Search Results (54)

Vascular smooth muscle cells (VSMCs) are a major cell type in the arterial wall responsible for regulating vascular homeostasis. Under physiological conditions, VSMCs reside in the medial layer of the arteries, express elevated levels of contractile proteins, regulate vascular tone, and provide mechanical strength and elasticity to the blood vessel. In response to obesity, hyperglycemia, and insulin resistance, critical pathogenic hallmarks of Type 2 diabetes (T2D), VSMCs undergo a phenotypic transformation, adopting new phenotypes with increased proliferative (synthetic), inflammatory (macrophage-like), or bone-like (osteogenic) properties. While crucial for normal repair and vascular adaptation, VSMC phenotypic plasticity is a key driver for the development and progression of macrovascular complications associated with T2D. Despite advances in lineage tracing and multi-omics profiling that have uncovered key molecular regulators of VSMC phenotypic switching in vasculopathy, our understanding of the cellular and molecular mechanisms underlying VSMC transformation into diseased phenotypes in T2D remains incomplete. This review will provide a holistic summary of research from the past 15 years, with a focus on the signaling pathways and transcriptional regulators that govern VSMC phenotypic transition in response to obesity, hyperglycemia, and insulin resistance. We examine the integrated molecular mechanisms that orchestrate VSMC fate reprogramming in T2D and highlight the dynamic interplay among diverse signaling and transcriptional networks. Emphasis is placed on how these interconnected pathways collectively influence VSMC behavior and contribute to the pathogenesis of T2D-associated atherosclerosis. Full article

Background: Persistent neuromuscular deficits following anterior cruciate ligament reconstruction (ACLR) are frequently attributed to arthrogenic muscle inhibition (AMI). The type of autologous graft used may influence the trajectory of neuromuscular recovery. Objective: To investigate the influence of graft type—bone–patellar tendon–bone (BPTB), hamstring tendon (HT), and quadriceps tendon (QT)—on the contractile properties of periarticular knee muscles over a 9-month post-operative period. Hypothesis: Each graft type would result in distinct recovery patterns of muscle contractility, as measured by tensiomyography (TMG). Methods: Thirty-one patients undergoing ACLR with BPTB (n = 8), HT (n = 12), or QT (n = 11) autografts were evaluated at 3, 6, and 9 months post-operatively. TMG was used to measure contraction time (Tc) and maximal displacement (Dm) in the rectus femoris, vastus medialis, vastus lateralis, and biceps femoris. Results: Significant within-group improvements in Tc and Dm were observed across all graft types from 3 to 9 months (Tc: p < 0.001 to p = 0.02; Dm: p < 0.001 to p = 0.01). The QT group showed the most pronounced Tc reduction in RF (from 30.16 ± 2.4 ms to 15.44 ± 1.6 ms, p < 0.001) and VM (from 31.05 ± 2.6 ms to 18.65 ± 1.8 ms, p = 0.004). In contrast, HT grafts demonstrated limited Tc recovery in BF between 6 and 9 months compared to BPTB and QT (p < 0.001), indicating a stagnation phase. BPTB exhibited persistent bilateral deficits in both quadriceps and BF at 9 months. Conclusions: Autograft type significantly influences neuromuscular recovery patterns after ACLR. TMG enables objective, muscle-specific monitoring of contractile dynamics and may support future individualized rehabilitation strategies. Full article

Polarized fluorescence microscopy of “ghost” muscle fibers, containing fluorescently labeled F-actin, tropomyosin, and myosin, has provided new insights into the molecular mechanisms underlying muscle contraction. At low Ca²⁺, the troponin-induced overtwisting of the actin filament alters the configuration of myosin binding sites, preventing actin–myosin interactions. As Ca²⁺ levels rise, the actin filament undergoes untwisting, while tropomyosin becomes overtwisted, facilitating the binding of myosin to actin. In the weakly bound state, myosin heads greatly increase both the internal twist and the bending stiffness of actin filaments, accompanied by the untwisting of tropomyosin. Following phosphate (Pi) release, myosin induces the untwisting of overtwisted actin filaments, driving thin-filament sliding relative to the thick filament during force generation. Point mutations in tropomyosin significantly alter the ability of actin and tropomyosin filaments to respond to Pi release, with coordinated changes in twist and bending stiffness. These structural effects correlate with changes in actomyosin ATPase activity. Together, these findings support a model in which dynamic filament twisting is involved in the molecular mechanisms of muscle contraction together with the active working stroke in the myosin motor, and suggest that impairment of this ability may cause contractile dysfunction. Full article

The functional status of the plantaris muscle (PM) remains controversial and is historically dismissed as vestigial; yet, it is increasingly recognized for its structural and clinical complexity. This narrative review synthesizes current evidence from embryological development, adult morphological studies, comparative mammalian anatomy, and clinical case reports to reassess the role of the PM in humans. Developmental data reveal that the PM is consistently present during fetal life, with tendon morphology and insertion patterns emerging early and resembling adult anatomical variants. Rather than indicating postnatal regression, it suggests a stable polymorphism rooted in prenatal development. Across mammalian species, the PM varies in presence and function, correlating with locomotor specialization from proprioception in primates to propulsion in carnivores, and absence in ungulates. In humans, high proprioceptive fiber density and anatomical variability support the hypothesis that the PM may be undergoing functional repurposing from a contractile to a sensorimotor role. Clinically, its relevance is evident in imaging interpretation, surgical tendon harvesting, and the pathophysiology of Achilles tendinopathy. Recent discoveries, including the identification of the plantaris ligamentous tendon (PLT), further underscore the complexity of this region and support the need to reassess its structural and clinical significance. We conclude that the PM should not be regarded as a regressing remnant but as a dynamically adapting structure with potential neuromechanical function. Future studies involving electromyography and neuroanatomical mapping are essential to elucidate its evolving role. Full article

Recent bioassay studies have unexpectedly supported the high (computationally predicted) binding affinities of angiotensin receptor blockers (ARBs) at α-adrenergic receptors (αARs) in isolated smooth muscle. Computational predictions from ligand docking studies are consistent with very low concentrations of ARBs (e.g., sartans or bisartans) that partially reduce (20–50%) the contractile response to phenylephrine, suggesting that some ARBs may function as partial inverse agonists at αARs. Virtual ligand screening (docking) and molecular dynamics (MD) simulations were carried out to explore the binding affinities and stabilities of selected non-peptide ligands (e.g., ARBs and small-molecule opioids) for several G-protein coupled receptor (GPCR) types, including angiotensin II (AngII) type 1 receptor (AT₁R), α1AR, α2AR, and μ-(µOR) and ժ-opioid receptors (ժOR). Results: All ligands docked preferentially to the binding pocket on the cell surface domain of the GPCR types investigated. Drug binding was characterized by weak interactions (hydrophobic, hydrogen bonding, pi-pi) and stronger ionic and salt-bridge interactions (cation-pi and cation-anion interactions). Ligands specific to each GPCR category showed considerable cross-binding with alternative GPCRs, with small-molecule medications appearing less selective than their peptide or ARB functional equivalents. ARBs that exhibit higher affinities for AT₁R also demonstrate higher affinities for µORs and ժORs than opiate ligands, such as fentanyl and naltrexone. Moreover, ARBs had a higher affinity for αARs than either alpha agonists (epinephrine and phenylephrine) or inhibitors (prazosin and doxazosin). MD simulations of membrane-embedded ARB-GPCR complexes proved stable over nanosecond time scales and suggested that some ARBs may behave as agonists or antagonists depending on the GPCR type. Based on the results presented in this and related investigations, we propose that agonists bind to the resting A-site of GPCRs, while inverse agonists occupy the desensitizing D-site, which partial agonists like morphine and fentanyl share, contributing to addiction. ARBs block both AngII and alpha receptors, suggesting that they are more potent antihypertensive drugs than ACE inhibitors. ARBs have the potential to inhibit morphine tolerance and appear to disrupt receptor desensitization processes, potentially by competing at the D-site. Our results suggest the possible therapeutic potential of ARBs in treating methamphetamine and opiate addictions. Full article

Chronic obstructive pulmonary disease (COPD), whose main risk factor is cigarette smoking, is among the most prevalent diseases worldwide. Previous studies have shown that cigarette smoke extract (CSE) can directly affect pulmonary artery function independently of hypoxia resulting from the airway obstruction. In addition, CSE also affects bronchial smooth muscle, leading to airway hyper-responsiveness. However, its specific impact on the contractile machinery of this compartment remains unclear. In this study, using in vitro experiments with human bronchial smooth muscle cells (hBSMCs), we found that CSE exposure disrupted calcium homeostasis, increased ROS and lipid peroxidation, and reduced cell antioxidant defenses. Furthermore, CSE exposure altered the cell contractile apparatus by decreasing key cytoskeletal proteins and impairing actin dynamics, potentially contributing to the dysregulated contractile response of cells. Notably, these effects were significantly attenuated by antioxidant drugs such as mitoTEMPO and N-acetylcysteine, as well as by the inhibition of the endoplasmic reticulum (ER) calcium channels with 2-aminoethoxydiphenyl borate (2-APB). More importantly, mitoTEMPO partially restored the contractile response of bronchus upon CSE challenge. Collectively, our findings give evidence that CSE-mediated increase in ROS and intracellular calcium contribute to cytoskeletal disruption and functional impairment in airway smooth muscle. Moreover, these results also point to potential therapeutical approaches for mitigating the harmful effects of cigarette smoke in the lung. Full article

A 40-year-old woman with a known diagnosis of Ehlers–Danlos syndrome (EDS) began experiencing progressive shortness of breath and reduced exercise tolerance following her second pregnancy. The patient underwent an unenhanced computed tomography (CT) scan that showed a marked elevation of the left diaphragm. Suspecting diaphragm dysfunction, the patient underwent Dynamic Digital Radiography (DDR) that confirmed a reduction in left diaphragm motility, indicative of impaired diaphragm function. Based on the DDR findings, which demonstrated reduced but preserved diaphragmatic motion without paradoxical movement or complete immobility, the thoracic surgeon decided that surgical intervention, such as plication, was not necessary. Instead, rehabilitation exercises, including breathing techniques and diaphragm strengthening, were recommended. EDS includes connective tissue disorders that vary in severity but are typically characterized by hypermobility of the joints, skin hyper-elasticity, and a predisposition to vascular fragility. One of the complications of EDS is weakened connective tissues, which can affect the diaphragm, impairing the contractility of the muscle and leading to impaired mobility and respiratory symptoms such as shortness of breath. Diaphragm dysfunction can manifest as reduced movement, potentially related to the underlying connective tissue weakness. This case highlights the clinical value of DDR as a non-invasive, low-dose, and dynamic imaging modality in the diagnosis of diaphragmatic dysfunction in EDS patients, enabling individualized treatment planning and potentially avoiding unnecessary surgical interventions. Full article

Background/Objectives: Under physiological conditions, vascular smooth muscle cells (VSMCs) are in a quiescent contractile state, but under pathological conditions, such as atherosclerosis, they change their phenotype to synthetic, characterized by increased proliferation, migration, and production of an extracellular matrix. Furthermore, VSMCs can undergo calcification, switching to an osteoblast-like phenotype, contributing to plaque instability. Methods: In this study, we analyzed the phenotypic changes in VSMCs during the transition from a physiological to a pathological state, a key process in the progression of atherosclerosis, using confocal and transmission electron microscopy, real-time PCR, and intracellular calcium quantification. Results: Confocal and transmission electron microscopy revealed a prominent remodeling of the actin cytoskeleton, increasing autophagic vacuoles in synthetic VSMCs and the deposition of calcium microcrystals in calcified cells. Immunofluorescence analysis revealed differential expression of α-SMA (contractile marker) and galectin-3 (synthetic marker), confirming the phenotypic changes. Real-time PCR further validated these changes, showing upregulation of RUNX-2, a marker of osteogenic transition, in calcified VSMCs. Conclusions: This study highlights the dynamic plasticity of VSMCs and their role in atherosclerosis progression. Understanding the characteristics of these phenotypic transitions can help develop targeted therapies to mitigate vascular calcification and plaque instability, potentially countering cardiovascular disease. Full article

Background: Three-dimensional skeletal muscle organoids (3D SkMO) are becoming of increasing interest for preclinical studies in Duchenne muscular dystrophy (DMD), provided that the used platform demonstrates the possibility to form functional and reproducible 3D SkMOs, to investigate on potential patient-related phenotypic differences. Methods: In this study, we employed fibrin-based 3D skeletal muscle organoids derived from immortalized myogenic precursors of DMD patients carrying either a stop codon mutation in exon 59 or a 48–50 deletion. We compared dystrophic lines with a healthy wild-type control (HWT) by assessing microtissue formation ability, contractile function at multiple timepoints along with intracellular calcium dynamics via calcium imaging, as well as expression of myogenic markers. Results: We found patient-specific structural and functional differences in the early stages of 3D SkMO development. Contractile force, measured as both single twitch and tetanic responses, was significantly lower in dystrophic 3D SkMOs compared to HWT, with the most pronounced differences observed at day 7 of differentiation. However, these disparities diminished over time under similar culturing conditions and in the absence of continuous nerve-like stimulation, suggesting that the primary deficit lies in delayed myogenic maturation, as also supported by gene expression analysis. Conclusions: Our results underline that, despite the initial maturation delay, DMD muscle precursors retain the capacity to form functional 3D SkMOs once this intrinsic lag is overcome. This suggests a critical role of dystrophin in early myogenic development, while contraction-induced stress and/or an inflammatory microenvironment are essential to fully recapitulate dystrophic phenotypes in 3D SkMOs. Full article

Background/Objectives: Thick-filament-based regulation in muscle is generally conceived as processes that modulate the number of myosin heads capable of force generation. It has been generally assumed that biochemical and structural assays of myosin active and inactive states provide equivalent measures of myosin recruitment, but recent studies indicate that this may not always be the case. Here, we studied the steady-state and dynamic mechanical changes in skinned porcine myocardium before and after treatment with omecamtiv mecarbil (OM) or piperine to help decipher how the biochemical and structural states of myosin separately affect contractile force. Methods: Force–Ca²⁺ relationships were obtained from skinned cardiomyocytes isolated from porcine myocardium before and after exposure to 1 μM OM and 7 μM piperine. Crossbridge kinetics were acquired using a step response stretch activation protocol allowing myosin attachment and detachment rates to be calculated. Results: OM augmented calcium-activated force at submaximal calcium levels that can be attributed to increased thick filament recruitment, increases in calcium sensitivity, an increased duty ratio, and from decelerated crossbridge detachment resulting in slowed crossbridge cycling kinetics. Piperine, in contrast, was able to increase activated force at submaximal calcium levels without appreciably affecting crossbridge cycling kinetics. Conclusions: Our study supports the notion that thick filament activation is primarily a process of myosin recruitment that is not necessarily coupled with the chemo-cycling of crossbridges. These new insights into thick filament activation mechanisms will need to be considered in the design of sarcomere-based therapies for treatment of myopathies. Full article

Vascular smooth muscle cells (VSMCs) can dynamically change their phenotype between contractile and synthetic forms in response to environmental stress, which is pivotal in maintaining vascular homeostasis and mediating pathological remodeling of blood vessels. We previously reported that suppression of canonical transient receptor potential 6 (TRPC6) channel-mediated cation entry sustains VSMCs contractile phenotype and promotes the blood flow recovery after hindlimb ischemia in mice. We also reported that Zn²⁺, a metal biomolecule mobilized by TRPC6 channel activation, exerts potential beneficial effects on cardiac contractility and remodeling. Therefore, we hypothesized that TRPC6-mediated Zn²⁺ influx participates in phenotype switching of VSMCs and vascular remodeling. We established rat aortic smooth muscle cells (RAoSMCs) stably expressing wild type (WT) and Zn²⁺ only impermeable TRPC6 (KYD) mutant. Although the resting phenotypes were similar in both RAoSMCs, pharmacological TRPC6 activation by PPZ2 prevented the transforming growth factor (TGF) β-induced reduction in the intracellular Zn²⁺ amount and contractile differentiation in RAoSMCs (WT), but failed to prevent them in RAoSMCs (KYD). There were no significant differences in TRPC6-dependent cation currents among all RAoSMCs pretreated with or without TGFβ and/or PPZ2, suggesting that TRPC6 channels are functionally expressed in RAoSMCs regardless of their phenotype. Treatment of mice with PPZ2 attenuated the progression of vascular remodeling caused by chronic angiotensin II infusion. These results suggest that Zn²⁺ influx through TRPC6 channels negatively regulates the TGFβ-induced contractile differentiation of VSMCs and the progression of vascular remodeling in rodents. Full article

Background: This paper aims to complement the latest contribution in the literature that provides estimates of physiological parameters of a dynamic model for the elbow time profile during walking while linking them to a neurodegenerative disorder (Parkinsons’s disease) characterized by motor symptoms. An upper limb model is here proposed in which an active contractile element is included within a model, viewing the arm as a double pendulum system and muscles as represented by a Kelvin–Voight system. All model parameters characterizing both the shoulder and the elbow of each subject are estimated via a gradient-like identifier whose exponential convergence properties are determined by a non-anticipative Lyapunov function, ensuring robustness features. Methods: Joint angle data from different walking subjects (healthy subjects and patients with Parkinson’s disease) have been recorded using an IMU sensor system and compared with the joint angles obtained by means of the proposed model, which was adapted to each subject using available anthropometric knowledge and relying on the estimated parameters. Results: Experiments show that the reconstruction of shoulder and elbow time profiles can be definitely achieved through the proposed procedure with the estimated stiffness parameters turning out to constitute objective and quantitative indices of muscle stiffness (as a pivotal symptom of the pathology), which are able to track changes due to the therapy. Conclusions: The same dynamic model is actually able to capture the main features of the upper limb movement of both (healthy and pathological) walking subjects, with its parameters, in turn, characterizing the nature and progress of the pathology. Full article

Pediatric dilated cardiomyopathy (DCM) is a rare heart muscle disorder leading to the enlargement of all chambers and systolic dysfunction. We identified a novel de novo variant, c.88A>G (p.Lys30Glu, K30E), in the TPM1 gene encoding the major cardiac muscle tropomyosin (Tpm) isoform, Tpm1.1. The variant was found in a proband with DCM and left ventricular non-compaction who progressed to terminal heart failure at the age of 3 years and 8 months. To study the properties of the mutant protein, we produced recombinant K30E Tpm and used various biochemical and biophysical methods to compare its properties with those of WT Tpm. The K30E substitution decreased the thermal stability of Tpm and its complex with actin and significantly reduced the sliding velocity of the regulated thin filaments over a surface covered by ovine cardiac myosin in an in vitro motility assay across the entire physiological range of Ca²⁺ concentration. Our molecular dynamics simulations suggest that the charge reversal of the 30th residue of Tpm alters the actin monomer to which it is bound. We hypothesize that this rearrangement of the actin–Tpm interaction may hinder the transition of a myosin head attached to a nearby actin from a weakly to a strongly bound, force-generating state, thereby reducing myocardial contractility. The impaired myosin interaction with regulated actin filaments and the decreased thermal stability of the actin–Tpm complex at a near physiological temperature likely contribute to the pathogenicity of the variant and its causative role in progressive DCM. Full article

Spinal muscular atrophy (SMA) is caused by a deficiency of the ubiquitously expressed survival motor neuron (SMN) protein. The main pathological hallmark of SMA is the degeneration of lower motor neurons (MNs) with subsequent denervation and atrophy of skeletal muscle. However, increasing evidence indicates that low SMN levels not only are detrimental to the central nervous system (CNS) but also directly affect other peripheral tissues and organs, including skeletal muscle. To better understand the potential primary impact of SMN deficiency in muscle, we explored the cellular, ultrastructural, and molecular basis of SMA myopathy in the SMNΔ7 mouse model of severe SMA at an early postnatal period (P0-7) prior to muscle denervation and MN loss (preneurodegenerative [PND] stage). This period contrasts with the neurodegenerative (ND) stage (P8-14), in which MN loss and muscle atrophy occur. At the PND stage, we found that SMN∆7 mice displayed early signs of motor dysfunction with overt myofiber alterations in the absence of atrophy. We provide essential new ultrastructural data on focal and segmental lesions in the myofibrillar contractile apparatus. These lesions were observed in association with specific myonuclear domains and included abnormal accumulations of actin-thin myofilaments, sarcomere disruption, and the formation of minisarcomeres. The sarcoplasmic reticulum and triads also exhibited ultrastructural alterations, suggesting decoupling during the excitation–contraction process. Finally, changes in intermyofibrillar mitochondrial organization and dynamics, indicative of mitochondrial biogenesis overactivation, were also found. Overall, our results demonstrated that SMN deficiency induces early and MN loss-independent alterations in myofibers that essentially contribute to SMA myopathy. This strongly supports the growing body of evidence indicating the existence of intrinsic alterations in the skeletal muscle in SMA and further reinforces the relevance of this peripheral tissue as a key therapeutic target for the disease. Full article

Background/Objectives: This study explores an optimization-based strategy for muscle force estimation by employing simplified cost functions integrated with physiologically relevant muscle models. Methods: Considering elbow flexion as a case study, we employ an inverse-dynamics approach to estimate muscle forces for the biceps brachii, brachialis, and brachioradialis, utilizing different combinations of cost functions and muscle constitutive models. Muscle force generation is modeled by accounting for active and passive contractile behavior to varying degrees using Hill-type models. In total, three separate cost functions (minimization of total muscle force, mechanical work, and muscle stress) are evaluated with each muscle force model to represent potential neuromuscular control strategies without relying on electromyography (EMG) data, thereby characterizing the interplay between muscle models and cost functions. Results: Among the evaluated models, the Hill-type muscle model that incorporates both active and passive properties, combined with the stress minimization cost function, provided the most accurate predictions of muscle activation and force production for all three arm flexor muscles. Our results, validated against existing biomechanical data, demonstrate that even simplified cost functions, when paired with detailed muscle models, can achieve high accuracy in predicting muscle forces. Conclusions: This approach offers a versatile, EMG-free alternative for estimating muscle recruitment and force production, providing a more accessible and adaptable tool for muscle force analysis. It has profound implications for enhancing rehabilitation protocols and athletic training, not only broadening the applicability of muscle force estimation in clinical and sports settings but also paving the way for future innovations in biomechanical research. Full article