Lih-Huei L. Liaw

Enhanced detection of early-stage oral cancer in vivo by optical coherence tomography using multimodal delivery of gold nanoparticles

Contrast in optical coherence tomography (OCT) images can be enhanced by utilizing surface plasmon resonant gold nanoparticles. To improve the poor in vivo transport of gold nanoparticles through biological barriers, an efficient delivery strategy is needed. In this study, the improved penetration and distribution of gold nanoparticles were achieved by microneedle and ultrasound, respectively, and it was demonstrated that this multimodal delivery of antibody-conjugated PEGylated gold nanoparticles enhanced the contrast in in vivo OCT images of oral dysplasia in a hamster model.

Root Canal Preparation Using the Second Harmonic KTP:YAG Laser: A Thermographic and Scanning Electron Microscopic Study

Thermal and microstructural events resulting from KTP laser use during root canal preparation were investigated in 30 extracted single-rooted human teeth. In the first section of this study, thermal events occurring on the root surfaces of 18 teeth during and after exposure of the root canal were measured using thermography. A variety of parameters were used to determine settings that would be effective without causing thermal damage to the periodontal ligament. In the second section of the study, root canals of 12 teeth exposed to KTP laser irradiation at parameters derived from section 1 were evaluated using Scanning electron microscopy. KTP laser application at a power setting of 3 W, an exposure time of 2 s, and a frequency of 5 Hz, applied five times, removed smear layer and debris from the root canal surface at temperatures below the thermal injury threshold for periodontal tissue.

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 µg of VEGF and 20 µg of bFGF was performed on adult male Dutch belted rabbits. In other animals either 20 µg or 50 µg bFGF-containing pellets was implanted intravitreally; also, either 20 µg VEGF or 50 µg VEGF-containing pellets was implanted. Control rabbits received either blank polymeric pellets or a pellet containing 30 µg 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.

Spatial frequency domain imaging of burn wounds in a preclinical model of graded burn severity

Abstract. Frequent monitoring of early-stage burns is necessary for deciding optimal treatment and management. Both superficial and full thickness burns are relatively easy to diagnose based on clinical observation. In between these two extremes are superficial-partial thickness and deep-partial thickness burns. These burns, while visually similar, differ dramatically in terms of clinical treatment and are known to progress in severity over time. The objective of this study was to determine the potential of spatial frequency domain imaging (SFDI) for noninvasively mapping quantitative changes in chromophore and optical properties that may be an indicative of burn wound severity. A controlled protocol of graded burn severity was developed and applied to 17 rats. SFDI data was acquired at multiple near-infrared wavelengths over a course of 3 h. Burn severity was verified using hematoxylin and eosin histology. From this study, we found that changes in water concentration (edema), deoxygenated hemoglobin concentration, and optical scattering (tissue denaturation) to be statistically significant at differentiating superficial partial-thickness burns from deep-partial thickness burns.

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

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.

Sealing of human dentinal tubules by XeCl 308-nm excimer laser

Root hypersensitivity occurs as a result of exposed dentinal tubules. Various methods and materials have been tried in an attempt to occlude these tubules. The purpose of this investigation was to study by scanning electron microscope the effects of XeCl excimer laser on exposed dentinal tubules of human extracted teeth. Fifteen 3-mm-thick slices were cut at the cementoenamel junction from 15 extracted human teeth by an electric saw. By using a diamond bur to remove the cementum layer the dentinal tubules were exposed. Each slice was scored by a permanent marker into four equal quadrants. Three of the quadrants were lased for 4 s by XeCl excimer laser with fluences ranging from 0.5 to 7.0 J/cm2 and pulse repetition of 25 Hz. The unlased quadrant served as control. The specimens were mounted on a stub, sputter coated by gold, and examined by scanning electron microscope. Nonlased surfaces showed numerous exposed dentinal tubules. In contrast, all specimens lased at fluences of up to 1 J/cm2 showed the presence of melted dentin which closed the dentinal tubules. At fluences of 4 J/cm2 and higher, rupture of molten materials and exposure of dentinal tubules were noted. The results indicate the application of XeCl excimer laser at specific fluences can cause melting of dentin and closure of exposed dentinal tubules.

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.

Analysis of DNA double-strand break response and chromatin structure in mitosis using laser microirradiation

In this study the femtosecond near-IR and nanosecond green lasers are used to induce alterations in mitotic chromosomes. The subsequent double-strand break responses are studied. We show that both lasers are capable of creating comparable chromosomal alterations and that a phase paling observed within 1–2 s of laser exposure is associated with an alteration of chromatin as confirmed by serial section electron microscopy, DAPI, γH2AX and phospho-H3 staining. Additionally, the accumulation of dark material observed using phase contrast light microscopy (indicative of a change in refractive index of the chromatin) ∼34 s post-laser exposure corresponds spatially to the accumulation of Nbs1, Ku and ubiquitin. This study demonstrates that chromosomes selectively altered in mitosis initiate the DNA damage response within 30 s and that the accumulation of proteins are visually represented by phase-dark material at the irradiation site, allowing us to determine the fate of the damage as cells enter G1. These results occur with two widely different laser systems, making this approach to study DNA damage responses in the mitotic phase generally available to many different labs. Additionally, we present a summary of most of the published laser studies on chromosomes in order to provide a general guide of the lasers and operating parameters used by other laboratories.

Effect of ArF-193 nm excimer laser on human dentinal tubules: A scanning electron microscopic study

The purpose of this study was to evaluate the effects of the ArF-193 nm excimer laser on the dentinal tubules of extracted human teeth under a scanning electron microscope. Fifteen 3 mm thick slices were cut with an electric saw at the cementoenamel junction from 15 extracted human teeth. A diamond bur was used to remove the cementum layer and expose the dentinal tubules. Each slice was scored by a permanent marker into four equal quadrants. The ArF excimer laser was applied for 5 seconds on three of the quadrants with fluences that ranged from 0.2 J/cm2 to 15 J/cm2 and pulse repetition of 25 Hz. The untouched quadrant served as a control. The specimens were mounted on stubs, sputter coated by gold, and examined by a scanning electron microscope. The effects of the ArF excimer laser irradiation varied. Laser fluences of 0.2, 0.5, and 1.0 J/cm2 had no effect. Although fluence of 15 J/cm2 caused significant removal of peritubular dentin, melting and resolidification of the dentinal smear layer was also observed under the scanning electron microscope with a laser fluence of 5 J/cm2.

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.