Tendon Function Research Articles

Age-dependent mathematical models of ligaments and tendons

Understanding the internal structure and the underlying physical mechanisms governing the mechanical properties of ligaments and tendons, particularly the elastic modulus, across different stages of life is critical for enhancing tissue strength during growth, maturation, and aging. This knowledge is essential not only for preventing tissue failure in older adults but also for advancing the development of biomaterials that can substitute or augment ligament and tendon function across all age groups. Despite the significance of this area, a comprehensive, mechanistic understanding of the relationship between structural changes and mechanical properties over time remains largely unexplored. To date, there is a lack of detailed studies that elucidate the physical mechanisms involved in these age-related changes. The absence of such mechanistic insights highlights a significant gap in the literature, necessitating further investigation. Therefore, this research delves into the age-dependent structural and mechanical property changes in ligaments and tendons, emphasizing both growth and mature phases. Utilizing a comprehensive approach, we have developed new mathematical models that directly correlate the growth of collagen in fibrils with the increasing elastic modulus in the fibers of ligaments and tendons over time. By integrating experimental data from mouse tail tendons in published work and conducting simulations, we have observed that the cross-sectional area of collagen in fibrils and the elastic modulus of a collagen fiber increase rapidly during the growth phase and stabilize during the mature phase. Our proposed models effectively describe the trends in collagen growth and the elastic modulus of fibers in ligaments and tendons over different ages, exhibiting consistency with experimental data. Through detailed analysis, we elucidate the mechanistic relationship between collagen growth and the elastic modulus of fibers as they age. This comprehensive approach significantly enhances our understanding of the age-related structural and mechanical property changes in connective tissues, providing a robust framework for future investigations.

Tendon-targeted knockout of collagen XI disrupts patellar and Achilles tendon structure and mechanical properties during murine postnatal development

ABSTRACT Background Collagen XI is a fibril-forming collagen typically associated with type II collagen tissues but is also expressed in type I collagen-rich tendons, especially during development. We previously showed that tendon-targeted (Scx-Cre) Col11a1 knockout mice have smaller tendons in adulthood with aberrant fibril structure and impaired mechanical properties. However, the manifestation of this phenotype is not clearly understood. Therefore, our objective is to define the spatiotemporal roles of collagen XI in tendon structure-function during postnatal development. Given the high expression of collagen XI during embryonic development, we hypothesized that collagen XI knockout leads to the deposition of weakened extracellular matrix during early postnatal timepoints, disrupting the establishment of tendon structure and function. Methods Patellar and Achilles tendons from postnatal (P) days 0, 10, 20, and 30 tendon-targeted scleraxis-Cre heterozygous and homozygous Col11a1 knockout mice were evaluated for morphology, nuclear organization, fibril morphology, mechanical properties, and gene expression. Results At P0, there were no differences in tendon length or fibril diameter of either tendon. By P10, striking structural and functional differences emerged, with collagen XI deficiency resulting in increased tendon length, a heterogeneous and larger diameter population of fibrils, and inferior mechanical properties in both patellar and Achilles tendons. Differences increased in magnitude through P30, supporting our hypothesis that impaired structure-function during postnatal development may drive tendon lengthening and reduced mechanical properties. Conclusions Though collagen XI is a quantitatively minor component of the tendon extracellular matrix, these results highlight the critical role of collagen XI in the acquisition of tendon structure-function.

Research progress on the repair of tendon injuries with Tissue Engineering Technology

Tendon injuries are frequently encountered in clinical practice, and traditional repair methods rarely achieve complete restoration of the tendon's original structure and functionality. The challenges of accelerating and optimizing the healing of injured tendons, enhancing the strength of the regenerated tendon, and preventing adhesion remain significant in clinical settings. Tendon tissue engineering, which combines material science, cell biology, and molecular biology, involves the synergistic application of multiple factors to create functional constructs and has become an emerging technique with promise for repair. The maximization of the regenerative potential of these elements is a critical research question. Future research should concentrate on discovering the optimal combinations of cells, biosignals, and scaffolds to produce tissue that emulates the characteristics of an undamaged, natural tendon. This review will cover various aspects, including the fabrication of tendon tissue scaffolds, the selection of seed cells, strategies for the modulation of healing-related biosignals, and provide a summary with prospective insights, aiming to enhance the comprehension of tissue engineering techniques in tendon injury repair and to inspire innovative applications in this domain.

Loss of Cochlin drives impairments in tendon structure and function.

Aging tendons undergo disruptions in homeostasis, increased susceptibility to injury, and reduced capacity for healing. Exploring the mechanisms behind this disruption in homeostasis is essential for developing therapeutics aimed at maintaining tendon health through the lifespan. We have previously identified that the extracellular matrix protein, Cochlin , which is highly expressed in healthy flexor tendon, is consistently lost during both natural aging and upon depletion of Scleraxis-lineage cells in young animals, which recapitulates many aging-associated homeostatic disruptions. Therefore, we hypothesized that loss of Cochlin would disrupt tendon homeostasis, including alterations in collagen fibril organization, and impaired tendon mechanics. By 3-months of age, Cochlin -/- flexor tendons exhibited altered collagen structure, with these changes persisting through at least 9-months. In addition, Cochlin -/- tendons demonstrated significant declines in structural and material properties at 6-months, and structural properties at 9-months. While Cochlin -/- did not drastically change the overall tendon proteome, consistent decreases in proteins associated with RNA metabolism, extracellular matrix production and the cytoskeleton were observed in Cochlin -/- . Interestingly, homeostatic disruption via Cochlin -/- did not impair the tendon healing process. Taken together, these data define a critical role for Cochlin in maintaining tendon homeostasis and suggest retention or restoration of Cochlin as a potential therapeutic approach to retain tendon structure and function through the lifespan.

Structural and Functional Properties of Lower Extremity Tendons in Men.

Comstock, BA, Flanagan, SD, Denegar, CR, Newton, RU, Häkkinen, K, Volek, JS, Maresh, CM, Kraemer, WJ. Structural and functional properties of lower extremity tendons in men. J Strength Cond Res XX(X): 000-000, 2024-The purpose of this study was to understand further patellar and Achilles tendon structure and function, body composition, and serum collagen turnover biomarkers in young men who performed heavy resistance training (RT, n = 13, age: 22.2 ± 1.4 years) compared with recreationally active men who were not resistance-trained (NR, n = 13, age: 22.8 ± 2.2 years). Tendon properties were measured at rest and during maximal voluntary isometric efforts using ultrasonography and dynamometry. Lean body mass (LBM) and bone mineral density (BMD) were assessed with dual X-ray absorptiometry. Serum collagen turnover markers were analyzed and related to tendon measures. Resistance-trained men had significantly (p ≤ 0.05) greater LBM and BMD compared with recreationally active men. Resistance-trained men also showed significantly greater patellar tendon (PT) stiffness (45%) and Young's modulus (36%), though the PT cross-sectional area (CSA) did not differ significantly between groups. Achilles tendon CSA was significantly larger in resistance-trained men. Still, other properties such as stiffness and modulus did not differ significantly between the groups. Serum collagen turnover markers showed no significant differences between groups and were not correlated to any tendon or bone biomarkers. The findings support that resistance-trained men have greater LBM and BMD. However now, it reveals that tendon adaptations differ, as not all measures were similarly affected in both tendons. The blood biomarkers did not show any obvious roles in explaining the differential changes in tendons. Heavy RT induces differential tendon changes potentially due to complex interactions of training variables.

Cellular senescence impairs tendon extracellular matrix remodeling in response to mechanical unloading.

Musculoskeletal injuries, including tendinopathies, present a significant clinical burden for aging populations. While the biological drivers of age-related declines in tendon function are poorly understood, it is well accepted that dysregulation of extracellular matrix (ECM) remodeling plays a role in chronic tendon degeneration. Senescent cells, which have been associated with multiple degenerative pathologies in musculoskeletal tissues, secrete a highly pro-inflammatory senescence-associated secretory phenotype (SASP) that has potential to promote ECM breakdown. However, the role of senescent cells in the dysregulation of tendon ECM homeostasis is largely unknown. To assess this directly, we developed an invitro model of induced cellular senescence in murine tendon explants. This novel technique enables us to study the isolated interactions of senescent cells and their native ECM without interference from age-related systemic changes. We document multiple biomarkers of cellular senescence in induced tendon explants including cell cycle arrest, apoptosis resistance, and sustained inflammatory responses. We then utilize this invitro senescence model to compare the ECM remodeling response of young, naturally aged, and induced-senescent tendons to an altered mechanical stimulus. We found that both senescence and aging independently led to alterations in ECM-related gene expression, reductions in protein synthesis, and tissue compositional changes. Furthermore, MMP activity was sustained, thus shifting the remodeling balance of aged and induced-senescent tissues towards degradation over production. Together, this demonstrates that cellular senescence plays a role in the altered mechano-response of aged tendons and likely contributes to poor clinical outcomes in aging populations.

Simultaneous targeting of TGF-β1/PD-L1 via a hydrogel-nanoparticle system to remodel the ECM and immune microenvironment for limiting adhesion formation

Adhesion seriously affects the recovery of tendon gliding function. Our group previously found that inhibition of TGF-β1, which is closely related to adhesion formation, effectively attenuated adhesions but did not eliminate them, suggesting that there may be other mechanisms involved in adhesion formation. In this study, we considered that uncontrolled and excessively proliferating fibroblasts undergo immune escape, which aggravates the deposition of extracellular matrix during the adhesion formation. We found that the expression of the immune checkpoint PD-L1 was significantly elevated after injury and may be involved in adhesion formation. Therefore, we intended to silence both TGF-β1 and PD-L1 to improve the immune advantage in the microenvironment after flexor tendon injury to further reduce adhesion. We constructed the nanoparticle/TGF-β1 or/and PD-L1 siRNAs complexes and verified their high biocompatibility and high transfection efficiency. We found that CD8+T cells had a greater killing effect on the excessively proliferating cells that were transfected with nanoparticle/TGF-β1 or/and PD-L1 siRNAs. The hydrogel-nanoparticle/TGF-β1 or/and PD-L1 siRNAs system could effectively improve the gliding function of the tendons without weakening the mechanical properties in injured rat FDL tendon and chicken FDP tendon models. In addition, the potential of CD8+T cells to encircle the adhesion cells on the tendon surface was observed, which resulted in increased levels of cell apoptosis. Thus, our study confirmed that combined knockdown of TGF-β1 and PD-L1 could activate immunodominance after flexor tendon repair and provided a potential treatment to limit adhesion formation and improve gliding function. Statement of SignificanceAdhesion seriously affects the recovery of tendon gliding function. TGF-β1 is related to adhesion formation as it regulates the production of extracellular matrix. We found that excessively proliferated fibroblasts might undergo immune escape, which aggravated the deposition of extracellular matrix. Therefore, we constructed a hydrogel-nanoparticle/TGF-β1 and PD-L1 siRNAs system for silencing TGF-β1 and PD-L1 to improve the immune advantage in the microenvironment after tendon injury. This system could improve the gliding function of tendons without weakening the mechanical property and increase the killing effect of CD8+T cells. Combined knockdown of TGF-β1 and PD-L1 could activate immunodominance after tendon repair and provide a potential treatment to limit adhesion formation.

Injury to the palmar supporting structures of the fetlock alters limb stiffness and fetlock angle.

In vivo measurement of limb stiffness and conformation provides a non-invasive proxy assessment of superficial digital flexor tendon (SDFT) and suspensory ligament (SL) function. Here, we compared it in fore and hindlimbs and after injury. To compare the limb stiffness and conformation in forelimbs and hindlimbs, changes with age, and following injury to the SDFT and SL. Retrospective cohort study. Limb stiffness was calculated using floor scales and an electrogoniometer taped to the dorsal fetlock. The fetlock angle and weight were simultaneously recorded five times with the limb weight-bearing and when the opposite limb was picked up (increased load). Limb stiffness of both limbs was calculated from the gradient of the regression line of angle versus load. Fetlock angle when the weight was zero was extrapolated from the graph and used as a measure of conformation. Limb stiffness was measured in uninjured forelimbs (n = 42 limbs), hindlimbs (n = 19 limbs), forelimbs with SDFT injury (n = 18) and hindlimbs with SL injury (n = 5). Limb stiffness correlated with weight in forelimbs as shown previously (p < 0.001) but also in hindlimbs (p = 0.006). When normalised to the horse's weight (503 kg, IQR 471.5-560), forelimb stiffness was significantly higher (22.3 [±4.5] × 10-3 degree-1) than for the hindlimb (16.4 [±4.0] × 10-3 degree-1; p < 0.001). While there were no significant differences between forelimb and hindlimb conformation in unaffected or SDFT injury, both limb stiffness and conformation was significantly greater in limbs with SL injury (p = 0.009 and p = 0.002, respectively). Small sample size, lack of clinical data including lameness and quantification of injuries. Injury to the forelimb SDFT does not alter limb stiffness or conformation in the long-term, while hindlimb SL injury simultaneously increases limb stiffness and fetlock angle, suggesting an increase in SL length following injury.

Focal adhesion kinase regulates tendon cell mechanoresponse and physiological tendon development.

Tendons enable locomotion by transmitting high tensile mechanical forces between muscle and bone via their dense extracellular matrix (ECM). The application of extrinsic mechanical stimuli via muscle contraction is necessary to regulate healthy tendon function. Specifically, applied physiological levels of mechanical loading elicit an anabolic tendon cell response, while decreased mechanical loading evokes a degradative tendon state. Although the tendon response to mechanical stimuli has implications in disease pathogenesis and clinical treatment strategies, the cell signaling mechanisms by which tendon cells sense and respond to mechanical stimuli within the native tendon ECM remain largely unknown. Therefore, we explored the role of cell-ECM adhesions in regulating tendon cell mechanotransduction by perturbing the genetic expression and signaling activity of focal adhesion kinase (FAK) through both invitro and invivo approaches. We determined that FAK regulates tendon cell spreading behavior and focal adhesion morphology, nuclear deformation in response to applied mechanical strain, and mechanosensitive gene expression. In addition, our data reveal that FAK signaling plays an essential role in invivo tendon development and postnatal growth, as FAK-knockout mouse tendons demonstrated reduced tendon size, altered mechanical properties, differences in cellular composition, and reduced maturity of the deposited ECM. These data provide a foundational understanding of the role of FAK signaling as a critical regulator of insitu tendon cell mechanotransduction. Importantly, an increased understanding of tendon cell mechanotransductive mechanisms may inform clinical practice as well as lead to the discovery of diagnostic and/or therapeutic molecular targets.

A Treatment Protocol for Achilles Tendinopathy with Extracorporeal Shockwave Therapy.

Achilles tendinopathy is a common musculoskeletal condition characterized by pain, lower muscle strength, gait abnormality, and reduced quality of life. There are two categories of Achilles tendinopathy: insertional Achilles tendinopathy and mid-portion Achilles tendinopathy. Currently, mechanical loading programs are considered the standard of care for the population with Achilles tendinopathy. Extracorporeal shockwave therapy (ESWT) is considered a secondary conservative treatment for tendinopathy as it is effective and safe. It can be used either as a monotherapy or as part of a multimodal treatment plan. ESWT has been extensively studied in orthopedics, where it was shown to intensify fracture healing and successfully treat overuse conditions of tendons and fascia. It is believed that shockwaves have both mechanical and cellular effects that ultimately result in the repair of damaged tendinous tissue and improved function of the Achilles tendon. However, there is a lack of consistency in the literature surrounding the effectiveness, especially the protocols. Therefore, we enrolled 36 patients with a diagnosis of Achilles tendinopathy, using radial ESWT (0.48 mJ/mm2, 2,000 shockwaves, 10 Hz, 1.6bars, 2 sessions once a week). Freedom from pain was experienced by 16.7% of these participants, and there was a significant decrease in pain in all of them.

Female Tendons are from Venus and Male Tendons are from Mars, But Does it Matter for Tendon Health?

Tendons play fundamental roles in the execution of human movement and therefore understanding tendon function, health and disease is important for everyday living and sports performance. The acute mechanical behavioural and physiological responses to short-term loading of tendons, as well as more chronic morphological and mechanical adaptations to longer term loading, differ between sexes. This has led some researchers to speculate that there may be a sex-specific injury risk in tendons. However, the link between anatomical, physiological and biomechanical sex-specific differences in tendons and their contributory role in the development of tendon disease injuries has not been critically evaluated. This review outlines the evidence surrounding the sex-specific physiological and biomechanical responses and adaptations to loading and discusses how this evidence compares to clinical evidence on tendon injuries and rehabilitation in the Achilles and patellar tendons in humans. Using the evidence available in both sports science and medicine, this may provide a more holistic understanding to improve our ability to enhance human tendon health and performance in both sexes.

Elemental and Structural Characterization of Heterotopic Ossification during Achilles Tendon Healing Provides New Insights on the Formation Process.

Heterotopic ossification (HO) in tendons can lead to increased pain and poor tendon function. Although it is believed to share some characteristics with bone, the structural and elemental compositions of HO deposits have not been fully elucidated. This study utilizes a multimodal and multiscale approach for structural and elemental characterization of HO deposits in healing rat Achilles tendons at 3, 6, 12, 16, and 20 weeks post transection. The microscale tomography and scanning electron microscopy results indicate increased mineral density and Ca/P ratio in the maturing HO deposits (12 and 20 weeks), when compared to the early time points (3 weeks). Visually, the mature HO deposits present microstructures similar to calcaneal bone. Through synchrotron-based X-ray scattering and fluorescence, the hydroxyapatite (HA) crystallites are shorter along the c-axis and become larger in the ab-plane with increasing healing time, while the HA crystal thickness remains within the reference values for bone. At the mineralization boundary, the overlap between high levels of calcium and prominent crystallite formation was outlined by the presence of zinc and iron. In the mature HO deposits, the calcium content was highest, and zinc was more present internally, which could be indicative of HO deposit remodeling. This study emphasizes the structural and elemental similarities between the calcaneal bone and HO deposits.

Intermittent cyclic stretch of engineered ligaments drives hierarchical collagen fiber maturation in a dose- and organizational-dependent manner

Hierarchical collagen fibers are the primary source of strength in tendons and ligaments; however, these fibers largely do not regenerate after injury or with repair, resulting in limited treatment options. We previously developed a static culture system that guides ACL fibroblasts to produce native-sized fibers and early fascicles by 6 weeks. These constructs are promising ligament replacements, but further maturation is needed. Mechanical cues are critical for development in vivo and in engineered tissues; however, the effect on larger fiber and fascicle formation is largely unknown. Our objective was to investigate whether intermittent cyclic stretch, mimicking rapid muscle activity, drives further maturation in our system to create stronger engineered replacements and to explore whether cyclic loading has differential effects on cells at different degrees of collagen organization to better inform engineered tissue maturation protocols. Constructs were loaded with an established intermittent cyclic loading regime at 5 or 10 % strain for up to 6 weeks and compared to static controls. Cyclic loading drove cells to increase hierarchical collagen organization, collagen crimp, and tissue tensile properties, ultimately producing constructs that matched or exceeded immature ACL properties. Further, the effect of loading on cells varied depending on degree of organization. Specifically, 10 % load drove early improvements in tensile properties and composition, while 5 % load was more beneficial later in culture, suggesting a shift in mechanotransduction. This study provides new insight into how cyclic loading affects cell-driven hierarchical fiber formation and maturation, which will help to develop better rehabilitation protocols and engineer stronger replacements. Statement of significanceCollagen fibers are the primary source of strength and function in tendons and ligaments throughout the body. These fibers have limited regenerate after injury, with repair, and in engineered replacements, reducing treatment options. Cyclic load has been shown to improve fibril level alignment, but its effect at the larger fiber and fascicle length-scale is largely unknown. Here, we demonstrate intermittent cyclic loading increases cell-driven hierarchical fiber formation and tissue mechanics, producing engineered replacements with similar organization and mechanics as immature ACLs. This study provides new insight into how cyclic loading affects cell-driven fiber maturation. A better understanding of how mechanical cues regulate fiber formation will help to develop better engineered replacements and rehabilitation protocols to drive repair after injury.

In Vivo Photoacoustic Ultrasound (PAUS) Assay for Monitoring Tendon Collagen Compositional Changes during Injury and Healing.

Tendon injury and healing involve significant changes to tissue biology and composition. Current techniques often require animal sacrifice or tissue destruction, limiting assessment of dynamic changes in tendons, including treatment response, disease development, rupture risk, and healing progression. Changes in tendon composition, such as altered collagen content, can significantly impact tendon mechanics and function. Analyses of compositional changes typically require ex vivo techniques with animal sacrifice or destruction of the tissue. In vivo evaluation of tendons is critical for longitudinal assessment. We hypothesize that photoacoustic ultrasound detects differences in collagen concentration throughout healing. We utilized photoacoustic ultrasound, a hybrid imaging modality that combines ultrasound and laser-induced photoacoustic signals to create detailed and high-resolution images of tendons, to identify its endogenous collagen composition. We correlated the photoacoustic signal to picrosirius red staining. The results show that the photoacoustic ultrasound-estimated collagen content in tendons correlates well with picrosirius red staining. This study demonstrates that photoacoustic ultrasound can assess injury-induced compositional changes within tendons and is the first study to image these targets in rat Achilles tendon in vivo.

Uni-directional release of ibuprofen from an asymmetric fibrous membrane enables effective peritendinous anti-adhesion

Drug-loaded porous membranes have been deemed to be effective physicochemical barriers to separate postoperative adhesion-prone tissues in tendon healing. However, cell viability and subsequent tissue regeneration might be severely interfered with the unrestricted release and the locally excessive concentration of anti-inflammatory drugs. Herein, we report a double-layered membrane with sustained and uni-directional drug delivery features to prevent peritendinous adhesion without hampering the healing outcome. A vortex-assisted electrospinning system in combination with ibuprofen (IBU)-in-water emulsion was utilized to fabricate IBU-loaded poly-ʟ-lactic-acid (PLLA) fiber bundle membrane (PFB-IBU) as the anti-adhesion layer. The resultant highly porous structure, oleophilic and hydrophobic nature of PLLA fibers enabled in situ loading of IBU with a concentration gradient across the membrane thickness. Aligned collagen nanofibers were further deposited at the low IBU concentration side of the membrane for regulating cell growth and achieving uni-directional release of IBU. Drug release kinetics showed that the release amount of IBU from the high concentration side reached 79.32% at 14 d, while it was only 0.35% at the collagen side. Therefore, fibroblast proliferation at the high concentration side was successfully inhibited without affecting the oriented growth of tendon-derived stem cells at the other side. In vivo evaluation of the rat Achilles adhesion model confirmed the successful peritendinous anti-adhesion of our double-layered membrane, in that the macrophage recruitment, the inflammatory factor secretion and the deposition of pathological adhesion markers such as α-SMA and COL-III were all inhibited, which greatly improved the peritendinous fibrosis and restored the motor function of tendon.

CatWalk XT Gait Parameters Associated with Mouse Achilles Tendon Injury and Healing.

Tendons are connective tissues with limited healing potential which results in permanently impaired function. Although direct mechanical testing of tendon remains the gold standard for functional analyses, this assay is terminal and tracking healing over time requires the use of many animals. An alternative method for quantifying tendon function is gait analysis, which is non-terminal and enables longitudinal tracking of the same animal. To date, commercial systems used to analyze gait are mostly applied to study neurobehavior, and applications for tendon research has been limited. Since these systems typically output many parameters, it is challenging to know which parameter is most relevant for a specific injury model. To address this challenge, we used a well-established rodent locomotion system (CatWalk XT) to measure longitudinal gait parameters in sham and Achilles tendon-injured mice (from PREOP to 56 days post-injury) and identified relevant and reproducible parameters specifically associated with Achilles tendon injury and healing. Micro computed tomography (micro-CT) and mechanical testing also confirmed persistent tendon impairment in the same animals at the terminal timepoint. Collectively, our results provide a useful reference and recommendation of CatWalk gait parameters and the control comparisons that both conserve the use of animals while maintaining high reproducibility standards for Achilles tendon injury studies.

Exploring the impact of vitamin D on tendon health: a comprehensive review.

Tendons are vital components of the musculoskeletal system, facilitating movement and supporting mechanical loads. Emerging evidence suggests that vitamin D, beyond its well-established role in bone health, exerts significant effects on tendon physiology. The aim of this manuscript is to review the impact of vitamin D on tendons, focusing on its mechanisms of action, clinical implications, and therapeutic applications. A comprehensive search of scientific electronic databases was conducted to identify articles on the effects of vitamin D ontendon health. Fourteen studies were included in this review. Five studies were performed invitro, and nine studies were conducted invivo. Despite some conflicting results, the included studies showed that vitamin D regulates collagen synthesis, inflammation, and mineralization within tendons through its interaction with vitamin D receptors. Epidemiological studies link vitamin D deficiency with tendon disorders, including tendinopathy and impaired healing. Supplementation with vitamin D shows promise in improving tendon strength and function, particularly in at-risk populations such as athletes and the elderly. Future research should address optimal supplementation strategies and explore the interplay between vitamin D and other factors influencing tendon health. Integrating vitamin D optimization into clinical practice could enhance tendon integrity and reduce the burden of tendon-related pathologies.

Lysyl Oxidase Production by Murine C3H10T1/2 Mesenchymal Stem Cells Is Increased by TGFβs and Differentially Modulated by Mechanical Stimuli.

Tendons are frequently injured and have limited regenerative capacity. This motivates tissue engineering efforts aimed at restoring tendon function through strategies to direct functional tendon formation. Generation of a crosslinked collagen matrix is paramount to forming mechanically functional tendon. However, it is unknown how lysyl oxidase (LOX), the primary mediator of enzymatic collagen crosslinking, is regulated by stem cells. This study investigates how multiple factors previously identified to promote tendon formation and healing (transforming growth factor [TGF]β1 and TGFβ2, mechanical stimuli, and hypoxia-inducible factor [HIF]-1α) regulate LOX production in the murine C3H10T1/2 mesenchymal stem cell (MSC) line. We hypothesized that TGFβ signaling promotes LOX activity in C3H10T1/2 MSCs, which is regulated by both mechanical stimuli and HIF-1α activation. TGFβ1 and TGFβ2 increased LOX levels as a function of concentration and time. Inhibiting the TGFβ type I receptor (TGFβRI) decreased TGFβ2-induced LOX production by C3H10T1/2 MSCs. Low (5 mPa) and high (150 mPa) magnitudes of fluid shear stress were applied to test impacts of mechanical stimuli, but without TGFβ2, loading alone did not alter LOX levels. Low loading (5 mPa) with TGFβ2 increased LOX at 7 days greater than TGFβ2 treatment alone. Neither HIF-1α knockdown (siRNA) nor activation (CoCl2) affected LOX levels. Ultimately, results suggest that TGFβ2 and appropriate loading magnitudes contribute to LOX production by C3H10T1/2 MSCs. Potential application of these findings includes treatment with TGFβ2 and appropriate mechanical stimuli to modulate LOX production by stem cells to ultimately control collagen matrix stiffening and support functional tendon formation.

Single nucleus and spatial transcriptomic profiling of healthy human hamstring tendon.

The molecular and cellular basis of health in human tendons remains poorly understood. Among human tendons, hamstring tendon has markedly low pathology and can provide a prototypic healthy tendon reference. The aim of this study was to determine the transcriptomes and location of all cell types in healthy hamstring tendon. Using single nucleus RNA sequencing, we profiled the transcriptomes of 10 533 nuclei from four healthy donors and identified 12 distinct cell types. We confirmed the presence of two fibroblast cell types, endothelial cells, mural cells, and immune cells, and identified cell types previously unreported in tendons, including different skeletal muscle cell types, satellite cells, adipocytes, and undefined nervous system cells. The location of these cell types within tendon was defined using spatial transcriptomics and imaging, and potential transcriptional networks and cell-cell interactions were analyzed. We demonstrate that fibroblasts have the highest number of potential cell-cell interactions in our dataset, are present throughout the tendon, and play an important role in the production and organization of extracellular matrix, thus confirming their role as key regulators of hamstring tendon homeostasis. Overall, our findings underscore the complexity of the cellular networks that underpin healthy human tendon function and the central role of fibroblasts as key regulators of hamstring tendon tissue homeostasis.

Biomechanical functions analysis of the Mallard webbed foot: A study of macroscopic and microscopic material assembly and tendon morphology

The mallard webbed foot represents an exemplary model of biomechanical efficiency in avian locomotion. This study delves into the intricate material assembly and tendon morphology of the mallard webbed foot, employing both macroscopic and microscopic analyses. Through histological slices and scanning electron microscopy (SEM), we scrutinized the coupling assembly of rigid and flexible materials such as skin, tendon, and bone, while elucidating the biomechanical functions of tendons across various segments of the tarsometatarsophalangeal joint (TMTPJ). The histological examination unveiled a complex structural hierarchy extending from the external integument to the skeletal framework. Notably, the bone architecture, characterized by compact bone and honeycombed trabeculae, showcases a harmonious blend of strength and lightweight design. Tendons, traversing the phalangeal periphery, surrounded by elastic fibers, collagen fibers, and fat tissue. Fat chambers beneath the phalanx, filled with adipocytes, provide effective buffering, enabling the phalanx to withstand gravity, provide support, and facilitate locomotion. Furthermore, SEM analysis provided insights into the intricate morphology and arrangement of collagen fiber bundles within tendons. Flexor tendons in proximal and middle TMTPJ segments adopt a wavy-type, facilitating energy storage and release during weight-bearing activities. In contrast, distal TMTPJ flexor tendons assume a linear-type, emphasizing force transmission across phalangeal interfaces. Similarly, extensor tendons demonstrate segment-specific arrangements tailored to their respective biomechanical roles, with wavy-type in proximal and distal segments for energy modulation and linear-type in middle segments for enhanced force transmission and tear resistance. Overall, our findings offer a comprehensive understanding of the mallard webbed foot's biomechanical prowess, underscoring the symbiotic relationship between material composition, tendon morphology, and locomotor functionality. This study not only enriches our knowledge of avian biomechanics but also provides valuable insights for biomimetic design and tissue engineering endeavors.