Joseph T. Keyes

Adaptation of a planar microbiaxial optomechanical device for the tubular biaxial microstructural and macroscopic characterization of small vascular tissues.

Murine models of disease are a powerful tool for researchers to gain insight into disease formation, progression, and therapies. The biomechanical indicators of diseased tissue provide a unique insight into some of these murine models, since the biomechanical properties in scenarios such as aneurysm and Marfan syndrome can dictate tissue failure and mortality. Understanding the properties of the tissue on the macroscopic scale has been shown to be important, as one can then understand the tissues ability to withstand the high stresses seen in the cardiac pulsatile cycle. Alterations in the biomechanical response can foreshadow prospective mechanical failure of the tissue. These alterations are often seen on the microstructural level, and obtaining detailed information on such changes can offer a better understanding of the phenomena seen on the macroscopic level. Unfortunately, mouse models present problems due to the size and delicate features in the mechanical testing of such tissues. In addition, some smaller arteries in large-animal studies (e.g., coronary and cerebral arteries) can present the same issues, and are sometimes unsuitable for planar biaxial testing. The purpose of this paper is to present a robust method for the investigation of the mechanical properties of small arteries and the classification of the microstructural orientation and degree of fiber alignment. This occurs through the cost-efficient modification of a planar biaxial tester that works in conjunction with a two-photon nonlinear microscope. This system provides a means to further investigate how microstructure and mechanical properties are modified in diseased transgenic animals where the tissue is in small tube form. Several other hard-to-test tubular specimens such as cerebral aneurysm arteries and atherosclerotic coronary arteries can also be tested using the described modular device.

Design and demonstration of a microbiaxial optomechanical device for multiscale characterization of soft biological tissues with two-photon microscopy.

The biomechanical response of tissues serves as a valuable marker in the prediction of disease and in understanding the related behavior of the body under various disease and age states. Alterations in the macroscopic biomechanical response of diseased tissues are well documented; however, a thorough understanding of the microstructural events that lead to these changes is poorly understood. In this article we introduce a novel microbiaxial optomechanical device that allows two-photon imaging techniques to be coupled with macromechanical stimulation in hydrated planar tissue specimens. This allows that the mechanical response of the microstructure can be quantified and related to the macroscopic response of the same tissue sample. This occurs without the need to fix tissue in strain states that could introduce a change in the microstructural configuration. We demonstrate the passive realignment of fibrous proteins under various types of loading, which demonstrates the ability of tissue microstructure to reinforce itself in periods of high stress. In addition, the collagen and elastin response of tissue during viscoelastic behavior is reported showing interstitial fluid movement and fiber realignment potentially responsible for the temporal behavior. We also demonstrate that nonhomogeneities in fiber strain exist over biaxial regions of assumed homogeneity.

Evaporation-induced assembly of biomimetic polypeptides

We report an evaporation assisted plasma lithography (EAPL) process for guided self-assembly of a biomimetic silk-elastinlike protein (SELP). We demonstrate the formation of SELP structures from millimeter to submicrometer range on plasma-treatment surface templates during an evaporation-induced self-assembly process. The self-assembly processes at different humidities and droplet volumes were investigated. The process occurs efficiently in a window of optimized operating conditions found to be at 70% relative humidity and 8μl volume of SELP solution. The EAPL approach provides a useful technique for the realization of functional devices and systems using these biomimetic materials.

Comparisons of Planar and Tubular Biaxial Tensile Testing Protocols of the Same Porcine Coronary Arteries

To identify the orthotropic biomechanical behavior of arteries, researchers typically perform stretch-pressure-inflation tests on tube-form arteries or planar biaxial testing of splayed sections. We examined variations in finite element simulations (FESs) driven from planar or tubular testing of the same coronary arteries to determine what differences exist when picking one testing technique vs. another. Arteries were tested in tube-form first, then tested in planar-form, and fit to a Fung-type strain energy density function. Afterwards, arteries were modeled via finite element analysis looking at stress and displacement behavior in different scenarios (e.g., tube FESs with tube- or planar-driven constitutive models). When performing FESs of tube inflation from a planar-driven constitutive model, pressure–diameter results had an error of 12.3% compared to pressure-inflation data. Circumferential stresses were different between tube- and planar-driven pressure-inflation models by 50.4% with the planar-driven model having higher stresses. This reduced to 3.9% when rolling the sample to a tube first with planar-driven properties, then inflating with tubular-driven properties. Microstructure showed primarily axial orientation in the tubular and opening-angle configurations. There was a shift towards the circumferential direction upon flattening of 8.0°. There was also noticeable collagen uncrimping in the flattened tissue.

Deformationally dependent fluid transport properties of porcine coronary arteries based on location in the coronary vasculature.

OBJECTIVE Understanding coronary artery mass transport allows researchers to better comprehend how drugs or proteins move through, and deposit into, the arterial wall. Characterizing how the convective component of transport changes based on arterial location could be useful to better understand how molecules distribute in different locations in the coronary vasculature. METHODS AND RESULTS We measured the mechanical properties and wall fluid flux transport properties of de-endothelialized (similar to post-stenting or angioplasty) left anterior descending (LADC) and right (RC) porcine coronary arteries along their arterial lengths. Multiphoton microscopy was used to determine microstructural differences. Proximal LADC regions had a higher circumferential stiffness than all other regions. Permeability decreased by 198% in the LADC distal region compared to other LADC regions. The RC artery showed a decrease of 46.9% from the proximal to middle region, and 51.7% from the middle to distal regions. The porosity increased in the intima between pressure states, without differences through the remainder of the arterial thickness. CONCLUSIONS We showed that the permeabilities and mechanical properties do vary in the coronary vasculature. With variations in mechanical properties, overexpansion of stents can occur more easily while variations in permeability may lead to altered transport based on location.

Location - Dependent Coronary Artery Diffusive and Convective Mass Transport Properties of a Lipophilic Drug Surrogate Measured Using Nonlinear Microscopy

ABSTRACTPurposeArterial wall mass transport properties dictate local distribution of biomolecules or locally delivered dugs. Knowing how these properties vary between coronary artery locations could provide insight into how therapy efficacy is altered between arterial locations.MethodsWe introduced an indocarbocyanine drug surrogate to the lumens of left anterior descending and right coronary (LADC; RC) arteries from pigs with or without a pressure gradient. Interstitial fluorescent intensity was measured on live samples with multiphoton microscopy. We also measured binding to porcine coronary SMCs in monoculture.ResultsDiffusive transport constants peaked in the middle sections of the LADC and RC arteries by 2.09 and 2.04 times, respectively, compared to the proximal and distal segments. There was no statistical difference between the average diffusivity value between LADC and RC arteries. The convection coefficients had an upward trend down each artery, with the RC being higher than the LADC by 3.89 times.ConclusionsThis study demonstrates that the convective and diffusive transport of lipophilic molecules changes between the LADC and the RC arteries as well as along their length. These results may have important implications in optimizing drug delivery for the treatment of coronary artery disease.

A Gimbal-Mounted Pressurization Chamber for Macroscopic and Microscopic Assessment of Ocular Tissues

The biomechanical model of glaucoma considers intraocular pressure-related stress and resultant strain on load bearing connective tissues of the optic nerve and surrounding peripapillary sclera as one major causative influence that effects cellular, vascular, and axonal components of the optic nerve. By this reasoning, the quantification of variations in the microstructural architecture and macromechanical response of scleral shells in glaucomatous compared to healthy populations provides an insight into any variations that exist between patient populations. While scleral shells have been tested mechanically in planar and pressure-inflation scenarios the link between the macroscopic biomechanical response and the underlying microstructure has not been determined to date. A potential roadblock to determining how the microstructure changes based on pressure is the ability to mount the spherical scleral shells in a method that does not induce unwanted stresses to the samples (for instance, in the flattening of the spherical specimens), and then capturing macroscopic and microscopic changes under pressure. Often what is done is a macroscopic test followed by sample fixation and then imaging to determine microstructural organization. We introduce a novel device and method, which allows spherical samples to be pressurized and macroscopic and microstructural behavior quantified on fully hydrated ocular specimens. The samples are pressurized and a series of markers on the surface of the sclera imaged from several different perspectives and reconstructed between pressure points to allow for mapping of nonhomogenous strain. Pictures are taken from different perspectives through the use of mounting the pressurization scheme in a gimbal that allows for positioning the sample in several different spherical coordinate system configurations. This ability to move the sclera in space about the center of the globe, coupled with an upright multiphoton microscope, allows for collecting collagen, and elastin signal in a rapid automated fashion so the entire globe can be imaged.

A Finite Element Study on Variations in Mass Transport in Stented Porcine Coronary Arteries Based on Location in the Coronary Arterial Tree

Drug-eluting stents have a significant clinical advantage in late-stage restenosis due to the antiproliferative drug release. Understanding how drug transport occurs between coronary arterial locations can better help guide localized drug treatment options. Finite element models with properties from specific porcine coronary artery sections (left anterior descending (LAD), right (RCA); proximal, middle, distal regions) were created for stent deployment and drug delivery simulations. Stress, strain, pore fluid velocity, and drug concentrations were exported at different time points of simulation (0-180 days). Tests indicated that the highest stresses occurred in LAD sections. Higher-than-resting homeostatic levels of stress and strain existed at upwards of 3.0 mm away from the stented region, whereas concentration of species only reached 2.7 mm away from the stented region. Region-specific concentration showed 2.2 times higher concentrations in RCA artery sections at times corresponding to vascular remodeling (peak in the middle segment) compared to all other segments. These results suggest that wall transport can occur differently based on coronary artery location. Awareness of peak growth stimulators and where drug accumulation occurs in the vasculature can better help guide local drug delivery therapies.

Retention and Transport of Hydrophobic and Hydrophilic Drug Surrogate Molecules in Coronary Arteries Measured Nondestructively With Photoacoustic Ultrasound

The design and implementation of local drug delivery mechanisms in cardiovascular applications provides a method by which localized action can occur without potentially problematic systemic effects. This has been especially relevant in the case of drug-eluting stents (DESs). It has been previously shown that the degree of chemical polarization can significantly change the degree of transport and the degree of vascular retention of drugs. Understanding how these differences occur in real-time, and nondestructively, can better help guide the design of such pharmaceuticals. Previous work by our laboratory has indicated differences in transport based on location within the coronary tree (Fig. 1) [1].Copyright

Alterations in Finite Element Results Given Constitutive Models From Tubular and Planar Biaxial Testing of the Same Porcine Coronary Arteries

The biomechanical characterization of tissue offers insight into items such as progression of disease and design parameters for implants1,2. To biomechanically evaluate the properties of blood vessels, biaxial testing is frequently performed because biological samples often exhibit anisotropy, and are most frequently under tension from the applied pressures and stretches3,4. Deciding whether to splay a tubular sample open to test in planar form versus performing pressure-inflation testing is a decision often determined by what testing equipment is available. The purpose of this abstract is to compare pressure-inflation behavior, stress distributions, and fiber architecture in planar versus tubular biaxial testing of the same arteries.Copyright