Targeting of prostate-specific membrane antigen for radio-ligand therapy of triple-negative breast cancer.

BACKGROUND: Triple-negative breast cancer has extremely high risk of relapse due to the lack of targeted therapies, intra- and inter-tumoral heterogeneity, and the inherent and acquired resistance to therapies. In this study, we evaluate the potential of prostate-specific membrane antigen (PSMA) as target for radio-ligand therapy (RLT). METHODS: Tube formation was investigated after incubation of endothelial HUVEC cells in tumor-conditioned media and monitored after staining using microscopy. A binding study with 68Ga-labeled PSMA-addressing ligand was used to indicate targeting potential of PSMA on tumor-conditioned HUVEC cells. For mimicking of the therapeutic application, tube formation potential and vitality of tumor-conditioned HUVEC cells were assessed following an incubation with radiolabeled PSMA-addressing ligand [177Lu]-PSMA-617. For in vivo experiments, NUDE mice were xenografted with triple-negative breast cancer cells MDA-MB231 or estrogen receptor expressing breast cancer cells MCF-7. Biodistribution and binding behavior of [68Ga]-PSMA-11 was investigated in both tumor models at 30 min post injection using µPET. PSMA- and CD31-specific staining was conducted to visualize PSMA expression and neovascularization in tumor tissue ex vivo. RESULTS: The triple-negative breast cancer cells MDA-MB231 showed a high pro-angiogenetic potential on tube formation of endothelial HUVEC cells. The induced endothelial expression of PSMA was efficiently addressed by radiolabeled PSMA-specific ligands. 177Lu-labeled PSMA-617 strongly impaired the vitality and angiogenic potential of HUVEC cells. In vivo, as visualized by µPET, radiolabeled PSMA-ligand accumulated specifically in the triple-negative breast cancer xenograft MDA-MB231 (T/B ratio of 43.3 ± 0.9), while no [68Ga]-PSMA-11 was detected in the estrogen-sensitive MCF-7 xenograft (T/B ratio of 1.1 ± 0.1). An ex vivo immunofluorescence analysis confirmed the localization of PSMA on MDA-MB231 xenograft-associated endothelial cells and also on TNBC cells. CONCLUSIONS: Here we demonstrate PSMA as promising target for two-compartment endogenous radio-ligand therapy of triple-negative breast cancer.

Vascular Effects of Photodynamic Therapy with Curcumin in a Chorioallantoic Membrane Model.

Photodynamic Therapy (PDT) is a treatment that requires light, a photosensitizing agent, and molecular oxygen. The photosensitizer is activated by light and it interacts with the oxygen that is present in the cellular microenvironment. The molecular oxygen is transformed into singlet oxygen, which is highly reactive and responsible for the cell death. Therefore, PS is an important element for the therapy happens, including its concentration. Curcumin is a natural photosensitizer and it has demonstrated its anti-inflammatory and anti-oxidant effects that inhibit several signal transduction pathways. PDT vascular effects of curcumin at concentrations varying from 0.1 to 10 mM/cm² and topical administration were investigated in a chick Chorioallantoic Membrane (CAM) model. The irradiation was performed at 450 nm, irradiance of 50 mW/cm² during 10 min, delivering a total fluence of 30 J/cm². The vascular effect was followed after the application of curcumin, with images being obtained each 30 min in the first 3 h, 12 h, and 24 h. Those images were qualitatively and quantitatively analyzed with a MatLAB®. Curcumin was expected to exhibit a vascular effect due to its angio-inhibitory effect. Using curcumin as photosensitizer, PDT induced a higher and faster vascular effect when compared to the use of this compound alone.

Role of the adjunctive antimicrobial photodynamic therapy to periodontal treatment at plasmatic oxidative stress and vascular behavior.

BACKGROUND: To evaluate for the first time in vivo the effects of methylene blue (MB) photosensitizer dissolved in ethanol in antimicrobial photodynamic therapy (aPDT) as adjuvant periodontal treatment, at plasmatic oxidative stress and vascular behavior in rat model. METHODS: Wistar rats were divided into negative control (NC, no periodontitis) and positive control (PC, with periodontitis, without any treatment). The other groups had periodontitis and were treated with scaling and root planing (SRP); SRP+aPDT+MB dissolved in water (aPDT I); SRP+aPDT+MB dissolved in ethanol (aPDT II). The periodontitis was induced by ligature at the mandibular right first molar. At 7/15/30days, rats were euthanized, the plasma was used to determine oxidative stress parameters and gingival tissue for histomorphometric analysis. RESULTS: PC showed higher thiobarbituric acid reactive substances levels in 7/15/30days. aPDT II was able to block the lipid peroxidation, especially between 15th and 30th days. Glutathione reduced levels were consumed in PC, aPDT I and II groups throughout the experiment. aPDT II increased the vitamin C levels which were restored in this group in the 30th day. aPDT II group showed the highest number of blood vessels. CONCLUSION: In summary, the aPDT with MB dissolved in ethanol provides better therapeutic responses in periodontitis treatment.

Experimental and numerical investigation on the transient vascular thermal response to multi-pulse Nd:YAG laser.

BACKGROUND AND OBJECTIVE: Port wine stains (PWS) are congenital vascular malformations that progressively darken and thicken with age. Laser therapy is currently the most effective way in clinical practice for PWS. A 1,064 nm Nd:YAG laser in the near-infrared band can achieve a deeper treatment depth compared to the current widely adopted pulsed dye laser. However, because of its relatively weak absorption by blood, single-pulse Nd:YAG laser requires high energy density to cause effective vessel damage, but may inflict undesirable burning to surrounding collagen. Multi-pulse laser has great potential in clinical treatment because it needs less energy density for each pulse. This paper presented an experimental and theoretical study of the transient thermal effects of low-energy multi-pulse Nd:YAG laser on blood vessels. STUDY DESIGN/MATERIALS AND METHODS: In vivo experiments were performed on dorsal skin chamber. By using a high speed camera (up to 2,000 fps), the complete and dynamic thermal response of blood vessels during laser irradiation and between pulse intervals was obtained. In vitro experiment in capillary tubes and Numerical simulations by two-scale heat transfer model were also conducted to further explore the in vivo experimental findings. RESULTS: The complete and dynamic response of blood vessels were obtained, including vessel dilation, thrombus formation, partial vessel constriction, thread-like constriction, cavitation and bubbles, and hemorrhage. Thread-like constriction is the desirable treatment end point, which will only occur after thrombus completely occludes the vessel lumen. Cavitation can cause hemorrhage when thrombus fails to occlude the vessel lumen. In vitro experiment found that vessel constriction was due to the constriction of thrombus induced by laser irradiation. Theoretical investigation revealed that the mechanism for the effective reduction of energy density by multi-pulse Nd:YAG laser was due to enhanced light absorption of the blood with thrombus formation. CONCLUSIONS: For multi-pulse treatment, laser parameters are recommended as repetition rate of 10 Hz and pulse number of 10. The incident energy in each pulse should be strong enough to induce blood coagulation through seven or eight pulses and should be lower than the threshold of blood cavitation. Lasers Surg. Med. 49:852-865, 2017. © 2017 Wiley Periodicals, Inc.

Combination of Gold Nanoparticle-Conjugated Tumor Necrosis Factor-α and Radiation Therapy Results in a Synergistic Antitumor Response in Murine Carcinoma Models.

PURPOSE: Although remarkable preclinical antitumor effects have been shown for tumor necrosis factor-α (TNF) alone and combined with radiation, its clinical use has been hindered by systemic dose-limiting toxicities. We investigated the physiological and antitumor effects of radiation therapy combined with the novel nanomedicine CYT-6091, a 27-nm average-diameter polyethylene glycol-TNF-coated gold nanoparticle, which recently passed through phase 1 trials. METHODS AND MATERIALS: The physiologic and antitumor effects of single and fractionated radiation combined with CYT-6091 were studied in the murine 4T1 breast carcinoma and SCCVII head and neck tumor squamous cell carcinoma models. RESULTS: In the 4T1 murine breast tumor model, we observed a significant reduction in the tumor interstitial fluid pressure (IFP) 24 hours after CYT-6091 alone and combined with a radiation dose of 12 Gy (P<.05 vs control). In contrast, radiation alone (12 Gy) had a negligible effect on the IFP. In the SCCVII head and neck tumor model, the baseline IFP was not markedly elevated, and little additional change occurred in the IFP after single-dose radiation or combined therapy (P>.05 vs control) despite extensive vascular damage observed. The IFP reduction in the 4T1 model was also associated with marked vascular damage and extravasation of red blood cells into the tumor interstitium. A sustained reduction in tumor cell density was observed in the combined therapy group compared with all other groups (P<.05). Finally, we observed a more than twofold delay in tumor growth when CYT-6091 was combined with a single 20-Gy radiation dose-notably, irrespective of the treatment sequence. Moreover, when hypofractionated radiation (12 Gy × 3) was applied with CYT-6091 treatment, a more than five-fold growth delay was observed in the combined treatment group of both tumor models and determined to be synergistic. CONCLUSIONS: Our results have demonstrated that TNF-labeled gold nanoparticles combined with single or fractionated high-dose radiation therapy is effective in reducing IFP and tumor growth and shows promise for clinical translation.

Experimental study on the vascular thermal response to visible laser pulses.

Port-wine stains (PWSs) are congenital vascular malformations that progressively darken and thicken with age, and laser therapy is the most effective in clinical practice. Using dorsal skin chamber (DSC), this study evaluated thermal response of blood vessel to a 595-nm pulsed dye laser (PDL) with controlled energy doses and pulse durations. Totally, 32 vessels (30∼300 µm in diameter) are selected from the dorsal skin of the mouse to match those in port-wine stain. The experimental results showed that the thermal response of the blood vessels to laser irradiation can be recognized as coagulation, constriction with diameter decrease, disappearance (complete constriction), hemorrhage, and collagen damage in the order of increasing laser radiant exposure. Blood vessels with small diameter would response poorly and survive from the laser heating because their thermal relaxation time is much shorter than the pulse duration. The optimalradiant exposure is from 10 to 12 J/cm(2) under 6 ms pulse duration without considering the epidermal light absorption. Numerical simulations were also conducted using a 1,000-µm deep Sprague-Dawley (SD) mouse skinfold. The light transportation and heat diffusion in dorsal skin were simulated with the Monte Carlo method and heat transfer equation, while the blood vessel photocoagulation was evaluated by Arrhenius-type kinetic integral. Both experimental observation and numerical simulation supported that hemorrhage is the dominant thermal response, which occurs due to preferential heating of the superior parts of large blood vessels. In clinical practice for 595 nm PDL, the consequent purpura caused by hemorrhage can be used as a treatment end point.

Histological analysis of the periodontal ligament and alveolar bone during dental movement in diabetic rats subjected to low-level laser therapy.

OBJECTIVE: The purpose of this research was to evaluate the histological changes of the periodontal ligament and alveolar bone during dental movement in diabetic rats subjected to low level laser therapy (LLLT). METHODS: The movement of the upper molar was performed in 60 male Wistar rats divided into four groups (n=15): CTR (control), DBT (diabetic), CTR/LT (irradiated control) and DBT/LT (irradiated diabetic). Diabetes was induced with alloxan (150 mg/kg, i.p.). LLLT was applied with GaAlAs laser at 780 nm (35 J/cm(2)). After 7, 13 and 19 days, the periodontal ligament and alveolar bone were histologically analyzed. RESULTS: The mean of osteoblasts (p<0.01) and blood vessels (p<0.05) were significantly decreased in DBT compared with CTR at 7 days, whereas the mean of osteoclasts was lower at 7 (p<0.001) and 13 days (p<0.05). In DBT/LT, only the mean of osteoclasts was lower than in CTR (p<0.05) at 7 days, but no difference was observed at 13 and 19 days (p>0.05). The collagenization of the periodontal ligament was impaired in DBT, whereas DBT/LLT showed density/disposition of the collagen fibers similar to those observed in CTR. CONCLUSIONS: LLLT improved the periodontal ligament and alveolar bone remodeling activity in diabetic rats during dental movement.

Thermal relaxation times: an outdated concept in photothermal treatments.

Thermal relaxation times were introduced into modern skin-laser science with the inception of selective photothermolysis. As a result, laser pulsewidths were determined according to the thermal relaxation times of the tissue targets. The Arrhenius Damage Integral shows that this approach is incorrect. The important parameter is the time required to induce irreversible protein denaturation within the target. This time is determined by the tissue's intrinsic structure, not its physical dimensions. This report explains why thermal relaxation times should not be considered when treating many skin conditions with lasers or IPL systems.

Influence of pulsing electromagnetic field therapy on resting blood pressure in aging adults.

This double-blind study tested the effects of pulsating electromagnetic field (PEMF) therapy sessions on the changes in peripheral cardiovascular function in a group of aging adults after 12 weeks of treatment. Each therapy session involved 15 min of exposure to low-frequency PEMF with asymmetrical waveforms emitted by the Impulser™ Pro mattress. The treatment was provided 5 days per week for a total of 60 sessions. Resting blood pressure and arterial stiffness index were determined for peripheral cardiovascular function. Fifty-four older men and women (mean age 59.8 ± 3.5 yrs) completed the entire protocol involving either the PEMF or a sham treatment. The results include statistically significant reductions in systolic and pulse blood pressure, while no significant difference in diastolic pressure or the index of arterial stiffness was observed. These findings suggest that the PEMF treatment might be linked to improvements in peripheral resistance or circulation.

Nutritional supplementation with L-arginine prevents pelvic radiation-induced changes in morphology, density, and regulating factors of blood vessels in the wall of rat bladder.

PURPOSE: To determine whether L-arginine has protective effects against radiation-induced alterations in the morphology and regulatory factors of vesical blood vessels in rats. METHODS: Male rats aged 3-4 months were divided into groups of 10 animals each: (a) controls, consisting of non-treated animals; (b) radiated-only rats; and (c) radiated rats receiving L-arginine supplementation. Radiation was in one session of 10 Gy and was aimed at the pelvic-abdominal region. L-arginine was administered once a day (0.65 g/kg body weight), starting 7 days before radiation and continuing until killing on the 16th day after radiation. The density, relative area, and wall thickness of blood vessels were measured in the vesical lamina propria using histological methods, and the expression of vascular endothelial growth factor (VEGF) and fibroblast growth factors (FGF) in the bladder wall was assessed by RT-PCR. RESULTS: Compared with controls, radiation alone decreased the density and relative area of blood vessels by 32 % (p < 0.01) and 25 % (p < 0.05), respectively, and reduced the arterial wall thickness by 42 % (p < 0.004). VEGF and FGF mRNA levels after radiation were diminished by 67 % (p < 0.002) and 56 % (p < 0.04), respectively. The radiated animals supplemented with L-arginine were not significantly different from controls. CONCLUSIONS: Pelvic radiation leads to significant vesical modifications, as in the morphology of blood vessels and in VEGF and FGF expression. All these changes, however, were prevented by L-arginine treatment. These results emphasize, therefore, the potential use of this amino acid as a radioprotective drug.

Time-dependent effects of low-level laser therapy on the morphology and oxidative response in the skin wound healing in rats.

This study aims to investigate the effect of different energy densities provided by low-level laser therapy (LLLT) on the morphology of scar tissue and the oxidative response in the healing of secondary intention skin wounds in rats. Twenty-four male adult Wistar rats were used. Skin wounds were made on the backs of the animals, which were randomized into three groups of eight animals each as follows, 0.9% saline (control); laser GaAsAl 30 J/cm(2) (L30); laser GaAsAl 90 J/cm(2) (L90). The experiment lasted 21 days. Every 7 days, the wound contraction index (WCI) was calculated and tissue from different wounds was removed to assess the proportion of cells and blood vessels, collagen maturation index (CMI), thiobarbituric acid reactive substance (TBARS) levels and catalase activity (CAT). On the 7th and 14th days, the WCI and the proportion of cells were significantly higher in groups L30 and L90 compared to the control (p < 0.05). At all the time points analyzed, there was a greater proportion of blood vessels and a higher CMI in group L90 compared to the other groups (p < 0.05). On the 7th and 14th days, lower TBARS levels and increased CAT activity were found in the L90 group compared to the control (p < 0.05). On the 7th day, a moderately negative correlation was found between TBARS levels and WCI, CMI and CAT in all the groups. LLLT may modulate the oxidative status of wounded tissue, constituting a possible mechanism through which the LLLT exerts its effects in the initial phases of tissue repair.

[Molecular-cellular mechanisms of light radiation and weak magnetic fields on the blood status and microcirculation system (at therapeutic doses) with using modern magnetolaser equipment (review of the literature)].

This review covers the molecular-cellular mechanisms of therapeutic action of light and magnetic field on blood components, blood vessels and the microcirculation system. Noted the role of the magnetic field as a trigger of vasodilation/vasoconstriction, depending on the initial vascular tone. Discussed the importance of NO-dependent effects of magnetic field on the microcirculatory response and angiogenesis.

[Use of alternating magnetic field with the frequency of spontaneous local vessels oscillation for rehabilitation of patients after dental implantation].

New software-hardware system for diagnostic and treatment (RF patents: number 75294, 2008 and number 75314, 2008) allowing to use the technique of microcirculatory bed regional pulse vessels control (RF patent number 2383369, 2008) for optimization of patients' rehabilitation process after dental implantation operation. It was shown the normalizing action of alternating magnetic field with the frequency of spontaneous local vessels oscillation by comparison of regional blood flow indices.

KTP/532 YAG laser treatment for allergic rhinitis.

BACKGROUND: The purpose of this study was to investigate the efficacy of the KTP/532 YAG laser to reduce nasal congestion and discharge in patients with allergic rhinitis. METHODS: Forty-eight patients with symptoms of allergic rhinitis were treated with the KTP/532 laser. All had positive skin tests for common allergens. Treatments were provided on an ambulatory basis in one to three sessions under local anesthesia with lidocaine nose spray. Outcome was determined by daily symptom reports and regular endoscopy examination and interviews for 12 months. RESULTS: Treatment was very well tolerated. There were no major side effects. At examination after 1 year, nasal obstruction was improved in 69% and nasal discharge in 40% of cases. CONCLUSION: The KTP/532 YAG laser is effective for the treatment of nasal obstruction and discharge. Comparison with other techniques showed it to be the most effective in reducing nasal discharge. It can be done as an office procedure and does not damage the nasal mucous membrane. The KTP/532 YAG laser is effective as an additional treatment for patients refractory to medication.

Treatment of resistant port-wine stains with a pulsed dual wavelength 595 and 1064 nm laser: a histochemical evaluation of the vessel wall destruction and selectivity.

BACKGROUND AND OBJECTIVES: Pulsed dye laser (PDL) is the current treatment of choice for port-wine stains (PWS), but 25-50% of treated lesions do not demonstrate a significant improvement. Hybrid lasers may improve treatment efficacy, especially those using the synergies between PDL and Nd:YAG 1064 nm laser. The objectives of this study were to assess vessel wall damage and epidermal sparing after a dual wavelength treatment with the two lasers, using different laser parameters. MATERIAL AND METHODS: Post-treatment biopsies, after using a laser platform that allows sequential pulses of PDL and Nd:YAG 1064 nm lasers, were performed in five patients with PWS resistant to PDL. The biopsies were stained with nitroblue-tetrazolium chloride (NBTC), using enzymatic activity that stops immediately after cell death and allows a better identification of viable cells. RESULTS: Five patients with PWS and a median age of 33 years were enrolled in this study. Selectivity and efficacy was observed with this dual wavelength approach, with the best results observed with PDL pulses shorter than 10 ms, use of the 10 mm spot, and a second pass with PDL only. CONCLUSIONS: Histochemical studies with NBTC stain can help the laser surgeon establish the best treatment parameters and understand some of the unwanted side effects. The dual wavelength used in this study showed efficacy, but better assessment of treatment parameters, such as the delay between the two lasers, is needed to avoid side effects.

Pulsed electrical stimulation for control of vasculature: temporary vasoconstriction and permanent thrombosis.

A variety of medical procedures is aimed to selectively compromise or destroy vascular function. Such procedures include cancer therapies, treatments of cutaneous vascular disorders, and temporary hemostasis during surgery. Currently, technologies such as lasers, cryosurgery and radio frequency coagulation, produce significant collateral damage due to the thermal nature of these interactions and corresponding heat exchange with surrounding tissues. We describe a non-thermal method of inducing temporary vasoconstriction and permanent thrombosis using short pulse (microseconds) electrical stimulation. The current density required for vasoconstriction increases with decreasing pulse duration approximately as t(-0.25). The threshold of electroporation has a steeper dependence on pulse duration-exceeding t(-0.5). At pulse durations shorter than 5 micros, damage threshold exceeds the vasoconstriction threshold, thus allowing for temporary hemostasis without direct damage to surrounding tissue. With a pulse repetition rate of 0.1 Hz, vasoconstriction is achieved approximately 1 min after the beginning of treatment in both arteries and veins. Thrombosis occurs at higher electric fields, and its threshold increases with vessel diameter. Histology demonstrated a lack of tissue damage during vasoconstriction, but vascular endothelium was damaged during thrombosis. The temperature increase does not exceed 0.1 degrees C during these treatments.

Laser surgery of port wine stains using local vacuum pressure: changes in skin morphology and optical properties (Part I).

BACKGROUND AND OBJECTIVES: In a recent case study, the use of a suction device to aid in port wine stain (PWS) laser treatments showed favorable results. It is our objective to further understand the mechanisms of vacuum-assisted laser therapy by analyzing the mechanical and optical changes of the skin and musculoskeletal tissues during the application of mild vacuum pressure from a suction cup. STUDY DESIGN/MATERIALS AND METHODS: A mathematical model of tissue deformation was used to determine the changes in tissue morphology that affect the underlying laser-tissue interactions, such as epidermal stretching and thinning, blood vessel dilation, and change in blood vessel depth. Video imaging experiments were used to verify the bulk tissue deformation and skin surface stretching computed by the mathematical model. Additionally, visible reflectance spectroscopy was used to determine the changes in the optical characteristics of tissue, including blood vessel dilation and epidermal absorption coefficient. RESULTS: At a vacuum pressure of 50 kP(a), the epidermis at the center of the suction cup was measured to stretch 4% and was calculated to be thinned approximately 6%. Blood vessels embedded in the dermis were measured to dilate up to two times their original size. However, these vessels were calculated to be displaced toward the skin surface by a very small amount, approximately 1-3 microm. The absorption coefficient of the epidermis was also measured to be reduced significantly by approximately 25% at a wavelength of 585 nm. CONCLUSIONS: Mild vacuum pressure applied to the skin surface causes considerable changes in the morphology and optical properties of the tissue. These changes may be used for more efficient photothermolysis of small PWS blood vessels.

The effects of intense pulsed light (IPL) on blood vessels investigated by mathematical modeling.

BACKGROUND AND OBJECTIVES: Intense pulsed light (IPL) sources have been successfully used for coagulation of blood vessels in clinical practice. However, the broadband emission of IPL hampers the clinical evaluation of optimal light parameters. We describe a mathematical model in order to visualize the thermal effects of IPL on skin vessels, which was not available, so far. STUDY DESIGN/MATERIALS AND METHODS: One IPL spectrum was shifted towards the near infrared range (near IR shifted spectrum: NIRSS) and the other was heavily shifted toward the visible range (visible shifted spectrum: VSS). The broadband emission was separated in distinct wavelengths with the respective relative light intensity. For each wavelength, the light and heat diffusion equations were simultaneously solved with the finite element method. The thermal effects of all wavelengths at the given radiant exposure (15 or 30 J/cm2) were added and the temperature in the vessels of varying diameters (60, 150, 300, 500 microm) was calculated for the entire pulse duration of 30 milliseconds. RESULTS: VSS and NIRSS both provided homogeneous heating in the entire vessel. With the exception of the small vessels (60 microm), which showed only a moderate temperature increase, all vessels exhibited a temperature raise within the vessel sufficient for coagulation with each IPL parameter. The time interval for effective temperature raise in larger vessels (diameter >60 microm) was clearly shorter than the pulse duration. In most instances, the vessel temperature was higher for VSS when compared to NIRSS. CONCLUSIONS: We presented a mathematical model capable of calculating the photon distribution and the thermal effects of the broadband IPL emission within cutaneous blood vessels.

Laser-induced thermal injury to dermal blood vessels: analysis of wavelength (585 nm vs. 595 nm), cryogen spray cooling, and wound healing effects.

BACKGROUND AND OBJECTIVES: Successful laser treatment of cutaneous hyper-vascular lesions requires appropriate laser irradiation parameters for selective photothermolysis of ectatic dermal blood vessels as well as appropriate cooling parameters for epidermal protection based on an individual patient basis. Using the rabbit ear as an in vivo model for dermal vasculature, we investigated the influences of laser wavelength (585 nm vs. 595 nm) and cryogen spray cooling with various spurt durations on the laser-induced thermal injury to dermal blood vessels. Wound healing response was also evaluated in 2 hours and 4 days. STUDY DESIGN/MATERIALS AND METHODS: Flashlamp-pumped pulsed dye laser ScleroPlus (operated at the wavelength of 585 or 595 nm) was used for the comparison between the influences of two wavelengths (585 nm vs. 595 nm). R134-a cryogen spurts with the durations from 50 to 300 milliseconds were sprayed onto the sites to be irradiated and terminated 20 milliseconds before the onset of the laser pulses. In vivo rabbit ear was used as the model for cutaneous hyper-vascular lesions. Totally 10 New Zealand Albino white rabbits were experimented and in each rabbit ear six to seven sites were irradiated. Five animals were sacrificed 2 hours after the irradiation, and the remaining five sacrificed 4 days after the irradiation. Thermal injury to the blood vessel was assessed by hematoxylin and eosin stained histological sections and confirmed by an apoptosis assay. RESULTS: When the radiant exposures were above 10 J/cm2, 595 nm wavelength induced equivalent or more severe thermal injury to dermal blood vessels than 585 nm. Cryogen spray cooling with the spurt durations above 100 milliseconds resulted in increased depth of the most superficial thermal injury to dermal blood vessels than without cooling, indicating that superficial blood vessels were non-specifically cooled by the cryogen spurts applied at these parameters. Laser-induced thermal injury was significantly healed in the rabbit ear vasculature at 4 days post irradiation. CONCLUSIONS: Given sufficient radiant exposure, 595 nm wavelength can induce equivalent or more severe vascular injury compared with 585 nm. Cryogen spray cooling with the spurt durations above 100 ms may impair the photocoagulation of superficial blood vessels. Irreversible thermal injury to blood vessel can be achieved only when the basement membrane of blood vessel wall is irreversibly damaged.

Comparison of diffusion approximation and Monte Carlo based finite element models for simulating thermal responses to laser irradiation in discrete vessels.

Both diffusion approximation (DA) and Monte Carlo (MC) models have been used to simulate light distribution in multilayered human skin with or without discrete blood vessels. However, no detailed comparison of the light distribution, heat generation and induced thermal damage between these two models has been done for discrete vessels. Three models were constructed: (1) MC-based finite element method (FEM) model, referred to as MC-FEM; (2) DA-based FEM with simple scaling factors according to chromophore concentrations (SFCC) in the epidermis and vessels, referred to as DA-FEM-SFCC; and (3) DA-FEM with improved scaling factors (ISF) obtained by equalizing the total light energy depositions that are solved from the DA and MC models in the epidermis and vessels, respectively, referred to as DA-FEM-ISF. The results show that DA-FEM-SFCC underestimates the light energy deposition in the epidermis and vessels when compared to MC-FEM. The difference is nonlinearly dependent on wavelength, dermal blood volume fraction, vessel size and depth, etc. Thus, the temperature and damage profiles are also dramatically different. DA-FEM-ISF achieves much better results in calculating heat generation and induced thermal damage when compared to MC-FEM, and has the advantages of both calculation speed and accuracy. The disadvantage is that a multidimensional ISF table is needed for DA-FEM-ISF to be a practical modelling tool.