Distinct myofibre domains of the human myotendinous junction revealed by single-nucleus RNA sequencing.

The myotendinous junction (MTJ) is a specialized domain of the multinucleated myofibre that is faced with the challenge of maintaining robust cell-matrix contact with the tendon under high mechanical stress and strain. Here, we profiled 24,124 nuclei in semitendinosus muscle-tendon samples from three healthy males by using single-nucleus RNA sequencing (snRNA-seq), alongside spatial transcriptomics, to gain insight into the genes characterizing this specialization in humans. We identified a cluster of MTJ myonuclei represented by 47 enriched transcripts, of which the presence of ABI3BP, ABLIM1, ADAMTSL1, BICD1, CPM, FHOD3, FRAS1 and FREM2 was confirmed at the MTJ at the protein level in immunofluorescence assays. Four distinct subclusters of MTJ myonuclei were apparent, comprising two COL22A1-expressing subclusters and two subclusters lacking COL22A1 expression but with differing fibre type profiles characterized by expression of either MYH7 or MYH1 and/or MYH2. Our findings reveal distinct myonuclei profiles of the human MTJ, which represents a weak link in the musculoskeletal system that is selectively affected in pathological conditions ranging from muscle strains to muscular dystrophies.

Gluteus Maximus Distal Myotendinous Junction Tear in a Pickleball Player: A Case Report.

ABSTRACT: A 72-year-old male presented for evaluation of a 2-wk history left buttock pain that began while playing pickleball. He sustained a left inversion ankle sprain while in a squatted position and landed on his left buttock. Four days after his injury, he developed extensive bruising involving his lower back, buttock, and left thigh. On examination, he had tenderness to palpation at the left side of the sacrum and in the region of the deep external rotators. Left hip range of motion was full in extension but limited to 90° of flexion, which reproduced left-sided buttock pain. External rotation provoked pain, but internal rotation was full and pain free. MRI of the pelvis demonstrated a grade 2 partial thickness tear of the left gluteus maximus muscle at its distal myotendinous junction with associated retraction and intramuscular hematoma. He was managed with compression with biking shorts, icing, acetaminophen, and physical therapy. He returned to pickleball approximately 4 wk after his injury, and at his 4-wk follow-up, he reported 99% improvement in his symptoms with the only remaining complaint being minimal discomfort with gluteal stretching.

Larger interface area at the human myotendinous junction in type 1 compared with type 2 muscle fibers.

The myotendinous junction (MTJ) is structurally specialized to transmit force. The highly folded muscle membrane at the MTJ increases the contact area between muscle and tendon and potentially the load tolerance of the MTJ. Muscles with a high content of type II fibers are more often subject to strain injury compared with muscles with type I fibers. It is hypothesized that this is explained by a smaller interface area of MTJ in type II compared with type I muscle fibers. The aim was to investigate by confocal microscopy whether there is difference in the surface area at the MTJ between type I and II muscle fibers. Individual muscle fibers with an intact MTJ were isolated by microscopic dissection in samples from human semitendinosus, and they were labeled with antibodies against collagen XXII (indicating MTJ) and type I myosin (MHCI). Using a spinning disc confocal microscope, the MTJ from each fiber was scanned and subsequently reconstructed to a 3D-model. The interface area between muscle and tendon was calculated in type I and II fibers from these reconstructions. The MTJ was analyzed in 314 muscle fibers. Type I muscle fibers had a 22% larger MTJ interface area compared with type II fibers (p < 0.05), also when the area was normalized to fiber diameter. By the new method, it was possible to analyze the structure of the MTJ from a large number of human muscle fibers. The finding that the interface area between muscle and tendon is higher in type I compared with type II fibers suggests that type II fibers are less resistant to strain and therefore more susceptible to injury.

Chronic intramuscular calcific tendinitis of the deltoid muscle.

Calcific tendinitis is a potentially symptomatic disorder characterized by calcium deposits in the substance of the tendon. Although this condition can occur in any tendinous tissue throughout the human body, calcium deposition commonly occurs at tendon insertions near the bone-tendon junction. The musculotendinous junction of the deltoid muscle has peculiarly dense intramuscular tendons to which muscle fibers attach obliquely to create muscular strength. Given that the intramuscular tendons themselves, which form the consecutive part from the insertion, are subjected to unpredictable stress load or microtrauma similar to tendon insertions, it is reasonable to assume that calcific tendinitis could also occur at the intramuscular tendons. Here we report a case of chronic symptomatic calcium deposition in the lateral part of the deltoid muscle between the origin and the insertion, which was eventually surgically removed and confirmed as intramuscular calcific tendinitis.

Effects of Myostatin on Nuclear Morphology at the Myotendinous Junction.

Myostatin (Myo) is known to suppress skeletal muscle growth, and was recently reported to control tendon homeostasis. The purpose of the present study was to investigate the regulatory involvement of Myo in the myotendinous junction (MTJ) in vivo and in vitro. After Achilles tendon injury in mice, we identified unexpected cell accumulation on the tendon side of the MTJ. At postoperative day 7 (POD7), the nuclei had an egg-like profile, whereas at POD28 they were spindle-shaped. The aspect ratio of nuclei on the tendon side of the MTJ differed significantly between POD7 and POD28 (p = 4.67 × 10-34). We then investigated Myo expression in the injured Achilles tendon. At the MTJ, Myo expression was significantly increased at POD28 relative to POD7 (p = 0.0309). To investigate the action of Myo in vitro, we then prepared laminated sheets of myoblasts (C2C12) and fibroblasts (NIH3T3) (a pseudo MTJ model). Myo did not affect the expression of Pax7 and desmin (markers of muscle development), scleraxis and temonodulin (markers of tendon development), or Sox9 (a common marker of muscle and tendon development) in the cell sheets. However, Myo changed the nuclear morphology of scleraxis-positive cells arrayed at the boundary between the myoblast sheet and the fibroblast sheet (aspect ratio of the cell nuclei, myostatin(+) vs. myostatin(-): p = 0.000134). Myo may strengthen the connection at the MTJ in the initial stages of growth and wound healing.

Rupture of the gastrocnemius muscle at its distal musculotendinous junction: conservative treatment and outcomes in 11 dogs.

CASE HISTORY: Medical records from three veterinary referral centres and a university veterinary teaching hospital in Australia and the USA were reviewed to identify dogs with a diagnosis of distal gastrocnemius musculotendinous junction rupture (DGMJR) that were treated without surgery between 2007 and 2020. CLINICAL AND IMAGING FINDINGS: All dogs (n = 11) presented with unilateral, pelvic limb lameness and bruising, swelling or pain on palpation at the distal musculotendinous junction. The diagnosis was confirmed with ultrasound or MRI in six dogs; radiographs were used to excluded stifle and tarsus pathology in four dogs; and five dogs were diagnosed on physical examination findings. TREATMENT AND OUTCOME: All dogs were managed conservatively, either with complete confinement alone (n = 10; median 9 weeks), external coaptation alone (n = 1), or a combination of both (n = 4). Sporting dogs (n = 7) were completely confined (median 22 weeks) for longer periods than companion dogs (n = 3; median 5 weeks).A good to excellent outcome was achieved for all cases in this cohort. The seven sporting dogs achieved an excellent outcome; returning to their previous level of sport, with complete resolution of lameness and recovery of a normal tibiotarsal stance. The four companion dogs achieved a good outcome; returning to their previous level of activity but with persistently increased tibiotarsal standing angle compared to the contralateral limb. CLINICAL RELEVANCE: Conservative treatment represents a viable treatment option for dogs with a rupture of the gastrocnemius muscle at its distal musculotendinous junction.

A Cadaveric Limb Analysis of the Posterior Tibial Musculotendinous Junction to determine the feasibility of Interosseous Membrane Tendon Transfer.

The transfer of the posterior tibial tendon through the interosseous membrane is potentially an effective treatment to correct the deformity of the foot and ankle. Our study aimed to evaluate the anatomical feasibility of anterior transfer of the posterior tibial tendon through the interosseous membrane route using the musculotendinous junction (MTJ). Eighteen adult cadavers were used. The width and thickness of the tibial posterior MTJ, width of the interosseous membrane at the corresponding level, and the window size of the interosseous membrane were measured. Additionally, the distance between the distal end of the MTJ and the tip of the medial malleolus were recorded. The mean length of the posterior tibial tendon was 83.60 mm, the mean distance of the posterior tibial MTJ to medial malleolus tip was 45.48 mm and the mean length of MTJ was 31.74 mm. The mean width of distal end of MTJ was 7.76 mm, thickness of distal end of MTJ was 4.07 mm and the mean width of the interosseous membrane at the distal end of MTJ was 4.76 mm. We found the mean width of the proximal end of MTJ was 20.68 mm, the mean thickness of proximal end of MTJ was 5.52 mm, and mean width of interosseous membrane at the proximal end of MTJ was 8.76 mm. Our study has demonstrated that a 31 mm length incision made at approximately 45 mm from the proximal end of the medial malleolus can safely reach the MTJ. We recommend an opening length of the interosseous membrane of at least 20 mm.

Is There a Nerve-free Zone in Which a Subscapularis Split Can Safely be Performed? An Anatomical Study Using Embalmed Specimens.

BACKGROUND: The subscapularis muscle, which is part of the rotator cuff, is located anterior to the shoulder. In anterior approaches to the shoulder, its supplying nerves are at risk of iatrogenic injury, which may cause postoperative complications. It is unclear whether there is any nerve-free zone in which a subscapularis split can be performed without risking nerve damage. QUESTIONS/PURPOSES: In an anatomical study, we asked: (1) With the arm abducted 60° and externally rotated, what are the median and shortest distances between the entrance point of the subscapular nerves into subscapularis muscle and the myotendinous junction of this muscle? (2) What are those distances in other positions of the shoulder? (3) Did those measurements differ between specimen sexes or sides? METHODS: In 84 shoulders of 66 embalmed anatomic specimens, the distance from the myotendinous junction of the subscapularis muscle to the entrance points of the subscapular nerves into the subscapularis muscle was measured using an inelastic thread and a millimeter gauge with the arm abducted 60° and rotated externally. In 16 of 84 shoulders, which were selected randomly, after taking the measurements with the arm abducted 60° and rotated externally, arm positions were changed and further measurements were taken with the arm abducted 60° and rotated neutrally, abducted 60° and rotated internally, and abducted 90° and rotated externally. The positions of the entrance points were described with statistical parameters and compared between different sides, sexes, and joint positions. Measurements were verified using eight fresh-frozen shoulders, showing no difference in distances compared with embalmed specimens and confirming reproducibility of measurements. Absolute distances were used to minimize possible distortion when using correlations and for straightforwardness and clinical applicability. RESULTS: The median (range) distance was 43 mm (24 to 64) for the upper subscapular nerve and 38 mm (23 to 59) for the lower subscapular nerve with the arm rotated externally and abducted 60°. In the 16 subsamples, internal rotation decreased the distance to 34 mm (24 to 49) and 31 mm (15 to 43), respectively, and maximal external rotation and 90° of abduction increased it up to 49 mm (30 to 64) and 41 mm (27 to 56). Comparison of left and right sides yielded no difference. Comparison of sexes showed distances for the lower subscapular nerve of 36 mm (23 to 54) in females versus 39 mm (24 to 60) in males. CONCLUSION: In no specimen did the nerve come closer than 23 mm medial to the myotendinous junction with the arm rotated externally and abducted. Therefore, not exceeding a distance of 20 mm medial to the myotendinous junction with the arm rotated externally seems to provide sufficient protection from nerve injury during surgery. CLINICAL RELEVANCE: Based on the described zone of 20 mm medial to the myotendinous junction, the risk of nerve injury in a subscapularis split approach can be minimized.

Regeneration process of myotendinous junction injury induced by collagenase injection between Achilles tendon and soleus muscle in mice.

Recently, it has become clear that peri-muscular tissues play a significant role in the deterioration of muscle function. Understanding the function and regeneration of muscle, as well as its surrounding tissues, is crucial to determining the causes of muscular illnesses. However, the regeneration process of the myotendinous junction (MTJ), the most closely related peri-muscular tissue, is still unknown. Therefore, we generated a mouse model of MTJ injury by collagenase injection and searched for the process of regeneration of the MTJ and its adjacent regions. The MTJ region was damaged by collagenase injection, which greatly increased the tendon cross sectional area. Collagenase injections increased the proportion of myofibers with a central nucleus, which is a characteristic of regenerating muscle. The collagenase injection group had myofibers with central nuclei and expressing MTJ markers. Additionally, we measured the length of MTJs using serial cross sections of the soleus muscle and discovered that MTJs at 2 weeks after collagenase injection were shorter compared to the control group, with a propensity to progressively recover their length over time. The results showed that MTJs undergo morphological regeneration even when severely damaged, and that this regeneration occurs in conjunction with muscle regeneration. We anticipate that these findings will be valuable in upcoming research on motor unit regeneration.

An engineered in vitro model of the human myotendinous junction.

The myotendinous junction (MTJ) is a vulnerable region at the interface of skeletal muscle and tendon that forms an integrated mechanical unit. This study presents a technique for the spatially restrictive co-culture of human embryonic stem cell (hESC)-derived skeletal myocytes and primary tenocytes for two-dimensional modeling of the MTJ. Micropatterned lanes of extracellular matrix and a 2-well culture chamber define the initial regions of occupation. On day 1, both lines occupy less than 20 % of the initially vacant interstitial zone, referred to henceforth as the junction. Myocyte-tenocyte interdigitations are observed by day 7. Immunocytochemistry reveals enhanced organization and alignment of patterned myocyte and tenocyte features, as well as differential expression of multiple MTJ markers. On day 24, electrically stimulated junction myocytes demonstrate negative contractile strains, while positive tensile strains are exhibited by mechanically passive tenocytes at the junction. Unpatterned tenocytes distal to the junction experience significantly decreased strains in comparison to cells at the interface. Unpatterned myocytes have impaired organization and uncoordinated contractile behavior. These findings suggest that this platform is capable of inducing myocyte-tenocyte junction formation and mechanical coupling similar to the native MTJ, showing transduction of force across the cell-cell interface. STATEMENT OF SIGNIFICANCE: The myotendinous junction (MTJ) is an integrated structure that transduces force across the muscle-tendon boundary, making the region vulnerable to strain injury. Despite the clinical relevance, previous in vitro models of the MTJ lack the structure and mechanical accuracy of the native tissue and have difficulty transmitting force across the cell-cell interface. This study demonstrates an in vitro model of the MTJ, using spatially restrictive cues to inform human myocyte-tenocyte interactions and architecture. The model expressed MTJ markers and developed anisotropic myocyte-tenocyte integrations that resemble the native tissue and allow for force transduction from contracting myocytes to passive tenocyte regions. As such, this study presents a system capable of investigating development, injury, and pathology in the human MTJ.

The telocytes relationship with satellite cells: Extracellular vesicles mediate the myotendinous junction remodeling.

The peripheral nerve injury (PNI) affects the morphology of the whole locomotor apparatus, which can reach the myotendinous junction (MTJ) interface. In the injury condition, the skeletal muscle satellite cells (SC) are triggered, activated, and proliferated to repair their structure, and in the MTJ, the telocytes (TC) are associated to support the interface with the need for remodeling; in that way, these cells can be associated with SC. The study aimed to describe the SC and TC relationship after PNI at the MTJ. Sixteen adult Wistar rats were divided into Control Group (C, n = 8) and PNI Group (PNI, n = 8), PNI was performed by the constriction of the sciatic nerve. The samples were processed for transmission electron microscopy and immunostaining analysis. In the C group was evidenced the arrangement of sarcoplasmic evaginations and invaginations, the support collagen layer with a TC inside it, and an SC through vesicles internally and externally to then. In the PNI group were observed the disarrangement of invaginations and evaginations and sarcomeres degradation at MTJ, as the disposition of telopodes adjacent and in contact to the SC with extracellular vesicles and exosomes in a characterized paracrine activity. These findings can determine a link between the TCs and the SCs at the MTJ remodeling. RESEARCH HIGHLIGHTS: Peripheral nerve injury promotes the myotendinous junction (MTJ) remodeling. The telocytes (TC) and the satellite cells (SC) are present at the myotendinous interface. TC mediated the SC activity at MTJ.

Bioactive fiber-reinforced hydrogel to tailor cell microenvironment for structural and functional regeneration of myotendinous junction.

Myotendinous junction (MTJ) injuries are prevalent in clinical practice, yet the treatment approaches are limited to surgical suturing and conservative therapy, exhibiting a high recurrence rate. Current research on MTJ tissue engineering is scarce and lacks in vivo evaluation of repair efficacy. Here, we developed a three-dimensional-printed bioactive fiber-reinforced hydrogel containing mesenchymal stem cells (MSCs) and Klotho for structural and functional MTJ regeneration. In a rat MTJ defect model, the bioactive fiber-reinforced hydrogel promoted the structural restoration of muscle, tendon, and muscle-tendon interface and enhanced the functional recovery of injured MTJ. In vivo proteomics and in vitro cell cultures elucidated the regenerative mechanisms of the bioactive fiber-reinforced hydrogel by modulating oxidative stress and inflammation, thus engineering an optimized microenvironment to support the survival and differentiation of transplanted MSCs and maintain the functional phenotype of resident cells within MTJ tissues, including tendon/muscle cells and macrophages. This strategy provides a promising treatment for MTJ injuries.

Optimizing the classification of biological tissues using machine learning models based on polarized data.

Polarimetric data is nowadays used to build recognition models for the characterization of organic tissues or the early detection of some diseases. Different Mueller matrix-derived polarimetric observables, which allow a physical interpretation of a specific characteristic of samples, are proposed in literature to feed the required recognition algorithms. However, they are obtained through mathematical transformations of the Mueller matrix and this process may loss relevant sample information in search of physical interpretation. In this work, we present a thorough comparative between 12 classification models based on different polarimetric datasets to find the ideal polarimetric framework to construct tissues classification models. The study is conducted on the experimental Mueller matrices images measured on different tissues: muscle, tendon, myotendinous junction and bone; from a collection of 165 ex-vivo chicken thighs. Three polarimetric datasets are analyzed: (A) a selection of most representative metrics presented in literature; (B) Mueller matrix elements; and (C) the combination of (A) and (B) sets. Results highlight the importance of using raw Mueller matrix elements for the design of classification models.

Mechanical loading is required for initiation of extracellular matrix deposition at the developing murine myotendinous junction.

The myotendinous junction (MTJ) contributes to the generation of motion by connecting muscle to tendon. At the adult MTJ, a specialized extracellular matrix (ECM) is thought to contribute to the mechanical integrity of the muscle-tendon interface, but the factors that influence MTJ formation during mammalian development are unclear. Here, we combined 3D imaging and proteomics with murine models in which muscle contractility and patterning are disrupted to resolve morphological and compositional changes in the ECM during MTJ development. We found that MTJ-specific ECM deposition can be initiated via static loading due to growth; however, it required cyclic loading to develop a mature morphology. Furthermore, the MTJ can mature without the tendon terminating into cartilage. Based on these results, we describe a model wherein MTJ development depends on mechanical loading but not insertion into an enthesis.

Model of calf muscle tear during a simulated eccentric contraction, comparison between ex-vivo experiments and discrete element model.

The tearing of the muscle-tendon complex (MTC) is one of the common sports-related injuries. A better understanding of the mechanisms of rupture and its location could help clinicians improve the way they manage the rehabilitation period of patients. A new numerical approach using the discrete element method (DEM) may be an appropriate approach, as it considers the architecture and the complex behavior of the MTC. The aims of this study were therefore: first, to model and investigate the mechanical elongation response of the MTC until rupture with muscular activation. Secondly, to compare results with experimental data, ex vivo tensile tests until rupture were done on human cadavers {triceps surae muscle + Achilles tendon}. Force/displacement curves and patterns of rupture were analyzed. A numerical model of the MTC was completed in DEM. In both numerical and experimental data, rupture appeared at the myotendinous junction (MTJ). Moreover, force/displacement curves and global rupture strain were in agreement between both studies. The order of magnitude of rupture force was close between numerical (858 N for passive rupture and 996 N-1032 N for rupture with muscular activation) and experimental tests (622 N ± 273 N) as for the displacement of the beginning of rupture (numerical: 28-29 mm, experimental: 31.9 mm ± 3.6 mm). These differences could be explained by choices of DEM model and mechanical properties of MTC's components or their rupture strain values. Here we show that he MTC was broken by fibers' delamination at the distal MTJ and by tendon disinsertion at the proximal MTJ in agreement with experimental data and literature.

Is Curcumine Useful in the Treatment and Prevention of the Tendinopathy and Myotendinous Junction Injury? A Scoping Review.

Physical activity in general and sports in particular, is a mechanism that produces stress and generates great force in the tendon and in the muscle-tendon unit, which increases the risk of injury (tendinopathies). Eccentric and repetitive contraction of the muscle precipitates persistent microtraumatism in the tendon unit. In the development of tendinopathies, the cellular process includes inflammation, apoptosis, vascular, and neuronal changes. Currently, treatments with oral supplements are frequently used. Curcumin seems to preserve, and even repair, damaged tendons. In this systematic review, we focus more especially on the benefits of curcumin. The biological actions of curcumin are diverse, but act around three systems: (a) inflammatory, (b) nuclear factor B (NF-κB) related apoptosis pathways, and (c) oxidative stress systems. A bibliographic search is conducted under the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) as a basis for reporting reliable systematic reviews to perform a Scoping review. After analysing the manuscripts, we can conclude that curcumin is a product that demonstrates a significant biological antialgic, anti-inflammatory, and antioxidant power. Therefore, supplementation has a positive effect on the inflammatory and regenerative response in tendinopathies. In addition, curcumin decreases and modulates the cell infiltration, activation, and maturation of leukocytes, as well as the production of pro-inflammatory mediators at the site of inflammation.

Rotura do peitoral maior: relato de caso de uma rotura incomum / Pectoralis major rupture: case report of an unusual rupture / Rotura del pectoral mayor: informe de un caso de rotura inusual

Apesar de não muito frequente, nos últimos 20 anos, houve um aumento significativo dos relatos sobre rotura do peitoral maior, normalmente associadas à prática de atividade física em que ocorre contração intensa e/ou uso de cargas pesadas exercendo resistência sobre o músculo. Neste relato de caso temos um paciente de 51 anos referindo dor no tórax à direita e no braço direito há 3 dias após tentar consertar o guidão da moto. Apresentava assimetria dos peitorais, perda de força do membro superior direito, dificuldade de movimentação e hematoma. A ressonância magnética demonstrou rotura completa da junção miotendínea do peitoral maior, com tendinopatia com fissuras insercionais e intrasubstanciais infraespinhal e tendinopatia com rotura parcial do tendão subescapular. Foi indicado por médico ortopedista o acompanhamento com o uso de medicação analgésica.

Although not very common, in the last 20 years, there has been a significant increase in reports of rupture of the pectoralis major, normally associated with the practice of physical activity in which intense contraction occurs and/or the use of heavy loads exerting resistance on the muscle. In this case report we have a 51-year-old patient reporting pain in his right chest and right arm for 3 days after trying to fix his motorcycle's handlebars. He had asymmetry of the pectorals, loss of strength in the right upper limb, difficulty moving and hematoma. Magnetic resonance imaging demonstrated complete rupture of the myotendinous junction of the pectoralis major, with tendinopathy with insertional and intrasubstantial infraspinatus fissures and tendinopathy with partial rupture of the subscapularis tendon. An orthopedic doctor recommended follow-up with the use of analgesic medication.

Aunque no es muy común, en los últimos 20 años se ha observado un aumento significativo en los reportes de rotura del pectoral mayor, normalmente asociado a la práctica de actividad física en la que se produce una contracción intensa y/o al uso de cargas pesadas ejerciendo resistencia sobre el mismo. el músculo. En este caso clínico tenemos un paciente de 51 años que refiere dolor en el pecho derecho y en el brazo derecho durante 3 días después de intentar arreglar el manillar de su motocicleta. Presentó asimetría de pectorales, pérdida de fuerza en miembro superior derecho, dificultad de movimiento y hematoma. La resonancia magnética demostró rotura completa de la unión miotendinosa del pectoral mayor, con tendinopatía con fisuras de inserción e intrasustancial del infraespinoso y tendinopatía con rotura parcial del tendón subescapular. Un médico ortopédico recomendó seguimiento con el uso de medicación analgésica.