This PDF file contains the front matter associated with SPIE Proceedings Volume 10066 including the Title Page, Copyright information, Table of Contents, Introduction, and Conference Committee listing.

The greatest challenge in image-guided thermal ablation (IGTA) of liver tumors is a relatively high recurrence rate (ca. 30%) due to incomplete ablation. To meet this challenge, we have developed a novel Thermal Accelerator (TA) to demonstrate its capability to, 1) augment microwave (MW) energy from a distance unattainable by antenna alone; 2) turn into a gel at body temperature; 3) act as a CT or US contrast. We have examined the TA efficiency using in vitro and ex vivo models: microwave power, TA dose, frequencies and TA-to-tip distance were varied, and temperature readings compared with and without TA. Using the in vitro model, it was established that both the rate and magnitude of increase in ablation zone temperature were significantly greater with TA under all tested conditions (p<0.0001). On ultrasound imaging, the TA was echogenic as gel. On CT, TA density was proportional to dose, with average values ranging from 329 HU to 3071 HU at 10 mg/mL and 1,000mg/mL, respectively. TA can be accurately deposited to a target area using CT or US as image-guidance and augment MW energy effectively so that ablation time is significantly reduced, which will contribute to complete ablation. The preliminary results obtained from in vivo experiments using swine as an animal model are consistent with the observations made in in vitro and en vivo studies.

Magnetic fluid hyperthermia (MFH) is a promising avenue for noninvasive or minimally invasive therapies including tissue ablation, hyperthermia, and drug delivery. Magnetic particle imaging (MPI) is a promising new medical imaging modality with wide-ranging applications including angiography, cell tracking, and cancer imaging. MFH and MPI are kindred technologies leveraging the same physics: Both MFH and MPI function by exciting iron oxide magnetic nanoparticles with AC magnetic fields. In this manuscript, we show that this can be leveraged for combined MPI-MFH. The gradient fields employed in MPI can benefit MFH by providing high resolution targeting anywhere in the body, and a dual system provides opportunities for real-time diagnostic imaging feedback. Here we experimentally quantify the spatial localization of MFH using MPI gradient fields with a custom MPI-MFH system, demonstrating approximately 3 mm heating resolution in phantoms. We show an ability to precisely target phantom components as desired and provide heating of approximately 150 W g^-1. We also show preliminary simultaneous MPI-MFH data.

Nanosecond electric pulses (nsEPs) are known to cause a variety of effects on mammalian cells, ranging from destabilization of cell membranes to changes in cytoskeleton and elastic moduli. Measurement of a cells mechanoelastic properties have previously been limited to only invasive and destructive techniques such as atomic force microscopy or application of optical tweezers. However, due to recent advances, Brillouin spectroscopy has now become viable as a non-contact, non-invasive method for measuring these properties in cells and other materials. Here, we present progress toward applying Brillouin spectroscopy using a unique microscopy system for measuring changes in CHO-K1 cells when exposed to nsEPs of 600ns pulse duration with intensity of 50kV/cm. Successful measurement of mechanoelastic changes in these cells will demonstrate Brillouin spectroscopy as a viable method for measuring changes in elastic properties of other cells and living organisms.

It has recently been shown that cancer treatments such as radiation and hyperthermia, which have conventionally been viewed to have modest immune based anti-cancer effects, may, if used appropriately stimulate a significant and potentially effective local and systemic anti-cancer immune effect (abscopal effect) and improved prognosis. Using eight spontaneous canine cancers (2 oral melanoma, 3 oral amelioblastomas and 1 carcinomas), we have shown that hypofractionated radiation (6 x 6 Gy) and/or magnetic nanoparticle hyperthermia (2 X 43°C / 45 minutes) and/or an immunogenic virus-like nanoparticle (VLP, 2 x 200 μg) are capable of delivering a highly effective cancer treatment that includes an immunogenic component. Two tumors received all three therapeutic modalities, one tumor received radiation and hyperthermia, two tumors received radiation and VLP, and three tumors received only mNP hyperthermia. The treatment regimen is conducted over a 14-day period. All patients tolerated the treatments without complication and have had local and distant tumor responses that significantly exceed responses observed following conventional therapy (surgery and/or radiation). The results suggest that both hypofractionated radiation and hyperthermia have effective immune responses that are enhanced by the intratumoral VLP treatment. Molecular data from these tumors suggest Heat Shock Protein (HSP) 70/90, calreticulin and CD47 are targets that can be exploited to enhance the local and systemic (abscopal effect) immune potential of radiation and hyperthermia cancer treatment.

Plasma medicine is a rapidly growing field of treatment, with the number and type of medical applications growing annually, such as dentistry, cancer treatment, wound treatment, Antimicrobial (bacteria, biofilm, virus, fungus, prions), and surface sterilization. Work promoting muscle and blood vessel regeneration and osteointegration is being investigated. This review paper will cover the latest treatments using gas-based plasmas in medicine. Disinfection of water and new commercial systems will also be reviewed, as well as vaccine deactivation. With the rapid increase in new investigators, development of new devices and systems for treatment, and wider clinical applications, Plasma medicine is becoming a powerful tool in in the field of medicine. There are a wide range of Plasma sources that allows customization of the effect. These variations include frequency (DC to MHz), voltage capacity (kV), gas source (He, Ar; O₂, N₂, air, water vapor; combinations), direct/indirect target exposure, and water targets.

Data were previously reported on studies of the effects of electrical discharges on the corrosion and wear of simple, single-wire test devices immersed in isotonic saline ¹ . This work showed that there are a wide variety of mechanisms that can explain various aspects of electrode mass loss, even with very simple electrode geometries and operating conditions. It was found that the electrode material composition played an important role. Subsequently, our studies were expanded to include more realistic device geometries and operating conditions. This paper shows the results of studies on wear characteristics of electrodes made from a variety of highly corrosion resistant metals and alloys, including Waspaloy, Hastelloy, Inconel, Havar, Monel, and other pure metals such as Hafnium. All of these metals underwent wear testing under clinically relevant conditions. Depending on the operating conditions, multiple discrete physical and chemical effects were observed at different locations on the surface of an individual millimeter-scale device electrode. Scanning electron microscope (SEM) micrographs, Energy-dispersive X-ray spectroscopy (EDS) and area loss data will be presented for a variety of test conditions and electrode materials.

Antibacterial studies of inorganic nanoparticles (nps) have become important due to the increased bacterial resistance against antibiotics. We used Zinc oxide nanoparticles (ZnO nps), which possess excellent photocatalytic properties with a wide band gap (Eg), are listed as “generally recognized as safe” by the Food and Drug Administration (FDA) and have shown antibacterial activity (AA) against many bacterial strains. The AA of ZnO nps is partly attributed to the production of Reactive Oxygen Species (ROS) by photocatalysis. When ZnO nps in aqueous media are illuminated with an energy <Eg, electron-hole pairs are generated on nps surface reacting with water and Oxygen molecules to generate hydroxyl-radical (OH• ), superoxide-radical (O₂ •- ) and hydrogen-peroxide (H₂O₂). These ROS induce cell membrane damage resulting in cell death. However, the application of inorganic nps in medical treatments is limited due to the possible long-term side effects of nps release. To prevent its release, ZnO nps were dispersed into Polycaprolactone (PCL) fibers obtained by electrospinning technique. To optimize the use of ZnO nps concentration, we developed coreshell coaxial electrospun fibers where the core corresponded to PCL and the shell to a mixture of ZnO nps/PCL. Thus, ZnO nps were only dispersed on the surface of the fibers increasing its superficial contact area. We evaluated the AA against E. coli of different electrospun ZnO nps/PCL fibers under two different conditions: UVA pre-illumination and darkness. Preliminary results suggest that the AA against E. coli is better when electrospun ZnO nps/PCL were preilluminated with UVA than under darkness conditions.

Discogenic back pain presents a major public health issue, with current therapeutic interventions limited to short-term symptom relief without providing regenerative remedies for diseased intervertebral discs (IVD). Many of these interventions are invasive and can diminish the biomechanical integrity of the IVDs. Low intensity pulsed ultrasound (LIPUS) is a potential treatment option that is both non-invasive and regenerative. LIPUS has been shown to be a clinically effective method for the enhancement of wound and fracture healing. Recent in vitro studies have shown that LIPUS stimulation induces an upregulation functional matrix proteins and downregulation of inflammatory factors in cultured IVD cells. However, we do not know the effects of LIPUS on an in vivo model for intervertebral disc degeneration. The objective of this study was to show technical feasibility of building a LIPUS system that can target the rat tail IVD and apply this setup to a model for acute IVD degeneration. A LIPUS exposimetry system was built using a 1.0 MHz planar transducer and custom housing. Ex vivo intensity measurements demonstrated LIPUS delivery to the center of the rat tail IVD. Using an established stab-incision model for disc degeneration, LIPUS was applied for 20 minutes daily for five days. For rats that displayed a significant injury response, LIPUS treatment caused significant upregulation of Collagen II and downregulation of Tumor Necrosis Factor – α gene expression. Our preliminary studies indicate technical feasibility of targeted delivery of ultrasound to a rat tail IVD for studies of LIPUS biological effects.

Recent failures in hysteroscopic female sterilization procedures have brought into question the implantation of nonresorbable metal devices into the fallopian tubes due to long-term risks such as migration, fragmentation, and tubal perforation. The goal of this study is to assess whether a porous, biodegradable implant can be deposited into the fallopian tube lumen with or without a local mild heat treatment to generate a safe and permanent fallopian tube occlusion/sterilization event. The technologies investigated included freeze-cast collagen-based scaffolds and magnetic nanoparticle (MNP) based scaffolds. In vitro assessment of iron oxide MNP-based scaffolds was performed to determine the absorption rate density (ARD); subsequent computational modeling quantified the thermal in vivo steady state temperature as a function of tubal radius for treatment planning. For collagen-based scaffolds, in vivo testing was performed to study the biocompatibility in a mouse flank model, followed by implantation into an in vivo anestrus feline uterine horn (animal model for the fallopian tube). Biological responses were studied histopathologically. Uterine horn patency was assessed via radiographic imaging. Preliminary studies suggest the MNP-impregnated scaffold and a safe, noninvasive AMF excitation field have potential to generate a sufficient focal fallopian tube thermal dose to create a fibrotic healing event and ultimately, permanent tubal occlusion.

A faster healing process was observed in superficial skin wounds after irradiation with a blue LED (EmoLED) photocoagulator. EmoLED is a compact handheld device, used to induce a thermal effect and thus coagulation in superficial abrasions. We present the results of an in vivo study, conducted in a mouse model, to analyze the induced wound healing. Two superficial abrasions were produced on the back of the mice: one area was treated with EmoLED (1.4 W/cm2, 30 s treatment time), while the other one was left naturally recovering. During the treatment, a temperature around 40-45°C was induced on the abrasion surface. Mice back healthy skin was used as a control. The animals underwent a follow up study and were sacrificed at 0, 1, 3, 6, 9, 12, 18, 21, 24 hours p.o. and 6 days p.o.. Samples from the two abraded areas were harvested and examined by histopathological and immunofluorescence analysis, SHG imaging and confocal microscopy. The aim of the study was to investigate the inflammatory infiltrate, mastocyte population, macrophage subpopulation, fibroblasts and myofibroblasts. Our results show that soon after the treatment, both the inflammatory infiltrate and the M1 macrophage subpopulation appear earlier in the treated, compared to a delayed appearance in the untreated samples. There was no alteration in collagen morphology in the recovered wound. This study confirms the preliminary results obtained in a previous study on a rat model: the selective photothermal effect we used for inducing immediate coagulation in superficial wounds seems to be associated to a faster and improved healing process.

Stroke is a devastating disease, which is the third leading cause of death and disability worldwide. Although the incidence of stroke increases progressively with age, morbidity among young and middle-aged adults is increasing annually. Medications nevertheless remain the bulwarks of stroke. The treatment is ineffective, speculative and has a long treatment cycle. The function of acupuncture and moxibustion, which are potential therapeutic tools for stroke, is still controversial. Recently, Low-level light therapy (LLLT) has been demonstrated potent in vivo efficacy for treatment of ischemic conditions of acute myocardial infraction and stroke in multiple validated animal models. Optimum LLLT treatment has a dominant influence on therapy of stroke. While more than a thousand clinical trials have been halted, only a few trials on animals have been reported. We addressed this issue by simulating near-infrared light propagation with accurate visible Chinese human head by Monte Carlo modeling. The visible human head embody region of atherosclerotic plaques in head. Through comparing the light propagation of different light illumination, we can get a precise, optimized and straightforward treatment. Here, we developed a LLLT helmet for treating stroke depend on near-infrared light. There are more than 30 LED arrays in in multi-layered 3D printed helmet. Each LED array has independent water-cooling module and can be adjusted to touch the head of different subjects based on Electro pneumatic module. Moreover, the software provides the setup of illumination parameters and 3D distribution of light fluence rate distribution in human brain.

It is now known that many tumors develop molecular signals (immune checkpoint modulators) that inhibit an effective tumor immune response. New information also suggest that even well-known cancer treatment modalities such as radiation and hyperthermia generate potentially beneficial immune responses that have been blocked or mitigated by such immune checkpoints, or similar molecules. The cancer therapy challenge is to; a) identify these treatment-based immune signals (proteins, antigens, etc.); b) the treatment doses or regimens that produce them; and c) the mechanisms that block or have the potential to promote them. The goal of this preliminary study, using the B6 mouse – B16 tumor model, clinically relevant radiation doses and fractionation schemes (including those used clinically in hypofractionated radiation therapy), magnetic nanoparticle hyperthermia (mNPH) and sophisticated protein, immune and tumor growth analysis techniques and modulators, is to determine the effect of specific radiation or hyperthermia alone and combined on overall treatment efficacy and immunologic response mechanisms. Preliminary analysis suggests that radiation dose (10 Gy vs. 2 Gy) significantly alters the mechanism of cell death (apoptosis vs. mitosis vs. necrosis) and the resulting immunogenicity. Our hypothesis and data suggest this difference is protein/antigen and immune recognition-based. Similarly, our evidence suggest that radiation doses larger than the conventional 2 Gy dose and specific hyperthermia doses and techniques (including mNP hyperthermia treatment) can be immunologically different, and potentially superior to, the radiation and heat therapy regimens that are typically used in research and clinical practice.

Alternating magnetic field (AMF) configurable at a range of frequencies is a critical need for optimization of magnetic nanoparticle based hyperthermia, and for their application in targeted drug delivery. Currently, most commercial AMF devices including induction heaters operate at one factory-fixed frequency, thereby limiting customized frequency configuration required for triggered drug release at mild hyperthermia (40-42°C) and ablations (>55°C). Most AMF devices run as an inductor-capacitor resonance network that could allow AMF frequencies to be changed by changing the capacitor bank or the coil looped with it. When developing AMF inhouse, the most expensive component is usually the RF power amplifier, and arguably the most critical step of building a strong AMF field is impedance-matched coupling of RF power to the coolant-cooled AMF coil. AMF devices running at 10KA/m strength are quite common, but generating AMF at that level of field strength using RF power less than 1KW has remained challenging. We practiced a few techniques for building 10KA/m AMFs at different frequencies, by utilizing a 0.5KW 80-800KHz RF power amplifier. Among the techniques indispensable to the functioning of these AMFs, a simple cost-effective technique was the tapping methods for discretely or continuously adjusting the position of an RF-input-tap on a single-layer or the outer-layer of a multi-layer AMF coil for maximum power coupling into the AMF coil. These in-house techniques when combined facilitated 10KA/m AMF at frequencies of 88.8 KHz and higher as allowed by the inventory of capacitors using 0.5KW RF power, for testing heating of 10-15nm size magnetic particles and on-going evaluation of drug-release by low-level temperature-sensitive liposomes loaded with 15nm magnetic nanoparticles.

The magneto-thermo-acoustic effect that we predicted in 2013 refers to the generation of acoustic-pressure wave from magnetic nanoparticle (MNP) when thermally mediated under an alternating magnetic field (AMF) at a pulsed or frequency-chirped application. Several independent experimental studies have since validated magneto-thermoacoustic effect, and a latest report has discovered acoustic-wave generation from MNP at the second-harmonic frequency of the AMF when operating continuously. We propose that applying two AMFs with differing frequencies to MNP will produce acoustic-pressure wave at the summation and difference of the two frequencies, in addition to the two second-harmonic frequencies. Analysis of the specific absorption dynamics of the MNP when exposed to two AMFs of differing frequencies has shown some interesting patterns of acoustic-intensity at the multiple frequency components. The ratio of the acoustic-intensity at the summation-frequency over that of the difference-frequency is determined by the frequency-ratio of the two AMFs, but remains independent of the AMF strengths. The ratio of the acoustic-intensity at the summation- or difference-frequency over that at each of the two second-harmonic frequencies is determined by both the frequency-ratio and the field-strength-ratio of the two AMFs. The results indicate a potential strategy for localization of the source of a continuous-wave magneto-thermalacoustic signal by examining the frequency spectrum of full-field non-differentiating acoustic detection, with the field-strength ratio changed continuously at a fixed frequency-ratio. The practicalities and challenges of this magnetic spatial localization approach for magneto-thermo-acoustic imaging using a simple envisioned set of two AMFs arranged in parallel to each other are discussed.

Although there is long association of medical hyperthermia and immune stimulation, the relative lack of a quantifiable and reproducible effect has limited the utility and advancement of this relationship in preclinical/clinical cancer and non-cancer settings. Recent cancer-based immune findings (immune checkpoint modulators etc.) including improved mechanistic understanding and biological tools now make it possible to modify and exploit the immune system to benefit conventional cancer treatments such as radiation and hyperthermia. Based on the prior experience of our research group including; cancer-based heat therapy, magnetic nanoparticle (mNP) hyperthermia, radiation biology, cancer immunology and Cowpea Mosaic Virus that has been engineered to over express antigenic proteins without RNA or DNA (eCPMV/VLP). This research was designed to determine if and how the intra-tumoral delivery of mNP hyperthermia and VLP can work together to improve local and systemic tumor treatment efficacy. Using the C3H mouse/MTG-B mammary adenocarcinoma cell model and the C57-B6 mouse/B-16-F10 melanoma cancer cell model, our data suggests the appropriate combination of intra-tumoral mNP heat (e.g. 43°C /30-60 minutes) and VLP (100 μg/200 mm³ tumor) not only result in significant primary tumor regression but the creation a systemic immune reaction that has the potential to retard secondary tumor growth (abscopal effect) and resist tumor rechallenge. Molecular data from these experiments suggest treatment based cell damage and immune signals such as Heat Shock Protein (HSP) 70/90, calreticulin, MTA1 and CD47 are potential targets that can be exploited to enhance the local and systemic (abscopal effect) immune potential of hyperthermia cancer treatment

Purpose: A ferritin-containing nanoparticle conjugated with a target-specific antibody was investigated as a MRI contrast agent for tumor detection. A genetically modified ferritin to markedly improve Fe (III) payload (up to 7,000 Fe ions), was chemically tethered to a monoclonal antibody against rat Nectin-like molecule 5 (Necl-5). Necl-5 is a cell surface glycoprotein that is highly expressed on the cell surface of many common epithelial cancers, including prostate cancer. It was previously demonstrated that this novel nanoconjugate agent exhibited effective in vitro targeting of Necl- 5 expressing tumor cells and exhibited strong MRI contrast characteristics via shortening of T₂. Here, we demonstrate that the nanoconjugate-Necl-5 interaction can be exploited to target and detect tumor in vivo by MRI. Procedure: Using an in vivo tumor model (i.e., tumor size 0.5^-1 cm, immunodeficient beige/nude/xid mouse, xenograft injection with transformed rat prostate cells), efficacy of the conjugate targeting the tumor was examined. We used two injection strategies, a direct and a tail vein injection (0.8 mg, 300 μL per subject). Pre-injection baseline and postinjection scans were performed with the following spin-echo sequence parameters: Field of view = 90x53mm, reconstruction matrix size = 192x114, slice thickness = 1mm (10 slices), repetition time (TR) = 2070 ms, echo times (TE) = 11-198 ms in 11ms steps (18 echoes), number of averages = 2, acquisition time per scan = 7min 56s. Results: All T₂ data obtained were converted to R₂ for demonstration purposes (R₂ = 1/T₂). The tail vein injected conjugate significantly increased R₂ response (22.9 ± 5.2 s^-1) as compared to control (13.5 ±1.7 s^-1) at 4 h. The weaker R₂ increase was noted (15.2 ± 2.0 s^-1) at 24 h. No notable changes in R₂ were observed in surrounding tissues regardless the stages of the measurement. We also measured the initial conjugate kinetics for both injection methods with respect to the ability of targeting the tumor. Direct injection of the nanoconjugate in to the center of the tumor showed a stronger and more rapid increase in R2 than the tail vein injection. Conclusion: The nanoconjugate interacts strongly and selectively in situ with Necl-5 overexpressing tumor cells. Direct injection of the nanoconjugate into the body of the tumor caused a more significant in situ R₂ increase in MRI than the tail vein injection. Varying degrees of R₂ increase within the tumor mass is likely to represent different distribution patterns of the conjugate, reflective of tumor heterogeneity.

Endoluminal high-intensity ultrasound offers spatially-precise thermal ablation of tissues adjacent to body lumens, but is constrained in treatment volume and penetration depth by the effective aperture of integrated transducers, which are limited in size to enable delivery through anatomical passages, endoscopic instrumentation, or laparoscopic ports. This study introduced and investigated three distinct endoluminal ultrasound applicator designs that can be delivered in a compact state then deployed or expanded at the target luminal site to increase the effective therapeutic aperture. The first design incorporated an array of planar transducers which could be unfolded at specific angles of convergence between the transducers. Two alternative designs consisted of fixed transducer sources surrounded by an expandable multicompartment balloon that contained acoustic reflector and dynamically-adjustable fluid lenses compartments. Parametric studies of acoustic output were performed across device design parameters via the rectangular radiator and secondary sources methods. Biothermal models were used to simulate resulting temperature distributions in three-dimensional heterogeneous tissue models. Simulations indicate that a deployable transducer array can increase volumetric coverage and penetration depth by ~80% and ~20%, respectively, while permitting more conformal thermal lesion shapes based on the degree of convergence between the transducers. The applicator designs incorporating reflector and fluid lenses demonstrated enhanced focal gain and penetration depth that increased with the diameter of the expanded reflector-lens balloon. Thermal simulations of assemblies with ~12 mm compact profiles and ~50 mm expanded balloon diameters demonstrated generation of localized thermal lesions at depths up to ~10 cm in liver tissue.

Penetrating thermal tissue damage/spread is an important aspect of many electrosurgical devices and correlates with effective tissue cutting, hemostasis, preservation of adjacent critical structures and tissue healing. This study compared the thermal damage/spread associated with the PhotonBlade, Valleylab Pencil, Valleylab EDGE Coated Pencil, PlasmaBlade 3.0S and PlasmaBlade 4.0, when performing a single pass dynamic tissue cut in fresh extirpated porcine longissimus muscle. These devices were used in a fashion that emulated their use in the clinical setting. Each device’s thermal damage/spread, at Minimum, Median and Maximum power input settings, was assessed with nitroblue tetrazolium viability staining in the WVU Pathology Laboratory for Translational Medicine. The thermal damage/spread associated with the PhotonBlade was compared with the other devices tested based on the individual treatment results (n=179 cuts combined). In summary, the PhotonBlade overall demonstrated the least penetrating thermal tissue damage/spread, followed by the PlasmaBlade 4.0, then Valleylab Pencil and PlasmaBlade 3.0S and then Valleylab EDGE Coated Pencil in order of increasing thermal damage/spread depths.

Thermal ablation is a dominant therapeutic option for minimally invasive treatment of menorrhagia. Compared to other energy modalities for ablation, microwaves offer the advantages of conformal energy delivery to tissue within short times. The objective of endometrial ablation is to destroy the endometrial lining of the uterine cavity, with the clinical goal of achieving reduction in bleeding. Previous efforts have demonstrated clinical use of microwaves for endometrial ablation. A considerable shortcoming of most systems is that they achieve ablation of the target by translating the applicator in a point-to-point fashion. Consequently, treatment outcome may be highly dependent on physician skill. Global endometrial ablation (GEA) not only eliminates this operator dependence and simplifies the procedure but also facilitates shorter and more reliable treatments. The objective of our study was to investigate antenna structures and microwave energy delivery parameters to achieve GEA. Another objective was to investigate a method for automatic and reliable determination of treatment end-point. A 3D-coupled FEM electromagnetic and heat transfer model with temperature and frequency dependent material properties was implemented to characterize microwave GEA. The unique triangular geometry of the uterus where lateral narrow walls extend from the cervix to the fundus forming a wide base and access afforded through an endocervical approach limit the overall diameter of the final device. We investigated microwave antenna designs in a deployed state inside the uterus. The impact of ablation duration on treatment outcome was investigated. Prototype applicators were fabricated and experimentally evaluated in ex vivo tissue to verify the simulation results and demonstrate proof-of-concept.

Expansions in minimally invasive medical devices and technologies with thermal mechanisms of action are continuing to advance the practice of medicine. These expansions have led to an increasing need for appropriate animal models to validate and quantify device performance. The planning of these studies should take into consideration a variety of parameters, including the appropriate animal model (test system - ex vivo or in vivo; species; tissue type), treatment conditions (test conditions), predicate device selection (as appropriate, control article), study timing (Day 0 acute to more than Day 90 chronic survival studies), and methods of tissue analysis (tissue dissection - staining methods). These considerations are discussed and illustrated using the fresh extirpated porcine longissimus muscle model for endometrial ablation.

Microwave ablation (MWA) is an emerging minimally invasive treatment option for malignant lung tumors. Compared to other energy modalities, such as radiofrequency ablation, MWA offers the advantages of deeper penetration within high impedance tissues such as aerated lung, shorter treatment times, and less susceptibility to the cooling heat-sink effects of air and blood flow. Previous studies have demonstrated clinical use of MWA for treating lung tumors; however, these procedures have relied upon the percutaneous application of rigid microwave antennas. The objective of our work was to develop and characterize a novel flexible microwave applicator which could be integrated with a bronchoscopic imaging and software guidance platform to expand the use of MWA as a treatment option for small (< 2cm) pulmonary tumors. This applicator would allow physicians an even less invasive, immediate treatment option for lung tumors identified within the scope of current medical procedures. It may also improve applicator placement accuracy and increase efficacy while minimizing the risk of procedural complications. A 2D-axisymmetric coupled FEM electromagnetic-heat transfer model was implemented to characterize expected antenna radiation patterns, ablation size and shape, and optimize antenna design for lung tissue. A prototype device was fabricated and evaluated in ex vivo tissues to verify simulation results and serve as proof-of-concept. Additional experiments were conducted in an in vivo animal model to further characterize the proposed system.

Background: Hyperthermia, i.e., raising tissue temperature to 40-45°C for 60 min, has been demonstrated to increase the effectiveness of radiation and chemotherapy for cancer. Although multi-element conformal heat applicators are under development to provide more adjustable heating of contoured anatomy, to date the most often used applicator to heat superficial disease is the simple microwave waveguide. With only a single power input, the operator must be resourceful to adjust heat treatment to accommodate variable size and shape tumors spreading across contoured anatomy. Methods: We used multiphysics simulation software that couples electromagnetic, thermal and fluid dynamics physics to simulate heating patterns in superficial tumors from commercially available microwave waveguide applicators. Temperature distributions were calculated inside homogenous muscle and layered skin-fat-muscle-tumor-bone tissue loads for a typical range of applicator coupling configurations and size of waterbolus. Variable thickness waterbolus was simulated as necessary to accommodate contoured anatomy. Physical models of several treatment configurations were constructed for comparison of simulation results with experimental specific absorption rate (SAR) measurements in homogenous muscle phantom. Results: Accuracy of the simulation model was confirmed with experimental SAR measurements of three unique applicator setups. Simulations demonstrated the ability to generate a wide range of power deposition patterns with commercially available waveguide antennas by controllably varying size and thickness of the waterbolus layer. Conclusion: Heating characteristics of 915 MHz waveguide antennas can be varied over a wide range by controlled adjustment of microwave power, coupling configuration, and waterbolus lateral size and thickness. The uniformity of thermal dose delivered to superficial tumors can be improved by cyclic switching of waterbolus thickness during treatment to proactively shift heat peaks and nulls around under the aperture, thereby reducing patient pain while increasing minimum thermal dose by end of treatment.

Thermal ablation is the use of heat to induce cell death through coagulative necrosis. Ideally, complete ablation of tumor cells with no damage to surrounding critical structures such as blood vessels, nerves or even organs is desired. Ablation monitoring techniques are often employed to ensure optimal tumor ablation. In thermal tissue ablation, tissue damage is known to be dependent on the temperature and time of exposure. Aptly, current methods for monitoring ablation rely profoundly on local tissue temperature and duration of heating to predict the degree of tissue damage. However, such methods do not take into account the microstructural and physiological changes in tissues as a result of thermocoagulation. Light propagation within biological tissues is known to be dependent on the tissue microstructure and physiology. During tissue denaturation, changes in tissue structure alter light propagations in tissue which could be used to directly assess the extent of thermal tissue damage. We report the use of a spectroscopic system for monitoring the tissue optical properties during heating of ex vivo liver tissues. We observed that during tissue denaturation, continuous changes in wavelength-averaged μ_a(λ) and μ’_s(λ) followed a sigmoidal trend and are correlated with damage predicted by Arrhenius model.

The speed of sound (SOS) for ultrasound devices used for imaging soft tissue is often calibrated to water, 1540 m/s¹ , despite in-vivo soft tissue SOS varying from 1450 to 1613 m/s² . Images acquired with 1540 m/s and used in conjunction with stereotactic external coordinate systems can thus result in displacement errors of several millimeters. Ultrasound imaging systems are routinely used to guide interventional thermal ablation and cryoablation devices, or radiation sources for brachytherapy³ . Brachytherapy uses small radioactive pellets, inserted interstitially with needles under ultrasound guidance, to eradicate cancerous tissue⁴ . Since the radiation dose diminishes with distance from the pellet as 1/r² , imaging uncertainty of a few millimeters can result in significant erroneous dose delivery^5,6. Likewise, modeling of power deposition and thermal dose accumulations from ablative sources are also prone to errors due to placement offsets from SOS errors⁷ . This work presents a method of mitigating needle placement error due to SOS variances without the need of ionizing radiation^2,8. We demonstrate the effects of changes in dosimetry in a prostate brachytherapy environment due to patientspecific SOS variances and the ability to mitigate dose delivery uncertainty. Electromagnetic (EM) sensors embedded in the brachytherapy ultrasound system provide information regarding 3D position and orientation of the ultrasound array. Algorithms using data from these two modalities are used to correct bmode images to account for SOS errors. While ultrasound localization resulted in >3 mm displacements, EM resolution was verified to <1 mm precision using custom-built phantoms with various SOS, showing ~1% accuracy in SOS measurement.

Large vessels can be reliably sealed with radio frequency current. High apposition pressures are necessary to ensure a high probability of a successful seal. However, the complex architecture of the vessels, particularly arteries, means that results can vary substantially even with similar thermal histories. The relative volume fractions and spatial distributions of collagen, elastin, and smooth muscle dominate the vessel function in vivo and can even vary from proximal to distal locations in the same vessel. We begin by reviewing the architectural features characteristic of porcine and canine large vessels and conclude with an experimental and numerical modeling demonstration of the reasons why cylindrical electrodes are a sub-optimal choice.

Surgical vessel sealing systems are widely used to achieve hemostasis and dissection in open surgery and minimally invasive, laparoscopic surgery. This enabling technology was developed about 17 years ago and continues to evolve with new devices and systems achieving improved outcomes. Histopathological assessment of thermally sealed tissues is a valuable tool for refining and comparing performance among surgical vessel sealing systems. Early work in this field typically assessed seal time, burst rate, and failure rate (in-situ). Later work compared histological staining methods with birefringence to assess the extent of thermal damage to tissues adjacent to the device. Understanding the microscopic architecture of a sealed vessel is crucial to optimizing the performance of power delivery algorithms and device design parameters. Manufacturers rely on these techniques to develop new products. A system for histopathological evaluation of vessels and sealing performance was established, to enable the direct assessment of a treatment’s tissue effects. The parameters included the commonly used seal time, pressure burst rate and failure rate, as well as extensions of the assessment to include its likelihood to form steam vacuoles, adjacent thermal effect near the device, and extent of thermally affected tissue extruded back into the vessel lumen. This comprehensive assessment method provides an improved means of assessing the quality of a sealed vessel and understanding the exact mechanisms which create an optimally sealed vessel.

We present the development of a 5 mm, piezo-actuated, ultrafast laser scalpel for microsurgery with a capability to deliver energies in excess of 1 μJ per pulse. Having previously established that the maximum energy deliverable was limited by cladding damage in photonic badgap fibers, we utilized a large, 35μm cored inhibited-coupling Kagome fiber that allowed the delivery of micro-Joule energy femtosecond pulses. To maintain diffraction limited performance over the entire scan range of the piezo-actuated fiber tip, special objective lenses were developed and manufactured out of a high-refractive index Zinc Sulfide (ZnS) crystal. The probe was packaged in hypodermic 304SS stainless steel with a form factor minimizing in-line configuration. The probe’s performance was tested via metal and tissue ablation studies, characterizing highspeed ablation parameters and uniformity of ablation over the scan area. Additionally, we studied the nonlinear performance of ZnS and Calcium Fluoride (CaF₂) as materials for refractive optics and determined the maximum energy deliverable through our probe using these optical materials. The high energy delivery through the probe system should allow for fast and effective tissue ablation.

A continuous-wave, 40 Watt, 1470 nm laser was explored for rapid and precise dissection of porcine mesentery fascia and liver tissues, ex vivo. Laser energy was delivered through a 550-μm-core optical fiber inside a 5-mm-OD, laparoscopic probe, with detachable, 2 mm, sapphire ball rolling tip. Fascia tissue was cleanly dissected with scanning rates from 2.0 - 4.5 mm/s using 16 - 31W. Fascia collateral thermal damage measured as low as 180 ± 50 μm at 4.5 mm/s scan speed. Porcine liver ablation crater depth measured up to 1010 ± 220 μm with 30 W at 2.0 mm/s or as shallow as 80 ± 30 μm with 10 W at 10 mm/s. Peak temperatures reached 130 °C at ball tip and 75 °C on metal jaws. The 1470-nm laser and probe show promise for laparoscopic tissue cutting and coagulation.

Nanosecond pulsed electric fields (nsPEF) have proven useful for transporting cargo across cell membranes and selectively activating cellular pathways. The chemistry and biophysics governing this cellular response, however, are complex and not well understood. Recent studies have shown that the conductivity of the solution cells are exposed in could play a significant role in plasma membrane permeabilization and, thus, the overall cellular response. Unfortunately, the means of detecting this membrane perturbation has traditionally been limited to analyzing one possible consequence of the exposure – diffusion of molecules across the membrane. This method has led to contradictory results with respect to the relationship between permeabilization and conductivity. Diffusion experiments also suffer from “saturation conditions” making multi-pulse experiments difficult. As a result, this method has been identified as a key stumbling block to understanding the effects of nsPEF exposure. To overcome these limitations, we recently developed a nonlinear optical imaging technique based on second harmonic generation (SHG) that allows us to identify nanoporation in live cells during the pulse in a wide array of conditions. As a result, we are able to explore and fully test whether lower conductivity extracellular solutions could induce more efficient nanoporation. This hypothesis is based on membrane charging and the relative difference between the extracellular solution and the cytoplasm. The experiments also allow us to test the noise floor of our methodology against the effects of ion leakage. The results emphasize that the electric field, not ionic phenomenon, are the driving force behind nsPEF-induced membrane nanoporation.

Irreversible electroporation therapy is utilized to remove cancerous tissues thru the delivery of rapid (250Hz) and high voltage (V) (1,500V/cm) electric pulses across microsecond durations. Clinical research demonstrated that bipolar (BP) high voltage microsecond pulses opposed to monophasic waveforms relieve muscle contraction during electroporation treatment. Our group along with others discovered that nanosecond electric pulses (nsEP) can activate second messenger cascades, induce cytoskeletal rearrangement, and depending on the nsEP duration and frequency, initiate apoptotic pathways. Of high interest across in vivo and in vitro applications, is how nsEP affects muscle physiology, and if nuances exist in comparison to longer duration electroporation applications. To this end, we exposed mature skeletal muscle cells to monopolar (MP) and BP nsEP stimulation across a wide range of electric field amplitudes (1-20 kV/cm). From live confocal microscopy, we simultaneously monitored intracellular calcium dynamics along with nsEP-induced muscle movement on a single cell level. In addition, we also evaluated membrane permeability with Yo-PRO-1 and Propidium Iodide (PI) across various nsEP parameters. The results from our findings suggest that skeletal muscle calcium dynamics, and nsEP-induced contraction exhibit exclusive responses to both MP and BP nsEP exposure. Overall the results suggest in vivo nsEP application may elicit unique physiology and field applications compared to longer pulse duration electroporation.

A new paradigm for strengthening of corneal tissue as well as permanent correction of refractive errors has been proposed. Ultrafast laser irradiation is confined to the levels below optical breakdown such that tissue damage is avoided while creating an ionization field responsible for subsequent photochemical modification of the stroma. The concept was assed using newly developed platform for precise application of a near-IR femtosecond laser irradiation to the cornea in in-vitro experiments. Targeted irradiation with tightly focused ultrafast laser pulses allows spatially resolved crosslinking in the interior of the porcine cornea in the absence of photosensitizers. The formation of intra- or interstromal covalent bonds in collagen matrix locally increases lamellar density. Due to high resolution, treatment is spatially resolved and therefore can be tailored to either enhance structure of corneal stroma or adjust corneal curvature towards correcting refractive errors. As the induced modification is primarily driven by nonlinear absorption, the treatment is essentially wavelength independent, and as such potentially less harmful than current method of choice, joint application of UVA light irradiation in conjunction with riboflavin. Potential applicability of a near-IR femtosecond laser for biomechanical stabilization of cornea and non-invasive refractive eye corrections is discussed.

This paper focuses on the evaluation of a prototype for a computer-based tutoring system for prostate cryosurgery, while reviewing its key building blocks and their benchmark performance. The tutoring system lists geometrical constraints of cryoprobe placement, displays a rendered shape of the prostate, simulates cryoprobe insertion, enables distance measurements, simulates the corresponding thermal history, and evaluates the mismatch between the target region shape and a pre-selected planning isotherm. The quality of trainee planning is measured in comparison with a computergenerated plan, created for each case study by a previously developed planning algorithm, known as bubble-packing. While the tutoring level in this study aims only at geometrical constraints on cryoprobe placement and the resulting thermal history, it creates a unique opportunity to gain insight into the process outside of the operation room. System validation of the tutor has been performed by collecting training data from surgical residents, having no prior experience or advanced knowledge of cryotherapy. Furthermore, the system has been evaluated by graduate engineering students having no formal education in medicine. In terms of match between a planning isotherm and the target region shape, results demonstrate medical residents’ performance improved from 4.4% in a pretest to 37.8% in a posttest over a course of 50 minutes of training (within 10% margins from a computer-optimized plan). Comparing those results with the performance of engineering students indicates similar results, suggesting that planning of the cryoprobe layout essentially revolves around geometric considerations.