[Show abstract][Hide abstract] ABSTRACT: The elastic properties of engineered biomaterials and tissues impact their post-implantation repair potential and structural integrity, and are critical to help regulate cell fate and gene expression. The measurement of properties (e.g., stiffness or shear modulus) can be attained using elastography, which exploits noninvasive imaging modalities to provide functional information of a material indicative of the regeneration state. In this review, we outline the current leading elastography methodologies available to characterize the properties of biomaterials and tissues suitable for repair and mechanobiology research. We describe methods utilizing magnetic resonance, ultrasound, and optical coherent elastography, highlighting their potential for longitudinal monitoring of implanted materials in vivo, in addition to spatiotemporal limits of each method for probing changes in cell-laden constructs. Micro-elastography methods now allow acquisitions at length scales approaching 5–100 μm in two and three dimensions. Many of the methods introduced in this review are therefore capable of longitudinal monitoring in biomaterials and tissues approaching the cellular scale. However, critical factors such as anisotropy, heterogeneity and viscoelasity—inherent in many soft tissues—are often not fully described and therefore require further advancements and future developments.
Full-text · Article · Jan 2016 · Annals of Biomedical Engineering
[Show abstract][Hide abstract] ABSTRACT: The in vivo measurement of articular cartilage deformation is essential to understand how mechanical forces distribute throughout the healthy tissue and change over time in the pathologic joint. Displacements or strain may serve as a functional imaging biomarker for healthy, diseased, and repaired tissues, but unfortunately intratissue cartilage deformation in vivo is largely unknown. Here, we directly quantified for the first time deformation patterns through the thickness of tibiofemoral articular cartilage in healthy human volunteers. Magnetic resonance imaging acquisitions were synchronized with physiologically relevant compressive loading and used to visualize and measure regional displacement and strain of tibiofemoral articular cartilage in a sagittal plane. We found that compression (of 1/2 body weight) applied at the foot produced a sliding, rigid-body displacement at the tibiofemoral cartilage interface, that loading generated subject- and gender-specific and regionally complex patterns of intratissue strains, and that dominant cartilage strains (approaching 12%) were in shear. Maximum principle and shear strain measures in the tibia were correlated with body mass index. Our MRI-based approach may accelerate the development of regenerative therapies for diseased or damaged cartilage, which is currently limited by the lack of reliable in vivo methods for noninvasive assessment of functional changes following treatment.
[Show abstract][Hide abstract] ABSTRACT: The lubricating proteoglycan, lubricin, facilitates the remarkable low friction and wear properties of articular cartilage in the synovial joints of the body. Lubricin lines the joint surfaces and plays a protective role as a boundary lubricant in sliding contact; decreased expression of lubricin is associated with cartilage degradation and the pathogenesis of osteoarthritis. An unmet need for early osteoarthritis treatment is the development of therapeutic molecules that mimic lubricin function and yet are also resistant to enzymatic degradation common in the damaged joint. Here, we engineered a lubricin mimic (mLub) that is less susceptible to enzymatic degradation and binds to the articular surface to reduce friction. mLub was synthesized using a chondroitin sulfate backbone with type II collagen and hyaluronic acid (HA) binding peptides to promote interaction with the articular surface and synovial fluid constituents. In vitro and in vivo characterization confirmed the binding ability of mLub to isolated type II collagen and HA, and to the cartilage surface. Following trypsin treatment to the cartilage surface, application of mLub, in combination with purified or commercially available hyaluronan, reduced the coefficient of friction, and adhesion, to control levels as assessed over macro-to micro-scales by rheometry and atomic force microscopy. In vivo studies demonstrate an mLub residency time of less than 1 week. Enhanced lubrication by mLub reduces surface friction and adhesion, which may suppress the progression of degradation and cartilage loss in the joint. mLub therefore shows potential for treatment in early osteoarthritis following injury.
[Show abstract][Hide abstract] ABSTRACT: The purpose of this study is to evaluate the ability of machine learning to discriminate between magnetic resonance images (MRI) of normal and pathological human articular cartilage obtained under standard clinical conditions.
An approach to MRI classification of cartilage degradation is proposed using pattern recognition and multivariable regression in which image features from MRIs of histologically scored human articular cartilage plugs were computed using weighted neighbor distance using compound hierarchy of algorithms representing morphology (WND-CHRM). The WND-CHRM method was first applied to several clinically available MRI scan types to perform binary classification of normal and osteoarthritic osteochondral plugs based on the OARSI histological system. In addition, the image features computed from WND-CHRM were used to develop a multiple linear least-squares regression model for classification and prediction of an OARSI score for each cartilage plug.
The binary classification of normal and osteoarthritic plugs yielded results of limited quality with accuracies between 36% and 70%. However, multiple linear least-squares regression successfully predicted OARSI scores and classified plugs with accuracies as high as 86%. The present results improve upon the previously-reported accuracy of classification using average MRI signal intensities and parameter values.
MRI features detected by WND-CHRM reflect cartilage degradation status as assessed by OARSI histologic grading. WND-CHRM is therefore of potential use in the clinical detection and grading of osteoarthritis.
Published by Elsevier Ltd.
Full-text · Article · Jun 2015 · Osteoarthritis and Cartilage
[Show abstract][Hide abstract] ABSTRACT: PurposeThis work evaluates the ability of MRIcontrasts to discriminate between normal and pathological human osteochondral plugscharacterized by the OARSI histological system.Methods
Normal and osteoarthritic human osteochondralplugswere scored using the OARSI histological system and imaged at 3T using MRI sequences measuring T1, T2, and T2* relaxation times, magnetization transfer, anddiffusion. The classification accuracies of quantitative MRI parameters and corresponding weighted image contrasts were evaluated.Classification models based on the Mahalanobis distance metric for each MRI measurement were trained and validated using leave-one-out cross-validation with plugsgrouped according to OARSI histological grade and score. MRI measurements used for classification were performed using a region-of-interest analysis which included superficial, deep, and full-thickness cartilage.ResultsThe best classifiers based on OARSI grade and score were T1- and T2-weighted image contrast, which yielded accuracies of 0.68 and 0.75, respectively. Classification accuracies using OARSI score-based group membership were generally higher compared with grade-based group membership.ConclusionsMRI-based classification—either using quantitative MRI parameters or weighted image contrast—is able to detect early osteoarthritic tissue changes as classified by the OARSI histological system.These findings suggest the benefit of incorporating quantitative MRI acquisitions in a comprehensive clinical evaluation of OA. This article is protected by copyright. All rights reserved
No preview · Article · Jan 2015 · Journal of Orthopaedic Research
[Show abstract][Hide abstract] ABSTRACT: Visualizing the three-dimensional morphology and spatial patterning of cells embedded deep within dense connective tissues of the musculoskeletal system has been possible only by utilizing destructive techniques. Here we utilize fructose-based clearing solutions to image cell connectivity and deep tissue-scale patterning in situ by standard confocal microscopy. Optical clearing takes advantage of refractive index matching of tissue and the embedding medium to visualize light transmission through a broad range of bovine and whole mount murine tissues, including cartilage, bone, and ligament, of the head and hindlimb. Using non-destructive methods, we show for the first time intercellular chondrocyte connections throughout the bulk of cartilage, and we reveal in situ patterns of osteocyte processes and the lacunar-canalicular system deep within mineralized cortical bone. Optical clearing of connective tissues is expected to find broad application for the study of cell responses in normal physiology and disease pathology.
[Show abstract][Hide abstract] ABSTRACT: Collagen fibrils form the structural basis for a broad range of complex biological tissues and materials. Collagen serves as an ideal natural polymer, formed as gels or matrices, for engineering solutions aimed at the regeneration of tissues following damage or disease. Recapitulation of native tissue hierarchical structure involves the careful consideration of the fibril-microstructure of the target tissue extracellular matrix and the choice of fibrillogenesis conditions that favor spatially-dependent fibril alignment. While magnetic fields non-destructively influence collagen fibrillogenesis and alignment, previous methods have demonstrated only limited control, especially when preparing large volume tissue constructs suitable for implantation. In this study, we investigate the use of temperature-controlled fibrillogenesis over a range of applicable collagen concentrations for improved magnetic alignment of polymerizable collagen-fibril gels. Magnetically aligned collagen gels show that bulk and microscale fibril alignment depend on both polymerization temperature and collagen concentration. The degree of fibril alignment at the microscale increased with decreasing polymerization temperature and collagen concentration. Further, computational simulations suggest that lower polymerization temperatures affect the internal gel temperature distribution and convective fluid velocity, potentially facilitating greater fibril alignment. This work demonstrates improvements in observed fibril anisotropy compared to previous work using similar magnetic field strengths, suggesting that temperature and collagen concentration may be utilized to achieve desired fibril alignment and structural properties. Improved control of collagen-based gel structure may better emulate native tissue structural (alignment) and physical properties, with enhanced potential for repair success in vivo.
[Show abstract][Hide abstract] ABSTRACT: Functional imaging refers broadly to the visualization of organ or tissue physiology using medical image modalities. In load-bearing tissues of the body, including articular cartilage lining the bony ends of joints, changes in strain, stress, and material properties occur in osteoarthritis (OA), providing an opportunity to probe tissue function through the progression of the disease. Here, biomechanical measures in cartilage and related joint tissues are discussed as key imaging biomarkers in the evaluation of OA. Emphasis will be placed on the (1) potential of radiography, ultrasound, and magnetic resonance imaging to assess early tissue pathomechanics in OA, (2) relative utility of kinematic, structural, morphological, and biomechanical measures as functional imaging biomarkers, and (3) improved diagnostic specificity through the combination of multiple imaging biomarkers with unique contrasts, including elastography and quantitative assessments of tissue biochemistry. In comparison to other modalities, magnetic resonance imaging provides an extensive range of functional measures at the tissue level, with conventional and emerging techniques available to potentially to assess the spectrum of preclinical to advance OA.
No preview · Article · Oct 2014 · Osteoarthritis and Cartilage
[Show abstract][Hide abstract] ABSTRACT: Engineered tissue microenvironments impart specialized cues that drive distinct cellular phenotypes and function. Microenvironments with defined properties, such as mechanical properties and fibril alignment, can elicit specific cellular responses that emulate those observed in vivo. Collagen- and glycosaminoglycan(GAG)- based tissue matrix have been popularized due to their biological ubiquity in a broad range of tissues and the ability to tune structure and mechanical properties through a variety of processes. Here, we investigate the combined effects of static magnetic fields, and GAG and cell encapsulation, on the structure (e.g. collagen fibril orientation) and material properties of collagen matrices. We found that magnetic fields align the collagen-GAG matrix, alter equilibrium mechanical properties, and provide a method for encapsulating cells within a 3D aligned matrix. Cells are encapsulated prior to polymerization, allowing for controlled cell density and eliminating the need for cell seeding. Increased relative GAG concentrations reduced the ability to magnetically align collagen fibrils, in part through a mechanism involving increased viscosity and polymerization time of the collagan-GAG solution. This work provides a functional design space for the development of pure collagen and hybrid collagen-GAG matrices in the presence of magnetic fields. Additionally, this work shows that magnetic fields are effective for the fabrication of collagen constructs with controlled fibril orientation, and can be coupled with GAG incorporation to modulate mechanical properties and the response of embedded cells.
[Show abstract][Hide abstract] ABSTRACT: The purpose of this study was to compare displacement behavior of cyclically loaded cadaveric human intervertebral discs as measured noninvasively on a clinical 3.0 T and a research 9.4 T MRI system. Intervertebral discs were cyclically compressed at physiologically relevant levels with the same MRI-compatible loading device in the clinical and research systems. Displacement-encoded imaging was synchronized to cyclic loading to measure displacements under applied loading with MRI (dualMRI). Displacements from the two systems were compared individually using linear regression and, across all specimens, using Bland–Altman analysis. In-plane displacement patterns measured at 3.0 T and 9.4 T were qualitatively comparable and well correlated. Bland–Altman analyses showed that over 90% of displacement values within the intervertebral disc regions of interest lay within the limits of agreement. Measurement of displacement using dualMRI using a 3.0 T clinical system is comparable to that of a 9.4 T research system. Additional refinements of software, technique implementation, and image processing have potential to improve agreement between different MRI systems. Despite differences in MRI systems in this initial implementation, this work demonstrates that dualMRI can be reliably implemented at multiple magnetic field strengths, permitting translation of dualMRI for a variety of applications in the study of tissue and biomaterial biomechanics.
[Show abstract][Hide abstract] ABSTRACT: The purpose of this study was to compare displacement behavior of cyclically loaded cadaveric human intervertebral discs as measured noninvasively on a clinical 3.0T and a research 9.4T MRI system. Intervertebral discs were cyclically compressed at physiologically relevant levels with the same MRI-compatible loading device in the clinical and research systems. Displacement-encoded imaging was synchronized to cyclic loading to measure displacements under applied loading with MRI (dualMRI). Displacements from the two systems were compared individually using linear regression and, across all specimens, using Bland-Altman analysis. In-plane displacement patterns measured at 3.0T and 9.4T were qualitatively comparable and well correlated. Bland-Altman analyses showed that over 90% of displacement values within the intervertebral disc regions of interest lay within the limits of agreement. Measurement of displacement using dualMRI using a 3.0T clinical system is comparable to that of a 9.4T research system. Additional refinements of software, technique implementation, and image processing have potential to improve agreement between different MRI systems. Despite differences in MRI systems in this initial implementation, this work demonstrates that dualMRI can be reliably implemented at multiple magnetic field strengths, permitting translation of dualMRI for a variety of applications in the study of tissue and biomaterial biomechanics.
No preview · Article · Jun 2014 · Journal of Biomechanics
[Show abstract][Hide abstract] ABSTRACT: Purpose:
Noninvasive assessment of tissue mechanical behavior could enable insights into tissue function in healthy and diseased conditions and permit the development of effective tissue repair treatments. Measurement of displacements under applied loading with MRI (dualMRI) has the potential for such biomechanical characterization on a clinical MRI system.
dualMRI was translated from high-field research systems to a 3T clinical system. Precision was calculated using repeated tests of a silicone phantom. dualMRI was demonstrated by visualizing displacements and strains in an intervertebral disc and compared to T2 measured during cyclic loading.
The displacement and strain precisions were 24 µm and 0.3% strain, respectively, under the imaging parameters used in this study. Displacements and strains were measured within the intervertebral disc, but no correlations were found with the T2 values.
The translation of dualMRI to a 3T system unveils the potential for in vivo studies in a myriad of tissue and organ systems. Because of the importance of mechanical behavior to the function of a variety of tissues, it's expected that dualMRI implemented on a clinical system will be a powerful tool in assessing the interlinked roles of structure, mechanics, and function in both healthy and diseased tissues.
No preview · Article · Mar 2014 · Magnetic Resonance in Medicine
[Show abstract][Hide abstract] ABSTRACT: Purpose:
Medical imaging has the potential to noninvasively diagnose early disease onset and monitor the success of repair therapies. Unfortunately, few reliable imaging biomarkers exist to detect cartilage diseases before advanced degeneration in the tissue.
In this study, we quantified the ability to detect osteoarthritis (OA) severity in human cartilage explants using a multicontrast magnetic resonance imaging (MRI) approach, inclusive of novel displacements under applied loading by MRI, relaxivity measures, and standard MRI.
Displacements under applied loading by MRI measures, which characterized the spatial micromechanical environment by 2D finite and Von Mises strains, were strong predictors of histologically assessed OA severity, both before and after controlling for factors, e.g., patient, joint region, and morphology. Relaxivity measures, sensitive to local macromolecular weight and composition, including T1ρ, but not T1 or T2, were predictors of OA severity. A combined multicontrast approach that exploited spatial variations in tissue biomechanics and extracellular matrix structure yielded the strongest relationships to OA severity.
Our results indicate that combining multiple MRI-based biomarkers has high potential for the noninvasive measurement of OA severity and the evaluation of potential therapeutic agents used in the treatment of early OA in animal and human trials.
Full-text · Article · Feb 2014 · Magnetic Resonance in Medicine
[Show abstract][Hide abstract] ABSTRACT: Nuclear structure and mechanics play a critical role in diverse cellular functions, such as organizing direct access of chromatin to transcriptional regulators. Here, we use a new, to our knowledge, hybrid method, based on microscopy and hyperelastic warping, to determine three-dimensional strain distributions inside the nuclei of single living cells embedded within their native extracellular matrix. During physiologically relevant mechanical loading to tissue samples, strain was transferred to individual nuclei, resulting in submicron distributions of displacements, with compressive and tensile strain patterns approaching a fivefold magnitude increase in some locations compared to tissue-scale stimuli. Moreover, nascent RNA synthesis was observed in the interchromatin regions of the cells studied and spatially corresponded to strain patterns. Our ability to measure large strains in the interchromatin space, which reveals that movement of chromatin in the nucleus may not be due to random or biochemical mechanisms alone, but may result from the transfer of mechanical force applied at a distant tissue surface.
Preview · Article · Nov 2013 · Biophysical Journal
[Show abstract][Hide abstract] ABSTRACT: Biomechanical factors play an important role in the growth, regulation, and maintenance of engineered biomaterials and tissues. While physical factors (e.g. applied mechanical strain) can accelerate regeneration, and knowledge of tissue properties often guide the design of custom materials with tailored functionality, the distribution of mechanical quantities (e.g. strain) throughout native and repair tissues is largely unknown. Here, we directly quantify distributions of strain using noninvasive magnetic resonance imaging (MRI) throughout layered agarose constructs, a model system for articular cartilage regeneration. Bulk mechanical testing, giving both instantaneous and equilibrium moduli, was incapable of differentiating between the layered constructs with defined amounts of 2% and 4% agarose. In contrast, MRI revealed complex distributions of strain, with strain transfer to softer (2%) agarose regions, resulting in amplified magnitudes. Comparative studies using finite element simulations and mixture (biphasic) theory confirmed strain distributions in the layered agarose. The results indicate that strain transfer to soft regions is possible in vivo as the biomaterial and tissue changes during regeneration and maturity. It is also possible to modulate locally the strain field that is applied to construct-embedded cells (e.g. chondrocytes) using stratified agarose constructs.
Full-text · Article · Oct 2013 · Journal of Biomechanics