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This PDF file contains the front matter associated with SPIE Proceedings Volume 10042, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.
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Cardiovascular diseases remain the greatest cause of death in the US and gene therapy has the potential to be an effective
therapy. In this study, we demonstrated MMP-9 based protease-activatable virus (PAV) for selective infection of
myocardial infarct (MI) that is associated with active MMP-9 expression. To test the specificity of PAV, we used
expression of a far-red fluorescence protein (iRFP) delivered by the PAV together with a dual PET/NIRF imaging agent
specific for active MMP-9 activity at the site of MI in a murine model. Calibrated fluorescence imaging employed a
highly-sensitive intensified camera, laser diode excitation sources, and filtration schemes based upon the spectra of iRFP
and the NIRF agent. One to two days after ligation of the left anterior descending artery, the PAV or WT AAV9 virus
encoding for iRFP (5x1010 genomic particles) and radiolabeled MMP-9 imaging agent (3 nmol) were injected
intravenously (i.v.). PET imaging showed MMP activity was associated with adverse tissue remodeling at the site of the
MI. One week after, animals were again injected i.v. with the MMP-9 agent (3 nmol) and 18-24 h later, the animals
were euthanized and the hearts were harvested, sliced, and imaged for congruent iRFP transgene expression and NIRF
signals associated with MMP-9 tissue activity. The fluorescent margins of iRFP and NIRF contrasted tissues were
quantified in terms Standard International units of mW/cm2/sr. The sensitivity, specificity, and accuracy of PAV and
WT targeting to sites of MI was determined from these calibrated fluorescence measurements. The PAV demonstrated
significantly higher delivery performance than that of the WT AAV9 virus.
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Atrial structure plays an important role in the mechanisms of atrial disease. However, detailed imaging of human atria
remains limited due to many imaging modalities lacking sufficient resolution. We propose the use of optical coherence
tomography (OCT), which has micrometer resolution and millimeter-scale imaging depth well-suited for the atria,
combined with image stitching algorithms, to develop large, detailed atria image maps. Human atria samples (n = 7) were
obtained under approved protocols from the National Disease Research Interchange (NDRI). One right atria sample was
imaged using an ultrahigh-resolution spectral domain OCT system, with 5.52 and 2.72 μm lateral and axial resolution in
air, respectively, and 1.78 mm imaging depth. Six left atria and five pulmonary vein samples were imaged using the spectral
domain OCT system, Telesto I (Thorlabs GmbH, Germany) with 15 and 6.5 μm lateral and axial resolution in air,
respectively, and 2.51 mm imaging depth. Overlapping image volumes were obtained from areas of the human left and
right atria and the pulmonary veins. Regions of collagen, adipose, and myocardium could be identified within the OCT
images. Image stitching was applied to generate fields of view with side dimensions up to about 3 cm. This study
established steps towards mapping large regions of the human atria and pulmonary veins in high resolution using OCT.
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Cardiovascular disease is a major contributor to US morbidity. Taking preventive action can greatly reduce or eliminate the impact on quality of life. However, many issues often go undetected until the patient presents a physical symptom. Non-intrusive continuous cardiovascular monitoring systems may make detecting and monitoring abnormalities earlier feasible. One candidate system is photoplethysmographic imaging (PPGI), which is able to assess arterial blood pulse characteristics in one or multiple individuals remotely from a distance. In this case study, we showed that PPGI can be used to detect cardiac arrhythmia that would otherwise require contact-based monitoring techniques. Using a novel system, coded hemodynamic imaging (CHI), strong temporal blood pulse waveform signals were extracted at a distance of 1.5 m from the participant using 850-1000 nm diffuse illumination for deep tissue penetration. Data were recorded at a sampling rate of 60 Hz, providing a temporal resolution of 17 ms. The strong fidelity of the signal allowed for both temporal and spectral assessment of abnormal blood pulse waveforms, ultimately to detect the onset of abnormal cardiac events. Data from a participant with arrhythmia was analyzed and compared against normal blood pulse waveform data to validate CHI’s ability to assess cardiac arrhythmia. Results indicate that CHI can be used as a non-intrusive continuous cardiac monitoring system.
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Atrial fibrillation (AF) occurs following myocardial infarction (MI) and is associated with left ventricular dysfunction, which promotes the development of atrial remodeling and permanent atrial fibrosis. The purpose of this study was determining the effects of MI on left atrial (LA) remodeling with and without therapy with an angiotensin converting enzyme inhibition (ACEi) utilizing optical coherence tomography (OCT). As the composition of the myocardial tissue changes during LA remodeling the optical attenuation of the light will also change providing a metric to quantify the structural remodeling process. Lewis rats (240-275 g) underwent either surgical ligation of left coronary artery creating chronic MI, or SHAM surgery. 13 weeks post-surgery, ex vivo OCT imaging was performed of the LA appendage. Depth-resolved, attenuation coefficient volumes were calculated and the resulting atrial wall attenuation values were analyzed for four experimental groups: SHAM, SHAM with ACEi, MI no ACEi, and MI with ACEi. Quantification of tissue attenuation was performed and shown to significantly increase with MI in association with increases in collagen as verified with corresponding histological sectioning. Fractal analysis of the LA wall trabeculation patterns, 100 µm below the surface, was performed to quantify wall thickening associated with LA remodeling. A significant increase in fractal dimension was determined post MI compared to SHAM corresponding to a loss of the trabeculation pattern and wall thickening. The results from this study demonstrate OCT as an imaging technique capable of investigate LA remodeling with high resolution and label-free optical contrast processing.
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We present an initial study to describe the potential of optical coherence tomography (OCT) for characterizing
endomyocardial tissue. We obtained ventricular OCT images from 15 fresh human hearts. Layer thickness
measurements and texture features were extracted from volumetric datasets with endocardial thickening, normal
myocardium, myocardium with interstitial fibrosis, normal endocardium, and adipose tissue. Within our datasets, we
observed that the thickness of endocardium was not different within samples with normal or myocardium with interstitial
fibrosis, however samples with endocardial thickening showed statistically increased endocardial thickness.
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The rupture of atherosclerotic plaques is the leading cause of acute coronary events, so accurate assessment of plaque is critical. A large lipid pool, thin fibrous cap, and inflammatory reaction are the crucial characteristics for identifying vulnerable plaques. In our study, a tri-modality imaging system for intravascular imaging was designed and implemented. The tri-modality imaging system with a 1-mm probe diameter is able to simultaneously acquire optical coherence tomography (OCT), intravascular ultrasound (IVUS), and fluorescence imaging. Moreover, for fluorescence imaging, we used the FDA-approved indocyanine green (ICG) dye as the contrast agent to target lipid-loaded macrophages. Firstly, IVUS is used as the first step for identifying plaque since IVUS enables the visualization of the layered structures of the artery wall. Due to low soft-tissue contrast, IVUS only provides initial identification of the lipid plaque. Then OCT is used for differentiating fibrosis and lipid pool based on its relatively higher soft tissue contrast and high sensitivity/specificity. Last, fluorescence imaging is used for identifying inflammatory reaction to further confirm whether the plaque is vulnerable or not. Ex vivo experiment of a male New Zealand white rabbit aorta was performed to validate the performance of our tri-modality system. H and E histology results of the rabbit aorta were also presented to check assessment accuracy. The miniature tri-modality probe, together with the use of ICG dye suggest that the system is of great potential for providing a more accurate assessment of vulnerable plaques in clinical applications.
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Visualization and characterization of inner arterial plaque depositions is of vital diagnostic interest. Established intravascular imaging techniques provide valuable morphological information, but cannot deliver information about the chemical composition of individual plaques. Probe based Raman spectroscopy offers the possibility for a biochemical characterization of atherosclerotic plaque formations during an intravascular intervention. From post mortem studies it is well known that the severity of a plaque and its stability are strongly correlated with its biochemical composition. Especially the identification of vulnerable plaques remains one of the most important and challenging aspects in cardiology. Thus, specific information about the composition of a plaque would greatly improve the risk assessment and management. Furthermore, knowledge about the composition can offer new therapeutic and medication strategies. Plaque calcifications as well as major lipid components such as cholesterol, cholesterol esters and triglycerides can be spectroscopically easily differentiated. Intravascular optical coherence tomography (OCT) is currently a prominent catheter based imaging technique for the localization and visualization of atherosclerotic plaque depositions. The high resolution of OCT with 10 to 15 µm allows for very detailed characterization of morphological features such as different plaque formations, thin fibrous caps and accurate measurements of lesion lengths. In combination with OCT imaging the obtained spectral information can provide substantial information supporting on on-site diagnosis of various plaque types and therefor an improved risk assessment. The potential and feasibility of combining OCT with Raman spectroscopy is demonstrated on excised plaque samples, as well as under in vivo conditions.
Acknowledgements: Financial support from the Carl Zeiss Foundation is greatly acknowledged.
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FL-IVUS combines intravascular ultrasound with fluorescence lifetime imaging to obtain morphologic and biochemical details from the arterial wall. Ultrasound measurements alone provide morphologic information (plaque burden, remodeling index and presence of calcium). Fluorescence lifetime can determine the presence of a thick fibrous cap, macrophage infiltration, and lipid cores beneath thin fibrous caps. These details are important to assess plaque vulnerability. In this study, we focused on the ability of FL-IVUS to differentiate between early and advanced lipid cores-advanced cores are vulnerable to rupture. We imaged N=12 ex vivo human coronary arteries and performed hematoxylin and eosin, Movat’s pentachrome and CD68 immunohistochemistry at 500 micron intervals throughout the length of the vessels. We found only N=1 thin-capped fibroatheroma (TCFA) with an advanced necrotic core and N=7 cases of foam cell infiltration, early lipid cores or deep necrotic cores. IVUS was able to observe the increased plaque burden and calcification of the advanced and deep necrotic cores, but could not identify early lipid cores, foam cell infiltration or discriminate between deep necrotic cores and TCFA. The addition of FLIm to IVUS allowed the TCFA to be discriminated from early lipid accumulation, particularly at 542±50 nm (355 nm pulsed excitation): 7.6 ± 0.5 ns compared to 6.6 ± 0.4 ns, respectively (P<0.001 by ANOVA analysis). These differences need to be validated in a larger cohort, but exist due to specific lipid content in the necrotic core as well as increased extracellular matrix in early lesions.
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Endovascular abdominal aortic aneurysms repair (EVAR) involves the minimally invasive implantation of a stent-graft
within the aorta to exclude the aneurysm from the circulation thus preventing its rupture. The feasibility of such
operation is highly dependent on the aorta morphology and in general the presence of one/both renal arteries emerging
from the aneurysm is the absolute limit for the implantation of a standard stent-graft. Consequently, classical intervention
methods involve the implantation of a custom-made graft with fenestrations, leading to extremely complicated surgeries
with high risks for the patient and high costs. Recent techniques introduced the use of standard grafts (i.e. without
fenestrations) in association with mechanical in-situ fenestration, but this procedure is limited principally by the
brittleness and low stability of the environment, in addition to the difficulty of controlling the guidance of the
endovascular tools due to the temporarily block of the blood flow. In this work we propose an innovative EVAR strategy,
which involves in-situ fenestration with a fiber guided laser tool, controlled via an electromagnetic navigation system.
The fiber is sensorized to be tracked by means of the driving system and, using a 3D model of the patient anatomy, the
surgeon can drive the fiber to the aneurysm, where the stent has been previously released, to realize the proper
fenestration(s). The design and construction of the catheter laser tool will be presented, togheter with preliminary
fenestration tests on graft-materials, including the effects due to the presence of blood and tissues.
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Cardiomyocytes derived from human induced pluripotent stem cells (hiPS-HCM) have the potential to provide individualized therapies for patients and to test drug candidates for cardiac toxicity. In order for hiPS-CM to be useful for such applications, there is a need for high-throughput technology to rapidly assess cardiac electrophysiology parameters. Here, we designed and tested a fully contactless optical mapping (OM) and optical pacing (OP) system capable of imaging and point stimulation of hiPS-CM in small wells. OM allowed us to characterize cardiac electrophysiological parameters (conduction velocity, action potential duration, etc.) using voltage-sensitive dyes with high temporal and spatial resolution over the entire well. To improve OM signal-to-noise ratio, we tested a new voltage-sensitive dye (Fluovolt) for accuracy and phototoxicity. Stimulation is essential because most electrophysiological parameters are rate dependent; however, traditional methods utilizing electrical stimulation is difficult in small wells. To overcome this limitation, we utilized OP (λ = 1464 nm) to precisely control heart rate with spatial precision without the addition of exogenous agents. We optimized OP parameters (e.g., well size, pulse width, spot size) to achieve robust pacing and minimize the threshold radiant exposure. Finally, we tested system sensitivity using Flecainide, a drug with well described action on multiple electrophysiological properties.
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For last two decades, endovenous laser therapy (EVLT) is one of the most widely accepted surgical options for treating incompetent great and small saphenous veins. However, due to excessive heating during EVLT, the major complications include pain and burning that often increase the risk of dermatitis disease. The aim of the current study was to quantitatively compare commercially-available radial fibers with newly-developed diffusing applicators for 1470 nm-EVLA in terms of temperature elevation and vein deformation. Rabbit veins were used as an ex vivo model for EVLA. A 5-W 1470 nm laser system in conjunction with the radial and diffusing fibers was employed to thermally coagulate the venous tissue. A goniometric measurement validated uniform and isotropic distribution of laser light in polar and longitudinal directions (i.e., normalized intensity = 0.84±0.08). The diffusing applicator induced a 20 % lower maximum temperature than the radial fiber did (maximum temperature = 79.2 °C for radial vs. 63.3 °C for diffusing). Due to higher irradiance, the radial fiber was associated with a transient temperature change of 5.9 °C/s, which was 1.5-fold faster than the diffusing applicator (i.e., 2.4 °C/s). However, the degree of cross-sectional area reduction in the veins was almost comparable for both the fibers (i.e., 53% for radial vs. 48% for diffusing). Due to longer irradiation length, the diffusing applicator demonstrated wider treatment coverage and less fiber speed-dependent. On account of easy pullback technique and uniform thermal effect, the proposed cylindrically diffusing applicator can be a feasible optical device to effectively treat varicose veins. Further in vivo studies will be performed to identify the complete removal of the vein disease and healing response of the venous tissue.
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Intravascular optical coherence tomography (IVOCT) is a new intravascular imaging modality which enables arterial
structures to be visualized at a microstructure level. The determination of these structures is currently performed
manually based on relative light intensities which is difficult because there are many factors, including the position
inside the artery and vendor of the catheter, which can influence these intensities. In this study we demonstrate how
optical attenuation and backscattering values can be computed and used as better characterizing features for different
types of atherosclerotic plaque such as fibro-atheroma, lipid-pools and calcified areas. To validate the method, different
plaque components are segmented in multiple IVOCT pullback runs using matching histology-data. The optical
attenuation, backscattering and light intensity features of the segmented regions are then automatically extracted and
analyzed for their entropy with regards to tissue characterization. The results of the validation analysis show that the
computed attenuation and backscattering measurements are in agreement with those published in literature and that
especially attenuation is a more robust feature than light intensity when it comes to tissue characterization. As a practical
application we show how attenuation and backscattering can be used to quickly determine the presence of lipid or
calcified plaques which can be important factors to determine patient treatment. Based on these findings we intend to
develop a fully automatic tissue characterization method for IVOCT.
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Background: Uric acid crystals have recently been identified as a possible therapeutic target for coronary artery disease. Being subcellular in size, it is difficult to identify these crystals in situ. Micro optical coherence tomography (Micro-OCT) allows one to image subcellular structures with 1–micron resolution. Even though Micro-OCT should be capable of resolving urate crystals, it’s difficult to differentiate these structures from other scattering particles within tissue. In this work we developed a novel polarization sensitive micro OCT (ps-Micro-OCT) system for identification of uric acid crystals.
Methods: A spectrometer based ps-Micro-OCT system was developed using a broadband light source. The broadband input light was divided into reference and sample signals using a beam splitter. The reference signal was further divided into two polarized signals with different polarization states. Reflected reference and sample signals were combined and sent to a spectrometer that recorded the interference signal.
Results: To test the performance of system, a mirror was used as sample and a quarter wave-plate was placed in the sample path. The measured quarter wave-plate angle values matched closely to actual angle values. Next we prepared uric acid crystals in our lab and imaged them using this system.We were able to image and identify these crystals based on polarization measurements.
Conclusion: In this work we imaged and identified uric acid crystals using a newly developed ps-Micro-OCT system. The proposed technique will enable imaging uric acid crystals in coronary artery.
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Cardiac hypertrophy, a process initiated by mechanical alterations, is hypothesized to cause long-term molecular-level alteration in the sarcomere lattice, which is the main force-generating component in the heart muscle. This molecular-level alteration is beyond the resolving capacity of common light microscopy. Second harmonic generation (SHG) microscopy has unique capability for visualizing ordered molecular structures in biological tissues without labeling. Combined with polarization imaging technique, SHG microscopy is able to extract structural details of myosin at the molecular level so as to reveal molecular-level alterations that occur during hypertrophy. The myosin filaments are believed to possess C6 symmetry; thus, the nonlinear polarization response relationship between generated second harmonic light I^2ωand incident fundamental light I^ω is determined by nonlinear coefficients, χ_15, χ_31 and χ_33. χ_31/χ_15 is believed to be an indicator of the molecular symmetry of myosin filament, whileχ_33/χ_15represents the intramyosin orientation angle of the double helix. By changing the polarization of the incident light and evaluating the corresponding SHG signals, the molecular structure of the myosin, reflected by the χ coefficients, can be revealed. With this method, we studied the structural properties of heart tissues in different conditions, including those in normal, physiologically hypertrophic (heart tissue from postpartum female rats), and pathologically hypertrophic (heart tissue from transverse-aorta constricted rats) conditions. We found that ratios of χ_31/χ_15 showed no significant difference between heart tissues from different conditions; their values were all close to 1, which demonstrated that Kleinman symmetry held for all conditions. Ratios of χ_33/χ_15 from physiologically or pathologically hypertrophic heart tissues were raised and showed significant difference from those from normal heart tissues, which indicated that the intramyosin orientation angle of the double helix was altered when heart tissues hypertrophied. Polarization-resolved SHG microscopy permitted us to study heart tissues at the molecular level and may serve as a diagnostic tool for cardiac hypertrophy.
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Electroanatomical mapping (EAM) is an invaluable tool for guiding cardiac radiofrequency ablation (RFA) therapy. The principle roles of EAM is the identification of candidate ablation sites by detecting regions of abnormal electrogram activity and lesion validation subsequent to RF energy delivery. However, incomplete lesions may present interim electrical inactivity similar to effective treatment in the acute setting, despite efforts to reveal them with pacing or drugs, such as adenosine. Studies report that the misidentification and recovery of such lesions is a leading cause of arrhythmia recurrence and repeat procedures. In previous work, we demonstrated spectroscopic characterization of cardiac tissues using a fiber optic-integrated RF ablation catheter. In this work, we introduce OSAM (optical spectroscopic anatomical mapping), the application of this spectroscopic technique to obtain 2-dimensional biodistribution maps. We demonstrate its diagnostic potential as an auxiliary method for lesion validation in treated swine preparations.
Endocardial lesion sets were created on fresh swine cardiac samples using a commercial RFA system. An optically-integrated catheter console fabricated in-house was used for measurement of tissue optical spectra between 600-1000nm. Three dimensional, Spatio-spectral datasets were generated by raster scanning of the optical catheter across the treated sample surface in the presence of whole blood. Tissue optical parameters were recovered at each spatial position using an inverse Monte Carlo method. OSAM biodistribution maps showed stark correspondence with gross examination of tetrazolium chloride stained tissue specimens. Specifically, we demonstrate the ability of OSAM to readily distinguish between shallow and deeper lesions, a limitation faced by current EAM techniques. These results showcase the OSAMs potential for lesion validation strategies for the treatment of cardiac arrhythmias.
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Using light-based catheters for radiofrequency ablation (RFA) therapies grants the ability to accurately derive tissue
properties such as lesion depth and overtreatment from spectroscopic information. However, this information is heavily
reliant on contact quality with the treatment area and the orientation of the catheter. Thus to improve assessments of
tissue properties, this work utilizes Bayesian modelling to classify whether the catheter is indeed in proper contact with
the tissue. Initially in-laboratory experiments were conducted with ten fresh swine hearts submerged in blood. A total of
1555 unique near infrared spectra were collected from a spectrometer using a light-based catheter and manually tagged
as “full perpendicular contact,” “angled contact,” and “no contact,” between the catheter and heart tissue. Three features
were prominent in all spectra for distinguishing purposes: area underneath the spectra, an intensity “valley” between 730
nm and 800 nm, along with the slope between 850 nm and 1150 nm. A classifier featuring bootstrapping, adaboost, and
k-means techniques was thus created and achieved a 96.05% accuracy in classifying full contact, 98.33% accuracy in
classifying angled contact, and 100% accuracy in classifying no contact.
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The aim of our study was to identify fluorescence excitation-emission pairs correlated with atherosclerotic
pathology in ex-vivo human aorta. Wide-field images of atherosclerotic human aorta were captured using UV and
visible excitation and emission wavelength pairs of several known fluorophores to investigate correspondence with
gross pathologic features. Fluorescence spectroscopy and histology were performed on 21 aortic samples. A matrix
of Pearson correlation coefficients were determined for the relationship between relevant histologic features and the
intensity of emission for 427 wavelength pairs. A multiple linear regression analysis indicated that elastin (370/460
nm) and tryptophan (290/340 nm) fluorescence predicted 58% of the variance in intima thickness (R-squared =
0.588, F(2,18) = 12.8, p=.0003), and 48% of the variance in media thickness (R-squared = 0.483, F(2,18) = 8.42,
p=.002), suggesting that endogenous fluorescence intensity at these wavelengths can be utilized for improved
pathologic characterization of atherosclerotic plaques.
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