No evidence for collateral effects of electromagnetic fields used to increase dissolved oxygen levels on the behavior and physiology of freshwater fishes.

Hypoxia in surface waters driven by warming climate and other anthropogenic stressors is a major conservation concern, and technological solutions for water quality remediation are sorely needed. One potential solution involves the use of low-intensity electromagnetic fields (EMFs) to increase dissolved oxygen levels, but potential collateral effects of the EMFs on aquatic animals have not been formally evaluated. We examined the effects of EMF exposure on wild-caught, captive sunfish (Lepomis spp.) over 8-day and 3-day exposures, with and without aeration in mesocosms and stock tanks (respectively). We also quantified ambient fish abundance in close proximity to EMF devices deployed in Opinicon Lake (ON). We found no significant differences in a suite of blood-based stress physiology biomarkers, behaviors, and putative aerobic capacities between EMF and control conditions over 8 days. Aerated mesocosms equipped with activated EMFs consistently had higher oxygen levels in the water than aerated controls. There were no differences in mortality during 3-day oxygen depletion trials under EMF or control conditions, and we detected no differences in fish abundance when the devices were activated in the lake. Our findings suggest that deploying EMF devices in field settings is not likely to exert negative effects on exposed fish populations. PRACTITIONER POINTS: Low-cost, low-energy technological solutions to remediate aquatic hypoxia are sorely needed Electromagnetic fields (EMFs) can increase oxygen flux across air/water interfaces and increase dissolved oxygen levels We found no evidence of negative effects of EMFs on fish physiology or behavior and our results support their use in alleviating hypoxic conditions.

Advances in the development of gene therapy, noncoding RNA, and exosome-based treatments for tendinopathy.

Tendinopathy is a common musculoskeletal disorder characterized by chronic low-grade inflammation and tissue degeneration. Tendons have poor innate healing ability and there is currently no cure for tendinopathy. Studies elucidating mechanisms underlying the pathogenesis of tendinopathy and mechanisms mediating the genesis of tendons during development have provided novel targets and strategies to enhance tendon healing and repair. This review summarizes the current understanding and treatments for tendinopathy. The review also highlights recent advances in gene therapy, the potential of noncoding RNAs, such as microRNAs, and exosomes, which are nanometer-sized extracellular vesicles secreted from cells, for the treatment of tendinopathy.

Cycle-dependent matrix remodeling gene expression response in fatigue-loaded rat patellar tendons.

Expression profiling of selected matrix remodeling genes was conducted to evaluate differences in molecular response to low-cycle (100) and high-cycle (7,200) sub-failure-fatigue loading of patellar tendons. Using our previously developed in vivo patellar tendon model, tendons were loaded for 100 or 7,200 cycles and expression of selected metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), and collagens were quantified by real-time RT-PCR at 1- and 7-day post-loading. Expression profiles were also obtained from lacerated tendons as an acute injury model. The high-cycle group showed upregulation of TIMP-1, -2, Col3a1, and Col5a1, and downregulation TIMP-4 at both time points, upregulation of MMP-2 at 7-day post-loading and downregulation of MMP-13 and -14 at 1-day post-loading, suggesting overall repair/remodeling. In contrast, the low-cycle loaded group showed upregulation of MMP-2, -3, -13, and Col12a1 at both time points, upregulation of TIMP-1, -2, -3, Col3a1, and integrin ß1 and downregulation of integrin α11 at 1-day post-loading and upregulation of Col1a1 at 7-day post-loading, consistent with a hypertrophic (adaptive) pattern. Lacerated tendons showed a typical acute wound response with upregulation of all examined remodeling genes. Differences found in tendon response to high- and low-cycle loading are suggestive of the underlying mechanisms associated with a healthy or damaging response.

A knee-specific finite element analysis of the human anterior cruciate ligament impingement against the femoral intercondylar notch.

This work presents a finite element analysis of anterior cruciate ligament (ACL) impingement against the intercondylar notch during tibial external rotation and abduction, as a mechanism of noncontact ACL injuries. Experimentally, ACL impingement was measured in a cadaveric knee in terms of impingement contact pressure and six degrees-of-freedom tibiofemoral kinematics. Three-dimensional geometries of the ACL, femur and tibia were incorporated into the finite element model of the individual knee specimen. A fiber-reinforced model was adopted, which accounts for the anisotropy, large deformation, nonlinearity and incompressibility of the ACL. With boundary conditions specified based on the experimental tibiofemoral kinematics, the finite element analysis showed that impingement between the ligament and the lateral wall of intercondylar notch could occur when qthe knee at 45 degrees was externally rotated at 29.1 degrees and abducted at 10.0 degrees . Strong contact pressure and tensile stress occurred at the impinging and nonimpinging sides of the ligament, respectively. The impingement force and contact area estimated from the model matched their counterparts from the corresponding cadaver experiment. The modeling and experimental approach provides a useful tool to characterize potential ACL impingement on a knee-specific basis, which may help elucidate the ACL injury mechanism and develop more effective treatments.

Second harmonic generation imaging and Fourier transform spectral analysis reveal damage in fatigue-loaded tendons.

Conventional histologic methods provide valuable information regarding the physical nature of damage in fatigue-loaded tendons, limited to thin, two-dimensional sections. We introduce an imaging method that characterizes tendon microstructure three-dimensionally and develop quantitative, spatial measures of damage formation within tendons. Rat patellar tendons were fatigue loaded in vivo to low, moderate, and high damage levels. Tendon microstructure was characterized using multiphoton microscopy by capturing second harmonic generation signals. Image stacks were analyzed using Fourier transform-derived computations to assess frequency-based properties of damage. Results showed 3D microstructure with progressively increased density and variety of damage patterns, characterized by kinked deformations at low, fiber dissociation at moderate, and fiber thinning and out-of-plane discontinuities at high damage levels. Image analysis generated radial distributions of power spectral gradients, establishing a "fingerprint" of tendon damage. Additionally, matrix damage was mapped using local, discretized orientation vectors. The frequency distribution of vector angles, a measure of damage content, differed from one damage level to the next. This study established an objective 3D imaging and analysis method for tendon microstructure, which characterizes directionality and anisotropy of the tendon microstructure and quantitative measures of damage that will advance investigations of the microstructural basis of degradation that precedes overuse injuries.

Early response to tendon fatigue damage accumulation in a novel in vivo model.

This study describes the development and application of a novel rat patellar tendon model of mechanical fatigue for investigating the early in vivo response to tendon subfailure injury. Patellar tendons of adult female Sprague-Dawley rats were fatigue loaded between 1-35N using a custom-designed loading apparatus. Patellar tendons were subjected to Low-, Moderate- or High-level fatigue damage, defined by grip-to-grip strain measurement. Molecular response was compared with that of a laceration-repair injury. Histological analyses showed that progression of tendon fatigue involves formation of localized kinked fiber deformations at Low damage, which increased in density with presence of fiber delaminations at Moderate damage, and fiber angulation and discontinuities at High damage levels. RT-PCR analysis performed at 1- and 3-day post-fatigue showed variable changes in type I, III and V collagen mRNA expression at Low and Moderate damage levels, consistent with clinical findings of tendon pathology and were modest compared with those observed at High damage levels, in which expression of all collagens evaluated were increased markedly. In contrast, only type I collagen expression was elevated at the same time points post-laceration. Findings suggest that cumulative fatigue in tendon invokes a different molecular response than laceration. Further, structural repair may not be initiated until reaching end-stage fatigue life, where the repair response may unable to restore the damaged tendon to its pre-fatigue architecture.

Subrupture tendon fatigue damage.

The mechanical and microstructural bases of tendon fatigue, by which damage accumulates and contributes to degradation, are poorly understood. To investigate the tendon fatigue process, rat flexor digitorum longus tendons were cyclically loaded (1-16 N) until reaching one of three levels of fatigue damage, defined as peak clamp-to-clamp strain magnitudes representing key intervals in the fatigue life: i) Low (6.0%-7.0%); ii) Moderate (8.5%-9.5%); and iii) High (11.0%-12.0%). Stiffness, hysteresis, and clamp-to-clamp strain were assessed diagnostically (by cyclic loading at 1-8 N) before and after fatigue loading and following an unloaded recovery period to identify mechanical parameters as measures of damage. Results showed that tendon clamp-to-clamp strain increased from pre- to post-fatigue loading significantly and progressively with the fatigue damage level (p

Coordinate regulation of IL-1beta and MMP-13 in rat tendons following subrupture fatigue damage.

Mechanical overloading is a major causative factor of tendinopathy; however, its underlying mechanisms are unclear. We hypothesized mechanical overloading would damage tendons and alter genes associated with tendinopathy in a load-dependent manner. To test this hypothesis, we fatigue loaded rat patellar tendons in vivo and measured expression of the matrix-degrading enzyme MMP-13 and the inflammatory cytokine IL-1beta. We also examined these responses in cultured tenocytes exposed to intermittent hydrostatic pressure in vitro. Additionally, we hypothesized load-induced changes in tenocyte MMP-13 expression would be dependent on expression of IL-1beta. In vivo fatigue loading at 1.7% strain caused overt microstructural damage and upregulated expression of MMP-13 and IL-1beta, while 0.6% strain produced only minor changes in matrix microstructure and downregulated expression of both MMP-13 and IL-1beta. Loading of cultured tenocytes at 2.5 and 7.5 MPa produced comparable changes in expression to those of in vivo tendon loading. Blocking IL-1beta expression with siRNA suppressed load-induced both MMP-13 mRNA expression and activity. The data suggest fatigue loading alters expression of MMP-13 and IL-1beta in tendons in vivo and tenocytes in vitro in a load-dependent manner. The data also suggest MMP-13 is regulated by both IL-1beta-dependent and IL-1beta-independent pathways.

ACL impingement prediction based on MRI scans of individual knees.

Although tibial external rotation and abduction do not load the ACL strongly in cadaver-based biomechanical studies, such knee positions are associated with ACL injuries in clinical practice. We hypothesized the ACL could be injured in such knee positions because of its impingement against the intercondylar notch. We developed a three-dimensional geometric ACL impingement model through segmentation of MR images of individual knees. We investigated impingement by determining the deformed geometry and elongation of the ligament as it wrapped around the notch surface during impingement. When impingement did not occur, the gap width separating the ligament and the notch surface was computed. Tibial external rotation/abduction could cause the ACL to impinge against the lateral notch wall and elongate as it wraps around the surface of the notch wall. The impingement occurred between the middle portion of the ligament (at 45% +/- 8% [mean +/- standard deviation] of the ligament length) and the convex surface of the lateral notch wall. Considering the multiband fiber architecture of the ligament, the anteromedial band of the ACL sustained greater elongation during impingement and showed a smaller gap width from the notch surface than the intermediate band or the posterolateral band.

Modeling of ACL impingement against the intercondylar notch.

OBJECTIVE: To develop a 3-D mathematical model that accurately evaluates anterior cruciate ligament impingement against the intercondylar notch. DESIGN: The model simulated physical interactions between the anterior cruciate ligament and the intercondylar notch in tibiofemoral movement. BACKGROUND: Anterior cruciate ligament impingement has been evaluated through planar radiographic images, which may not characterize the complex 3-D notch shape associated with impingement. METHODS: After examining potential anterior cruciate ligament impingement in five cadaver knee specimens, the model was implemented using data from an individual cadaveric knee with representative impingement. The knee was loaded passively in various patterns to induce impingement, and the impingement force and six degrees-of-freedom tibiofemoral kinematics were measured. The femur, tibia, and anterior cruciate ligament were digitized. Spatial data points representing the notch surfaces were surface-fitted using bicubic splines. The model detected for impingement during the tibiofemoral movement and used a "crawling algorithm" to determine the deformed geometry of the impinging ligament. RESULTS: The model detected the impingement accurately and the ligament strain determined by the model was highly correlated with the recorded impingement force when impingement occurred during the tibiofemoral movement. Distance between the anterior cruciate ligament and the notch wall was determined when impingement was not detected. CONCLUSION: The model quantitatively characterized impingement of the anterior cruciate ligament against the intercondylar notch in 3-D space. RELEVANCE: The approach helps us better understand anterior cruciate ligament injury mechanisms in individual knees. Clinically, the model could potentially be used to analyze subject-specific potential/actual anterior cruciate ligament impingement based on the subject's MRI scans.