skip to main content

Title: Dynamic Loading and Tendon Healing Affect Multiscale Tendon Properties and ECM Stress Transmission

The extracellular matrix (ECM) is the primary biomechanical environment that interacts with tendon cells (tenocytes). Stresses applied via muscle contraction during skeletal movement transfer across structural hierarchies to the tenocyte nucleus in native uninjured tendons. Alterations to ECM structural and mechanical properties due to mechanical loading and tissue healing may affect this multiscale strain transfer and stress transmission through the ECM. This study explores the interface between dynamic loading and tendon healing across multiple length scales using living tendon explants. Results show that macroscale mechanical and structural properties are inferior following high magnitude dynamic loading (fatigue) in uninjured living tendon and that these effects propagate to the microscale. Although similar macroscale mechanical effects of dynamic loading are present in healing tendon compared to uninjured tendon, the microscale properties differed greatly during early healing. Regression analysis identified several variables (collagen and nuclear disorganization, cellularity, and F-actin) that directly predict nuclear deformation under loading. Finite element modeling predicted deficits in ECM stress transmission following fatigue loading and during healing. Together, this work identifies the multiscale response of tendon to dynamic loading and healing, and provides new insight into microenvironmental features that tenocytes may experience following injury and after cell delivery therapies.

; ; ; ; ; ; ; ;
Publication Date:
Journal Name:
Scientific Reports
Nature Publishing Group
Sponsoring Org:
National Science Foundation
More Like this
  1. Dynamic loading is a shared feature of tendon tissue homeostasis and pathology. Tendon cells have the inherent ability to sense mechanical loads that initiate molecular-level mechanotransduction pathways. While mature tendons require physiological mechanical loading in order to maintain and fine tune their extracellular matrix architecture, pathological loading initiates an inflammatory-mediated tissue repair pathway that may ultimately result in extracellular matrix dysregulation and tendon degeneration. The exact loading and inflammatory mechanisms involved in tendon healing and pathology is unclear although a precise understanding is imperative to improving therapeutic outcomes of tendon pathologies. Thus, various model systems have been designed to help elucidate the underlying mechanisms of tendon mechanobiology via mimicry of the in vivo tendon architecture and biomechanics. Recent development of model systems has focused on identifying mechanoresponses to various mechanical loading platforms. Less effort has been placed on identifying inflammatory pathways involved in tendon pathology etiology, though inflammation has been implicated in the onset of such chronic injuries. The focus of this work is to highlight the latest discoveries in tendon mechanobiology platforms and specifically identify the gaps for future work. An interdisciplinary approach is necessary to reveal the complex molecular interplay that leads to tendon pathologies and will ultimatelymore »identify potential regenerative therapeutic targets.« less
  2. INTRODUCTION: Quadriceps tendon autografts have experienced a rapid rise in popularity for anterior cruciate ligament (ACL) reconstruction due to advantages in graft sizing and potential improvement in biomechanics. While there is a growing body of literature on use of quadriceps tendon grafts, deeper investigation into the biomechanical properties of stitch techniques in this construct has been limited. The purpose of this study was to evaluate the performance of a novel suture needle against different conventional suture needles by comparing the biomechanical properties of two commonly used stitch methods, a whip stitch, and a locking stitch in quadriceps tendon. It was hypothesized that the new device would be capable of creating both whip stitches and locking stitches that are biomechanically equivalent to similar stitch techniques performed with conventional needle products. METHODS: This was a controlled biomechanical study. A total of 24 matched pair cadaveric knees were dissected and a total of 48 quadriceps tendons were harvested and tested. All tendon grafts were standardized to the same size. Samples were then randomized into the following groups, keeping the matched pairs together: (Group 1, n=16) consisted of Company W’s novel two-part suture needle design, (Group 2, n=16) consisted of Company A suture, andmore »(Group 3, n=16) consisted of Company B suture. For each group, the matched pairs were categorized into subgroups to be instrumented with either a whip stitch or a locking stitch. Two fellowship-trained surgeons performed all stitching, where they each instrumented 8 tendon grafts per group. For instrumentation, the grafts were clamped to a preparation stand in accordance with the manufacturer’s recommendations for passing each suture needle. A skin marker was used to identify and mark five evenly spaced points, 0.5 cm apart, as a guide to create a 5-stitch series. For Group 1, the whip stitch as well as the locking whip stitch were performed with a novel 2-part needle. For Group 2, the whip stitch was performed with loop suture needle and the locking stitch was krackow with a curved needle. Similarly, for Group 3, the whip stitch was performed with loop suture needle and the locking stitch was krackow with a curved needle (Figure 1). Cyclical testing was performed using a servohydraulic testing machine (MTS Bionix) equipped with a 5kN load cell. A standardized length of tendon, 7 cm, was coupled to the MTS actuator by passing it through a cryoclamp cooled by dry ice to a temperature of -5°C (Figure 2). All testing samples were then pre-conditioned to normalize viscoelastic effects and testing variability through application of cyclical loading to 25-100 N for three cycles. The samples were then held at 89 N for 15 minutes. Thereafter, the samples were loaded to 50-200 N for 500 cycles at 1 Hz. If samples survived, they were ramped to failure at 20 mm/min. Displacement and force data was collected throughout testing. Metrics of interest were total elongation (mm), stiffness (N/mm), ultimate failure load (N) and failure mode. Data are presented as averages plus/minus standard deviation. A one-way analysis of variance (ANOVA) with a Tukey pairwise comparison post hoc analysis was used to evaluate differences between the various stitching methods. Statistical significance was set at P = .05. RESULTS SECTION: For the whip stitch methods, the total elongation was found to be equivalent across all methods (W: 36 ± 10 mm; A: 32 ± 18 mm; B: 33 ± 8 mm). The stiffness of Company A (103 ± 11 N/mm) method was significantly larger than Company W (64 ± 8 N/mm; p=.001), whereas stiffness of whip stitch by Company W was equivalent to Company B (80 ± 32 N/mm). The ultimate failure load was equivalent across all whip stitch methods (W: 379 ± 31 mm; A: 412 ± 103 mm; B: 438 ± 63 mm). For the locking stitch method, the total elongation (W: 26 ± 10 mm; A: 14 ± 2 mm; B: 29 ± 5 mm), stiffness (W: 75 ± 11 N/mm; A: 104 ± 23 N/mm; B: 79 ± 10 N/mm) and ultimate load (W: 343 ± 22 N; A: 369 ± 30 N; B: 438 ± 63 N) were found to be equivalent across all methods. The failure mode for all groups is in Table 1. The common mode of failure across study groups and stitch configuration was suture breakage. However, the whip stitch from Company A and Company B had varied failure modes. DISCUSSION: Products from the three manufacturers were found to produce biomechanically equivalent whip stitches and locking stitches with respect to elongation and ultimate failure load. The only significant difference observed was that the whip stitch created with Company A’s product had a higher stiffness than Company W’s product, which could have been due to differences in the suture material. In this cadaveric quadriceps tendon model, it was shown that when using Company W’s novel two-part suture needle, users were capable of creating whip stitches and locking stitches that achieved equivalent biomechanical performance compared to similar stitch techniques performed with conventional needle products. A failure mode limited solely to suture breakage for methods completed with Company W’s needle product suggest a reliable suture construct with limited tissue damage. SIGNIFICANCE/CLINICAL RELEVANCE: Having a suture needle device with the versatility to easily perform different stitching constructs may provide surgeons an advantage needed to improve clinical outcomes. The data presented illustrates a strong new suture technique that has equivalent performance when compared to conventional needle devices and has promising applications in graft preparation for ligament and tendon reconstruction.« less
  3. Abstract Pregnant women experience weight gain, gait changes, and biochemical fluctuations that impair joint function and alter the maternal skeleton. Hormonal changes increase pelvic ligament laxity in preparation for childbirth and affect peripheral joint laxity. Calcium demands also rise during pregnancy and lactation, resulting in reduced bone mineral density (BMD) and maternal bone loss. Altered tendon properties and bone loss during pregnancy and lactation may impact tendon insertion sites, such as rotator cuff tendons where insertion site ruptures are common. However, the effects of pregnancy and lactation at the tendon-to-bone interface have not been investigated. Therefore, the objective of this study was to evaluate supraspinatus tendon mechanical properties and insertion site microstructure during pregnancy, lactation, and postweaning recovery in female rats. We hypothesized that pregnancy and lactation would compromise supraspinatus tendon mechanical properties and subchondral bone microstructure. Female rats were divided into virgin, pregnancy, lactation, and recovery groups, and supraspinatus tendons were mechanically evaluated. Surprisingly, tendon mechanics was unaffected by pregnancy and lactation. However, tendon modulus decreased two-weeks postweaning. Additionally, tendons failed by bony avulsion at the insertion site, and the lactation group exhibited reduced failure properties corresponding to decreased subchondral bone mineralization. Lactation also resulted in dramatic bone lossmore »at the epiphysis, but trabecular bone microarchitecture recovered postweaning. In conclusion, lactation following pregnancy impaired trabecular bone microstructure and subchondral bone mineralization, leading to reduced supraspinatus tendon-to-bone insertion site failure properties. These findings will contribute toward understanding the pathogenesis of tendon-to-bone disorders.« less
  4. Diabetes mellitus (DM) is associated with musculoskeletal complications—including tendon dysfunction and injury. Patients with DM show altered foot and ankle mechanics that have been attributed to tendon dysfunction as well as impaired recovery post-tendon injury. Despite the problem of DM-related tendon complications, treatment guidelines specific to this population of individuals are lacking. DM impairs tendon structure, function, and healing capacity in tendons throughout the body, but the Achilles tendon is of particular concern and most studied in the diabetic foot. At macroscopic levels, asymptomatic, diabetic Achilles tendons may show morphological abnormalities such as thickening, collagen disorganization, and/or calcific changes at the tendon enthesis. At smaller length scales, DM affects collagen sliding and discrete plasticity due to glycation of collagen. However, how these alterations translate to mechanical deficits observed at larger length scales is an area of continued investigation. In addition to dysfunction of the extracellular matrix, tendon cells such as tenocytes and tendon stem/progenitor cells show significant abnormalities in proliferation, apoptosis, and remodeling capacity in the presence of hyperglycemia and advanced glycation end-products, thus contributing to the disruption of tendon homeostasis and healing. Improving our understanding of the effects of DM on tendons—from molecular pathways to patients—will progress toward targetedmore »therapies in this group at high risk of foot and ankle morbidity.« less
  5. Abstract

    Mesenchymal stem cells (MSCs) respond to environmental forces with both cytoskeletal re-structuring and activation of protein chaperones of mechanical information, β-catenin, and yes-associated protein 1 (YAP1). To function, MSCs must differentiate between dynamic forces such as cyclic strains of extracellular matrix due to physical activity and static strains due to ECM stiffening. To delineate how MSCs recognize and respond differently to both force types, we compared effects of dynamic (200 cycles × 2%) and static (1 × 2% hold) strain on nuclear translocation of β-catenin and YAP1 at 3 hours after force application. Dynamic strain induced nuclear accumulation of β-catenin, and increased cytoskeletal actin structure and cell stiffness, but had no effect on nuclear YAP1 levels. Critically, both nuclear actin and nuclear stiffness increased along with dynamic strain-induced β-catenin transport. Augmentation of cytoskeletal structure using either static strain or lysophosphatidic acid did not increase nuclear content of β-catenin or actin, but induced robust nuclear increase in YAP1. As actin binds β-catenin, we considered whether β-catenin, which lacks a nuclear localization signal, was dependent on actin to gain entry to the nucleus. Knockdown of cofilin-1 (Cfl1) or importin-9 (Ipo9), which co-mediate nuclear transfer of G-actin, prevented dynamic strain-mediated nuclear transfer of bothmore »β-catenin and actin. In sum, dynamic strain induction of actin re-structuring promotes nuclear transport of G-actin, concurrently supporting nuclear access of β-catenin via mechanisms used for actin transport. Thus, dynamic and static strain activate alternative mechanoresponses reflected by differences in the cellular distributions of actin, β-catenin, and YAP1.

    « less