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1.  Delivery of paclitaxel from cobalt–chromium alloy surfaces without polymeric carriers 
Biomaterials  2010;31(20):5372-5384.
Polymer-based carriers are commonly used to deliver drugs from stents. However, adverse responses to polymer coatings have raised serious concerns. This research is focused on delivering drugs from stents without using polymers or any carriers. Paclitaxel (PAT), an anti-restenotic drug, has strong adhesion towards a variety of material surfaces. In this study, we have utilized such natural adhesion property of PAT to attach these molecules directly to cobalt–chromium (Co–Cr) alloy, an ultra-thin stent strut material. Four different groups of drug coated specimens were prepared by directly adding PAT to Co–Cr alloy surfaces: Group-A (PAT coated, unheated, and ethanol cleaned); Group-B (PAT coated, heat treated, and ethanol cleaned); Group-C (PAT coated, unheated, and not ethanol cleaned); and Group-D (PAT coated, heat treated and not ethanol cleaned). In vitro drug release of these specimens was investigated using high performance liquid chromatography. Groups A and B showed sustained PAT release for up to 56 days. A simple ethanol cleaning procedure after PAT deposition can remove the loosely bound drug crystals from the alloy surfaces and thereby allowing the remaining strongly bound drug molecules to be released at a sustained rate. The heat treatment after PAT coating further improved the stability of PAT on Co–Cr alloy and allowed the drug to be delivered at a much slower rate, especially during the initial 7 days. The specimens which were not cleaned in ethanol, Groups C and D, showed burst release. PAT coated Co–Cr alloy specimens were thoroughly characterized using scanning electron microscopy, atomic force microscopy, and X-ray photoelectron spectroscopy. These techniques were collectively useful in studying the morphology, distribution, and attachment of PAT molecules on Co–Cr alloy surfaces. Thus, this study suggests the potential for delivering paclitaxel from Co–Cr alloy surfaces without using any carriers.
doi:10.1016/j.biomaterials.2010.03.043
PMCID: PMC4076912  PMID: 20398928
Drug-eluting stents; Drug delivery; Cobalt–hromium alloy; Surface treatment; Surface modification
2.  Intravascular optical coherence tomography measurement of size and apposition of metallic stents 
Biomedical Optics Express  2013;4(10):1876-1882.
Effect of beam size and catheter position on the apparent size and apposition of metallic stent struts in IVOCT images were examined. Micro-CT data was employed to determine light - stent strut interactions. Simulated results suggest that location of the reflecting regions depend on relative orientation and position of stent struts to the IVOCT beam. Erroneous stent apposition measurements can occur when the IVOCT catheter is at an eccentric position. A method that mitigates stent strut apposition measurement errors is proposed.
doi:10.1364/BOE.4.001876
PMCID: PMC3799652  PMID: 24156050
(110.0110) Imaging systems; (110.4500) Optical coherence tomography
3.  Combined Two-Photon Luminescence Microscopy and OCT for Macrophage Detection in the Hypercholesterolemic Rabbit Aorta Using Plasmonic Gold Nanorose 
Lasers in surgery and medicine  2012;44(1):49-59.
Background and Objectives
The macrophage is an important early cellular marker related to risk of future rupture of atherosclerotic plaques. Two-channel two-photon luminescence (TPL) microscopy combined with optical coherence tomography (OCT) was used to detect, and further characterize the distribution of aorta-based macrophages using plasmonic gold nanorose as an imaging contrast agent.
Study Design/Materials and Methods
Nanorose uptake by macrophages was identified by TPL microscopy in macrophage cell culture. Ex vivo aorta segments (8 × 8 × 2 mm3) rich in macrophages from a rabbit model of aorta inflammation were imaged by TPL microscopy in combination with OCT. Aorta histological sections (5 µm in thickness) were also imaged by TPL microscopy.
Results
Merged two-channel TPL images showed the lateral and depth distribution of nanorose-loaded macrophages (confirmed by RAM-11 stain) and other aorta components (e.g., elastin fiber and lipid droplet), suggesting that nanorose-loaded macrophages are diffusively distributed and mostly detected superficially within 20 µm from the luminal surface of the aorta. Moreover, OCT images depicted detailed surface structure of the diseased aorta.
Conclusions
Results suggest that TPL microscopy combined with OCT can simultaneously reveal macrophage distribution with respect to aorta surface structure, which has the potential to detect vulnerable plaques and monitor plaque-based macrophages overtime during cardiovascular interventions.
doi:10.1002/lsm.21153
PMCID: PMC3696498  PMID: 22246984
atherosclerosis; macrophage; nanorose; two-photon luminescence microscopy; optical coherence tomography; photothermal wave imaging
4.  Two-photon luminescence properties of gold nanorods 
Biomedical Optics Express  2013;4(4):584-595.
Gold nanorods can be internalized by macrophages (an important early cellular marker in atherosclerosis and cancer) and used as an imaging contrast agent for macrophage targeting. Objective of this study is to compare two-photon luminescence (TPL) properties of four aspect ratios of gold nanorods with surface plasmon resonance at 700, 756, 844 and 1060 nm respectively. TPL from single nanorods and Rhodamine 6G particles was measured using a laser-scanning TPL microscope. Nanorod TPL emission spectrum was recorded by a spectrometer. Quadratic dependence of luminescence intensity on excitation power (confirming a TPL process) was observed below a threshold (e.g., <1.6 mW), followed by photobleaching at higher power levels. Dependence of nanorod TPL intensity on excitation wavelength indicated that the two-photon action cross section (TPACS) is plasmon-enhanced. Largest TPACS of a single nanorod (12271 GM) was substantially larger than a single Rhodamine 6G particle (25 GM) at 760 nm excitation. Characteristics of nanorod TPL emission spectrum can be explained by plasmon-enhanced interband transition of gold. Comparison results of TPL brightness, TPACS and emission spectrum of nanorods can guide selection of optimal contrast agent for selected imaging applications.
doi:10.1364/BOE.4.000584
PMCID: PMC3617720  PMID: 23577293
(160.4236) Nanomaterials; (190.1900) Diagnostic applications of nonlinear optics; (190.4180) Multiphoton processes; (160.4760) Optical properties; (170.6280) Spectroscopy, fluorescence and luminescence
5.  Bio-telemetric device for measurement of left ventricular pressure-volume loops using the admittance technique in conscious, ambulatory rats 
Physiological measurement  2011;32(6):701-715.
This paper presents the design, construction and testing of a device to measure pressure volume loops in the left ventricle of conscious, ambulatory rats. Pressure is measured with a standard sensor, but volume is derived from data collected from a tetrapolar electrode catheter using a novel admittance technique. There are two main advantages of the admittance technique to measure volume. First, the contribution from the adjacent muscle can be instantaneously removed. Second, the admittance technique incorporates the nonlinear relationship between the electric field generated by the catheter and the blood volume. A low power instrument weighing 27 g was designed, which takes pressure-volume loops every 2 minutes and runs for 24 hours. Pressure-volume data are transmitted wirelessly to a base station. The device was first validated in thirteen rats with an acute preparation with 2-D echocardiography used to measure true volume. From an accuracy standpoint, the admittance technique is superior to both the conductance technique calibrated with hypertonic saline injections, and calibrated with cuvettes. The device was then tested in six rats with a 24-hour chronic preparation. Stability of the animal preparation and careful calibration are important factors affecting the success of the device.
doi:10.1088/0967-3334/32/6/007
PMCID: PMC3176664  PMID: 21606560
admittance; impedance; conductance; wireless; left ventricular pressure; left ventricular volume; rats; pressure volume loop
6.  Imaging flow dynamics in murine coronary arteries with spectral domain optical Doppler tomography 
Biomedical Optics Express  2012;3(4):701-714.
Blood flow in murine epicardial and intra-myocardial coronary arteries was measured in vivo with spectral domain optical Doppler tomography (SD-ODT). Videos at frame rates up to 180 fps were collected and processed to extract phase shifts associated with moving erythrocytes in the coronary arteries. Radial averaging centered on the vessel lumen provided spatial smoothing of phase noise in a single cross-sectional frame for instantaneous peak velocity measurement without distortion of the flow profile. Temporal averaging synchronized to the cardiac cycle (i.e., gating) was also performed to reduce phase noise, although resulting in lower flow profiles. The vessel angle with respect to incident imaging beam was measured with three-dimensional raster scans collected from the same region as the high speed cross-sectional scans. The variability in peak phase measurement was 10-15% from cycle to cycle on a single animal but larger for measurements among animals. The inter-subject variability is attributed to factors related to real physiological and anatomical differences, instrumentation variables, and measurement error. The measured peak instantaneous flow velocity in a ~40-µm diameter vessel was 23.5 mm/s (28 kHz Doppler phase shift). In addition to measurement of the flow velocity, we observed several dynamic features of the vessel and surrounding myocardium in the intensity and phase sequences, including asymmetric vessel deformation and rapid flow reversal immediately following maximum flow, in confirmation of known coronary artery flow dynamics. SD-ODT is an optical imaging tool that can provide in vivo measures of structural and functional information on cardiac function in small animals.
doi:10.1364/BOE.3.000701
PMCID: PMC3345800  PMID: 22574259
(170.4500) Optical coherence tomography; (170.3880) Medical and biological imaging
8.  Characterization of Nanoporous Surfaces as Templates for Drug Delivery Devices 
The AAPS Journal  2009;11(4):758-761.
doi:10.1208/s12248-009-9152-x
PMCID: PMC2782086  PMID: 19876743
cyanoacrylate; drug delivery; methyl orange; nanopores; volume calculation
9.  Small Multifunctional Nanoclusters (Nanoroses) for Targeted Cellular Imaging and Therapy 
ACS nano  2009;3(9):2686-2696.
The ability of 20–50 nm nanoparticles to target and modulate the biology of specific types of cells will enable major advancements in cellular imaging and therapy in cancer and atherosclerosis. A key challenge is to load an extremely high degree of targeting, imaging, and therapeutic functionality into small, yet stable particles. Herein we report ~30 nm stable uniformly sized near-infrared (NIR) active, superparamagnetic nanoclusters formed by kinetically controlled self-assembly of gold-coated iron oxide nanoparticles. The controlled assembly of nanocomposite particles into clusters with small primary particle spacings produces collective responses of the electrons that shift the absorbance into the NIR region. The nanoclusters of ~70 iron oxide primary particles with thin gold coatings display intense NIR (700–850 nm) absorbance with a cross section of ~10−14 m2. Because of the thin gold shells with an average thickness of only 2 nm, the r2 spin–spin magnetic relaxivity is 219 mM−1 s−1, an order of magnitude larger than observed for typical iron oxide particles with thicker gold shells. Despite only 12% by weight polymeric stabilizer, the particle size and NIR absorbance change very little in deionized water over 8 months. High uptake of the nanoclusters by macrophages is facilitated by the dextran coating, producing intense NIR contrast in dark field and hyperspectral microscopy, both in cell culture and an in vivo rabbit model of atherosclerosis. Small nanoclusters with optical, magnetic, and therapeutic functionality, designed by assembly of nanoparticle building blocks, offer broad opportunities for targeted cellular imaging, therapy, and combined imaging and therapy.
doi:10.1021/nn900440e
PMCID: PMC2841963  PMID: 19711944
gold; iron oxide; nanocluster; near-infrared; macrophage targeted imaging; MRI; atherosclerosis; cancer
10.  Depth resolved photothermal OCT detection of macrophages in tissue using nanorose 
Biomedical Optics Express  2010;1(1):2-16.
Application of photothermal Optical Coherence Tomography (OCT) to detect macrophages in ex vivo rabbit arteries which have engulfed nanoclusters of gold coated iron oxide (nanorose) is reported. Nanorose engulfed by macrophages associated with atherosclerotic lesions in rabbit arteries absorb incident laser (800nm) energy and cause optical pathlength (OP) variation which is measured using photothermal OCT. OP variation in polydimethyl siloxane tissue phantoms containing varying concentrations of nanorose match values predicted from nanoparticle and material properties. Measurement of OP variation in rabbit arteries in response to laser excitation provides an estimate of nanorose concentration in atherosclerotic lesions of 2.5x109 particles/ml. OP variation in atherosclerotic lesions containing macrophages taking up nanorose has a different magnitude and profile from that observed in control thoracic aorta without macrophages and is consistent with macrophage presence as identified with RAM-11 histology staining. Our results suggest that tissue regions with macrophages taking up nanorose can be detected using photothermal OCT.
doi:10.1364/BOE.1.000002
PMCID: PMC3005175  PMID: 21258441
(170.4500) Optical Coherence Tomography; (170.3880) Medical and Biological Imaging
11.  Interleukin-18 Knockout Mice Display Maladaptive Cardiac Hypertrophy in Response to Pressure Overload 
Interleukin (IL)-18 is a cardiotropic proinflammatory cytokine chronically elevated in the serum of patients with cardiac hypertrophy (LVH). The purpose of this study was to examine the role of IL-18 in pressure-overload hypertrophy using wild type (WT) and IL-18 −/− (null) mice. Adult male C57Bl/6 mice underwent transaortic constriction (TAC) for 7 days or sham surgery. Heart weight/body weight ratios showed blunted hypertrophy in IL-18 null TAC mice compared to WT TAC animals. Microarray analyses indicated differential expression of hypertrophy-related genes in WT versus IL-18 nulls. Northern, Western, and EMSA analyses showed Akt and GATA4 were increased in WT but unchanged in IL-18 null mice. Our results demonstrate blunted hypertrophy with reduced expression of contractile-, hypertrophy-, and remodeling-associated genes following pressure overload in IL-18 null mice, and suggest that IL-18 plays a critical role in the hypertrophic response.
doi:10.1016/j.bbrc.2007.01.030
PMCID: PMC1847636  PMID: 17250807
Interleukins; myocardial hypertrophy; Akt; signal transduction; mouse; pressure overload

Results 1-11 (11)