Reliability of Continuous Shear Wave Elastography in the Pathological Patellar Tendon.

OBJECTIVES: Patellar tendon injuries occur via various mechanisms such as overuse, or due to surgical graft harvest for anterior cruciate ligament reconstruction (ACLR). Quantified patellar tendon stiffness after injury may help guide clinical care. Continuous shear wave elastography (cSWE) allows for the assessment of viscosity and shear modulus in tendons. The reliability of the measure, however, has not been established in the patellar tendon. The purpose of this study was to investigate the interrater reliability, intrarater reliability, and between-day stability of cSWE in both healthy and pathological patellar tendons. METHODS: Participants with patellar tendinopathy (n = 13), history of ACLR using bone-patellar tendon-bone autograft (n = 9), and with no history of patellar tendon injury (n = 13) were recruited. cSWE was performed 4 times by multiple raters over 2 days. Intraclass correlations (ICC) and minimum detectable change (MDC95% ) were calculated. RESULTS: Good to excellent between-day stability were found for viscosity (ICC = 0.905, MDC95%  = 8.3 Pa seconds) and shear modulus (ICC = 0.805, MDC95%  = 27.4 kPa). The interrater reliability measures, however, were not as reliable (ICC = 0.591 and 0.532). CONCLUSIONS: cSWE is a reliable assessment tool for quantifying patellar tendon viscoelastic properties over time. It is recommended, however, that a single rater performs the measure as the interrater reliability was less than ideal.

Multifidus Denervation After Radiofrequency Ablation of the Medial Nerve Alters the Biomechanics of the Spine-A Computational Study.

Radiofrequency ablation of the medial branch is commonly used to treat chronic low back pain involving facet joints, which accounts for 12% to 37% of the total cases of chronic low back pain. An adverse effect of this procedure is the denervation of the multifidus muscle, which may lead to its atrophy which can affect the spine and possibly disc degeneration. This study aims to quantify changes in joint angles and loading caused by multifidus denervation after radiofrequency ablation. AnyBody model of the torso was used to evaluate intervertebral joints in flexion, lateral bending, and torsion. Force-dependent kinematics was used to calculate joint angles and forces. These dependent variables were investigated in intact multifidus, unilateral, and bilateral ablations of L3L4, L4L5, and L5S1 joints. The results showed pronounced angular joint changes, especially in bilateral ablations in flexion, when compared with other cases. The same changes' trend from intact to unilaterally then bilaterally ablated multifidus occurred in joint angles of lateral bending. Meanwhile, joint forces were not adversely affected. These results suggest that multifidus denervation after radiofrequency ablation affects spinal mechanics. Such changes may be associated with abnormal tissue deformations and stresses that can potentially alter their mechanobiology and homeostasis, thereby possibly affecting the health of the spine.

Tendon loading in runners with Achilles tendinopathy: Relations to pain, structure, and function during return-to-sport.

We aimed to (1) compare pain, tendon structure, lower limb function, and Achilles tendon loads while running between limbs in runners with Achilles tendinopathy, and (2) explore the relations of pain, tendon structure, and lower limb function to Achilles tendon loads while running. Twenty runners with Achilles tendinopathy participated in this pilot study. Pain was assessed with questionnaires, quantitative sensory testing, and functional testing. Tendon morphology and mechanical properties were evaluated with ultrasound imaging, continuous shear wave elastography, and ultrasound imaging combined with dynamometry. Lower limb function was assessed with an established test battery. Achilles tendon loads were estimated from biomechanical data acquired during running. Compared to the least symptomatic limb, the most symptomatic limb had lower scores on the Victorian Institute of Sports Assessment - Achilles questionnaire and worse pain during drop countermovement jumping, hopping, and running. Tendon thickness and cross-sectional area were greater, and Young's modulus, drop countermovement jump height, and plyometric quotient during hopping were lower on the most symptomatic limb. Side-to-side differences in drop countermovement jump height were significantly associated with side-to-side differences in Achilles tendon peak forces and average loading rates during running. Various measures of pain, structure, and function differ between limbs in runners with Achilles tendinopathy during return-to-sport. Tendon forces, however, do not differ between limbs during comfortable running. In addition to measures that differ between limbs, measures of performance during drop countermovement jumping may aid in clinical decision-making during return-to-sport because they are associated with tendon forces while running.

Echography analysis of musculoskeletal, heart and liver alterations associated with endothelial dysfunction in obese rats.

BACKGROUND: Modern imaging plays a central role in the care of obese patients, and there is an integral focus on its use and accessibility in individuals who have alterations of various in various organs. The objective in this study was to perform an echographic analysis of musculoskeletal system disorders, endothelial dysfunction and the left ventricle (LV) in obese rats. METHODS: Sprague Dawley rats (250 ± 5 g) were obtained and divided into two groups: the control (C) group was fed with a standard diet, and the obese (Ob) group was fed hyper caloric diet with a high fructose-fat content for 4 months. Body weight, cholesterol, triglycerides, glucose, inflammatory cytokines and adhesion molecules (ICAM-1, VCAM-1) were measured. Additionally, two-dimensional echocardiography, abdominal ultrasound and musculoskeletal system studies were performed in the lower extremities. RESULTS: The body weight in the Ob group was increased compared to that in the control group, (p < 0.001); in addition, increased glucose, cholesterol and triglyceride concentrations (p < 0.05) as well as increased levels of the adhesion molecules ICAM-1 and, VCAM-1 (p < 0.01) were found in the Ob group vs the C group. On ultrasound, 75% of the Ob group presented fatty liver and distal joint abnormalities. CONCLUSION: Obese rats exhibit endothelial dysfunction and musculoskeletal changes, also, fatty liver and articular cysts in the posterior region of the distal lower- extremity joints.

Effectiveness of Saccharomyces boulardii and Metronidazole for Small Intestinal Bacterial Overgrowth in Systemic Sclerosis.

BACKGROUND: Small intestinal bacterial overgrowth (SIBO) affects up to 60% of patients with systemic sclerosis (SSc), and it improves with antibiotics. The addition of probiotics could lead to better results. AIMS: To evaluate the efficacy and safety of Saccharomyces boulardii (SB) versus metronidazole (M) versus M + SB for 2 months, to reduce gastrointestinal symptoms and SIBO assessed with hydrogen breath test in SSc. METHODS: An open pilot clinical trial performed in forty patients with SIBO and SSc (ACR-EULAR 2013) who signed informed consent. Three groups were assigned: M, SB, and M + SB, for 2 months. Hydrogen was measured in parts per million with a hydrogen breath test to evaluate SIBO. The National Institutes of Health Patient-Reported Outcomes Measurement Information System (NIH-PROMIS) questionnaire was applied to quantify gastrointestinal symptoms with a raw score of eight symptoms. This study is registered in ClinicalTrials.gov with the following ID: NCT03692299. RESULTS: Baseline characteristics were similar between groups. The average age was 53.2 ± 9.3 years, and the evolution of SSc was 13.5 (1-34) years. After 2 months of treatment, SIBO was eradicated in 55% of the M + SB group: 33% of SB, and 25% of M. The SB and M + SB groups had decreased diarrhea, abdominal pain, and gas/bloating/flatulence, but M remained unchanged. Reductions in expired hydrogen at 45 to 60 min were as follows: M + SB 48% and 44%, M 18% and 20%, and SB 53% and 60% at the first and second months, respectively (p < 0.01). Adverse effects were epigastric burning and constipation in M (53%) and M + SB (36%), and flatulence/diarrhea in SB (22%). CONCLUSIONS: Metronidazole treatment is partially effective in SIBO, but S. boulardii in monotherapy or in combination improves the gastrointestinal outcomes in SSc.

Achilles tendon cross-sectional area at 12 weeks post-rupture relates to 1-year heel-rise height.

PURPOSE: Achilles tendon rupture leads to long-term plantar flexor deficits, but some patients recover functional performance better than others. Early indicators of tendon healing could be helpful in establishing patient prognosis and making individualized decisions regarding rehabilitation progression. The purpose of this study was to investigate relationships between early tendon morphology and mechanical properties to long-term heel-rise and jumping function in individuals after Achilles tendon rupture. METHODS: Individuals after Achilles tendon rupture were assessed at 4, 8, 12, 24, and 52 weeks post-injury. Tendon cross-sectional area, length, and mechanical properties were measured using ultrasound. Heel-rise and jump tests were performed at 24 and 52 weeks. Correlation and regression analysis were used to identify relationships between tendon structural variables in the first 12 weeks to functional outcomes at 52 weeks, and determine whether the addition of tendon structural characteristics at 24 weeks strengthened relationships between functional performance at 24 and 52 weeks. Functional outcomes of individuals with < 3 cm of elongation were compared to those with > 3 cm of elongation using a Mann-Whitney U test. RESULTS: Twenty-two participants [mean (SD) age = 40 (11) years, 17 male] were included. Tendon cross-sectional area at 12 weeks was the strongest predictor of heel-rise height (R2 = 0.280, p = 0.014) and work symmetry (R2 = 0.316, p = 0.008) at 52 weeks. Jumping performance at 52 weeks was not significantly related to any of the tendon structural measures in the first 12 weeks. Performance of all functional tasks at 24 weeks was positively related to performance on the same task at 52 weeks (r = 0.456-0.708, p < 0.05). The addition of tendon cross-sectional area improved the model for height LSI (R2 = 0.519, p = 0.001). Tendon elongation > 3 cm significantly reduced jumping symmetry (p < 0.05). CONCLUSION: Tendon cross-sectional area and excessive elongation related to plantar flexor performance on functional testing after Achilles tendon rupture. Once an individual is able to perform function-based testing, tendon structural measures may inform long-term prognosis. Ultrasound-based measures of tendon structure early in recovery seem to relate to later performance on functional testing. Clinically, assessing tendon structure has the potential to be used as a biomarker of tendon healing early in recovery and better predict patients at risk of negative functional outcome. LEVEL OF EVIDENCE: II.

Changes in Shear Modulus of the Lumbar Multifidus Muscle During Different Body Positions.

Multifidus function is important for active stabilization of the spine, but it can be compromised in patients with chronic low back pain and other spine pathologies. Force production and strength of back muscles are often evaluated using isometric or isokinetic tests, which lack the ability to quantify multifidi contribution independent of the erector spinae and adjacent hip musculature. The objective of this study is to evaluate localized force production capability in multifidus muscle using ultrasound shear wave elastography (SWE) in healthy individuals. Three different body positions were considered: lying prone, sitting up, and sitting up with the right arm lifted. These positions were chosen to progressively increase multifidus contraction and to minimize body motion during measurements. Shear modulus was measured at the superficial and deeper layers of the multifidus. Repeatability and possible sources of error of the shear modulus measurements were analyzed. Multifidus shear modulus (median (interquartile range)) increased from prone, i.e., 16.15 (6.69) kPa, to sitting up, i.e., 27.28 (15.72) kPa, to sitting up with the right arm lifted position, i.e., 45.02 (25.27) kPa. Multifidi shear modulus in the deeper layer of the multifidi was lower than the superficial layer, suggesting lower muscle contraction. Intraclass correlation coefficients (ICCs) for evaluation of shear modulus by muscle layer were found to be excellent (ICC = 0.76-0.80). Results suggest that the proposed protocol could quantify local changes in spinal muscle function in healthy adults; further research in patients with spine pathology is warranted.

Tendon morphology and mechanical properties assessed by ultrasound show change early in recovery and potential prognostic ability for 6-month outcomes.

PURPOSE: Optimizing tendon structural recovery during the first 12 weeks after Achilles tendon rupture is a prime target to improve patient outcomes, but a comprehensive understanding of biomarkers is needed to track early healing. The purpose of this study was to observe healing of tendon structure over time using ultrasound-based, tendon-specific measures and to identify relationships between tendon structural characteristics and clinical measures of gait and strength. METHODS: Twenty-seven participants (21 males, mean (SD) age 39 (11) years) were assessed at 4, 8, 12, and 24 weeks after injury or surgery using ultrasound imaging techniques. Gait analysis and strength testing were added at the later time points. RESULTS: Ruptured tendons had significantly lower dynamic shear modulus (p < 0.001), greater tendon cross-sectional area (p < 0.001), and greater length (p < 0.001) than the uninjured side. Dynamic shear modulus, cross-sectional area, and length were found to increase over time (p < 0.01). Tendon structure at 4 weeks post-injury [cross-sectional area symmetry (r = 0.737, p = 0.002) and dynamic shear modulus (r = 0.518, p = 0.040)] related to stance phase walking symmetry at 24 weeks. CONCLUSIONS: Tendon structure assessed by ultrasound imaging changes over the first 24 weeks of healing after Achilles tendon rupture, suggesting it could be used as a biomarker to track tendon healing early in recovery. Additionally, tendon structure within the first 12 weeks relates to later walking gait and heel-rise symmetry, which may indicate that tendon structure could have prognostic value in the care of these patients. This study's clinical relevance is in its support for using ultrasound imaging to assess early patient healing and prognosticate later patient outcomes after Achilles tendon rupture. LEVEL OF EVIDENCE: Level 2, prospective cohort prognostic study.

Early Detection of Vascular Obstruction in Microvascular Flaps Using a Thermographic Camera.

BACKGROUND: In microsurgical reconstruction, vascular obstruction occurs in approximately 20% of patients. Close monitoring is central to their care. Clinical/Doppler detection of vascular obstruction could be enhanced by thermography. METHODS: A diagnostic test design included consecutive cases of hospitalized patients, ≥18 years old, who underwent surgery with free flaps. Two researchers separately evaluated patients with clinical/Doppler methods and thermographic camera hourly for 24 hours, every 2 hours for the next 24 hours, and then every 3 hours until discharge. The gold standard was visualization of thrombus or vascular obstruction during surgical reintervention. Sensitivity, specificity, positive/negative predictive value (PPV/NPV), and a delta temperature receiver operating characteristic (ROC) curve were calculated. RESULTS: A total of 2,364 tests were performed with a thermographic camera in 40 patients (31 females, 9 males) aged 50.12 ± 9.7 years. There were 28 deep inferior epigastric perforator, 5 anterolateral thigh, 3 radial, 2 scapular, 1 fibular, and 1 anteromedial thigh flaps included. Six (15%) had postoperative vascular obstruction (5 venous and 1 arterial). One flap developed partial necrosis and one total necrosis (overall survival 97.5%). ROC curve (area 0.97) showed the best results at ≥ 1.8°C of difference to the surrounding skin. Considering two consecutive positive evaluations, the sensitivity was 93%, specificity 96%, PPV 57%, and NPV 99%. The thermal imaging camera allows to identify the obstruction between 2 and 12 hours before the clinical method. CONCLUSION: Utilizing a thermographic camera can reduce detection time of vascular obstruction by several hours in microvascular free flaps that include the cutaneous island. This method proves useful for early diagnosis of postoperative vascular obstruction.

Impact of intestinal mannitol on hyperammonemia, oxidative stress and severity of hepatic encephalopathy in the ED.

Hyperammonemia results from hepatic inability to remove nitrogenous products generated by protein metabolism of intestinal microbiota, which leads to hepatic encephalopathy (HE) in chronic liver disease (CLD). In ammonium neurotoxicity, oxidative stress (OxS) plays a pathogenic role. Our objective was to evaluate if intestinal mannitol is as effective and safe as conventional treatment for diminishing hyperammonemia, OxS, and HE in patients with CLD. MATERIAL AND METHODS: We included 30 patients with HE classified by "Haven Criteria for Hepatic Encephalopathy". They were randomized into two groups: 1) Mannitol Group (MG) with mannitol 20% administered into the intestine by an enema, 2) conventional group (CG) with lactulose 40 g enema both substances were diluted in 800 mL of double distilled solution every 6 h; all patients received neomycin. We evaluated ammonia concentration, plasma oxidative stress, HE severity, intestinal discomfort and adverse effects. RESULTS: Hyperammonemia (171 ±â€¯104 vs 79 ±â€¯49 µmol ammonia/L, p < 0.01), and oxidative stress (MDA 29 vs 27%, formazan 15 vs 11%, carbonyls 16 vs 9% and dityrosines 10 vs 5%) were reduced in MG and CG respectively. The HE severity decreased by two degrees compared to baseline values in both groups. Intestinal discomfort and electrolyte plasma alterations were less frequent (p < 0.05) in MG than CG. CONCLUSIONS: Intestinal mannitol is as effective and safe as conventional treatment for reducing hyperammonemia, oxidative stress, and hepatic encephalopathy of CLD patients in the emergency room. Likewise, mannitol is better tolerated than conventional treatment.

Malnutrition is an independent risk factor for mortality in Mexican patients with systemic sclerosis: a cohort study.

Factors for mortality in systemic sclerosis (SSc) vary in different cohorts around the world. Case-control study nested in a cohort. We included patients ≥16 years of age with SSc (ACR/EULAR 2013), from 2005 to 2015. Demographic and clinical variables and causes of mortality were recorded. We calculated Crude Mortality Rate (CMR), Standardized Mortality Ratio (SMR), and Kaplan-Meier survival analysis was performed. A Cox proportional hazard (HR) regression analysis of the potential risk factors associated with mortality was also performed. A total of 220 patients with SSc were included. During follow-up, 28 deaths occurred. The sum of total time contributed by all subjects was 1074 years-person, the CMR was 12.72%, the overall SMR was 4.5, in women 3.7, and in men 4.7. The survival rate at 5 and 10 years was 83 and 70%, respectively. The causes of death were definitively attributed to SSc in 21.4% of the cases, probably in 28.7%, unrelated in 35.6%, and unknown in 14.3%. The direct cause of death of the patients was infection in 25% of cases, cardiovascular disease in 14%, lung involvement in 14%, pulmonary embolism in 11%, and neoplasia in 11%. The Cox regression analysis showed that the factors associated with mortality were: male gender (HR 5.84, CI 95% 1.31-26, p = 0.013), severe Medsger's score for general symptoms (HR 5.12, CI 95% 1.74-14.97, p = 0.021) and severe malnutrition (HR 3.77, CI 95% 1.23-11.06, p = 0.008). Infections, cardiovascular disease, and lung involvement were the leading cause of death. Male gender and severe general affection and malnutrition were associated with a poorer prognosis of SSc.

Expression of NLRP3 inflammasome, cytokines and vascular mediators in the skin of systemic sclerosis patients.

BACKGROUND: The activated NLRP3 inflammasome is associated with the etiology of fibrotic diseases. The role of inflammasomes in SSc is still poorly understood. OBJECTIVES: To determine the expression of NLRP3 (nucleotide-binding domain, leucine-rich-repeat-containing family, pyrin domain-containing 3) in the skin of patients with systemic sclerosis (SSc) and its relationship with pro-inflammatory cytokines and vascular mediators expression. METHODS: Skin biopsies were taken from 42 patients with either limited or diffuse SSc (21 lcSSc and 21 dcSSc), and from 13 healthy individuals. Using real-time polymerase chain reaction (PCR), the relative expression of caspase-1, IL-1ß, IL-18, IL-33, TGF-ß, ET-1, iNOS and eNOS genes, were measured. The location of NLRP3 and IL-1ß were also determined by immunohistochemistry. Clinical characteristics were evaluated. RESULTS: The mean age of the patients was 49.3 ± 12.9 (lcSSc), 44.6 ± 1 3.8 (dcSSc), and 45 ± 14.1 (healthy individuals). Compared to healthy individuals, the skin of both subtypes of SSc showed a significant increase (P < 0.05) in NLRP3, caspase-1, IL-1ß, IL-18 and ET-1. Samples of lcSSc also showed a significant increase of eNOS (P < 0.029), iNOS (P < 0.04) and TGF-ß (P < 0.05). Dermal fibrosis evaluated by modified Rodnan skin score (MRSS) had significant correlation with NLRP3, IL-1ß, IL-18, and ET-1. Immunohistochemical analysis showed stronger staining of NLRP3 and IL-1ß cytoplasmic expression in the keratinizing squamous epithelium of skin from SSc patients compared to controls. CONCLUSIONS: This study identified NLRP3 over-expression in skin of patients with SSc. Skin thickness correlates positively with the NLRP3 inflammasome gene expression and with the vascular mediator and pro-fibrotic ET-1, suggesting that NLRP3 inflammasome plays a role in the pathophysiology of skin fibrosis in human SSc.

The shear modulus of the nucleus pulposus measured using magnetic resonance elastography: a potential biomarker for intervertebral disc degeneration.

PURPOSE: This study aims to: (1) measure the shear modulus of nucleus pulposus (NP) in intact human vertebra-disc-vertebra segments using a magnetic resonance elastography setup for a 7T whole-body scanner, (2) quantify the effect of disc degeneration on the NP shear modulus measured using magnetic resonance elastography, and (3) compare the NP shear modulus to other magnetic resonance-based biomarkers of dis degeneration. METHODS: Thirty intact human disc segments were classified as normal, mild, or severely degenerated. The NP shear modulus was measured using a custom-made setup that included a novel inverse method less sensitive to noisy displacements. T2 relaxation time was measured at 7T. The accuracy of these parameters to classify different degrees of degeneration was evaluated using receiver operating characteristic curves. RESULTS: The magnetic resonance elastography measure of shear modulus in the NP was able to differentiate between normal, mild degeneration, and severe degeneration. The T2 relaxation time was able to differentiate between normal and mild degeneration, but it could not distinguish between mild and severe degeneration. CONCLUSIONS: This study shows that the NP shear modulus measured using magnetic resonance elastography is sensitive to disc degeneration and has the potential of being used as a clinical tool to quantify the mechanical integrity of the intervertebral disc.

Accurate Prediction of Stress in Fibers with Distributed Orientations Using Generalized High-Order Structure Tensors.

The orientation of collagen fibers plays an important role on the mechanics of connective tissues. Connective tissues have fibers with different orientation distributions. The angular integration formulation used to model the mechanics of fibers with distributed orientation is accurate, but computationally expensive for numerical methods such as finite elements. This study presents a formulation based on pre-integrated Generalized High-Order Structure Tensors (GHOST) which greatly improves the accuracy of the predicted stress. Simplifications of the GHOST formulation for transversely-isotropic and planar fiber distributions are also presented. Additionally, the GHOST and the angular integration formulations are compared for different loading conditions, fiber orientation functions, strain energy functions and degrees of fiber non-linearity. It was found that the GHOST formulation predicted the stress of the fibers with an error lower than 10% for uniaxial and biaxial tension. Fiber non-linearity increased the error of the GHOST formulation; however, the error was reduced to negligible values by considering higher order structure tensors. The GHOST formulation produced lower errors when used with an elliptical fiber density function and a binomial strain energy function. In conclusion, the GHOST formulation is able to accurately predict the stress of fibers with distributed orientation without requiring numerous integral calculations. Consequently, the GHOST formulation may reduce the computational effort needed to analyze the mechanics of fibrous tissues with distributed orientations.

Regional healing trajectory of the patellar tendon after bone-patellar tendon-bone autograft harvest for anterior cruciate ligament reconstruction.

Graft site morbidities after bone-patellar tendon-bone (BPTB) autograft harvest for anterior cruciate ligament reconstruction (ACLR) negatively impacts rehabilitation. The purpose of this study was to establish tendon structural properties 1-month after BPTB autograft harvest compared to the uninvolved patellar tendon, and subsequently to quantify the healing trajectory of the patellar tendon over the course of rehabilitation. Patellar tendon morphology (ultrasound) and mechanical properties (continuous shear wave elastography) from 3 regions of the tendon (medial, lateral, central) were measured in 34 participants at 1 month, 3-4 months, and 6-9 months after ACLR. Mixed models were used to compare tendon structure between limbs at 1 month, and quantify healing over 3 timepoints. The involved patellar tendon had increased cross-sectional area and thickness in all regions 1-month after ACLR. Thickness reduced uniformly over time. Possible tendon elongation was observed and remained stable over time. Tendon viscosity was uniform across the three regions in the involved limb while the medial region had higher viscosity in the uninvolved limb, and shear modulus was elevated in all three regions at 1 month. Viscosity and shear modulus in only the central region reduced over time. Statement of Clinical Significance: The entire patellar tendon, and not just the central third, is altered after graft harvest. Tendon structure starts to normalize over time, but alterations remain especially in the central third at the time athletes are returning to sport. Early rehabilitation consisting of tendon loading protocols may be necessary to optimize biologic healing at the graft site tendon.

Biaxial tension of fibrous tissue: using finite element methods to address experimental challenges arising from boundary conditions and anisotropy.

Planar biaxial tension remains a critical loading modality for fibrous soft tissue and is widely used to characterize tissue mechanical response, evaluate treatments, develop constitutive formulas, and obtain material properties for use in finite element studies. Although the application of tension on all edges of the test specimen represents the in situ environment, there remains a need to address the interpretation of experimental results. Unlike uniaxial tension, in biaxial tension the applied forces at the loading clamps do not transmit fully to the region of interest (ROI), which may lead to improper material characterization if not accounted for. In this study, we reviewed the tensile biaxial literature over the last ten years, noting experimental and analysis challenges. In response to these challenges, we used finite element simulations to quantify load transmission from the clamps to the ROI in biaxial tension and to formulate a correction factor that can be used to determine ROI stresses. Additionally, the impact of sample geometry, material anisotropy, and tissue orientation on the correction factor were determined. Large stress concentrations were evident in both square and cruciform geometries and for all levels of anisotropy. In general, stress concentrations were greater for the square geometry than the cruciform geometry. For both square and cruciform geometries, materials with fibers aligned parallel to the loading axes reduced stress concentrations compared to the isotropic tissue, resulting in more of the applied load being transferred to the ROI. In contrast, fiber-reinforced specimens oriented such that the fibers aligned at an angle to the loading axes produced very large stress concentrations across the clamps and shielding in the ROI. A correction factor technique was introduced that can be used to calculate the stresses in the ROI from the measured experimental loads at the clamps. Application of a correction factor to experimental biaxial results may lead to more accurate representation of the mechanical response of fibrous soft tissue.

Challenging the assumption of uniformity in patellar tendon structure: Regional patellar tendon morphology and mechanical properties in vivo.

Patellar tendons are assumed to be uniform in morphology and mechanical properties despite a higher prevalence of tendinopathies observed in the medial region. The purpose of this study was to compare the thickness, length, viscosity, and shear modulus of the medial, central, and lateral regions of healthy patellar tendons of young males and females in vivo. B-mode ultrasound and continuous shear wave elastography were performed on 35 patellar tendons (17 females, 18 males) over three regions of interest. A linear mixed-effects model (α = 0.05) was used to determine differences between the three regions and sexes followed by pairwise comparisons for significant findings. The lateral region (mean [95% confidence interval] = 0.34 [0.31-0.37] cm) was thinner compared with the medial (0.41 [0.39-0.44] cm, p < 0.001), and central (0.41 [0.39-0.44] cm, p < 0.001) regions regardless of sex. Viscosity was lower in the lateral (19.8 [16.9-22.7] Pa-s) versus medial region (27.4 [24.7-30.2] Pa-s, p = 0.001). Length had a region-by-sex interaction (p = 0.003) characterized by a longer lateral (4.83 [4.54-5.13] cm) versus medial (4.42 [4.12-4.72] cm) region in males (p < 0.001), but not females (p = 0.992). Shear modulus was uniform between regions and sexes. The thinner, and less viscous lateral patellar tendon may reflect the lower load the tendon experiences explaining the differences in regional prevalence of developing tendon pathology. Statement of Clinical Significance: Healthy patellar tendons are not uniform in morphology or mechanical properties. Considering regional tendon properties may help guide targeted interventions for patellar tendon pathologies.

Biaxial tensile testing and constitutive modeling of human supraspinatus tendon.

The heterogeneous composition and mechanical properties of the supraspinatus tendon offer an opportunity for studying the structure-function relationships of fibrous musculoskeletal connective tissues. Previous uniaxial testing has demonstrated a correlation between the collagen fiber angle distribution and tendon mechanics in response to tensile loading both parallel and transverse to the tendon longitudinal axis. However, the planar mechanics of the supraspinatus tendon may be more appropriately characterized through biaxial tensile testing, which avoids the limitation of nonphysiologic traction-free boundary conditions present during uniaxial testing. Combined with a structural constitutive model, biaxial testing can help identify the specific structural mechanisms underlying the tendon's two-dimensional mechanical behavior. Therefore, the objective of this study was to evaluate the contribution of collagen fiber organization to the planar tensile mechanics of the human supraspinatus tendon by fitting biaxial tensile data with a structural constitutive model that incorporates a sample-specific angular distribution of nonlinear fibers. Regional samples were tested under several biaxial boundary conditions while simultaneously measuring the collagen fiber orientations via polarized light imaging. The histograms of fiber angles were fit with a von Mises probability distribution and input into a hyperelastic constitutive model incorporating the contributions of the uncrimped fibers. Samples with a wide fiber angle distribution produced greater transverse stresses than more highly aligned samples. The structural model fit the longitudinal stresses well (median R(2) ≥ 0.96) and was validated by successfully predicting the stress response to a mechanical protocol not used for parameter estimation. The transverse stresses were fit less well with greater errors observed for less aligned samples. Sensitivity analyses and relatively affine fiber kinematics suggest that these errors are not due to inaccuracies in measuring the collagen fiber organization. More likely, additional strain energy terms representing fiber-fiber interactions are necessary to provide a closer approximation of the transverse stresses. Nevertheless, this approach demonstrated that the longitudinal tensile mechanics of the supraspinatus tendon are primarily dependent on the moduli, crimp, and angular distribution of its collagen fibers. These results add to the existing knowledge of structure-function relationships in fibrous musculoskeletal tissue, which is valuable for understanding the etiology of degenerative disease, developing effective tissue engineering design strategies, and predicting outcomes of tissue repair.

Nucleus pulposus cells synthesize a functional extracellular matrix and respond to inflammatory cytokine challenge following long-term agarose culture.

Intervertebral disc degeneration is characterized by a cascade of cellular, biochemical and structural changes that may lead to functional impairment and low back pain. Interleukin-1 beta (IL-1ß) is strongly implicated in the etiology of disc degeneration, however there is currently no direct evidence linking IL-1ß upregulation to downstream biomechanical changes. The objective of this study was to evaluate long-term agarose culture of nucleus pulposus (NP) cells as a potential in vitro model system to investigate this. Bovine NP cells were cultured in agarose for 49 days in a defined medium containing transforming growth factor-beta 3, after which both mechanical properties and composition were evaluated and compared to native NP. The mRNA levels of NP cell markers were compared to those of freshly isolated NP cells. Glycosaminoglycan (GAG) content, aggregate modulus and hydraulic permeability of mature constructs were similar to native NP, and aggrecan and SOX9 mRNA levels were not significantly different from freshly isolated cells. To investigate direct links between IL-1ß and biomechanical changes, mature agarose constructs were treated with IL-1ß, and effects on biomechanical properties, extracellular matrix composition and mRNA levels were quantified. IL-1ß treatment resulted in upregulation of a disintegrin and metalloproteinase with thrombospondin motifs 4, matrix metalloproteinase-13 and inducible nitric oxide sythase, decreased GAG and modulus, and increased permeability. To evaluate the model as a test platform for therapeutic intervention, co-treatment with IL-1ß and IL-1 receptor antagonist (IL-1ra) was evaluated. IL-1ra significantly attenuated degradative changes induced by IL-1ß. These results suggest that this in vitro model represents a reliable and cost-effective platform for evaluating new therapies for disc degeneration.

Measurement of the shear modulus in thin-layered tissues using numerical simulations and shear wave elastography.

Measurement of mechanical properties of thin-layered tissues has broad applications in the diagnosis of several pathologies. Ultrasound shear wave elastography (SWE) measures the shear wave speed as a means of estimating the mechanical properties of tissues. However, the wave speed in thin-layered tissues is affected by their thickness and the properties of surrounding tissues. The objective of this study is to introduce a method that combines numerical simulations and SWE measurements to provide a more accurate calculation of shear modulus in layered tissues. In the proposed method, the spatial distribution of the acoustic radiation force (ARF) emitted by the transducer was first computed. The ARF was then used as input for simulating the guided wave propagation in the thin layer with its surroundings. The simulations were repeated for several values of the shear modulus of the layer to obtain the corresponding simulated wave speed. By comparing the measured and simulated wave speeds, a more accurate (corrected) shear modulus can be obtained. The proposed method was validated using experiments in agarose gels. In-vivo SWE measurements were also performed for the fascia of the tibialis anterior (TA) muscle and the aponeurosis of musculotendinous junction (MTJ) in medial gastrocnemius (MG) head in a group of healthy individuals. The simulated and measured wave speed in gel constructs were in good agreement with a maximum error of 7.22%. The average of measured wave speed of fascia and aponeurosis was 3.90 ±â€¯0.16 m/s and 2.33 ±â€¯0.60 m/s, while the corresponding corrected shear modulus was 95.63 ±â€¯17.89 kPa and 6.36 ±â€¯8.98 kPa, respectively. Thickness had a substantial effect on the wave speed in thin-layered tissues with decreasing speed for thinner tissues. The SWE-based simulation method presented in this study has the potential of enhancing clinical assessment for several musculoskeletal conditions involving thin-layered tissues.