopy after surgery at 24 hrs, 48 hrs, 4 days, 7 days, 14 days, 21 days, and 28 days. Findings were documented by color fundus photography and fluorescein angiography (FA). Eyes were enucleated and prepared for histologic analysis at 28 days following intravitreal implantation of the VEGF/bFGF-containing pellets. RESULTS: In all eyes implanted with VEGF/bFGF pellets, dilation and tortuosity of existing blood vessels were observed within 48 hrs after pellet implantation. The progression of retinal vascular changes was rapid and occurred over the entire optic disk and medullary rays between 4 and 7 days. Hemorrhage occurred as early as 14 days after VEGF/bFGF pellet implantation. In eyes with massive hemorrhage, total traction retinal detachment developed after the second week. The presence of abnormal tissues at the vitreo-retinal interface within 28 days was demonstrated by light microscopy while FA showed profuse leakage of dye from anomalous vessels within the first week. Neither bFGF-exposed eyes nor control eyes showed any vascular changes. Eyes that received only VEGF-containing pellets exhibited tortuosity of existing vessels, but neither hemorrhaging nor retinal detachment occurred. CONCLUSIONS: These results demonstrate that retinal vascular changes leading to hemorrhaging is produced rapidly in the rabbit by simultaneous intravitreal release of both VEGF and bFGF. Understanding how these growth factors induce retinal NV may suggest novel therapeutic treatment strategies.

Angioresistance of thermally modified cartilage grafts in the chick chorioallantoic membrane model.

BACKGROUND: The chick chorioallantoic membrane (CAM) model allows direct observation of vascularization acutely in explanted or cultured tissues in an immunologically isolated environment. In vivo, angioinvasion of the tissue matrix does not occur in viable cartilage tissue, whereas denatured or nonviable grafts are readily vascularized and/or resorbed. OBJECTIVE: To determine, using the CAM model, whether angioinvasion of thermally altered cartilage explants occurs acutely. MATERIALS AND METHODS: Porcine septal cartilage specimens were removed from freshly killed animals and divided into 3 groups (n = 10): an untreated control group, a group in which cartilage was boiled in isotonic sodium chloride solution (normal saline) for 1 hour, and a laser-irradiated group (Nd:YAG, lambda = 1.32 microm, 30.8 W/cm2, irradiation time = 10 seconds). Tissue specimens were then washed in antibiotic solutions, cut into small cubes (approximately 1.5 mm3), placed on the surface of 30 CAMs (7 days after fertilization), and allowed to incubate for an additional 7 days. After incubation, the membranes and specimens were fixed in situ with formaldehyde and then photographed using a dissection microscope. RESULTS: Examination with a dissecting microscope showed no obvious vascular invasion of the cartilage or loss of gross tissue integrity in any of the 3 experimental groups, although all specimens were completely enveloped by the CAM vascular network. No vascular invasion of the tissue matrix was observed histologically. CONCLUSION: These experiments demonstrate that cartilage specimens remain acutely resistant to angioinvasion or metabolism by the immunologically immature CAM whether native unmodified tissue, completely denatured (boiled), or thermally modified following laser irradiation.

Morphological study of the effects of CO2 laser emitted at 9.3 microm on human dentin.

OBJECTIVE: The purpose of this study was to investigate the effects of dentin ablation using a carbon dioxide (CO2) laser emitted at 9.3 microm by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). BACKGROUND DATA: There have been no reports on effects of CO2 laser irradiation emitted at 9.3 microm on dentin by SEM and CLSM. METHODS: Thirty extracted human teeth showing no clinical signs of caries were used. All teeth were horizontally sectioned to approximately 200 microm thickness and sections were irradiated using a 9.3 microm CO2 laser at different parameters as follows: 26 mJ [energy density (ED) 53.0 J/cm2] and 30 mJ (ED 61.1 J/cm2). After laser irradiation, samples were treated with sodium hypochlorite, stained using rhodamine-123, and observed with CLSM followed by SEM procedures. RESULTS: No craters or cracks were observed, but many small molten and rehardened particles were documented on the sample surface using SEM. Some small cracks were seen in the subsurface layer, and some patent dentinal tubules were detected using CLSM. CONCLUSION: These results suggest that laser irradiation at these parameters affected the sample surface only (less than 20 microm) and would be less harmful to thermal damage of dental pulp for dentin ablation.

Scanning electron microscopy comparison of corneal epithelial removal techniques before photorefractive keratectomy.

PURPOSE: To evaluate and compare the most common epithelial removal techniques by scanning electron microscopy. SETTINGS: Beckman Laser Institute, University of California, Irvine, California, USA. METHODS: The epithelium was removed in both eyes of 5 Dutch belted rabbits by 1 of 5 techniques: blunt mechanical debridement (with a blunt spatula), sharp mechanical debridement (with a sharp scalpel blade), application of topical alcohol, and transepithelial removal with the excimer laser and with the rotary epithelial scrubber. The animals were immediately killed and corneal specimens prepared for scanning electron microscopy. The degree of corneal trauma and the completeness of epithelial cell removal with the 5 techniques were evaluated by comparing the regularity and smoothness of the resulting stromal surfaces. RESULTS: Epithelial removal with alcohol, the excimer beam, and the rotary epithelial scrubber produced a smoother, more uniform anterior stromal surface than blunt and sharp mechanical debridement. CONCLUSIONS: The smooth and uniform anterior stromal surface produced with the rotary epithelial scrubber, alcohol, and the laser beam may be preferable prior to photorefractive keratectomy.

Systemic application of photosensitizers in the chick chorioallantoic membrane (CAM) model: photodynamic response of CAM vessels and 5-aminolevulinic acid uptake kinetics by transplantable tumors.

The aim of this study is to modify the chick chorioallantoic membrane (CAM) model into a whole-animal tumor model for photodynamic therapy (PDT). By using intraperitoneal (i.p.) photosensitizer injection of the chick embryo, use of the CAM for PDT has been extended to include systemic delivery as well as topical application of photosensitizers. The model has been tested for its capability to mimic an animal tumor model and to serve for PDT studies by measuring drug fluorescence and PDT-induced effects. Three second-generation photosensitizers have been tested for their ability to produce photodynamic response in the chick embryo/CAM system when delivered by i.p. injection: 5-aminolevulinic acid (ALA), benzoporphyrin derivative monoacid ring A (BPD-MA), and Lutetium-texaphyrin (Lu-Tex). Exposure of the CAM vasculature to the appropriate laser light results in light-dose-dependent vascular damage with all three compounds. Localization of ALA following i.p. injections in embryos, whose CAMs have been implanted with rat ovarian cancer cells to produce nodules, is determined in real time by fluorescence of the photoactive metabolite protoporphyrin IX (PpIX). Dose-dependent fluorescence in the normal CAM vasculature and the tumor implants confirms the uptake of ALA from the peritoneum, systemic circulation of the drug, and its conversion to PpIX.

In vitro and in vivo photosensitizing capabilities of 5-ALA versus photofrin in vascular endothelial cells.

BACKGROUND AND OBJECTIVE: The objective of the present study was to evaluate the feasibility of photodynamic therapy (PDT) for complicated hemangiomas. The photosensitizing activities of 5-aminolevulinic acid (5-ALA) and Photofrin were evaluated in vitro with human dermal microvascular endothelial cells (MEC) and in vivo with the chicken cox comb. STUDY DESIGN/MATERIALS AND METHODS: The in vitro absorption and photosensitizing activities of 5-ALA and Photofrin were examined in a MEC culture system. The percentages of MEC killed by different drug concentrations at a wavelength of 630 nm were measured by either live/dead or lactate dehydrogenase-released assays. Similarly, the in vivo biological activities of 5-ALA and Photofrin exposed to different total light dosages at 630 nm were studied by determining the amount of necrosis produced in chicken combs. RESULTS: MEC incubated with 5-ALA at a concentration of 35 microg/ml and exposed to laser light at 630 nm at a power density of 100 mW/cm2 showed a 50% cell kill. MEC incubated with Photofrin at a concentration of 3.5 microg/ml and exposed to laser light at 630 nm at a power density of 100 mW/cm2 showed a 50% cell kill. Chicken combs that received 200 mg/kg of 5-ALA exposed to laser light at 630 nm at a power density of 100 mW/cm2 had an injury depth of 362.5+/-27.6 microm at histologic examination. Combs exposed to a power density of 100 or 120 mW/cm2 showed injury depths of 732.5+/-29.1 and 792.5+/-36.0 microm, respectively. Chicken combs that received 2.5 mg/kg of Photofrin exposed to laser light at 630 nm at a power density of 80 mW/cm2 had an injury depth of 535.6+/-22.3 microm at histologic examination. Combs exposed to a power density of 100 or 120 mW/cm2 showed injury depths of 795.8+/-32.5 and 805.2+/-49.1 microm, respectively. CONCLUSION: Both 5-ALA and Photofrin have the capability to destroy MEC in vitro and vasculature in vivo. However, Photofrin achieved a higher degree of cell kill and tissue destruction at lower drug concentrations and at lower power densities.

Human corneal ablation threshold using the 193-nm ArF excimer laser.

PURPOSE: To determine the human corneal threshold ablation energy density for the 193-nm ArF excimer laser, approximating clinical conditions. METHODS: The VISX Star (Santa Clara, CA) 193-nm argon fluoride excimer laser was used to ablate the cornea in human eye bank eyes under clinical conditions. Corneas were exposed to energy densities of 10, 20, 30, 35, 40, 45, and 140 to 160 mJ/cm2. Corneas were fixed for light and transmission electron microscopy immediately after laser exposure. RESULTS: Different ablation thresholds for various corneal structural elements were observed. The ablation threshold for the collagen in the corneal stroma was determined to be 30 mJ/cm2. Keratocytes had ablation thresholds of 40 mJ/cm2. These different ablation thresholds accounted for the production of stromal peaks and valleys, with the keratocytes atop the peaks. CONCLUSIONS: Different corneal structural elements have different ablation threshold energy densities.

Effects of a 532 nm Q-switched nanosecond pulsed laser on dentin.

The purpose of this study was to determine whether a nanosecond-pulsed, frequency-doubled Nd:YAG laser emitting at 532 nm can be used as an alternative to mechanical methods of root canal treatment or as an adjunct to conventional endodontic preparation. Laser parameters whose thermal effects did not exceed safety thresholds for adjacent periodontal tissues were selected in a preliminary study. In 27 extracted human teeth, root canals were irradiated for 30 to 60 s at fluences of 2 to 2.2 J/cm2, and 10 Hz. Samples were observed using SEM. Laser irradiation could achieve smear layer removal after minimal manual preparation. However, results were inhomogeneous, and at higher energy densities thermal damage was observed, especially in the fully manually prepared samples. Nanosecond-pulsed irradiation at 532 nm can achieve complete smear layer removal. However, mechanisms must be developed to monitor laser effects and avoid potential damage to collateral structures.

In vivo detection of metastatic ovarian cancer by means of 5-aminolevulinic acid-induced fluorescence in a rat model.

STUDY OBJECTIVES: To determine the feasibility of macroscopic visualization of small ovarian cancer metastases in vivo by fluorescence after intravenous administration of 5-aminolevulinic acid (ALA); to assess the time after drug injection when fluorescence of small metastases is maximum; and to correlate macroscopic in vivo fluorescence with both microscopic ex vivo fluorescence and histologic findings. DESIGN: Controlled animal study (Canadian Task Force classification I). SETTING: University-based facility. SUBJECTS: Twenty-four healthy, female Fischer rats. INTERVENTION: Diffuse peritoneal metastatic cancer was induced in Fischer 344 rats by intraperitoneal injection of 1 million syngeneic ovarian cancer cells (NuTu-19). Four weeks after induction ALA100 mg/kg was injected intravenously, and diagnostic laparotomy was performed 1, 3, 6, or 9 hours thereafter. MEASUREMENTS AND MAIN RESULTS: The peritoneal cavity was illuminated with the Wood's lamp (ultraviolet light). Fluorescence was determined by direct visualization and compared with a calibrated fluorescent disk. Tissues were collected, sectioned, and examined by fluorescence and conventional light microscopy. Within 1 to 3 hours after intravenous injection of ALA, in vivo fluorescence of tumor nodules (diameter 0.4-5.0 mm) was macroscopically visible. Tumor-free peritoneum did not show fluorescence and was significantly distinguishable from cancer nodules. Fluorescence from intestinal tissues was comparable with tumor nodules. Microscopic fluorescence analysis showed similar values for tumor nodules and peritoneum. Stained histologic specimens of peritoneal surface revealed a superficial layer of cancer cells responsible for fluorescence. The time course of the fluorescence curve in the intestine peaked twice, at 1 and 6 hours after ALA injection. Macroscopically fluorescing nodules were histology confirmed as malignant. CONCLUSIONS: Fluorescence detection of small cancer nodules after intravenous injection of ALA is feasible for nodules smaller than 0.5 mm on the peritoneum. One to 3 hours after drug injection is optimal for diagnosis of metastases.

XeCl laser surgery of the vocal cords: a histologic comparison with CO2 laser in a porcine model.

Fresh cadaveric pig larynxes were ablated with a CO2 (lambda = 10.6 microm) and a XeCl excimer (lambda = 308 nm) laser. Histologic comparison of the ablation craters created by the two lasers was performed, and ablation crater depth and marginal tissue damage were measured. Crater depth for both laser treatments is correlated with energy deposition and exposure time. The CO2 laser creates three times more nonspecific, marginal tissue damage than the XeCl laser at the ranges of total energy and exposure times used. This study demonstrates the potential of the XeCl laser as an alternative to the CO2 laser in microlaryngeal surgery.

Photodynamic therapy of the ciliary body with tin ethyl etiopurpurin and tin octaethyl benzochlorin in pigmented rabbits.

BACKGROUND AND OBJECTIVES: The authors used a pigmented rabbit model to investigate two photosensitizers, tin ethyl etiopurpurin (SnET2) and tin octaethyl benzochlorin (BNZ 203), to determine their potential for creating ciliary body injuries during photodynamic therapy (PDT). MATERIALS AND METHODS: The biodistribution of SnET2 (n = 10) and BNZ 203 (n = 9) was studied by fluorescence microscopy using a low light detection system, based on charged-coupled device photography, with digital image processing at 1 and 24 hours after injection. PDT with SnET2 (n = 8; 664 +/- 7-nm light; 75 mW/cm2; 50 or 100 J/cm2; 1-mm spot size) and BNZ 203 (n = 6; 689 nm; 75 mW/cm2; 50 or 100 J/cm2; 1-mm spot size) was performed at 24 hours post-injection. The control subjects for SnET2 (n = 5) and BNZ 203 (n = 3) were given a maximal light dose (100 J/cm2). RESULTS: Both photosensitizers demonstrated an intravascular distribution at 1 hour that shifted to a ciliary body distribution at 24 hours (SnET2 much greater than BNZ 203). In addition, the SnET2 demonstrated suborgan localization to the nonpigmented ciliary body epithelium. Both photosensitizing agents were able to produce selective injury to the rabbit ciliary body (SnET2 much greater than BNZ 203), with evidence of a small component of thermal damage (SnET2 greater than BNZ 203). CONCLUSIONS: PDT with SnET2 or BNZ 203 can produce selective injury to the pigmented rabbit ciliary body. The nonpigmented ciliary body epithelium exhibits selective retention of SnET2. This finding warrants further investigation.

Cryogen spray cooling during Nd:YAG laser treatment of hemangiomas. A preliminary animal model study.

BACKGROUND: Successful laser treatment of hemangiomas requires selective photothermal destruction of dilated cutaneous vessels without damaging the overlying epidermis. Delivering a short cryogen spurt, on the order of milliseconds, has been shown to result in localized cooling of the superficial skin structures during laser irradiation. OBJECTIVE: The purpose of this study was to examine the effectiveness of cryogen spray cooling (CSC) in protecting superficial tissue structures during continuous Nd:YAG laser irradiation of an in vivo model hemangioma. METHODS: The highly vascularized chicken comb was selected as the animal model for hemangiomas. The Nd:YAG laser irradiation ranged from 2.6 to 35.1 J/mm2. A feedback system utilizing infrared radiometry monitored the comb surface temperature and controlled delivery time of the cryogen spurt. When comb surface temperature during laser irradiation reached 36-42 degrees C, a 30-100 msec cryogen spurt was delivered. Animals were euthanized 1 hour to 21 days following each experiment. Gross and histologic analyses were performed. RESULTS: Nd:YAG laser irradiation resulted in deep (up to 6.1 mm) tissue photocoagulation, while CSC preserved the overlying epidermis and papillary dermis. CONCLUSION: The results demonstrate that CSC is effective in protecting the epidermis and papillary dermis, while achieving deep tissue photocoagulation during Nd:YAG laser irradiation. Further pilot studies in humans appear warranted.

Photodynamic therapy for antifibrosis in a rabbit model of filtration surgery.

BACKGROUND AND OBJECTIVE: The purpose of this pilot study was to investigate the use of photodynamic therapy (PDT) using tin ethyl etiopurpurin (SnET2) as an adjunctive antifibrotic therapy for filtering surgery in a rabbit model. MATERIALS AND METHODS: The pharmacokinetics of SnET2 were established by intravenous (1 mg/kg) and subconjunctival (25, 50, or 75 micrograms) injections and compared with controls. Intravenous and subconjunctival SnET2 injections were given prior to posterior lip sclerectomies followed by postoperative laser irradiation (664 +/- 7 nm; 100 mW/cm2; 30 J/cm2). Antifibrotic efficacy was established by clinical response and histologic examination. RESULTS: After subconjunctival injections, large areas of avascular conjunctiva were produced and filtering bleb survival was prolonged. No effect was found for intravenously administered photosensitizer followed by light irradiation. CONCLUSIONS: PDT may be an alternative antifibrotic therapy for filtration surgery that does not use chemotherapeutic agents or ionizing radiation. Multiple parameters (light, drug dose, irradiation area) may be manipulated to improve predictability of the antifibrotic effect.

Nd:YAG laser irradiation in conjunction with cryogen spray cooling induces deep and spatially selective photocoagulation in animal models.

Successful laser treatment of haemangiomas requires selective photocoagulation of subsurface targeted blood vessels without thermal damage to the overlying epidermis. We present an in vivo experimental procedure, using a chicken comb animal model, and an infrared feedback system to deliver repetitive cryogen spurts (of the order of milliseconds) during continuous Nd:YAG laser irradiation. Gross and histologic observations show deep-tissue photocoagulation is achieved, while superficial structures are protected from thermal injury due to cryogen spray cooling. Experimental observation of epidermis protection in chicken comb animal models suggests selective photocoagulation of subsurface targeted blood vessels for successful treatment of haemangiomas can be achieved by repetitive applications of a cryogen spurt during continuous Nd:YAG laser irradiation.

Morphological effects of ArF excimer laser irradiation on enamel and dentin.

BACKGROUND AND OBJECTIVE: The aim of this investigation to determine the range of morphological and ablative effects that can be achieved on dental enamel and dentin using ArF excimer laser irradiation at a wavelength of 193nm. STUDY DESIGN/MATERIALS AND METHODS: Caries-free coronal enamel and dentin surfaces of 20 extracted human teeth were subjected to irradiation at 193nm using a Lamda-Physik model EMG 103 MSC and ArF fill. Morphology of cavity floors and walls were assessed by light microscopy and SEM. RESULTS: Morphological surface effects and ablation could be controlled effectively and reliably by choice of parameter combination, allowing the operator to achieve either a smooth, flat, or increasingly rough surface with differing degrees of selective ablation. No signs of thermal damage were apparent. CONCLUSION: Excimer laser irradiation at 193nm provided clinically useful cavity preparations and surface morphological effects.

Effects of nanosecond pulsed Nd: YAG laser irradiation on dentin resistance to artificial caries-like lesions.

BACKGROUND AND OBJECTIVE: Previous investigations have demonstrated improved enamel caries resistance after laser irradiation. In this study, effects of nanosecond pulsed Nd:YAG laser irradiation on crown/root dentin susceptibility to caries-like lesions were investigated. STUDY DESIGN/MATERIALS AND METHODS: Extracted human molar teeth were irradiated using a Q-switched nanosecond pulsed Nd:YAG laser. All teeth except controls were irradiated at: fluence, 1 or 5 J/cm2; spot size, 3 mm; and then subjected to demineralization. Measurements of caries lesion depth using scattering light microscopy and SEM were performed. RESULTS: Lesion depth measurements did not differ significantly between controls and irradiated samples (P < 0.01), but SEM results showed some irradiation-induced alterations on crown and root dentin surfaces. Irradiated surfaces were partially melted, with sometimes narrowed or occluded tubules. CONCLUSION: No consistent caries-protective effect of Q-switched nanosecond pulsed Nd:YAG laser irradiation on crown and root dentin was determined, but laser-induced morphological changes were observed.