Infrared neural stimulation of human spinal nerve roots in vivo.

Infrared neural stimulation (INS) is a neurostimulation modality that uses pulsed infrared light to evoke artifact-free, spatially precise neural activity with a noncontact interface; however, the technique has not been demonstrated in humans. The objective of this study is to demonstrate the safety and efficacy of INS in humans in vivo. The feasibility of INS in humans was assessed in patients ([Formula: see text]) undergoing selective dorsal root rhizotomy, where hyperactive dorsal roots, identified for transection, were stimulated in vivo with INS on two to three sites per nerve with electromyogram recordings acquired throughout the stimulation. The stimulated dorsal root was removed and histology was performed to determine thermal damage thresholds of INS. Threshold activation of human dorsal rootlets occurred in 63% of nerves for radiant exposures between 0.53 and [Formula: see text]. In all cases, only one or two monitored muscle groups were activated from INS stimulation of a hyperactive spinal root identified by electrical stimulation. Thermal damage was first noted at [Formula: see text] and a [Formula: see text] safety ratio was identified. These findings demonstrate the success of INS as a fresh approach for activating human nerves in vivo and providing the necessary safety data needed to pursue clinically driven therapeutic and diagnostic applications of INS in humans.

Nanoparticles in Medicine: Selected Observations and Experimental Caveats.

Medically useful nanoparticles measure 1-100 nm in at least one dimension and are engineered and manufactured for specific diagnostic and treatment applications. Most nanoparticles used currently used in medicine are engineered and manufactured for specific purposes. Medically significant nanoparticles are composed of a 1) central core that is usually the medically active component, 2) one or more layers of organic or inorganic materials that forms a capsule (corona) covering the core and 3) an outer surface layer that interacts with the environment and/or targeted cells and tissues. Effective nanoparticle function in the living, intact animal or human requires electrochemical stability necessary to bypass the reticuloendothelial system (RES) and avoid filtration through the renal glomerulus into the urine. Nanoparticles are present in "natural" as well as the manufacturing and clinical environments thus could pose as significant toxins because of their small sizes, their chemical and drug content and potential effect of causing long term disease including allergies, chronic inflammation and cancer. Currently published studies have focused on the effects of nanoparticles on cells in the extremely artificial environments of cell cultures. More clinical and preclinical studies documenting the short term and long term effects nanoparticle in the intact experimental animal and human are needed.

Micro-patterned drug delivery device for light-activated drug release.

BACKGROUND: The primary goal of this study was the fabrication, long-term stability, and measured release of a marker dye from a micro-patterned drug delivery device using (i) mechanical puncture and (ii) photodisruption with an ophthalmic Nd:YAG laser. MATERIALS AND METHODS: A drug delivery device was made from a transparent bio-compatible polymer. The device consisted of two 2.6 mm diameter reservoirs containing 10% Na fluorescein dye. The device was implanted in the rabbit's eye (n = 2) with the cap of the device facing toward the exterior of the eye. Once the animals recovered from the implant surgery, 100% anhydrous glycerol was topically applied to the eye at the implantation site to decrease light scattering in the conjunctiva and sclera. The dye was released from one of the reservoir either using a 28 G ½ needle or an ophthalmic Q-switched Nd:YAG laser. A fluorescence spectrophotometer (FS) with fiber optic probe was used to measure the half-life of the dye in the eye. Measurements of fluorescence intensity were collected until the measurements return to baseline and histology was done on the tissue surrounded the device. RESULTS: None of the devices leaked of 10% Na fluorescein dye after implant. The ablation threshold of the drug delivery device was between 6 and 10 mJ to create 100-500 µm holes. The half-life measurement of the dye was found to be 13 days at the vitreous chamber after measuring the fluorescence intensity through the dilated cornea. Histology study showed minimal immune and foreign body response such as mild inflammation. CONCLUSION: This study established that the drug delivery device seemed to elicit minimal inflammatory response and retained its fluidic content until it was released with relatively longer retention time (half-life). Thus, similar device could be used for controlled release of drugs for certain ocular diseases.

Ultrasound guidance and monitoring of laser-based fat removal.

BACKGROUND AND OBJECTIVES: We report on a study to investigate feasibility of utilizing ultrasound imaging to guide laser removal of subcutaneous fat. Ultrasound imaging can be used to identify the tissue composition and to monitor the temperature increase in response to laser irradiation. STUDY DESIGN/MATERIALS AND METHODS: Laser heating was performed on ex vivo porcine subcutaneous fat through the overlying skin using a continuous wave laser operating at 1,210 nm optical wavelength. Ultrasound images were recorded using a 10 MHz linear array-based ultrasound imaging system. RESULTS: Ultrasound imaging was utilized to differentiate between water-based and lipid-based regions within the porcine tissue and to identify the dermis-fat junction. Temperature maps during the laser exposure in the skin and fatty tissue layers were computed. CONCLUSIONS: Results of our study demonstrate the potential of using ultrasound imaging to guide laser fat removal.

Histological and modeling study of skin thermal injury to 2.0 microm laser irradiation.

BACKGROUND AND OBJECTIVE: Qualitative and quantitative gross histopathologic studies of skin damage were performed at 48 hours after irradiation with a 2.0 microm thulium CW laser to determine the mechanisms of laser effects in the skin under various exposure conditions. STUDY DESIGN/MATERIALS AND METHODS: Pig skin lesions were created at, below and beyond the threshold irradiation conditions for grossly apparent thermal lesions. Histological sections of these lesions were studied. For each threshold lesion, four quantitative histopathological parameters were measured: the widths of (1) epidermal necrosis at the surface, (2) the outer boundary of the thrombosis zone, (3) the depth of vascular thrombosis, and (4) the depth of perivascular inflammation (increased infiltrates of inflammatory cells) and edema. The quantitative histopathologic data were compared with predictions using an optical-thermal-damage model. RESULTS: Histologically, the thermal damage mechanisms for grossly apparent threshold lesions of persistent redness at 48 hours included necrosis of the epidermal cells, intravascular thrombosis and perivascular inflammation and edema in dermal blood vessels. At irradiation levels just below 'gross threshold', non-lethal thermal effects, such as perivascular inflammation and edema were found in the histological sections. When the radiation reached about 1.5-2.5 times beyond the threshold, decrease of dermal collagen birefringence was observed. CONCLUSIONS: A sequence of damage endpoints was defined in the skin as power increased. By choosing rate process coefficients to match specific mechanisms of lethal thermal damage, the optical-thermal-damage model is capable of predicting various types of thermal injury in the skin, such as epidermal necrosis, vascular thrombosis, and dermal collagen coagulation.

Corneal minimal visible lesion thresholds for 2.0 microm laser radiation.

To support refinement of the ANSI Maximum Permissible Exposure safety limits, a series of experiments were conducted in vivo on Dutch Belted rabbit corneas to determine corneal minimum visible lesion thresholds for 2.0 microm continuous-wave laser irradiation. Single pulse radiant exposures were made at specified pulse durations of 0.1, 0.25, 0.5, 1.0, 2.0, and 4.0 s for spot 1/e(2) diameters of 1.17 mm and 4.02 mm. Threshold lesions were defined as the presence of a superficial surface whitening one hour after irradiation. Temperature measurements indicated that threshold peak temperatures were dependent on spot size and exposure duration. The exposure duration dependence of threshold average radiant exposure was described by an empirical power law equation: threshold radiant exposure[J/cm(2)]=a x exposure duration[s](b).

Effect of pigmentation density upon 2.0 microm laser irradiation thermal response.

Yucatan mini-pigs with predominantly dark skin have been used to determine skin safety standards for infrared (IR) wavelength irradiation due to its anatomical similarity to all human skin. It has generally been argued that water is the principle absorber in the IR-B band and melanin has relatively low absorbance. To accept dark pigmented damage thresholds for skin with various melanin densities, it is necessary to investigate the potential role of melanin in producing skin injury as characterized by an erythermal response. A Yucatan mini-pig covered with lightly pigmented pink and darkly pigmented brown skin was used in this study. The significance of skin pigmentation was investigated by comparing the transient thermal response, absorption coefficient, and the threshold damage of instant redness within 1 min and persistent redness at 48 h post exposure for dark and light skin areas at 2.0 microm wavelength. The density of melanin granules did not significantly alter the thermal and optical properties of in vivo skin exposed to 2.0 microm laser irradiation. For Gaussian shaped beam radiation at 1 s exposure duration and 4.83 mm 1/e spot diameter, the average radiant exposures at instant and persistent redness thresholds were 3.88 J cm and 5.08 J cm for dark skin, respectively, as well as 4.09 J cm and 4.09 J cm for light colored skin. Subjectively speaking, however, lightly pigmented mini-pig skin was more suitable for damage threshold estimation because of the increased contrast for visual determination of redness on light skin.

Optically mediated nerve stimulation: Identification of injury thresholds.

BACKGROUND AND OBJECTIVE: Transient optical nerve stimulation is a promising new non-contact, spatially precise, artifact-free neural excitation technique useful in research and clinical settings. This study evaluates safety of this pulsed infrared laser technique by histopathologic examination of stimulated peripheral nerves. STUDY DESIGN/MATERIALS AND METHODS: Exposed rat sciatic nerves were functionally stimulated with the pulsed Holmium:YAG laser, previously validated as an effective tool for optical stimulation. Nerves were removed immediately and up to 2 weeks after stimulation and assessed histologically for thermal damage. Laser parameters studied include upper limits for radiant exposure, repetition rate, and duration of stimulation. RESULTS: Radiant exposures with <1% probability of thermal tissue damage (0.66-0.70 J/cm(2)) are significantly greater than radiant exposures required for reliable stimulation (0.34-0.48 J/cm(2)). The upper limit for safe laser stimulation repetition rate occurs near 5 Hz. Maximum duration for constant low repetition rate stimulation (2 Hz) is approximately 4 minutes with adequate tissue hydration. CONCLUSION: Results confirm that optical stimulation has the potential to become a powerful non-contact clinical and research tool for brief nerve stimulation with low risk of nerve thermal damage.

Magnetic resonance-guided focused ultrasound surgery of breast cancer: reliability and effectiveness.

BACKGROUND: Magnetic resonance-guided focused ultrasound surgery (MRgFUS) is a noninvasive technique that has been shown to coagulate benign and malignant tumors. The purpose of this study was to evaluate MRgFUS safety and effectiveness for the ablation of breast carcinomas. STUDY DESIGN: Thirty women with biopsy-proved breast cancer underwent MRgFUS treatment. Gadolinium-enhanced MR images were used for treatment planning and posttreatment radiologic assessment of treated tissue, and temperature-sensitive MR images provided real-time treatment monitoring. After MRgFUS, all 30 women underwent wide excision or mastectomy. The extent of thermal ablation was assessed with tumor histology. RESULTS: Treatment was well tolerated, with a minimum of adverse effects, especially when performed under local anesthesia. On pathologic examination, mean (+/-SD) necrosis of the targeted breast tumors was 96.9 +/- 4% (median 100%, range 78% to 100%) of tumor volume. Fifteen (53.5%) of 28 evaluable patients had 100% necrosis of the ablated tumor; only 3 patients (10.7%) had less than 95% necrosis. In 28 (93.3%) patients, 100% of the malignancy was within the treatment field, and 98% and 95% of tumor lay within the treatment field in 2 remaining patients. Retrospective analysis in two patients with residual tumor showed treatment was not delivered to the full recommended area, reaffirming the need for precise localization and the value of contrast-enhanced images for treatment planning. CONCLUSIONS: MRgFUS has great potential to become a viable noninvasive replacement for lumpectomy. Additional studies focusing on posttreatment image-based evaluation are needed.

Impact of physiological variables and genetic background on myocardial frequency-resistivity relations in the intact beating murine heart.

Conductance measurements for generation of an instantaneous left ventricular (LV) volume signal in the mouse are limited, because the volume signal is a combination of blood and LV muscle, and only the blood signal is desired. We have developed a conductance system that operates at two simultaneous frequencies to identify and remove the myocardial contribution to the instantaneous volume signal. This system is based on the observation that myocardial resistivity varies with frequency, whereas blood resistivity does not. For calculation of LV blood volume with the dual-frequency conductance system in mice, in vivo murine myocardial resistivity was measured and combined with an analytic approach. The goals of the present study were to identify and minimize the sources of error in the measurement of myocardial resistivity to enhance the accuracy of the dual-frequency conductance system. We extended these findings to a gene-altered mouse model to determine the impact of measured myocardial resistivity on the calculation of LV pressure-volume relations. We examined the impact of temperature, timing of the measurement during the cardiac cycle, breeding strain, anisotropy, and intrameasurement and interanimal variability on the measurement of intact murine myocardial resistivity. Applying this knowledge to diabetic and nondiabetic 11- and 20- to 24-wk-old mice, we demonstrated differences in myocardial resistivity at low frequencies, enhancement of LV systolic function at 11 wk and LV dilation at 20-24 wk, and histological and electron-microscopic studies demonstrating greater glycogen deposition in the diabetic mice. This study demonstrated the accurate technique of measuring myocardial resistivity and its impact on the determination of LV pressure-volume relations in gene-altered mice.

Detection of vulnerable plaque in a murine model of atherosclerosis with optical coherence tomography.

OBJECTIVES: The aim of this study was to evaluate the feasibility of optical coherence tomography (OCT) to identify the components of vulnerable plaques in a well-established murine model of human atherosclerosis. BACKGROUND: Although the pathologic features that predict plaque rupture at autopsy are well known, the development of a technology to identify these high risk features in vivo is lacking. OCT uses reflected light to provide histology-like images of plaque with higher resolution than competing imaging modalities. Whether OCT can reliably identify the features of an atherosclerotic plaque that define it as vulnerable-thin fibrous cap, large lipid core, and high percent of lipid in the artery-requires further study. METHODS: OCT images of the atherosclerotic innominate artery segments from the apolipoprotein E knockout (apoE(-/-)) mice were recorded and correlated with histology in both in vivo (n = 7) and well as in ex vivo experiments (n = 12). RESULTS: Excellent correlation between the OCT and histology measurements for fibrous cap thickness, lipid core size, and percentage lipid content was found. The fibrous cap thicknesses examined span those of human fibrous caps known to rupture (< 65 microm). Regions of greatest light reflection in OCT images were observed when calcium hydroxy-apatite was scattered in lipid, less in fibrous tissue, and least in lipid. CONCLUSIONS: These findings suggest that OCT holds promise for the identification of features defining vulnerable plaque including fibrous cap thickness, lipid core size, and the percentage of lipid content.

Modeling thermal damage in skin from 2000-nm laser irradiation.

An optical-thermal-damage model of the skin under laser irradiation is developed by using finite-element modeling software (FEMLAB 3.1, Comsol, Incorporated, Burlington, Massachusetts). The general model simulates light propagation, heat generation, transient temperature response, and thermal damage produced by a radically symmetric laser beam of normal incidence. Predictions from the model are made of transient surface temperatures and the thermal damage on a pigskin surface generated by 2000-nm laser irradiation, and these predictions are compared to experimental measurements. The comparisons validate the model predictions, boundary conditions, and optical, thermal, and rate process parameters. The model enables the authors to verify the suitability of the American National Standards Institute (ANSI) maximum permissible exposure (MPE) standard for a wavelength of 2000 nm with exposure duration from 0.1 to 1 s and 3.5-mm beam diameter. Compared with the ANSI MPE standard, however, the MPE values predicted by the model are higher for exposure durations less than 0.1 s. The model indicates that it may be necessary to modify the ANSI MPE standard for cases in which the laser-beam diameter is larger than 3.5 mm when a "safety factor" of ten is used. A histopathological analysis of the skin damage is performed to determine the mechanisms of laser-induced damage in the skin.

Porcine skin ED50 damage thresholds for 2,000 nm laser irradiation.

BACKGROUND AND OBJECTIVES: To gain refinement in safe-exposure limits, indicated by the maximum permissible exposure (MPE) limits, the minimum visible lesion thresholds for three spot sizes (5-15 mm) and four exposure durations (0.25-2.5 seconds) were determined for the skin at 2,000 nm continuous wave laser irradiation. STUDY DESIGN/MATERIALS AND METHODS: A series of experiments were conducted in vivo on female Yucatan mini-pigs to determine the ED50 damage thresholds for 2,000 nm continuous wave laser irradiation. The study employed Gaussian laser beam exposures with spot diameters (1/e2) of 4.83, 9.65, and 14.65 mm and exposure durations of 0.25, 0.5, 1.0, and 2.5 seconds as a function of laser power. The effect of each irradiation was evaluated within 1 minute after irradiation and the final determination was made at 48 hours post-exposure. Probit analysis was conducted to estimate the dose for 50% probability of laser-induced damage (ED50), defined as persistent redness at the site of irradiation for the mini-pig skin after 48 hours. RESULTS: The MPE spot size and exposure duration trends for 2,000 nm laser exposure is consistent for exposure diameters less than 3.5 mm. However, for larger exposure diameters of 4.83, 9.65, and 14.65 mm and exposure duration longer than 0.25 second, the current MPEs are bigger than one tenth of our damage thresholds. For Gaussian laser profile, which is common for many laser output irradiance distributions, lower energy is required to generate a lesion on skin for smaller spot sizes and shorter exposure duration. On the other hand, for spot sizes greater than 4.83 mm and exposure duration over 0.25 second, the average radiant exposure at threshold is inversely proportional to spot size. The irradiance-time and temperature-time power law at the threshold were investigated as well and showed that the irradiance-time power law was a close approximation to estimate laser irradiance at ED50 damage threshold. CONCLUSIONS: The thresholds study shows that consideration for lowering the MPE standards should be explored as the laser beam diameter becomes larger than 3.5 mm. Based on the limited experimental data, the duration and size dependences of the ED50 damage thresholds could be described by an empirical equation: Irradiance at the threshold = (5.669-1.81xspot diameter)xexposure duration -0.794.

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.

Effects of cryogen spray cooling and high radiant exposures on selective vascular injury during laser irradiation of human skin.

BACKGROUND: Increasing radiant exposure offers a means to increase treatment efficacy during laser-mediated treatment of vascular lesions, such as port-wine stains; however, excessive radiant exposure decreases selective vascular injury due to increased heat generation within the epidermis and collateral damage to perivascular collagen. OBJECTIVE: To determine if cryogen spray cooling could be used to maintain selective vascular injury (ie, prevent epidermal and perivascular collagen damage) when using high radiant exposures (16-30 J/cm2). DESIGN: Observational study. SETTING: Academic hospital and research laboratory. PATIENTS: Twenty women with normal abdominal skin (skin phototypes I-VI). INTERVENTIONS: Skin was irradiated with a pulsed dye laser (wavelength = 585 nm; pulse duration = 1.5 milliseconds; 5-mm-diameter spot) using various radiant exposures (8-30 J/cm2) without and with cryogen spray cooling (50- to 300-millisecond cryogen spurts). MAIN OUTCOME MEASURE: Hematoxylin-eosin-stained histologic sections from each irradiated site were examined for the degree of epidermal damage, maximum depth of red blood cell coagulation, and percentage of vessels containing perivascular collagen coagulation. RESULTS: Long cryogen spurt durations (>200 milliseconds) protected the epidermis in light-skinned individuals (skin phototypes I-IV) at the highest radiant exposure (30 J/cm2); however, epidermal protection could not be achieved in dark-skinned individuals (skin phototypes V-VI) even at the lowest radiant exposure (8 J/cm2). The red blood cell coagulation depth increased with increasing radiant exposure (to >2.5 mm for skin phototypes I-IV and to approximately 1.2 mm for skin phototypes V-VI). In addition, long cryogen spurt durations (>200 milliseconds) prevented perivascular collagen coagulation in all skin types. CONCLUSIONS: Cryogen spurt durations much longer than those currently used in therapy (>200 milliseconds) may be clinically useful for protecting the epidermis and perivascular tissues when using high radiant exposures during cutaneous laser therapies. Additional studies are necessary to prove clinical safety of these protocols.

CT-guided radiofrequency ablation of a pulmonary metastasis followed by surgical resection.

Outpatient CT-guided radiofrequency ablation (RFA) of a pulmonary metastasis followed by surgical resection and histopathological analysis was performed in a 72-year-old lady suffering from a peritoneal leiomyosarcoma. Histological workup 3 weeks post-ablation showed complete devitalization of the metastasis. This case report demonstrates that complete thermal destruction of a pulmonary metastasis by percutaneous image-guided RFA is possible.