Rate- and Region-Dependent Mechanical Properties of Göttingen Minipig Brain Tissue in Simple Shear and Unconfined Compression.

Traumatic brain injury (TBI), particularly from explosive blasts, is a major cause of casualties in modern military conflicts. Computational models are an important tool in understanding the underlying biomechanics of TBI but are highly dependent on the mechanical properties of soft tissue to produce accurate results. Reported material properties of brain tissue can vary by several orders of magnitude between studies, and no published set of material parameters exists for porcine brain tissue at strain rates relevant to blast. In this work, brain tissue from the brainstem, cerebellum, and cerebrum of freshly euthanized adolescent male Göttingen minipigs was tested in simple shear and unconfined compression at strain rates ranging from quasi-static (QS) to 300 s-1. Brain tissue showed significant strain rate stiffening in both shear and compression. Minimal differences were seen between different regions of the brain. Both hyperelastic and hyper-viscoelastic constitutive models were fit to experimental stress, considering data from either a single loading mode (unidirectional) or two loading modes together (bidirectional). The unidirectional hyper-viscoelastic models with an Ogden hyperelastic representation and a one-term Prony series best captured the response of brain tissue in all regions and rates. The bidirectional models were generally able to capture the response of the tissue in high-rate shear and all compression modes, but not the QS shear. Our constitutive models describe the first set of material parameters for porcine brain tissue relevant to loading modes and rates seen in blast injury.

Material Properties of Rat Middle Cerebral Arteries at High Strain Rates.

Traumatic brain injury (TBI), resulting from either impact- or nonimpact blast-related mechanisms, is a devastating cause of death and disability. The cerebral blood vessels, which provide critical support for brain tissue in both health and disease, are commonly injured in TBI. However, little is known about how vessels respond to traumatic loading, particularly at rates relevant to blast. To better understand vessel responses to trauma, the objective of this project was to characterize the high-rate response of passive cerebral arteries. Rat middle cerebral arteries (MCAs) were isolated and subjected to high-rate deformation in the axial direction. Vessels were perfused at physiological pressures and stretched to failure at strain rates ranging from approximately 100 to 1300 s-1. Although both in vivo stiffness and failure stress increased significantly with strain rate, failure stretch did not depend on rate.

Umbilical cord artery mechanical properties at various gestational ages.

OBJECTIVE: Umbilical cord tissue is naturally available after birth and may provide insight into the health of a newborn. Intraventricular hemorrhage (IVH) is a common complication of prematurity that is suspected to be associated with structural deficiency of the vasculature. We are interested in determining whether umbilical vessel properties could be used to indicate increased risk for IVH. As a first step toward this, we investigated umbilical artery properties as a function of gestational age. STUDY DESIGN: A total of 31 umbilical cord specimens were collected from births ranging from 24 to 40 weeks gestation. Specimens were grouped according to gestational age (less than 25, 26-30, 31-35, and 36-40 weeks). Tension tests were performed on axial and circumferential strips obtained from umbilical arteries. Stiffness, corresponding stretch values, and cross-sectional tissue areas were compared using analysis of variance. RESULTS: Stress-stretch curves displayed no apparent differences across the gestational age range. Statistical analysis of stiffness and stretch values suggested no differences between groups (p > 0.05). Significance was shown between cross-sectional areas of some groups. CONCLUSIONS: Mechanical characterization of umbilical arteries suggests that no significant changes in material properties occur in the range of 24 to 40 week gestational age.

Region specific anisotropy and rate dependence of Göttingen minipig brain tissue.

Traumatic brain injury is a major cause of morbidity in civilian as well as military populations. Computational simulations of injurious events are an important tool to understanding the biomechanics of brain injury and evaluating injury criteria and safety measures. However, these computational models are highly dependent on the material parameters used to represent the brain tissue. Reported material properties of tissue from the cerebrum and cerebellum remain poorly defined at high rates and with respect to anisotropy. In this work, brain tissue from the cerebrum and cerebellum of male Göttingen minipigs was tested in one of three directions relative to axon fibers in oscillatory simple shear over a large range of strain rates from 0.025 to 250 s-1. Brain tissue showed significant direction dependence in both regions, each with a single preferred loading direction. The tissue also showed strong rate dependence over the full range of rates considered. Transversely isotropic hyper-viscoelastic constitutive models were fit to experimental data using dynamic inverse finite element models to account for wave propagation observed at high strain rates. The fit constitutive models predicted the response in all directions well at rates below 100 s-1, after which they adequately predicted the initial two loading cycles, with the exception of the 250 s-1 rate, where models performed poorly. These constitutive models can be readily implemented in finite element packages and are suitable for simulation of both conventional and blast injury in porcine, especially Göttingen minipig, models.

Strain-induced collagen denaturation is rate dependent in failure of cerebral arteries.

While soft tissues are commonly damaged by mechanical loading, the manifestation of this damage at the microstructural level is not fully understood. Specifically, while rate-induced stiffening has been previously observed in cerebral arteries, associated changes in microstructural damage patterns following high-rate loading are largely undefined. In this study, we stretched porcine middle cerebral arteries to failure at 0.01 and >150 s-1, both axially and circumferentially, followed by probing for denatured tropocollagen using collagen hybridizing peptide (CHP). We found that collagen fibrils aligned with the loading direction experienced less denaturation following failure tests at high than low rates. Others have demonstrated similar rate dependence in tropocollagen denaturation during soft tissue failure, but this is the first study to quantify this behavior using CHP and to report it for cerebral arteries. These findings may have significant implications for traumatic brain injury and intracranial balloon angioplasty. We additionally observed possible tropocollagen denaturation in vessel layers primarily composed of fibrils transversely aligned to the loading axis. To our knowledge, this is the first observation of collagen denaturation due to transverse loading, but further research is needed to confirm this finding. STATEMENT OF SIGNIFICANCE: Previous work shows that collagen hybridizing peptide (CHP) can be used to identify collagen molecule unfolding and denaturation in mechanically overloaded soft tissues, including the cerebral arteries. But experiments have not explored collagen damage at rates relevant to traumatic brain injury. In this work, we quantified collagen damage in cerebral arteries stretched to failure at both high and low rates. We found that the collagen molecule is less damaged at high than at low rates, suggesting that damage mechanisms of either the collagen molecule or other elements of the collagen superstructure are rate dependent. This work implies that arteries failed at high rates, such as in traumatic brain injury, will have different molecular-level damage patterns than arteries failed at low rates. Consequently, improved understanding of damage characteristics may be expanded in the future to better inform clinically relevant cases of collagen damage such as angioplasty and injury healing.

Deformations and end effects in isolated blood vessel testing.

Blood vessels are commonly studied in isolation to define their mechanical and biological properties under controlled conditions. While sections of the wall are sometimes tested, vessels are most often attached to needles and examined in their natural cylindrical configuration where combinations of internal pressure and axial force can be applied to mimic in vivo conditions. Attachments to needles, however, constrain natural vessel response, resulting in a complex state of deformation that is not easily determined. As a result, measurements are usually limited to the midsection of a specimen where end effects do not extend and the deformation is homogeneous. To our knowledge, however, the boundaries of this uninfluenced midsection region have not been explored. The objective of this study was to define the extent of these end effects as a function of vessel geometry and material properties, loading conditions, and needle diameter. A computational fiber framework was used to model the response of a nonlinear anisotropic cylindrical tube, constrained radially at its ends, under conditions of axial extension and internal pressure. Individual fiber constitutive response was defined using a Fung-type strain energy function. While quantitative results depend on specific parameter values, simulations demonstrate that axial stretch is always highest near the constraint and reduces to a minimum in the uninfluenced midsection region. Circumferential stretch displays the opposite behavior. As a general rule, the length of the region disturbed by a needle constraint increases with the difference between the diameter of the needle and the equilibrium diameter of the blood vessel for the imposed loading conditions. The reported findings increase the understanding of specimen deformation in isolated vessel experiments, specifically defining considerations important to identifying a midsection region appropriate for measurement.

Biaxial softening of isolated cerebral arteries following axial overstretch.

Arteries play a critical role in carrying essential nutrients and oxygen throughout the brain; however, vessels can become damaged in traumatic brain injury (TBI), putting neural tissue at risk. Even in the absence of hemorrhage, large deformations can disrupt both the physiological and mechanical behavior of the cerebral vessels. Our group recently reported the effect of vessel overstretch on axial mechanics; however, that work did not address possible changes in circumferential mechanics that are critical to the regulation of blood flow. In order to address this in the present work, ovine middle cerebral arteries were isolated and overstretched axially to 10, 20, or 40% beyond the in vivo configuration. Results showed a statistically significant decrease in circumferential stiffness and strain energy, as well as an increase in vessel diameter following 40% overstretch (p < 0.05). These passive changes would lead to a decrease in vascular resistance and likely play a role in previous reports of cellular dysfunction. We anticipate that our findings will both increase understanding of vessel softening phenomena and also promote improved modeling of cerebrovascular mechanics following head trauma.

Stretch-Induced Intimal Failure in Isolated Cerebral Arteries as a Function of Development.

Recent clinical studies have shown that traumatic brain injury is a significant risk factor for stroke. Motivated to better understand possible mechanisms of this association, we studied subfailure disruption of the intima in overstretched sheep cerebral arteries, as this has been implicated in the increased risk of stroke following blunt cerebrovascular injury. Middle cerebral arteries from four age groups (ranging from fetal to adult) were stretched axially to failure, and intimal disruption was captured with a video camera. All vessels demonstrated intimal disruption prior to catastrophic failure, with nearly all incurring disruption at stretch values well below those at ultimate stress (means of 1.56 and 1.73, respectively); the lowest stretch associated with intimal disruption was 1.29. The threshold of intimal failure was independent of age. Additional analysis showed that disruption included failure of both the endothelium and internal elastic lamina. Although our experiments were conducted at quasi-static rates, the results likely have important implications for vessel function following trauma. Future work should seek to identify subfailure disruption of the cerebrovasculature in head trauma.

Cerebral blood vessel damage in traumatic brain injury.

Traumatic brain injury is a devastating cause of death and disability. Although injury of brain tissue is of primary interest in head trauma, nearly all significant cases include damage of the cerebral blood vessels. Because vessels are critical to the maintenance of the healthy brain, any injury or dysfunction of the vasculature puts neural tissue at risk. It is well known that these vessels commonly tear and bleed as an immediate consequence of traumatic brain injury. It follows that other vessels experience deformations that are significant though not severe enough to produce bleeding. Recent data show that such subfailure deformations damage the microstructure of the cerebral vessels, altering both their structure and function. Little is known about the prognosis of these injured vessels and their potential contribution to disease development. The objective of this review is to describe the current state of knowledge on the mechanics of cerebral vessels during head trauma and how they respond to the applied loads. Further research on these topics will clarify the role of blood vessels in the progression of traumatic brain injury and is expected to provide insight into improved strategies for treatment of the disease.

Molecular-level collagen damage explains softening and failure of arterial tissues: A quantitative interpretation of CHP data with a novel elasto-damage model.

The present experimental-modelling study provides a quantitative interpretation of mechanical data and damage measurements obtained from collagen hybridizing peptide (CHP) techniques on overstretched sheep cerebral arterial tissues. To this aim, a structurally-motivated constitutive model is developed in the framework of continuum damage mechanics. The model includes two internal variables for describing the effects of collagen triple-helical unfolding via interstrand delamination: one governs plastic mechanisms in collagen fibers, leading to a stress softening response of the tissue at the macroscale; the other one describes the loss of fiber structural integrity, leading to tissue final failure. The proposed model is calibrated using the obtained mechanical experimental data, showing excellent fitting capabilities. The predicted evolution of internal variables agree well with independent measurements of molecular-level CHP-based damage data, obtaining an independent a posteriori validation of damage predictions. Moreover, available data on inelastic tissue elongation following supraphysiological loads are successfully reproduced. These outcomes further the hypothesis that the accumulation of interstrand delamination is a primary cause for the evolution of inelastic mechanisms in tissues, and in particular of stress softening up to failure.

Detection and characterization of molecular-level collagen damage in overstretched cerebral arteries.

It is well established that overstretch of arteries alters their mechanics and compromises their function. However, the underlying structural mechanisms behind these changes are poorly understood. Utilizing a recently developed collagen hybridizing peptide (CHP), we demonstrate that a single mechanical overstretch of an artery produces molecular-level unfolding of collagen. In addition, imaging and quantification of CHP binding revealed that overstretch produces damage (unfolding) among fibers aligned with the direction of loading, that damage increases with overstretch severity, and that the onset of this damage is closely associated with tissue yielding. These findings held true for both axial and circumferential loading directions. Our results are the first to identify stretch-induced molecular damage to collagen in blood vessels. Furthermore, our approach is advantageous over existing methods of collagen damage detection as it is non-destructive, readily visualized, and objectively quantified. This work opens the door to revealing additional structure-function relationships in arteries. We anticipate that this approach can be used to better understand arterial damage in clinically relevant settings such as angioplasty and vascular trauma. Furthermore, CHP can be a tool for the development of microstructurally-based constitutive models and experimentally validated computational models of arterial damage and damage propagation across physical scales. STATEMENT OF SIGNIFICANCE: Arteries play a critical role by carrying oxygen and essential nutrients throughout the body. However, trauma to the head and neck, as well as surgical interventions, can overstretch arteries and alter their mechanics. In order to better understand the cause of these changes, we employ a novel collagen hybridizing peptide (CHP) to study collagen damage in overstretched arteries. Our approach is unique in that we go beyond the fiber- and fibril-level and characterize molecular-level disruption. In addition, we image and quantify fluorescently-labeled CHP to reveal a new structure-property relationship in arterial damage. We anticipate that our approach can be used to better understand arterial damage in clinically relevant settings such as angioplasty and vascular trauma.

Cerebrovascular dysfunction following subfailure axial stretch.

Cerebral blood vessels are vital to maintaining the health of the brain. Traumatic brain injury (TBI) commonly results in autoregulatory dysfunction and associated failure of cerebral vessels to maintain homeostasis in the brain. While post-injury changes to brain biochemistry are known to contribute to this dysfunction, tissue deformation may also directly alter vascular smooth muscle cell (SMC) function. As a first step toward understanding stretch-induced dysfunction, this study investigates the effect of overstretch on the contractile behavior of SMCs in middle cerebral arteries (MCAs). We hypothesized that vessel function is altered above a threshold of stretch and strain rate. Twenty-four MCAs from Sprague Dawley rats were tested. Following development of basal SMC tone, vessels were subjected to increasing levels of isosmotic extracellular potassium (K+). Samples were then subjected to an axial overstretch of either 1.2*λIV or 1.3*λIV at strain rates of 0.2 or 20s-1. Following overstretch, SMC contractile behavior was measured again, both immediately and 60min after overstretch. Control vessels were subjected to the same protocol but without overstretch. SMC contractile behavior was characterized using both percent contraction (%C) relative to the fully dilated inner diameter and the K+ dose required to evoke the half maximal contractile response (EC50). Control vessels exhibited increased sensitivity to K+ in successive characterization tests, so all effects were quantified relative to the time-matched control response. Samples exhibited the typical biphasic response to extracellular K+, dilating and contracting in response to small and large K+ concentrations, respectively. As hypothesized, axial overstretch altered SMC contractile behavior, as seen in a decrease in %C for sub-maximal contractile K+ doses (p<0.05) and an increase in EC50 (p<0.01), but only for the test group stretched rapidly to 1.3*λIV. While the change in %C was only significantly different immediately after overstretch, the change to EC50 persisted for 60min. These results indicate that deformation can alter SMC contractile behavior and thus potentially play a role in cerebrovascular autoregulatory dysfunction independent of the pathological chemical environment in the brain post-TBI.

Development of Mechanical and Failure Properties in Sheep Cerebral Arteries.

Traumatic brain injury (TBI) is a devastating problem for people of all ages, but the nature of the response to such injury is often different in children than in adults. Cerebral vessel damage and dysfunction are common following TBI, but age-dependent, large-deformation vessel response has not been characterized. Our objective was to investigate the mechanical properties of cerebral arteries as a function of development. Sheep middle cerebral arteries from four age groups (fetal, newborn, juvenile, and adult) were subjected to biaxial loading around physiological conditions and then to failure in the axial direction. Results show little difference among age groups under physiological loading conditions, but response varied significantly with age in response to large axial deformation. Vessels from all age groups reached the same ultimate stretch level, but the amount of stress carried at a given level of stretch increased significantly with age through the developmental period (fetal to juvenile). Our results are the first to identify changes in cerebral vessel response to large deformations with age and may lead to new insights regarding differences in response to TBI with age.

The state of head injury biomechanics: past, present, and future part 2: physical experimentation.

This presentation is the continuation of the article published in Critical Reviews of Biomedical Engineering, 29(5-6), 2001. That issue contained topics dealing with components and geometry of the human head, classification of head injuries, some early experimental studies, and tolerance considerations. It then dealt with head motion and load characterization, investigations during the period from 1939 to 1966, injury causation and early modeling efforts, the 1966 Head Injury Conference and its sequels, mechanical properties of solid tissues, fluid characterization, and early investigation of the mechanical properties of cranial materials. It continued with a description of the systematic investigations of solid cranial components and structural properties since 1966, fetal cranial properties, analytical head modeling, and numerical solutions of head injury. The paper concluded with experimental dynamic loading of human living and cadaver heads, dynamic loading of surrogate heads, and head injury mechanics. This portion of the paper describes physical head injury experimentation involving animals, primarily primates, human cadavers, volunteers, and inanimate physical models. In order to address the entire domain of head injury biomechanics in the two-part survey, it was intended that this information be supplemented by discussions of head injury tolerance and criteria, automotive and sports safety considerations, and the design of protective equipment, but Professor Goldsmith passed away before these sections could be completed. It is nevertheless anticipated that this attenuated installment will provide, in conjunction with the first part of the survey, a valuable resource for students and practitioners of head injury biomechanics.

Significance of source and size in the mechanical response of human cerebral blood vessels.

Cerebral blood vessels are frequently damaged in traumatic brain injury. Mechanical properties of fresh human cerebral vessels obtained through surgeries have been reported. Because surgical sources of human specimens are rare and produce a limited amount of material, we sought to compare the properties of more readily available cerebral arteries and veins obtained from cadavers to fresh vessel data. Additionally, because the previous study was limited to small vessels available in surgery, it was unknown how generally applicable the results were to larger cerebral arteries and veins. In the current study, large and small cerebral vessels from autopsy were stretched axially. Data from these and similar tests on fresh vessels were combined to determine the significance of source and size on mechanical properties. Structural comparisons of histological samples were additionally utilized to characterize differences. Results indicate that specimens from autopsy and surgery behave similarly except that vessels from autopsy tend to be less extensible. While tests on large vessels were limited, small arteries obtained from autopsy tended to be slightly stiffer than large arteries. In contrast, bridging veins from cadavers were typically stiffer and stretched less before structural failure than cortical veins from the same source. These effects are, however, secondary to differences identified between arteries and veins in the previous study.

Subfailure overstretch induces persistent changes in the passive mechanical response of cerebral arteries.

Cerebral blood vessels are critical in maintaining the health of the brain, but their function can be disrupted by traumatic brain injury (TBI). Even in cases without hemorrhage, vessels are deformed with the surrounding brain tissue. This subfailure deformation could result in altered mechanical behavior. This study investigates the effect of overstretch on the passive behavior of isolated middle cerebral arteries (MCAs), with the hypothesis that axial stretch beyond the in vivo length alters this response. Twenty nine MCA sections from 11 ewes were tested. Vessels were subjected to a baseline test consisting of an axial stretch from a buckled state to 1.05* in vivo stretch (λIV) while pressurized at 13.3 kPa. Specimens were then subjected to a target level of axial overstretch between 1.05*λIV (λz = 1.15) and 1.52*λIV (λz = 1.63). Following overstretch, baseline tests were repeated immediately and then every 10 min, for 60 min, to investigate viscoelastic recovery. Injury was defined as an unrecoverable change in the passive mechanical response following overstretch. Finally, pressurized MCAs were pulled axially to failure. Post-overstretch response exhibited softening such that stress values at a given level of stretch were lower after injury. The observed softening also generally resulted in increased non-linearity of the stress-stretch curve, with toe region slope decreasing and large deformation slope increasing. There was no detectable change in reference configuration or failure values. As hypothesized, the magnitude of these alterations increased with overstretch severity, but only once overstretch exceeded 1.2*λIV (p < 0.001). These changes were persistent over 60 min. These changes may have significant implications in repeated TBI events and in increased susceptibility to stroke post-TBI.

Biaxial and failure properties of passive rat middle cerebral arteries.

Rodents are commonly used as test subjects in research on traumatic brain injury and stroke. However, study of rat cerebral vessel properties has largely been limited to pressure-diameter response within the physiological loading range. A more complete, multiaxial description is needed to guide experiments on rats and rat vessels and to appropriately translate findings to humans. Accordingly, we dissected twelve rat middle cerebral arteries (MCAs) and subjected them to combined inflation and axial stretch tests around physiological loading conditions while in a passive state. The MCAs were finally stretched axially to failure. Results showed that MCAs under physiological conditions were stiffer in the axial than circumferential direction by a mean (±standard deviation) factor of 1.72 (±0.73), similar to previously reported behavior of human cerebral arteries. However, the stiffness for both directions was lower in rat MCA than in human cerebral arteries (p<0.01). Failure stretch values were higher in rat MCA (1.35±0.08) than in human vessels (1.24±0.09) (p=0.003), but corresponding 1st Piola Kirchhoff stress values for rats (0.42±0.09 MPa) were considerably lower than those for humans (3.29±0.64 MPa) (p<0.001). These differences between human and rat vessel properties should be considered in rat models of human cerebrovascular injury and disease.

Distribution of blood-brain barrier disruption in primary blast injury.

Traumatic brain injury (TBI) resulting from explosive-related blast overpressure is a topic at the forefront of neurotrauma research. Compromise of the blood-brain barrier (BBB) and other cerebral blood vessel dysfunction is commonly reported in both experimental and clinical studies on blast injury. This study used a rifle primer-driven shock tube to investigate cerebrovascular injury in rats exposed to low-impulse, pure primary blast at three levels of overpressure (145, 232, and 323 kPa) and with three survival times (acute, 24, and 48 h). BBB disruption was quantified immunohistochemically by measuring immunoglobulin G (IgG) extravasation with image analysis techniques. Pure primary blast generated small lesions scattered throughout the brain. The number and size of lesions increased with peak overpressure level, but no significant difference was seen between survival times. Despite laterally directed blast exposure, equal numbers of lesions were found in each hemisphere of the brain. These observations suggest that cerebrovascular injury due to primary blast is distinct from that associated with conventional TBI.

Abrupt increase in rat carotid blood flow induces rapid alteration of artery mechanical properties.

Vascular remodeling is essential to proper vessel function. Dramatic changes in mechanical environment, however, may initiate pathophysiological vascular remodeling processes that lead to vascular disease. Previous work by some of our group has demonstrated a dramatic rise in matrix metalloproteinase (MMP) expression shortly following an abrupt increase in carotid blood flow. We hypothesized that there would be a corresponding change in carotid mechanical properties. Unilateral carotid ligation surgery was performed to produce an abrupt, sustained increase in blood flow through the contralateral carotid artery of rats. The flow-augmented artery was harvested after sham surgery or 1, 2, or 6 days after flow augmentation. Vessel mechanical response in the circumferential direction was then evaluated through a series of pressure-diameter tests. Results show that the extent of circumferential stretch (normalized change in diameter) at in vivo pressure levels was significantly different (p<0.05) from normo-flow controls at 1 and 2 days following flow augmentation. Measurements at 1, 2, and 6 days were not significantly different from one another, but a trend in the data suggested that circumferential stretch was largest 1 day following surgery and subsequently decreased toward baseline values. Because previous work with this model indicated a similar temporal pattern for MMP-9 expression, an exploratory set of experiments was conducted where vessels were tested 1 day following surgery in animals treated with broad spectrum MMP inhibitors (either doxycycline or GM6001). Results showed a trend for the inhibitors to minimize changes in mechanical properties. Observations demonstrate that vessel mechanical properties change rapidly following flow augmentation and that alterations may be linked to expression of MMPs.

Traumatic spinal cord injury: accidental versus nonaccidental injury.

A 21-month-old boy with steroid-dependent asthma presented to the emergency room with Glascow Coma Score (GCS) 3 and retinal hemorrhages. He was found to have subdural and subarachnoid hemorrhage on computed tomography plus findings of hypoxic-ischemic encephalopathy (HIE). The caretaker history was thought to be inconsistent with the clinical and imaging features, and the patient was diagnosed with nonaccidental injury (NAI) and "shaken baby syndrome." The autopsy revealed a cranial impact site and fatal injury to the cervicomedullary junction. Biomechanical analysis provided further objective support that, although NAI could not be ruled out, the injuries could result from an accidental fall as consistently described by the caretaker.