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Background and Objective: Spatial Frequency Domain Imaging (SFDI) is a non-contact wide-field optical imaging technology currently being developed to investigate the feasibility of quantitative non-invasive evaluation of burn wound severity in a rat model. Our objective is to determine the potential of SFDI for mapping quantitative changes in spatially resolved tissue oxygen saturation and water concentration may be indicative of burn wound severity, healing, and further complications. In this portion of the investigation, we focus on the development of a rat burn model and the acute response of tissue to burn wounds. Study Design/Materials and Methods: A controlled burn protocol involving a heated brass comb was applied to 6 rats. Imaging was acquired at 17 evenly spaced wavelengths in the near-infrared from 650 to 970 nm. Over the course of the 3 hour post-burn period, we were able to map quantitative changes in spatially resolved chromophores. Burn severities were verified post-experiment using standard H and E histology and optical microscopy. Results/Conclusion: In total, we were able to induce 12 superficial-partial thickness burns, 8 deep-partial thickness burns, and 4 full thickness burns in our rat models. While several tissue chromophores were tracked, we found that changes in oxygen saturation and water concentration to be sensitive indicators of burn severity. Future work will include additional longitudinal studies over a period of days in order to investigate which parameters are correlated to tissue healing. © 2013 SPIE.

Acquired subglottic stenosis is a narrowing of the airway caused by prolonged endotracheal intubation. Currently, there are no non-invasive means to diagnose the disease. A previous study by this same group introduced optical coherence tomography (OCT) as a means of monitoring the progression of stenosis. The aim of the current study was to qualitatively and quantitatively analyze OCT images obtained from a subglottic stenosis model of the rabbit airway. 15 rabbits were used throughout the study, and a MEMs based OCT probe was utilized. The OCT images obtained were analyzed using a free software program, 3D Slicer. The region of scarred tissue was grown out and measured quantitatively. This study demonstrated the feasibility of using a program to quantify the progression of scarring in OCT images, in addition to qualitatively correlating between histology, endoscopic, and OCT images. Future works may include utilization of a long-range probe and use of a pressure necrosis model to better emulate the actual onset of neonatal subglottic stenosis. © 2013 SPIE.

BACKGROUND AND OBJECTIVE: Photodynamic therapy (PDT)-induced disruption of the blood-brain barrier (BBB) has been investigated as a technique for the delivery of therapeutic agents to selective regions of the brain. The purpose of this study was to determine the effects of PDT on the migration of systemically administered exogenous macrophages (Ma) loaded with iron oxide nanoparticles in non-tumor bearing rats. MATERIALS AND METHODS: A control group consisting of three Sprague-Dawley rats was injected with iron oxide-loaded rat alveolar Ma via jugular vein catheter while two animals were subjected to intracranial injection of iron oxide-loaded Ma. PDT-treated animals were injected with photosensitizer (AlPcS2a ; 1 mg/kg i.p.) followed by light irradiation (wavelength = 670 nm; light dose = 2.5 J) 48 hours later. Light irradiation was performed through the skull. Prior to light irradiation, iron oxide-loaded Ma were administered to each animal. Animals in all groups were imaged in a 7 Tesla (T) magnetic resonance (MR) imager to determine the extent of PDT-induced edema and to evaluate for the presence of iron oxide nanoparticles. Animals were sacrificed 7 days post-Ma administration and their brains analyzed for the presence of iron oxide using Perls staining. RESULTS: Significant uptake of iron oxide nanoparticles by rat alveolar Ma was observed thus providing the rationale for their use as delivery vectors. Histopathological analyses failed to find evidence of iron oxide in normal rat brain. Accumulations of iron oxide-loaded Ma were observed in both MR images and histological sections of non-tumor bearing rat brain following PDT-induced disruption of the BBB. CONCLUSIONS: MR imaging was shown to be useful for localizing iron-oxide loaded Ma in rat brains. Exogenous Ma are incapable of traversing the normal BBB and therefore, the use of Ma as delivery vehicles into the brain requires selective disruption of the BBB.

One common component of otolaryngological surgeries is the reshaping of cartilage. Previous studies have demonstrated the efficient achievement of this procedure through electromechanical reshaping (EMR), a technique that involves the direct application of voltage to cartilage that is mechanically deformed in a jig. Two main parameters, voltage and application time, may be regulated to achieve varying degrees of shape change. Although prior research has correlated these EMR parameters with degree of shape change, it remains necessary to correlate the same parameters with the degree of change in the mechanical properties of tissue. Once this is accomplished, an ideal balance may be determined, in which shape change is maximized while intrinsic tissue damage is minimized This study satisfies this need by providing comprehensive data on the pre- and post-EMR stiffness of both septal and auricular cartilage over a range of voltages (2-8V) with constant application time (2 min for septal, 3 min for auricular). EMR was applied using flat platinum electrodes to one of two 15 mm X 5 mm samples obtained from the same cartilage specimen, while the second sample was maintained as a control. Following a 15 min re-hydration period, the Young's modulus of the tissue was calculated for both the control and experimental sample from data obtained through a uniaxial tension test. A general reduction in stiffness was observed beginning at 3V, with the magnitude of reduction increasing at 6V. © 2010 Copyright SPIE - The International Society for Optical Engineering.

An emerging clinical treatment option for articular cartilage injury includes bone marrow stimulation techniques, such as microfracture, which has grown increasingly popular among athletes. During the microfracture procedure, the surgeon penetrates the subchondral bone with an awl and creates "microholes" deep enough to ensure bleeding from the bone marrow. This procedure triggers a spontaneous repair response that results in the formation of fibrocartilaginous repair tissue. This preliminary study aimed to evaluate the potential use of femtosecond lasers and Erbium:YAG lasers as alternatives to microfracture surgery of the knee by assessing the effects of ablation on bovine femoral condyles. Bovine femoral condyles were obtained and 8mm cube blocks were extracted. The specimen were ablated with various laser dosimetry parameters and observed using a high power dissecting microscope to examine the effects of the lasers. Further imaging with conventional histology (hematoxylin and eosin staining) was done to provide more accurate information. Preliminary results show some carbonization but demonstrate little thermal damage to surrounding tissues. The femtosecond laser offers a more precise and efficient ablation than the Erbium:YAG laser, but both are demonstrated to be possible alternatives to the surgical-skill dependent microfracture procedure. © 2010 Copyright SPIE - The International Society for Optical Engineering.

Electromechanical reshaping (EMR) of cartilage is a promising noninvasive technique with potential for broad application in reconstructive surgery. EMR involves applying direct current electrical fields to localized stress regions and initiating a series of oxidation-reduction reactions, thus effecting a shape change. Previous EMR studies have focused on macroscopic structural measurements of the shape change effect or monitoring of electrical current flow. Only limited investigation of structural changes in the tissue at the histologic level have been performed, and not in real time. This study is the first to use optical coherence tomography (OCT) to examine structural changes in cartilage during EMR. Two platinum needle electrodes were inserted into fixed rectangular rabbit nasal septal cartilage specimens. The spectral domain OCT probe was then positioned above the section of cartilage in which the anode needle was inserted. A constant voltage of 6V was applied for 3 minutes, and images were obtained (8 frames/second). OCT was also performed in specimens undergoing dehydration under ambient conditions and during pH changes produced by the addition of HCl, as both processes accompany EMR. The OCT data identified distinct findings among the three conditions, suggesting that EMR causes a much greater degree of reshaping on a molecular level than dehydration or a change in pH alone. OCT provides a means to gauge structural changes in the tissue matrix during EMR. The application of OCT to image the EMR process will add to our understanding of the mechanisms of action involved and potentially facilitate optimization of this process. © 2010 Copyright SPIE - The International Society for Optical Engineering.

Cryopreservation of cartilage has been investigated for decades and is currently an established protocol. However, the reliability and applicability of cartilage cryopreservation for the use in electromechanical reshaping (EMR) has not been studied exclusively. A system to cryopreserve large numbers of tissue specimens provides a steady source of cartilage of similar quality for experimentation at later dates. This will reduce error that may arise from different cartilage stock, and has the potential to maximize efficiency under time constraints. Our study utilizes a unique methodology to cryopreserve septal cartilage for use in EMR studies. Rabbit septal cartilage specimens were harvested and standardized to 20 x 8 x 1 mm, and placed in one of three solutions (normal saline, PBS, 10% DMSO in PBS) for four hours in a cold storage room at 4 degrees Celsius. Then, each cartilage specimen was vacuumed and sealed in an anti-frost plastic bag and stored in a freezer at -80 degrees Celsius for 1 to 3 weeks duration. EMR was performed using 2 and 6 volts for 2 minutes application time. Bend angle measurements of the cryopreserved cartilage specimens were compared to bend angles of fresh cartilage which underwent EMR using the same parameters. Results demonstrate that normal saline, phosphate buffered saline (PBS), and PBS with DMSO were effective in cryopreservation, and indicated no significant differences in bend angle measurements when compared to no cryopreservation. Our methodology to cryopreserve cartilage specimens provides a successful approach for use in conducting large-scale EMR studies. © 2010 Copyright SPIE - The International Society for Optical Engineering.

During laser therapy of port wine stain (PWS) birthmarks, regions of persistent perfusion may exist. We hypothesize that such regions, which are not readily visible, exist even during laser surgery performed by highly experienced clinicians. The objective of this study was to use objective feedback to assess the acute vascular response to laser therapy. We have developed a clinic-friendly laser speckle imaging (LSI) instrument to provide the clinician with real-time images of blood flow during laser therapy. We acquired images from patients undergoing laser therapy of PWS birthmarks at Scripps Clinic and Beckman Laser Institute and Medical Clinic. We extracted blood flow maps from the acquired imaging data. Collectively, we have observed two regimes of patient response to therapy: 1) an immediate increase in perfusion within minutes after laser therapy; and 2) an overall decrease in blood perfusion approximately one hour after laser therapy, with distinct regions of persistent perfusion apparent in the majority of post-treatment blood-flow images. A comparison of blood flow in PWS and adjacent normal skin demonstrated that PWS blood flow can be greater than or sometimes equivalent to that of normal skin. Regions of persistent perfusion frequently exist immediately after laser therapy of PWS birthmarks. Existence of these regions may be correlated to the need for multiple treatment sessions to improve substantially PWS skin appearance. With the use of intraoperative LSI, immediate retreatment of these regions may improve the outcome of each session. © 2009 SPIE.

Electromechanical reshaping (EMR) of cartilage is a novel technique that has significant potential for use in facial reconstructive surgery. EMR achieves permanent shape change by initiating electrochemical redox reactions in the vicinity of stress concentrations, thereby altering mechanical properties of tissue matrix. This study reports the use of a six electrode needle-based geometric configuration to reshape cartilage. Rectangular samples (24 × 12 × 1 mm) of rabbit nasal septal cartilages were bent at a right angle in a precision-machined reshaping jig. Two parallel arrays of three platinum needle electrodes were each inserted into cartilage along the bend at 3 mm from the bend line. One array served as an anode and the other as cathode. Constant voltage at 1, 2, 4, 6, and 8 volts was applied to the arrays for 2 minutes. The specimens were then removed from the jig and rehydrated for 15 minutes in phosphate buffered saline. Following rehydration, bend angles and thicknesses were measured. Bend angle increased with increasing voltage and application time. No statistically significant bending was observed below 6 volts for 2 minutes application time. Maximum bend angle of 33 ± 8 degrees or reshaping degree of 33% was observed at 8 volts applied for 2 minutes. Current flow was small (< 0.1 A) for each case. Sample thickness was 0.9 ± 0.2 mm. ANOVA analysis showed that cartilage thickness had no significant impact on the extent of reshaping at given voltage and application time. The six needle electrode geometric configuration conforms to the voltage- and time-dependent trends predicted by previous EMR studies. In the future, the reshaping properties of other geometric configurations will be explored. © 2009 SPIE.

Optical coherence tomography (OCT) is an evolving noninvasive imaging modality and has been used to image the human larynx during surgical endoscopy. The design of a long GRIN lens based probe capable of capturing images of the human larynx by use of swept-source OCT during a typical office-based laryngoscopy examination is presented. In vivo OCT imaging of the human larynx is demonstrated with 40 fame/second. Dynamic vibration of the vocal folds is recorded to provide not only high-resolution cross-sectional tissue structures but also vibration parameters, such as the vibration frequency and magnitude of the vocal cord, which provide important information for clinical diagnosis and treatment, as well as in fundamental research of the voice. Office-based OCT is a promising imaging modality to study the larynx. © 2009 SPIE.

Current methods for analysis of sebum excretion have limitations, such as irreproducible results in repeatable measurements due to the point measurement method, user-dependent artifact due to contact measurement or qualitative evaluation of the image, and long measurement time. A UV-induced fluorescent digital imaging system was developed to acquire facial images so that the distribution of sebum excretion on the face could be analyzed. The imaging system consisted of a constant UV-A light source, digital color camera, and head-positioning device. We describe the system characterization for acquisition of a fluorescent facial image and the image analysis method. The imaging modality provided uniform light distribution on the facial mannequin model and presented a discernible color fluorescent image. Valuable parameters of sebum excretion were obtained after image analysis. The imaging system, which provides a non-contact method, was proven to be a useful tool to evaluate sebum excretion and to characterize the pattern of sebum excretion. When compared to conventional "Wood's lamp" and "Sebutape" methods that provide similar parameters for sebum excretion, the method described herein is simpler and more reliable to evaluate the dynamics of sebum excretion in nearly real-time.

Neurologic complications have been reported with spinal transforaminal injections. Causes include intraneural injection, plus embolization occlusion of the radicular artery with subsequent spinal cord infarction. 1 Optical coherence tomography (OCT) is a non-invasive imaging modality, which is used to image tissue microstructure with very high resolution (less than 20 microns) in real-time. With a view toward needle tip OCT visualization of the spinal neuroforamen, we conducted animal studies to explore OCT imaging of paraspinal neurovascular structures. With institutional animal care committee approval, we performed ex-vivo and in situ OCT studies in a euthanized dog, pig, and rabbit. Image data was gathered on spinal nerve roots, dura, and brachial plexus. Two systems were used: frequency domain OCT imaging system developed at California Institute of Technology, and time domain Imalux NIRIS system with a 2.7 mm diameter probe. In a euthanized pig, excised dura was punctured with a 17-gauge Tuohy needle. FDOCT dural images of the puncture showed a subsurface cone-shaped defect. In a rabbit in situ study, puncture of the dura with a 26-gauge needle is imaged as a discontinuity. FDOCT imaging of both small artery and large arteries will be presented, along with H&E and OCT images of the brachial plexus.

The effects of 5-aminolevulinic acid (ALA)-mediated photodynamic therapy (PDT) and gamma radiation, and PDT and hyperthermia (HT) are investigated in human glioma spheroids. In the case of ionizing radiation and PDT, the degree of interaction between the two modalities depends in a complex manner on a number of factors, including light fluence, fluence rate and radiation dose. It is shown that, under certain conditions, PDT and gamma radiation interact synergistically to produce enhanced cell kill. The degree of interaction appears to be independent of both sequence and time intervals investigated. TUNEL assays show that low fluence-rate ALA-PDT is a very efficient inducer of apoptosis, whereas neither high-fluence rate PDT nor ionizing radiation results in significant apoptosis in this system. Synergistic interactions between HT and PDT are observed when the two modalities are given concurrently. The degree of synergism increases with increasing temperature and light fluence. It is shown that apoptosis is the primary mode of cell death following combined HT and low-fluence rate ALA-PDT.

Although many results on the near infrared spectroscopy and imaging of the human brain have been already published, the signal-to-noise ratio (SNR) of the instrumentation used in these studies has never been systematically optimized for the anatomy of the human head. Assuming a typical anatomical structure of the adult human head, we have studied the SNR of the frequency-domain instrument as a function of the modulation frequency and the source-detector distance. The study was performed both experimentally (using a pulsed laser system) and numerically (using the diffusion approximation and Monte Carlo simulations). For the specified geometry we have found that changes caused by non-homogeneities in the phase and in the ratio of the modulation amplitude to the mean intensity significantly increase with the modulation frequency and exhibit peaks between 400 and 1100 MHz, depending on the source-detector distance. Assuming the shot noise, we have found that the corresponding SNR peaks between 400 and 600 MHz. Increase in the source-detector distance results in the SNR increase only for distances below 30 mm. At larger source-detector distances the higher noise decreases the SNR. Our results show that increasing modulation frequency only to 300 MHz offers a very significant improvement in the functional near-infrared spectro-imaging of the adult human brain compared to the 100 MHz instrument.

Near-infrared wavelengths are absorbed less by epidermal melanin mainly located at the basal layer of epidermis (dermo-epidermal junction), and penetrate deeper into human skin dermis and blood than visible wavelengths. Therefore, laser irradiation using near-infrared wavelength may improve the therapeutic outcome of cutaneous hyper-vascular malformations in moderately to heavily pigmented skin patients and those with large-sized blood vessels or blood vessels extending deeply into the skin. A mathematical model composed of a Monte Carlo algorithm to estimate the distribution of absorbed light followed by numerical solution of a bio-heat diffusion equation was utilized to investigate the thermal response of human skin to near-infrared laser irradiation, and compared it with that to visible laser irradiation. Additionally, the effect of skin surface cooling on epidermal protection was theoretically investigated. Simulation results indicated that 940 nm wavelength is superior to 810 and 1064 nm in terms of the ratio of light absorption by targeted blood vessel to the absorption by the basal layer of epidermis, and is more efficient than 595 nm wavelength for the treatment of patients with large-sized blood vessels and moderately to heavily pigmented skin. Dermal blood content has a considerable effect on the laser-induced peak temperature at the basal layer of epidermis, while the effect of blood vessel size is minimum.