Recruitment Of Collagen Fibers Research Articles

Direct measurements of collagen fiber recruitment in the posterior pole of the eye

Collagen is the main load-bearing component of the peripapillary sclera (PPS) and lamina cribrosa (LC) in the eye. Whilst it has been shown that uncrimping and recruitment of the PPS and LC collagen fibers underlies the macro-scale nonlinear stiffening of both tissues with increased intraocular pressure (IOP), the uncrimping and recruitment as a function of local stretch have not been directly measured. This knowledge is crucial to understanding their functions in bearing loads and maintaining tissue integrity. In this project we measured local stretch-induced collagen fiber bundle uncrimping and recruitment curves of the PPS and LC. Thin coronal samples of PPS and LC of sheep eyes were mounted and stretched biaxially quasi-statically using a custom system. At each step, we imaged the PPS and LC with instant polarized light microscopy and quantified pixel-level (1.5 μm/pixel) collagen fiber orientations. We used digital image correlation to measure the local stretch and quantified collagen crimp by the circular standard deviation of fiber orientations, or waviness. Local stretch-recruitment curves of PPS and LC approximated sigmoid functions. PPS recruited more fibers than the LC at the low levels of stretch. At 10% stretch the curves crossed with 75% bundles recruited. The PPS had higher uncrimping rate and waviness remaining after recruitment than the LC: 0.9º vs. 0.6º and 3.1º vs. 2.7º. Altogether our findings support describing fiber recruitment of both PPS and LC with sigmoid curves, with the PPS recruiting faster and at lower stretch than the LC, consistent with a stiffer tissue. Statement of significancePeripapillary sclera (PPS) and lamina cribrosa (LC) collagen recruitment behaviors are central to the nonlinear mechanical behavior of the posterior pole of the eye. How PPS and LC collagen fibers recruit under stretch is crucial to develop constitutive models of the tissues but remains unclear. We used image-based stretch testing to characterize PPS and LC collagen fiber bundle recruitment under local stretch. We found that fiber-level stretch-recruitment curves of PPS and LC approximated sigmoid functions. PPS recruited more fibers at a low stretch, but at 10% bundle stretch the two curves crossed with 75% bundles recruited. We also found that PPS and LC fibers had different uncrimping rates and non-zero waviness's when recruited.

Who bears the load? IOP-induced collagen fiber recruitment over the corneoscleral shell

Collagen is the main load-bearing component of cornea and sclera. When stretched, both of these tissues exhibit a behavior known as collagen fiber recruitment. In recruitment, as the tissues stretch the constitutive collagen fibers lose their natural waviness, progressively straightening. Recruited, straight, fibers bear substantially more mechanical load than non-recruited, wavy, fibers. As such, the process of recruitment underlies the well-established nonlinear macroscopic behavior of the corneoscleral shell. Recruitment has an interesting implication: when recruitment is incomplete, only a fraction of the collagen fibers is actually contributing to bear the loads, with the rest remaining “in reserve”. In other words, at a given intraocular pressure (IOP), it is possible that not all the collagen fibers of the cornea and sclera are actually contributing to bear the loads.To the best of our knowledge, the fraction of corneoscleral shell fibers recruited and contributing to bear the load of IOP has not been reported. Our goal was to obtain regionally-resolved estimates of the fraction of corneoscleral collagen fibers recruited and in reserve. We developed a fiber-based microstructural constitutive model that could account for collagen fiber undulations or crimp via their tortuosity. We used experimentally-measured collagen fiber crimp tortuosity distributions in human eyes to derive region-specific nonlinear hyperelastic mechanical properties. We then built a three-dimensional axisymmetric model of the globe, assigning region-specific mechanical properties and regional anisotropy. The model was used to simulate the IOP-induced shell deformation. The model-predicted tissue stretch was then used to quantify collagen recruitment within each shell region. The calculations showed that, at low IOPs, collagen fibers in the posterior equator were recruited the fastest, such that at a physiologic IOP of 15 mmHg, over 90% of fibers were recruited, compared with only a third in the cornea and the peripapillary sclera. The differences in recruitment between regions, in turn, mean that at a physiologic IOP the posterior equator had a fiber reserve of only 10%, whereas the cornea and peripapillary sclera had two thirds. At an elevated IOP of 50 mmHg, collagen fibers in the limbus and the anterior/posterior equator were almost fully recruited, compared with 90% in the cornea and the posterior sclera, and 70% in the peripapillary sclera and the equator. That even at such an elevated IOP not all the fibers were recruited suggests that there are likely other conditions that challenge the corneoscleral tissues even more than IOP. The fraction of fibers recruited may have other potential implications. For example, fibers that are not bearing loads may be more susceptible to enzymatic digestion or remodeling. Similarly, it may be possible to control tissue stiffness through the fraction of recruited fibers without the need to add or remove collagen.

Modeling planar response of vascular tissues using quadratic functions of effective strain.

Simulation-based studies of the cardiovascular structure such as aorta have become increasingly popular for many biomedical problems such as predictions of aneurysm rupture. A critical step in these simulations is the development of constitutive models that accurately describe the tissue's mechanical behavior. In this work, we present a new constitutive model, which explicitly accounts for the gradual recruitment of collagen fibers. The recruitment is considered using an effective stretch, which is a continuum-scale kinematic variable measuring the uncrimped stretch of the tissue in an average sense. The strain energy of a fiber bundle is described by a quadratic function of the effective strain. Constitutive models formulated in this manner are applied to describe the responses of ascending thoracic aortic aneurysm and porcine thoracic aorta tissues. The heterogeneous properties of the ATAA tissue are extracted from bulge inflation test data, and then used in finite element analysis to simulate the inflation test. The descriptive and predictive capabilities are further assessed using planar testing data of porcine thoracic aortic tissues. It is found that the constitutive model can accurately describe the stress-strain relations. In particular, the finite element simulation replicates the displacement, strain, and stress distributions with excellent fidelity.

On strain-based rupture criterion for ascending aortic aneurysm: The role of fiber waviness

We propose a new approach for constructing strain-based rupture criterion for ascending thoracic aortic aneurysm. The rupture metric is formulated using an effective strain, which is a measure of net strain that the collagen bundles experience after fiber uncrimping. The effective strain is a function of the total strain and the waviness properties of the collagen fibers. In the present work, the waviness properties are obtained from fitting biaxial response data to constitutive models that explicitly consider the collagen waviness and fiber recruitment. Inflation test data from 10 ascending thoracic aortic aneurysm specimens are analyzed. For each specimen, tension-strain data at ∼2300 material points are garnered. The effective strain fields in the configuration immediately before rupture are computed. It is found that the hotspots of the effective strain match the rupture sites very well in all 10 samples. More importantly, the values of effective strain at the hotsopts are closely clustered around 0.1, in contrast to a much wider distribution of the total strain. The study underscores the importance of considering the fiber recruitment in formulating strain-based rupture metric, and suggests that ϵ¯≈0.1, where ϵ¯ is the effective strain metric defined in this work, can be considered as a criterion for assessing the imminent rupture risk of ascending aortic aneurysms. Statement of significanceWe advocate to use effective strain in ATAA rupture assessment. The effective strain is a measure of net strain in the collagen network after waviness uncrimping. We analyzed bulge inflation data of ATAA samples. It was found that, while the total strains at rupture varied from sample to sample, the effective strains were closely clustered around 0.1. And the hotspots of effective strain matched the rupture sites well. The work underlines the importance of considering collagen fiber waviness and recruitment when evaluating the rupture risk using strain, and suggests a new direction for developing sharper rupture metrics.

Explicit consideration of fiber recruitment in vascular constitutive formulation using beta functions

Constitutive models are of fundamental importance for describing the mechanical behavior of vascular tissues. In this article, we present a constitutive model that explicitly accounts for collagen fiber recruitment. The formulation, developed based two experimental observations, consists in a systematic use of beta functions. The distribution of random waviness is modeled by beta distribution, and the fiber stresses are represented by incomplete beta functions which are shown to be flexible in capturing different types of responses data. A four-point dispersion model, derived in the context of planar von Mises distribution, is developed to take into account of the orientation distribution of fibers. The dispersion weights appear as Bernstein polynomials of a single dispersion parameter. The descriptive and predictive capabilities are assessed using three groups of experimental data encompassing the responses of porcine thoracic aorta, human intracranial aneurysm, and human ascending thoracic aortic aneurysm tissues. The descriptive performance is evaluated by the quality of constitutive fitting. The predictive ability is tested based on the accuracy in predicting unfitted responses as well as in finite element simulations. The assessment indicates that the model has excellent descriptive and predictive capabilities.

A mathematical model for viscoelastic properties of biological soft tissue.

A quaternary viscoelastic structure model with two characteristic times is presented to describe the viscoelastic properties of parallel-fibered collagen tissue. The comparison results of model prediction and experimental data of rabbit medial collateral ligaments show that the model could accurately describe viscoelastic behavior such as stress-relaxation, strain-strengthening and creep of bio-soft-tissue within a small scope of errors. To study the biomechanical mechanism of viscoelasticity that biological soft tissue shows, the influence of model parameters on viscoelastic behavior of bio-soft-tissue is analyzed and researched, which indicated that the major influential elements of stress-relaxation in bio-soft-tissue are elastic modulus, relaxation time and strain rate of proteoglycan-rich matrix. The influence of elastic modulus of collagen fibers on stress-relaxation is not significant. However, the nonlinearity of stress-strain curve and viscoelastic behavior of bio-soft-tissue mostly depends on recruitment and reorientation of collagen fibers under external loading.

Uniaxial properties of ascending aortic aneurysms in light of effective stretch

A constitutive model that explicitly considers the gradual recruitment of collagen fibers is applied to investigate the uniaxial properties of human ascending aortic aneurysms. The model uses an effective stretch, which is a continuum scale kinematic variable measuring the true stretch of the tissue, to formulate the fiber stress. The constitutive equation contains two shape parameters characterizing the stochastic distribution of fiber waviness, and two elastic parameters accounting for, respectively, the elastic properties of ground substance and the straightened collagen fibers. The model is applied to 156 sets of uniaxial stress-stretch data obtained from 52 aneurysm samples. Major findings include (1) the uniaxial response can be well described by a quadratic strain energy function of the effective strain; (2) the ultimate stretches, when measured in terms of the effective stretch, are closely clustered around 1.1, in contrast to a much wider range in the original stretch; and (3) the ultimate stress correlates positively with the fiber stiffness. The age dependence and directional differences of constitutive parameters are also investigated. Results indicate that only the waviness depends strongly on age; no clear alterations occur in elastic parameters. Further, the fibers are wavier and stiffer in the circumferential direction than in the longitudinal direction. No other parameters exhibit significant direction difference. Statement of significanceWe introduced a constitutive model which explicitly accounts for collagen fiber recruitment to investigate the uniaxial properties of human ascending aortic aneurysm tissues. Uniaxial response data from 156 specimens were considered in the study. It was found that the seemingly dissimilar response curves are, in fact, similar if we measure the deformation using an effective stretch which factors out the uncrimping deformation. The rupture stretches in terms of the effective stretch are closely packed around 1.1. And the stress-stretch curves collapse to a canonical curve after a transformation.

Simulating the time evolving geometry, mechanical properties, and fibrous structure of bioprosthetic heart valve leaflets under cyclic loading

Currently, the most common replacement heart valve design is the ‘bioprosthetic’ heart valve (BHV), which has important advantages in that it does not require permanent anti-coagulation therapy, operates noiselessly, and has blood flow characteristics similar to the native valve. BHVs are typically fabricated from glutaraldehyde-crosslinked pericardial xenograft tissue biomaterials (XTBs) attached to a rigid, semi-flexible, or fully collapsible stent in the case of the increasingly popular transcutaneous aortic valve replacement (TAVR). While current TAVR assessments are positive, clinical results to date are generally limited to <2 years. Since TAVR leaflets are constructed using thinner XTBs, their mechanical demands are substantially greater than surgical BHV due to the increased stresses during in vivo operation, potentially resulting in decreased durability. Given the functional complexity of heart valve operation, in-silico predictive simulations clearly have potential to greatly improve the TAVR development process. As such simulations must start with accurate material models, we have developed a novel time-evolving constitutive model for pericardial xenograft tissue biomaterials (XTB) utilized in BHV (doi: 10.1016/j.jmbbm.2017.07.013). This model was able to simulate the observed tissue plasticity effects that occur in approximately in the first two years of in vivo function (50 million cycles). In the present work, we implemented this model into a complete simulation pipeline to predict the BHV time evolving geometry to 50 million cycles. The pipeline was implemented within an isogeometric finite element formulation that directly integrated our established BHV NURBS-based geometry (doi: 10.1007/s00466-015-1166-x). Simulations of successive loading cycles indicated continual changes in leaflet shape, as indicated by spatially varying increases in leaflet curvature. While the simulation model assumed an initial uniform fiber orientation distribution, anisotropic regional changes in leaflet tissue plastic strain induced a complex changes in regional fiber orientation. We have previously noted in our time-evolving constitutive model that the increases in collagen fiber recruitment with cyclic loading placed an upper bound on plastic strain levels. This effect was manifested by restricting further changes in leaflet geometry past 50 million cycles. Such phenomena was accurately captured in the valve-level simulations due to the use of a tissue-level structural-based modeling approach. Changes in basic leaflet dimensions agreed well with extant experimental studies. As a whole, the results of the present study indicate the complexity of BHV responses to cyclic loading, including changes in leaflet shape and internal fibrous structure. It should be noted that the later effect also influences changes in local mechanical behavior (i.e. changes in leaflet anisotropic tissue stress-strain relationship) due to internal fibrous structure resulting from plastic strains. Such mechanism-based simulations can help pave the way towards the application of sophisticated simulation technologies in the development of replacement heart valve technology.

Incorporating fiber recruitment in hyperelastic modeling of vascular tissues by means of kinematic average.

We present a framework for considering the gradual recruitment of collagen fibers in hyperelastic constitutive modeling. An effective stretch, which is a response variable representing the true stretch at the tissue-scale, is introduced. Properties of the effective stretch are discussed in detail. The effective stretch and strain invariants derived from it are used in selected hyperelastic constitutive models to describe the tissue response. This construction is investigated in conjunction with Holzapfel-Gasser-Ogden family strain energy functions. The ensuing models are validated against a large body of uniaxial and bi-axial stress-strain response data from human aortic aneurysm tissues. Both the descriptive and the predictive capabilities are examined. The former is evaluated by the quality of constitutive fitting, and the latter is assessed using finite element simulation. The models significantly improve the quality of fitting, and reproduce the experiment displacement, stress, and strain distributions with high fidelity in the finite element simulation.

Mechanical and structural contributions of elastin and collagen fibers to interlamellar bonding in the arterial wall.

The artery relies on interlamellar structural components, mainly elastin and collagen fibers, for maintaining its integrity and resisting dissection propagation. In this study, the contribution of arterial elastin and collagen fibers to interlamellar bonding was studied through mechanical testing, multiphoton imaging and finite element modeling. Steady-state peeling experiments were performed on porcine aortic media and the purified elastin network in the circumferential (Circ) and longitudinal (Long) directions. The peeling force and energy release rate associated with mode-I failure are much higher for aortic media than for the elastin network. Also, longitudinal peeling exhibits a higher energy release rate and strength than circumferential peeling for both the aortic media and elastin. Multiphoton imaging shows the recruitment of both elastin and collagen fibers within the interlamellar space and points to in-plane anisotropy of fiber distributions as a potential mechanism for the direction-dependent phenomena of peeling tests. Three-dimensional finite element models based on cohesive zone model (CZM) of fracture were created to simulate the peeling tests with the interlamellar energy release rate and separation distance at damage initiation obtained directly from peeling test. Our experimental results show that the separation distance at damage initiation is 80μm for aortic media and 40μm for elastin. The damage initiation stress was estimated from the model for aortic media (Circ: 60kPa; Long: 95kPa) and elastin (Circ: 9kPa; Long: 14kPa). The interlamellar separation distance at complete failure was estimated to be 3 - 4mm for both media and elastin. Furthermore, elastin and collagen fibers both play an important role in bonding of the arterial wall, while collagen has a higher contribution than elastin to interlamellar stiffness, strength and toughness. These results on microstructural interlamellar failure shed light on the pathological development and progression of aortic dissection.

The toughness of porcine skin: Quantitative measurements and microstructural characterization

An exceptional tear resistance is required of the skin to protect the body from external attacks, environmental damage, and other forms of aggression. To estimate the toughness of juvenile porcine skin, we conduct two types of experiments on pre-notched specimens, placing the tissue under shear (Mode III) by using the classical trouser test with a 25 mm long pre-notch, and opening (Mode I) with an experimental setup with the same pre-notch length. We obtain two distinct average toughness values of JIIIc≈20.4kJ/m2 and JIc=30.4kJ/m2, as a result of differences between these two modes of crack-tip loading and propagation, and collagen alignment. Digital image correlation coupled with single edge notch tests of 10 mm × 30 mm skin samples enables the mapping of the local strains around the tip of the crack. Effects of sample orientation and initial notch size ratio on the strain profile and on the net-section failure stress are discussed. The evaluation of the structure at the crack tip and regions undergoing more uniform states of deformation is conducted by ex situ transmission electron microscopy and in situ environmental scanning electron microscopy. Prior to crack propagation, the stress concentration is decreased by redistributing loads away from the crack tip, illustrated by gradual recruitment of collagen fibers ahead of the crack tip, thus delaying crack growth. After the crack has propagated, collagen fibers are substantially damaged, marked by delamination and recoil of the collagen fibrils.

Contributions of Glycosaminoglycans to Collagen Fiber Recruitment in Constitutive Modeling of Arterial Mechanics

The contribution of glycosaminoglycans (GAGs) to the biological and mechanical functions of biological tissue has emerged as an important area of research. GAGs provide structural basis for the organization and assembly of extracellular matrix (ECM). The mechanics of tissue with low GAG content can be indirectly affected by the interaction of GAGs with collagen fibers, which have long been known to be one of the primary contributors to soft tissue mechanics. Our earlier study showed that enzymatic GAG depletion results in straighter collagen fibers that are recruited at lower levels of stretch, and a corresponding shift in earlier arterial stiffening (Mattson et al., 2016). In this study, the effect of GAGs on collagen fiber recruitment was studied through a structure-based constitutive model. The model incorporates structural information, such as fiber orientation distribution, content, and recruitment of medial elastin, medial collagen, and adventitial collagen fibers. The model was first used to study planar biaxial tensile stress-stretch behavior of porcine descending thoracic aorta. Changes in elastin and collagen fiber orientation distribution, and collagen fiber recruitment were then incorporated into the model in order to predict the stress-stretch behavior of GAG depleted tissue. Our study shows that incorporating early collagen fiber recruitment into the model predicts the stress-stretch response of GAG depleted tissue reasonably well (rms = 0.141); considering further changes of fiber orientation distribution does not improve the predicting capability (rms = 0.149). Our study suggests an important role of GAGs in arterial mechanics that should be considered in developing constitutive models.

Numerical simulation of arterial remodeling in pulmonary autografts

AbstractThe Ross procedure is a surgical procedure where a diseased aortic valve is replaced by the person's own pulmonary valve. The proximal segment of the pulmonary artery is thereby placed in aortic position and therefore suddenly exposed to a sevenfold increase in blood pressure. Excessive dilatation of this pulmonary autograft is a common complication and has excited interest about mechanobiological adaptation in such situations. Mathematical modelling of growth and remodelling is a relevant approach to improve insights into this phenomenon.We introduced an algorithm that models the continuous degradation and deposition of extracellular matrix in an artery according to the constrained mixture theory. To compute temporal variations of collagen and elastin mass and to deduce related mechanical properties of the remodelled artery, we discretized time and defined a finite number of cohorts for each collagen fiber family. The degradation and production rates of each cohort were mediated by the difference between ambient and homeostatic stress within each fiber cohort.We applied the algorithm to predict the adaptation of a pulmonary autograft over an extended period and compared the results to experimental data obtained in sheep. We were able to consistently reproduce the experimentally observed remodeling effects such as dilatation and delayed collagen fiber recruitment. Our simulations revealed how elastin takes up the excessive stress in overstretched regions of the tissue.In conclusion, the algorithm yields very promising results regarding autograft adaptation in overstretched conditions. Future work will focus on other situations of vascular adaptation where growth‐related deformations will be considered.

Collagen fiber recruitment: A microstructural basis for the nonlinear response of the posterior pole of the eye to increases in intraocular pressure

Our goal was to quantify and characterize how the collagen fiber crimp waviness of the lamina cribrosa (LC) and peripapillary sclera (PPS) changes with intraocular pressure (IOP). Thirteen sheep (ovine) eyes were immersion and perfusion fixed while maintaining IOP at 0, 10, 15, 20, or 50 mmHg. Coronal optic nerve head (ONH) sections (30 µm) were imaged with polarized light microscopy (PLM) and analyzed for collagen fiber orientation and waviness (SD of fiber orientation). In the LC, the waviness of every LC beam was measured. In the PPS, at least 900 collagen bundles were measured per eye. Using the waviness at 50 mmHg IOP, we defined tissue-specific thresholds to determine the fraction of loaded or recruited fibers. We found that fiber waviness decreased with IOP (P < 0.001). At every IOP, the waviness of the collagen fibers, and the fraction of fibers recruited in the LC were smaller or equal than those of the PPS (P < 0.001). At 15 mmHg IOP, both LC and PPS had ¾ recruited fibers and ¼ left in reserve. The decreased waviness with IOP and associated fiber recruitment is experimental evidence of fiber-based nonlinear biomechanical behavior of the ONH. At all IOPs the PPS had an equal or larger fraction of fibers recruited than the LC. That both LC and PPS had the same fraction of recruited and reserve fibers at normal IOP suggests that this may be an optimal fraction of recruitment for healthy eyes. Whether this extends to human eyes remains unknown. Statement of significanceCollagen fibers exhibit a natural waviness or crimp that largely determine the nonlinear biomechanics of soft tissue. Experimental measurements of crimp morphology in the sheep eye, and how it changes with intraocular pressure (IOP), however, are exceedingly sparse. We quantified how posterior eye crimp changes with increasing IOP. We found that the lamina cribrosa and peripapillary sclera have fundamentally different crimp, and with increasing IOP, have different proportions of fibers that straighten, or get recruited, versus remaining wavy, or in reserve. Interestingly, at physiologic IOP of 15 mmHg, both tissues had about ¾ fibers recruited and ¼ fibers in reserve, indicating there may be an optimal fraction of fibers.

Native Bovine and Porcine Pericardia Respond to Load With Additive Recruitment of Collagen Fibers.

Bovine and porcine pericardia are currently used for manufacturing prosthetic heart valves: their design has become an increasingly important area of investigation in parallel with progressively expanding indications for the transcutaneous approach to heart valves replacement. Before being cut and shaped, pericardial tissues are expected to be properly characterized. Actually, the mechanical assessment of these biomaterials lacks standardized protocols. In particular, the role of preconditioning for achieving a constant mechanical response of tissue samples is still controversial. In the present work, the mechanical response to uniaxial load of native bovine and porcine pericardia, with and without preconditioning was assessed; moreover, the mechanical behavior of pericardia was investigated and explained. It was demonstrated that: (i) pericardial tissue samples hold memory of the loading history but just within the extent of the deformation applied; (ii) the behavior of native bovine and porcine pericardia in response to load is explained by a mechanism based on the additive recruitment of collagen fibers; (iii) the current concept that plasticity is absent in pericardium has to be at least in part reconsidered.

Transmural remodeling of right ventricular myocardium in response to pulmonary arterial hypertension.

Pulmonary arterial hypertension (PAH) imposes substantial pressure overload on the right ventricular free wall (RVFW), leading to myofiber hypertrophy and remodeling of its collagen fiber architecture. The transmural nature of these adaptations and their effects on the macroscopic mechanical behavior of the RVFW remain largely unexplored. In the present work, we extended our constitutive model for RVFW myocardium to investigate the transmural mechanical and structural remodeling post-PAH. Recent murine experimental studies provided us with comprehensive histomorphological and biaxial mechanical data for viable, passive myocardium for normal and post hypertensive cases. Multiple fiber-level remodeling events were found to be localized in the midwall region (40% < depth < 60%): (i) reorientation and alignment of both myo- and collagen fibers towards longitudinal (apex-to-outflow tract) direction, (ii) substantial increase in the rate of the recruitment of collagen fibers with strain, and (iii) a corresponding increase in the mechanical interactions between the collagen and myofibers. These adaptations suggest a denser and more fibrous connective tissue in the midwall region, and led to a substantially stiffer mechanical response along the longitudinal direction in post-PAH tissues. Moreover, using a Laplace-type mechanical equilibrium analysis of the right ventricle to approximate the wall stress state, we estimated that the longitudinal component of stress remained higher in the hypertensive state while the circumferential component approximately maintained homeostasis values. This result was consistent with our observation from the fiber- and tissue-level remodeling that longitudinally oriented collagen fibers, localized in the midwall region, dominated the remodeling process. The findings of this study highlight the need for more integrated cellular-tissue-organ analysis to better understand the remodeling events during PAH and design interventions.

Layer-dependent role of collagen recruitment during loading of the rat bladder wall.

In this work, we re-evaluated long-standing conjectures as to the source of the exceptionally large compliance of the bladder wall. Whereas these conjectures were based on indirect measures of loading mechanisms, in this work we take advantage of advances in bioimaging to directly assess collagen fibers and wall architecture during biaxial loading. A custom biaxial mechanical testing system compatible with multiphoton microscopy was used to directly measure the layer-dependent collagen fiber recruitment in bladder tissue from 9 male Fischer rats (4 adult and 5 aged). As for other soft tissues, the bladder loading curve was exponential in shape and could be divided into toe, transition and high stress regimes. The relationship between collagen recruitment and loading curves was evaluated in the context of the inner (lamina propria) and outer (detrusor smooth muscle) layers. The large extensibility of the bladder was found to be possible due to folds in the wall (rugae) that provide a mechanism for low resistance flattening without any discernible recruitment of collagen fibers throughout the toe regime. For more extensible bladders, as the loading extended into the transition regime, a gradual coordinated recruitment of collagen fibers between the lamina propria layer and detrusor smooth muscle layer was found. A second important finding was that wall extensibility could be lost by premature recruitment of collagen in the outer wall that cut short the toe region. This change was correlated with age. This work provides, for the first time, a mechanistic understanding of the role of collagen recruitment in determining bladder extensibility and capacitance.

Modeling the response of exogenously crosslinked tissue to cyclic loading: The effects of permanent set.

Bioprosthetic heart valves (BHVs), fabricated from exogenously crosslinked collagenous tissues, remain the most popular heart valve replacement design. However, the life span of BHVs remains limited to 10-15 years, in part because the mechanisms that underlie BHV failure remain poorly understood. Experimental evidence indicates that BHVs undergo significant changes in geometry with in vivo operation, which lead to stress concentrations that can have significant impact on structural damage. These changes do not appear to be due to plastic deformation, as the leaflets only deform in the elastic regime. Moreover, structural damage was not detected by the 65 million cycle time point. Instead, we found that this nonrecoverable deformation is similar to the permanent set effect observed in elastomers, which allows the reference configuration of the material to evolve over time. We hypothesize that the scission-healing reaction of glutaraldehyde is the underlying mechanism responsible for permanent set in exogenously crosslinked soft tissues. The continuous scission-healing process of glutaraldehyde allows a portion of the exogenously crosslinked matrix, which is considered to be the non-fibrous part of the extra-cellular matrix, to be re-crosslinked in the loaded state. Thus, this mechanism for permanent set can be used to explain the time evolving mechanical response and geometry of BHVs in the early stage. To model the permanent set effect, we assume that the exogenously crosslinked matrix undergoes changes in reference configurations over time. The changes in the collagen fiber architecture due to dimensional changes allow us to predict subsequent changes in mechanical response. Results show that permanent set alone can explain and, more importantly, predict how the mechanical response of the biomaterial change with time. Furthermore, we found is no difference in permanent set rate constants between the strain controlled and the stress controlled cyclic loading studies. An important finding we have is that the collagen fiber architecture has a limiting effect on the maximum changes in geometry that the permanent set effect can induce. This is due to the recruitment of collagen fibers as the changes in geometry due to permanent set increase. This means we can potentially optimize the BHV geometry based on the predicted the final BHV geometry after permanent set has largely ceased. Thus, we have developed the first structural constitutive model for the permanent set effect in exogenously crosslinked soft tissue, which can help to simulate BHV designs and reduce changes in BHV geometry during cyclic loading and thus potentially increasing BHV durability.

Characterization of mechanical properties of lamellar structure of the aortic wall: Effect of aging

Arterial wall tissues are sensitive to their mechanical surroundings and remodel their structure and mechanical properties when subjected to mechanical stimuli such as increased arterial pressure. Such remodeling is evident in hypertension and aging. Aging is characterized by stiffening of the artery wall which is assigned to disturbed elastin function and increased collagen content. To better understand and provide new insight on microstructural changes induced by aging, the lamellar model of the aortic media was utilized to characterize and compare wall structure and mechanical behavior of the young and old human thoracic aortic samples. Such model regards arterial media as two sets of alternating concentric layers, namely sheets of elastin and interlamellar layers. Histological and biaxial tests were performed and microstructural features and stress–strain curves of media were evaluated in young and old age groups. Then using optimization algorithms and hyperelastic constitutive equations the stress–strain curves of layers were evaluated for both age groups. Results indicated slight elevation in the volume fraction of interlamellar layer among old subjects most probably due to age related collagen deposition. Aging indicated substantial stiffening of interlamellar layers accompanied by noticeable softening of elastic lamellae. The general significant stiffening of old samples were attributed to both increase of volume fraction of interlamellar layers and earlier recruitment of collagen fibers during load bearing due to functional loss of elastin within wall lamellae. Mechanical characterization of lamellar structure of wall media is beneficial in study of arterial remodeling in response to alternated mechanical environment in aging and clinical conditions through coupling of wall microstructure and mechanical behavior.

Characteristics of thoracic aortic aneurysm rupture in vitro

Ascending thoracic aortic aneurysms (ATAAs) are focal dilatations in the aorta that are prone to rupture or dissection. To accurately evaluate the rupture risk, one must know the local mechanical conditions at the rupture site and understand how rupture is triggered in a layered fibrous media. A challenge facing experimental studies of ATAA rupture is that the ATAA tissue is highly heterogeneous; experimental protocols that operate under the premise of tissue homogeneity will have difficulty delineating the location conditions. In this work, we employed a previously established pointwise identification method to characterize wall stress, strain, and property distributions to a sub-millimeter resolution. Based on the acquired field data, we obtained the local mechanical properties at the rupture site in nine ATAA tissue samples. The rupture stress, ultimate strain, energy density, and the toughness of the tested samples were also reported. Our results show that the direction of the rupture is aligned with the direction of maximum stiffness, indicating that higher stiffness is not always related to higher strength. It was also found that the rupture generally occurs at a location of highest stored energy. As a higher stiffness and higher strain energy indicate a larger recruitment of collagen fibers in the tissue at the location and along the direction of rupture, the recruitment of collagen fibers in the deformation of the tissue is probably essential in ATAA rupture. Statement of SignificanceA major challenge in the experimental study of aneurysm properties is that the tissues are heterogeneous. When the specimens are not reasonably homogeneous, traditional tests that work under the premise of tissue homogeneity cannot reliably delineate the local conditions at the rupture site. In this work, we investigated the local characteristics of rupture of human ascending aortic aneurysm tissue. We identified the stress, strain, and elastic properties to a submillimeter resolution. Based on the field values, we determined the local conditions – elastic properties, direction of maximum stiffness, stress, strain, energy consumption – at the rupture site. It was found that the tissues consistently cleave in the direction of the maximum stiffness, and generally occurs at the location of highest energy. Since a higher stiffness and higher strain energy indicate a larger recruitment of collagen fibers in the tissue at the location and along the direction of rupture, the work suggests that the recruitment of collagen fibers in the deformation of the tissue is probably essential in aneurysm rupture.