epithelial-cell-repopulation-preparation-rodent-extracellular-matrix

Epithelial Cell Repopulation and Preparation of Rodent Extracellular Matrix Scaffolds for Renal Tissue Development

This protocol details the generation of acellular, yet biofunctional, renal extracellular matrix (ECM) scaffolds that are useful as small-scale model ...

Northwestern University

Falls that occur during walking are a significant health problem. One of the greatest impediments to solve this problem is that there is no single obviously "correct" way to quantify walking stability. While many people use variability as a proxy for stability, measures of variability do not quantify how the locomotor system responds to perturbations. The purpose of this study was to determine how changes in walking surface variability affect changes in both locomotor variability and stability. We modified an irreducibly simple model of walking to apply random perturbations that simulated walking over an irregular surface. Because the model's global basin of attraction remained fixed, increasing the amplitude of the applied perturbations directly increased the risk of falling in the model. We generated ten simulations of 300 consecutive strides of walking at each of six perturbation amplitudes ranging from zero (i.e., a smooth continuous surface) up to the maximum level the model could tolerate without falling over. Orbital stability defines how a system responds to small (i.e., "local") perturbations from one cycle to the next and was quantified by calculating the maximum Floquet multipliers for the model. Local stability defines how a system responds to similar perturbations in real time and was quantified by calculating short-term and long-term local exponential rates of divergence for the model. As perturbation amplitudes increased, no changes were seen in orbital stability (r(2)=2.43%; p=0.280) or long-term local instability (r(2)=1.0%; p=0.441). These measures essentially reflected the fact that the model never actually "fell" during any of our simulations. Conversely, the variability of the walker's kinematics increased exponentially (r(2)>or=99.6%; p<0.001) and short-term local instability increased linearly (r(2)=88.1%; p<0.001). These measures thus predicted the increased risk of falling exhibited by the model. For all simulated conditions, the walker remained orbitally stable, while exhibiting substantial local instability. This was because very small initial perturbations diverged away from the limit cycle, while larger initial perturbations converged toward the limit cycle. These results provide insight into how these different proposed measures of walking stability are related to each other and to risk of falling.

Dynamic stability of passive dynamic walking on an irregular surface.

When humans walk, the time duration of each stride varies from one stride to the next. These temporal fluctuations exhibit long-range correlations. It has been suggested that these correlations stem from higher nervous system centers in the brain that control gait cycle timing. Existing proposed models of this phenomenon have focused on neurophysiological mechanisms that might give rise to these long-range correlations, and generally ignored potential alternative mechanical explanations. We hypothesized that a simple mechanical system could also generate similar long-range correlations in stride times. We modified a very simple passive dynamic model of bipedal walking to incorporate forward propulsion through an impulsive force applied to the trailing leg at each push-off. Push-off forces were varied from step to step by incorporating both "sensory" and "motor" noise terms that were regulated by a simple proportional feedback controller. We generated 400 simulations of walking, with different combinations of sensory noise, motor noise, and feedback gain. The stride time data from each simulation were analyzed using detrended fluctuation analysis to compute a scaling exponent, a. This exponent quantified how each stride interval was correlated with previous and subsequent stride intervals over different time scales. For different variations of the noise terms and feedback gain, we obtained short-range correlations (alpha < 0.5), uncorrelated time series (alpha = 0.5), long-range correlations (0.5 < alpha < 1.0), or Brownian motion (alpha > 1.0). Our results indicate that a simple biomechanical model of walking can generate long-range correlations and thus perhaps these correlations are not a complex result of higher level neuronal control, as has been previously suggested.

Possible Biomechanical Origins of the Long-Range Correlations in Stride Intervals of Walking.

To detect macrophages in atherosclerotic plaques, plasmonic gold nanoparticles are introduced as a contrast agent for intravascular photoacoustic imaging. The phantom and ex vivo tissue studies show that the individual spherical nanoparticles, resonant at 530 nm wavelength, produce a weak photoacoustic signal at 680 nm wavelength while photoacoustic signal from nanoparticles internalized by macrophages is very strong due to the plasmon resonance coupling effect. These results suggest that intravascular photoacoustic imaging can assess the macrophage-mediated aggregation of nanoparticles and therefore identify the presence and the location of nanoparticles associated with macrophage-rich atherosclerotic plaques.

Plasmonic intravascular photoacoustic imaging for detection of macrophages in atherosclerotic plaques.

There is a need for an imaging technique that can reliably identify and characterize the vulnerability of atherosclerotic plaques. Catheter-based intravascular ultrasound (IVUS) is one of the imaging tools of the clinical evaluation of atherosclerosis. However, histopathological information obtained with IVUS imaging is limited. We present and discuss the applicability of a combined intravascular photoacoustic (IVPA) and intravascular ultrasound (IVUS) imaging approach to assess both vessel structure and tissue composition thus identifying rupture-prone atherosclerotic plaques. Photoacoustic (or optoacoustic and, generally, thermoacoustic) imaging relies on the absorption of electromagnetic energy, such as light, and the subsequent emission of an acoustic wave. Therefore, the amplitude and temporal characteristics of the photoacoustic signal is primarily determined by optical absorption properties of different types of tissues and can be used to differentiate the lipid, fibrous and fibro-cellular components of an inflammatory lesion. Simultaneous IVUS and IVPA imaging studies were conducted using 40 MHz clinical IVUS imaging catheter interfaced with a pulsed laser system. The performance of the IVPA/IVUS imaging was assessed using phantoms with point targets and vessel-mimicking phantoms. To detect the lipids in the plaque, ex-vivo IVPA imaging studies of a normal and an atherosclerotic rabbit aorta were performed at a 532 nm wavelength. To assess plaque composition, multi-wavelength (680-950 nm) spectroscopic IVPA imaging studies were carried out. Finally, molecular and cellular IVPA imaging was demonstrated using plasmonic nanoparticles. Overall, our studies suggest that plaque detection and characterization can be improved using the combined IVPA/IVUS imaging.

Intravascular ultrasound and photoacoustic imaging.

Large lipid core is common in rupture-prone atherosclerotic plaques. Detection of the location and distribution of lipid in the atherosclerotic plaques can greatly benefit the diagnosis and treatment of vulnerable plaques. Recently introduced intravascular photoacoustic (IVPA) imaging - a technique to image the optical absorption property of tissue - can be used to detect and differentiate atherosclerotic plaques. In this work, we further investigated the ability of using spectroscopic IVPA imaging to visualize the lipid in atherosclerotic plaques. IVPA imaging was performed on an ex-vivo rabbit aorta in the 1200 - 1230 nm wavelength range. In the lipid-rich plaques, the photoacoustic signal strength within this spectral range behaved similar to the optical absorption spectrum of fatty tissue. To distinguish lipid from other types of tissue, correlation analysis was used. Specifically, intraclass correlation between the IVPA signals and the absorption spectrum of lipid reconstructed from multi-wavelength IVPA images was conducted on a pixel-by-pixel basis. The resulted correlation map showed the distribution of lipid in the atherosclerotic plaques. The distribution of lipid is further confirmed by histopathological analysis of tissue. The results of our study suggest that spectroscopic IVPA imaging, together with correlation analysis, may be used to detect lipid in atherosclerotic plaques.

On the possibility to detect lipid in atherosclerotic plaques using intravascular photoacoustic imaging.

Coronary stents are the most commonly used treatment in the United States to repair narrowed or weakened arteries. The ability to visualize the stent during the stenting procedure and post-surgery is crucial to correctly place the stent with respect to the vessel stenosis, and to identify its position within the vessel wall. Current imaging modalities suffer from low contrast, resolution and/or unfavorable artifacts that can inhibit correct visualization of the stent in the artery. We demonstrated the effectiveness of a combined intravascular photoacoustic and intravascular ultrasound imaging method for high resolution and sufficient contrast imaging of commercial stents with respect to the vessel wall.

Photoacoustic imaging of coronary artery stents.

Lipid is a common constituent in atherosclerotic plaques. The location and area of the lipid region is closely related to the progression of the disease. Intravascular photoacoustic (IVPA) imaging, a minimally invasive imaging modality, can spatially resolve the optical absorption property of arterial tissue. Based on the distinct optical absorption spectrum of fat in the near infrared wavelength range, spectroscopic IVPA imaging may distinguish lipid from other water-based tissue types in the atherosclerotic artery. In this study, a bench-top spectroscopic IVPA imaging system was used to ex-vivo image both atherosclerotic and normal rabbit aortas. By combing the spectroscopic IVPA image with the intravascular ultrasound (IVUS) image, lipid regions in the aorta were identified. The results demonstrated that IVUS-guided spectroscopic IVPA imaging is a promising tool to differentiate lipid in atherosclerosis.

Detection of lipid in atherosclerotic vessels using ultrasound-guided spectroscopic intravascular photoacoustic imaging.

The ability to visualize and track temporarily or permanently implanted metal devices is important in many applications ranging from diagnosis to therapy. Specifically, reliable imaging of metal needles is required in today's clinical settings. Currently, ultrasound is utilized to image a needle inserted into tissue in real time. However, the diagnostic value and tracking ability of these images depends highly on the orientation of the needle, and also its proximity to regions of interest in the tissue. We examine the use of photoacoustic imaging combined with current ultrasound imaging methods to obtain high-contrast images of commonly used needles in the body. Experiments were performed using 21 G and 30 G needles inserted into ex vivo porcine tissue and tissue-mimicking phantoms. The needles and surrounding tissue were imaged using an ultrasound imaging system interfaced with the pulsed laser source necessary for photoacoustic imaging. The results suggest that photoacoustic imaging, combined with ultrasound imaging, is capable of real-time, high-contrast, and high-spatial-resolution visualization of metal implants within anatomical landmarks of the background tissue.

Photoacoustic imaging of clinical metal needles in tissue.

IMPORTANCE OF THE FIELD: Photoacoustic imaging is an imaging modality that derives image contrast from the optical absorption coefficient of the tissue being imaged. The imaging technique is able to differentiate between healthy and diseased tissue with either deeper penetration or higher resolution than other functional imaging modalities currently available. From a clinical standpoint, photoacoustic imaging has demonstrated safety and effectiveness in diagnosing diseased tissue regions using either endogenous tissue contrast or exogenous contrast agents. Furthermore, the potential of photoacoustic imaging has been demonstrated in various therapeutic interventions ranging from drug delivery and release to image-guided therapy and monitoring. AREAS COVERED IN THIS REVIEW: This article reviews the current state of photoacoustic imaging in biomedicine from a technological perspective, highlights various biomedical and clinical applications of photoacoustic imaging, and gives insights on future directions. WHAT THE READER WILL GAIN: Readers will learn about the various applications of photoacoustic imaging, as well as the various contrast agents that can be used to assist photoacoustic imaging. This review will highlight both pre-clinical and clinical uses for photoacoustic imaging, as well as discuss some of the challenges that must be addressed to move photoacoustic imaging into the clinical realm. TAKE HOME MESSAGE: Photoacoustic imaging offers unique advantages over existing imaging modalities. The imaging field is broad with many exciting applications for detecting and diagnosing diseased tissue or processes. Photoacoustics is also used in therapeutic applications to identify and characterize the pathology and then to monitor the treatment. Although the technology is still in its infancy, much work has been done in the pre-clinical arena, and photoacoustic imaging is fast approaching the clinical setting.

Advances in Clinical and Biomedical Applications of Photoacoustic Imaging.

Intravascular photoacoustic (IVPA) imaging is a catheter-based, minimally invasive, imaging modality capable of providing high-resolution optical absorption map of the arterial wall. Integrated with intravascular ultrasound (IVUS) imaging, combined IVPA and IVUS imaging can be used to detect and characterize atherosclerotic plaques building up in the inner lining of an artery. In this paper, we present and discuss various representative applications of combined IVPA/IVUS imaging of atherosclerosis, including assessment of the composition of atherosclerotic plaques, imaging of macrophages within the plaques, and molecular imaging of biomarkers associated with formation and development of plaques. In addition, imaging of coronary artery stents using IVPA and IVUS imaging is demonstrated. Furthermore, the design of an integrated IVUS/IVPA imaging catheter needed for in vivo clinical applications is discussed.

Intravascular Photoacoustic Imaging.

Brachytherapy seed therapy is an increasingly common way to treat prostate cancer through localized radiation. The current standard of care relies on transrectal ultrasound (TRUS) for imaging guidance during the seed placement procedure. As visualization of individual metallic seeds tends to be difficult or inaccurate under TRUS guidance, guide needles are generally tracked to infer seed placement. In an effort to improve seed visualization and placement accuracy, the use of photoacoustic (PA) imaging, which is highly sensitive to metallic objects in soft tissue, was investigated for this clinical application. The PA imaging properties of bare (i.e., embedded in pure gelatin) and tissue-embedded (at depths of up to 13 mm) seeds were investigated with a multi-wavelength (750 to 1090 nm) PA imaging technique. Results indicate that, much like ultrasonic (US) imaging, an angular dependence (i.e., seed orientation relative to imaging transducer) of the PA signal exists. Despite this shortcoming, however, PA imaging offers improved contrast, over US imaging, of a seed in prostate tissue if sufficient local fluence is achieved. Additionally, although the PA signal of a bare seed is greatest for lower laser wavelengths (e.g., 750 nm), the scattering that results from tissue tends to favor the use of higher wavelengths (e.g., 1064 nm, which is the primary wavelength of Nd:YAG lasers) when the seed is located in tissue. A combined PA and US imaging approach (i.e., PAUS imaging) shows strong potential to visualize both the seed and the surrounding anatomical environment of the prostate during brachytherapy seed placement procedures.

Photoacoustic imaging of prostate brachytherapy seeds.

Histopathological classification of human prostate cancer (PCA) relies on the morphological assessment of tissue specimens but has limited prognostic value. To address this deficiency, we performed comparative transcriptome analysis of human prostatic acini generated in a three-dimensional basement membrane that recapitulates the differentiated morphological characteristics and gene expression profile of a human prostate glandular epithelial tissue. We then applied an acinar morphogenesis-specific gene profile to two independent cohorts of patients with PCA (total n = 79) and found that those with tumors expressing this profile, which we designated acini-like tumors, had a significantly lower risk of postoperative relapse compared with those tumors with a lower correlation (hazard ratio, 0.078; log-rank test P = 0.009). Multivariate analyses showed superior prognostic prediction performance using this classification system compared with clinical criteria and Gleason scores. We prioritized the genes in this profile and identified programmed cell death protein 4 (PDCD4) and Kruppel-like factor 6 (KLF6) as critical regulators and surrogate markers of prostatic tissue architectures, which form a gene signature that robustly predicts clinical prognosis with a remarkable accuracy in several large series of PCA tumors (total n = 161; concordance index, 0.913 to 0.951). Thus, by exploiting the genomic program associated with prostate glandular differentiation, we identified acini-like PCA and related molecular markers that significantly enhance prognostic prediction of human PCA.

Molecular profiling of prostatic acinar morphogenesis identifies PDCD4 and KLF6 as tissue architecture-specific prognostic markers in prostate cancer.

Many patients with pancreatic ductal adenocarcinoma (PDAC) develop recurrent or metastatic diseases after surgery, so it is important to identify those most likely to benefit from aggressive therapy. Disruption of tissue microarchitecture is an early step in pancreatic tumorigenesis and a parameter used in pathology grading of glandular tumors. We investigated whether changes in gene expression during pancreatic epithelial morphogenesis were associated with outcomes of patients with PDAC after surgery.

A gene expression signature of epithelial tubulogenesis and a role for ASPM in pancreatic tumor progression.

Transcatheter aortic valve implantation is a novel treatment for severe aortic valve stenosis. Due to the recent use of this technology and the procedural variability, there is very little data that quantify the hemodynamic consequences of variations in valve placement. Changes in aortic wall stresses and fluid retention in the sinuses of Valsalva can have a significant effect on the clinical response a patient has to the procedure. By comprehensively characterizing complex flow in the sinuses of Valsalva using digital particle image velocimetry and an advanced heart-flow simulator, various positions of a deployed transcatheter valve with respect to a bioprosthetic aortic valve (valve-in-valve) were tested in vitro. Displacements of the transcatheter valve were axial and directed below the simulated native valve annulus. It was determined that for both blood residence time and aortic Reynolds stresses, it is optimal to have the annulus of the transcatheter valve deployed as close to the aortic valve annulus as possible.

Article	Year
Dynamic stability of passive dynamic walking on an irregular surface. Journal of biomechanical engineering\| PubMed ID: 18067383	2007
Possible Biomechanical Origins of the Long-Range Correlations in Stride Intervals of Walking. Physica A\| PubMed ID: 18335005	2007
Plasmonic intravascular photoacoustic imaging for detection of macrophages in atherosclerotic plaques. Nano letters\| PubMed ID: 18844426	2009
Intravascular ultrasound and photoacoustic imaging. Conference proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference\| PubMed ID: 19162578	2008
On the possibility to detect lipid in atherosclerotic plaques using intravascular photoacoustic imaging. Conference proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Annual Conference\| PubMed ID: 19964847	2009
Photoacoustic imaging of coronary artery stents. Optics express\| PubMed ID: 19997212	2009
Detection of lipid in atherosclerotic vessels using ultrasound-guided spectroscopic intravascular photoacoustic imaging. Optics express\| PubMed ID: 20389501	2010
Photoacoustic imaging of clinical metal needles in tissue. Journal of biomedical optics\| PubMed ID: 20459231	2010
Advances in Clinical and Biomedical Applications of Photoacoustic Imaging. Expert opinion on medical diagnostics\| PubMed ID: 21344060	2010
Intravascular Photoacoustic Imaging. IEEE journal of quantum electronics\| PubMed ID: 21359138	2010
Photoacoustic imaging of prostate brachytherapy seeds. Biomedical optics express\| PubMed ID: 21833361	2011
Molecular profiling of prostatic acinar morphogenesis identifies PDCD4 and KLF6 as tissue architecture-specific prognostic markers in prostate cancer. The American journal of pathology\| PubMed ID: 23219426	2013
A gene expression signature of epithelial tubulogenesis and a role for ASPM in pancreatic tumor progression. Gastroenterology\| PubMed ID: 23896173	2013
The effects of positioning of transcatheter aortic valves on fluid dynamics of the aortic root. ASAIO journal (American Society for Artificial Internal Organs : 1992)\| PubMed ID: 25010918	2014

Jimmy Su

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