Skeletal Model Research Articles

The effect of biocide chloromethylisothiazolinone/methylisothiazolinone (CMIT/MIT) mixture on C2C12 muscle cell damage attributed to mitochondrial reactive oxygen species overproduction and autophagy activation

ABSTRACT The mixture of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one (CMIT/MIT) is a biocide widely used as a preservative in various commercial products. This biocide has also been used as an active ingredient in humidifier disinfectants in South Korea, resulting in serious health effects among users. Recent evidence suggests that the underlying mechanism of CMIT/MIT-initiated toxicity might be associated with defects in mitochondrial functions. The aim of this study was to utilize the C2C12 skeletal muscle model to investigate the effects of CMIT/MIT on mitochondrial function and relevant molecular pathways associated with skeletal muscle dysfunction. Data demonstrated that exposure to CMIT/MIT during myogenic differentiation induced significant mitochondrial excess production of reactive oxygen species (ROS) and a decrease in intracellular ATP levels. Notably, CMIT/MIT significantly inhibited mitochondrial oxidative phosphorylation (Oxphos) and reduced mitochondrial mass at a lower concentration than the biocide amount, which diminished the viability of myotubes. CMIT/MIT induced activation of autophagy flux and decreased protein expression levels of myosin heavy chain (MHC). Taken together, CMIT/MIT exposure produced damage in C2C12 myotubes by impairing mitochondrial bioenergetics and activating autophagy. Our findings contribute to an increased understanding of the underlying mechanisms associated with CMIT/MIT-induced adverse skeletal muscle health effects.

Mechanical properties and restoring force model of Squeezing Rub-type Particle Damper

The sliding effect of isolation bearings has been proven to effectively mitigate the seismic effects on large-span bridges. However, it also results in excessive relative displacement between the main girder and piers during earthquakes. In this study, a pioneering Squeezing Rub-type Particle Damper (SRPD) is developed to address this issue. In order to verify the damping characteristics of SRPDs and investigate the working principle of squeezing friction energy dissipation, five quasi-static tests were conducted on SRPDs using two particle damping materials with different compactness levels. In order to compare the energy dissipation performance, comparative tests were conducted on Non-Squeezing-type Particle Dampers (NSPDs) and SRPDs. Utilizing regression analysis, two skeletal curve models for SRPDs were proposed, alongside a detailed restoring force model that incorporates considerations for both the pinching effect and stiffness degradation phenomena observed during the loading and unloading phases. The experimental results demonstrate that the maximum damping force of SRPDs can reach 16 times that of NSPDs, and the energy dissipation of each loading cycle can reach 21.06 times that of NSPDs. SRPDs exhibits improved energy dissipation performance when using high-density blended quartz sand. After sand replenishment repairs,damping force and energy dissipation per cycle of SRPDs were significantly enhanced. The proposed restoring force models of SRPDs aligns well with the experimental curves, underscoring its efficacy in accurately characterizing the hysteresis performance of SRPDs.

Effects of cannabidiol on AMPKα2 /HIF-1α/BNIP3/NIX signaling pathway in skeletal muscle injury

Cannabidiol: (CBD) is a non-psychoactive natural active ingredient from cannabis plant, which has many pharmacological effects, including neuroprotection, antiemetic, anti-inflammatory and anti-skeletal muscle injury. However, the mechanism of its effect on skeletal muscle injury still needs further research. In order to seek a scientifically effective way to combat skeletal muscle injury during exercise, we used healthy SD rats to establish an exercise-induced skeletal muscle injury model by treadmill training, and systematically investigated the effects and mechanisms of CBD, a natural compound in the traditional Chinese medicine Cannabis sativa L., on combating skeletal muscle injury during exercise. CBD effectively improved the fracture of skeletal muscle tissue and reduced the degree of inflammatory cell infiltration. Biochemical indexes such as CK, T, Cor, LDH, SOD, MDA, and GSH-Px in serum of rats returned to normal. Combining transcriptome and network analysis results, CBD may play a protective role in exercise-induced skeletal muscle injury through HIF-1 signaling pathway. The experimental results implied that CBD could down-regulate the expression of IL-6, NF-κB, TNF-α, Keap1, AMPKα2, HIF-1α, BNIP3 and NIX, and raised the protein expression of IL-10, Nrf2 and HO-1. These results indicate that the protective effect of CBD on exercise-induced skeletal muscle injury may be related to the inhibition of oxidative stress and inflammation, thus inhibiting skeletal muscle injury through AMPKα2/HIF-1α/BNIP3/NIX signal pathways.

Interferon-β-Induced Injury During Pediatric Muscle Differentiation: Insight Into Juvenile Dermatomyositis Pathogenesis.

Juvenile dermatomyositis (JDM) involves up-regulated type I interferons (IFNs), including IFNβ, yet pathologic mechanisms remain poorly understood. We aimed to characterize the functional and structural effects of IFNβ on in vitro human pediatric myoblast growth and differentiation in a three-dimensional skeletal muscle model (myobundles). Myobundles fabricated from myoblasts of a healthy pediatric donor were exposed to IFNβ at 0 to 5,600 IU/mL during growth (days 1-4), differentiation (days 4-11), and/or mature (days 11-18) periods. To assess myobundle structure and function, contractile force, kinetics, and fatigue were measured at day 18 with subsequent immunohistochemistry. Myobundles were not functionally affected by IFNβ exposure during growth period alone. However, when IFNβ exposure continued through differentiation, myobundles became dysfunctional (P < 0.0001). IFNβ during differentiation or mature periods alone resulted in dose-dependent decreases in contractility, with greater decrease in the differentiation alone group (P < 0.0001). Twitch kinetics and fatigue remained largely unchanged when myobundles were exposed to IFNβ only during growth, yet twitch time slowed (P < 0.005) and fatigue decreased (P < 0.002) when myobundles were exposed during differentiation or mature stages alone. Nuclei density and myofiber size and organization also decreased when IFNβ was added during differentiation period alone. IFNβ decreases pediatric myobundle contractile function most significantly during differentiation of myoblasts to myotubes. Function is not affected when IFNβ exposure is limited to myoblast proliferation alone. These findings implicate a pathologic role for IFNβ in JDM by impairing myoblast differentiation, leading to subsequent loss of function and ongoing need for muscle regeneration and repair.

Home-based guidance training system with interactive visual feedback using kinect on stroke survivors with moderate to severe motor impairment

The home-based training approach benefits stroke survivors by providing them with an increased amount of training time and greater feasibility in terms of their training schedule, particularly for those with severe motor impairment. Computer-guided training systems provide visual feedback with correct movement patterns during home-based training. This study aimed to investigate the improvement in motor performance among stroke survivors with moderate to severe motor impairment after 800 min of training using a home-based guidance training system with interactive visual feedback. Twelve patients with moderate to severe stroke underwent home-based training, totaling 800 min (20–40 min per session, with a frequency of 3 sessions per week). The home-based guidance training system uses Kinect to reconstruct the 3D human body skeletal model and provides real-time motor feedback during training. The training exercises consisted of six core exercises and eleven optional exercises, including joint exercises, balance control, and coordination. Pre-training and post-training assessments were conducted using the Fugl-Meyer Assessment-Upper Limb (FMA-UE), Fugl-Meyer Assessment-Lower Limb (FMA-LE), Functional Ambulation Categories (FAC), Berg Balance Scale (BBS), Barthel Index (BI), Modified Ashworth Scale (MAS), as well as kinematic data of joint angles and center of mass (COM). The results indicated that motor training led to the attainment of the upper limit of functional range of motion (FROM) in hip abduction, shoulder flexion, and shoulder abduction. However, there was no improvement in the active range of motion (AROM) in the upper extremity (U/E) and lower extremity (L/E) joints, reaching the level of the older healthy population. Significant improvements were observed in both left/right and superior/inferior displacements, as well as body sway in the mediolateral axis of the COM, after 800 min of training. In conclusion, the home-based guidance system using Kinect aids in improving joint kinematics performance at the level of FROM and balance control, accompanied by increased mediolateral body sway of the COM for stroke survivors with moderate to severe stroke. Additionally, spasticity was reduced in both the upper and lower extremities after 800 min of home-based training.

Establishment of a dexamethasone-induced zebrafish skeletal muscle atrophy model and exploration of its mechanisms

BackgroundSkeletal muscle atrophy is one of the main side effects of high-dose or continuous use of glucocorticoids (such as dexamethasone). However, there are limited studies on dexamethasone-induced skeletal muscle atrophy in zebrafish and even fewer explorations of the underlying molecular mechanisms. This study aimed to construct a model of dexamethasone-induced skeletal muscle atrophy in zebrafish and to investigate the molecular mechanisms. MethodsZebrafish soaked in 0.01 % dexamethasone solution for 10 days. Loli Track (Denmark) and Loligo Swimming Respirometer were used to observe the effect of dexamethasone on swimming ability. The effects of dexamethasone on zebrafish skeletal muscle were observed by Transmission electron microscopy, H&E, and wheat germ agglutinin techniques. Enriched genes and signaling pathways were analyzed using Transcriptome sequencing. Further, the levels of mitochondrial and endoplasmic reticulum-related proteins were examined to investigate possible mechanisms. Results0.01 % dexamethasone reduced zebrafish skeletal muscle mass (p < 0.05), myofibre size and cross-sectional area (p < 0.001), and increased protein degradation (ubiquitination and autophagy) (p < 0.05). In addition, 0.01 % dexamethasone reduced the swimming ability of zebrafish, as evidenced by the reluctance to move, fewer movement trajectories, decreased total distance traveled (p < 0.001), average velocity of movement (p < 0.001), oxygen consumption (p < 0.001), critical swimming speed (p < 0.01) and increased exhaustive swimming time (p < 0.001). Further, 0.01 % dexamethasone-induced mitochondrial dysfunction (decreased mitochondrial biogenesis, disturbs kinetic homeostasis, increased autophagy) and endoplasmic reticulum stress. Conclusions0.01 % dexamethasone induces skeletal muscle atrophy and impairs the swimming ability of zebrafish through mitochondrial dysfunction and endoplasmic reticulum stress.

666P MyoScreen™ infantile and late-onset Pompe human skeletal muscle disease models for drug discovery and development of next-generation gene therapies

666P MyoScreen™ infantile and late-onset Pompe human skeletal muscle disease models for drug discovery and development of next-generation gene therapies

Kinematic analysis of engagement and bending capabilities of a point-of-care, incremental skeletal fixation plate bending system

The current standard of care for skeletal reconstruction surgery is to join skeletal disunions with fixation plates and these plates are most commonly fit to bones through manual bending. The bending procedure is often performed in the surgical operating theatre with specialized pliers or, if available, lengthy pre-operative bending to fit a 3D printed skeletal model prepared on computer as part of a virtual surgical plan. Manual bending of fixation plates by eye or to a model can take considerable time. Repetitive bending at a single location can result in work hardening that increases the subsequent risk of fatigue failure. However, incremental forming systems may provide a solution to automatically bend fixation plates accurately, rapidly, and as little at any one location as possible. This paper is an investigation of the kinematics and manipulability of two versions of an incremental fixture plate bending system for Point of Care Manufacturing (POCM) of craniomaxillofacial (CMF) skeletal fixation hardware. The Automatic Plate Bender (APB) v1 is a minimal POCM system that is designed for simple straight plates with a constant incremental pitch. The APB v2 is a more complex POCM that is designed to accommodate a larger variety of standard plates. Kinematic and manipulability analysis demonstrate the engagement and bend classes that each system can accomplish, and identifies a critical singularity in the APB v2 mechanism. The paper concludes with two case studies in which a path planning algorithm uses a manipulability analysis to avoid singularities during incremental bending.

Histological Study on Possible Regenerative Effect of Bone Marrow Derived Mesenchymal Stem Cells versus Platelet Rich Plasma in Skeletal Muscle Injury Model in Adult Male Albino Rat

Histological Study on Possible Regenerative Effect of Bone Marrow Derived Mesenchymal Stem Cells versus Platelet Rich Plasma in Skeletal Muscle Injury Model in Adult Male Albino Rat

High mobility group box1 (HMGB1) is a potential disease biomarker in cell and mouse models of Duchenne muscular dystrophy.

Duchenne muscular dystrophy (DMD) is a progressive muscle wasting disorder affecting 1:3500 male births and is associated with myofiber degeneration, regeneration, and inflammation. Glucocorticoid treatments have been the standard of care due to immunomodulatory/immunosuppressive properties but novel genetic approaches, including exon skipping and gene replacement therapy, are currently being developed. The identification of additional biomarkers to assess DMD-related inflammatory responses and the potential efficacy of these therapeutic approaches are thus of critical importance. The current study uses RNA sequencing of skeletal muscle from two mdx mouse models to identify high mobility group box1 (HMGB1) as a candidate biomarker potentially contributing to DMD-related inflammation. HMGB1 protein content was increased in a human iPSC-derived skeletal myocyte model of DMD and microdystrophin treatment decreased HMGB1 back to control levels. In vivo, HMGB1 protein levels were increased in vehicle treated B10-mdx skeletal muscle compared to B10-WT and significantly decreased in B10-mdx animals treated with adeno-associated virus (AAV)-microdystrophin. However, HMGB1 protein levels were not increased in D2-mdx skeletal muscle compared to D2-WT, demonstrating a strain-specific difference in DMD-related immunopathology.

Involvement of lysophosphatidic acid-LPA1-YAP signaling in healthy and pathological FAPs migration

Skeletal muscle fibrosis is defined as the excessive accumulation of extracellular matrix (ECM) components and is a hallmark of muscular dystrophies. Fibro-adipogenic progenitors (FAPs) are the main source of ECM, and thus have been strongly implicated in fibrogenesis. In skeletal muscle fibrotic models, including muscular dystrophies, FAPs undergo dysregulations in terms of proliferation, differentiation, and apoptosis, however few studies have explored the impact of FAPs migration.Here, we studied fibroblast and FAPs migration and identified lysophosphatidic acid (LPA), a signaling lipid central to skeletal muscle fibrogenesis, as a significant migration inductor. We identified LPA receptor 1 (LPA1) mediated signaling as crucial for this effect through a mechanism dependent on the Hippo pathway, another pathway implicated in fibrosis across diverse tissues. This cross-talk favors the activation of the Yes-associated protein 1 (YAP) and Transcriptional coactivator with PDZ-binding motif (TAZ), leading to increased expression of fibrosis-associated genes. This study reveals the role of YAP in LPA-mediated fibrotic responses as inhibition of YAP transcriptional coactivator activity hinders LPA-induced migration in fibroblasts and FAPs.Moreover, we found that FAPs derived from the mdx4cv mice, a murine model of Duchenne muscular dystrophy, display a heightened migratory phenotype due to enhanced LPA signaling compared to wild-type FAPs. Remarkably, we found that the inhibition of LPA1 or YAP transcriptional coactivator activity in mdx4cv FAPs reverts this phenotype. In summary, the identified LPA-LPA1-YAP pathway emerges as a critical driver of skeletal muscle FAPs migration and provides insights into potential novel targets to mitigate fibrosis in muscular dystrophies.

Rehabilitation management of sports athletes’ muscle injury based on OpenSim technology

Due to the rapid development of sports and people’s love for sports, athletes often perform resistance training beyond the body to achieve better results, resulting in frequent muscle injuries. After the injury, the athlete cannot return to the state before the injury in time, which destroys the previous training results and hinders the athlete from improving to the professional level. Therefore, the research of athletes’ post-injury rehabilitation is the central subject of modern sports science research. This paper used OpenSim technology to study the rehabilitation management of sports athletes’ muscle injury. In this paper, the OpenSim skeletal muscle model was first established, followed by muscle modeling and characteristic analysis, and then a muscle rehabilitation system was constructed. The experimental part used the limb rehabilitation device model of this paper to carry out muscle rehabilitation training for patients with muscle injury, and compared it with conventional rehabilitation training. The experimental results showed that the limb rehabilitation device in this paper had a better rehabilitation effect. Compared with the traditional rehabilitation training, the excellent rate of the total active range of motion of the joints and the satisfaction of rehabilitation nursing were both increased by 10%.

Exploring the underlying mechanism by transcriptome sequencing in rats with high-voltage electrical burns and the role of iron metabolism

BackgroundClinically, the condition of skeletal muscle injury is the key to the process of high voltage electrical burn (HVEB) wound repair. The aim of this study was to identify the potential mechanisms and intervention targets of skeletal muscle injury after HVEB. MethodsA skeletal muscle injury model in SD rats with HVEB was made. Pathological examination and transcriptome sequencing of injured skeletal muscles were performed, and the expression levels of key proteins and genes in related signaling pathways were verified. ResultsSkeletal muscle injury was progressively aggravated within 48 h, then the injury was gradually repaired with scar formation occurring within 1 week. The mechanism of skeletal muscle injury is complex and varied, and ferroptosis is one of the mechanisms. The ferrous iron content in the injured skeletal muscle tissue of model rats increased significantly at 24 h after injury. After 24 h, damage to injured skeletal muscle tissue could be alleviated by increasing iron storage and blocking lysosomal phagocytosis of autophagy. ConclusionsSkeletal muscle injury caused by HVEB is characterized by adjacent endangered tissue progression after injury. Ferroptosis is involved in the mechanism of HVEB, and iron metabolism-related proteins may be potential targets for preventing progressive skeletal muscle injury.

Human-Exoskeleton Coupling Simulation for Lifting Tasks with Shoulder, Spine, and Knee-Joint Powered Exoskeletons.

In this study, we introduce a two-dimensional (2D) human skeletal model coupled with knee, spine, and shoulder exoskeletons. The primary purpose of this model is to predict the optimal lifting motion and provide torque support from the exoskeleton through the utilization of inverse dynamics optimization. The kinematics and dynamics of the human model are expressed using the Denavit-Hartenberg (DH) representation. The lifting optimization formulation integrates the electromechanical dynamics of the DC motors in the exoskeletons of the knee, spine, and shoulder. The design variables for this study include human joint angle profiles and exoskeleton motor current profiles. The optimization objective is to minimize the squared normalized human joint torques, subject to physical and task-specific lifting constraints. We solve this optimization problem using the gradient-based optimizer SNOPT. Our results include a comparison of predicted human joint angle profiles, joint torque profiles, and ground reaction force (GRF) profiles between lifting tasks with and without exoskeleton assistance. We also explore various combinations of exoskeletons for the knee, spine, and shoulder. By resolving the lifting optimization problems, we designed the optimal torques for the exoskeletons located at the knee, spine, and shoulder. It was found that the support from the exoskeletons substantially lowers the torque levels in human joints. Additionally, we conducted experiments only on the knee exoskeleton. Experimental data indicated that using the knee exoskeleton decreases the muscle activation peaks by 35.00%, 10.03%, 22.12%, 30.14%, 16.77%, and 25.71% for muscles of the erector spinae, latissimus dorsi, vastus medialis, vastus lateralis, rectus femoris, and biceps femoris, respectively.

Design-informed generative modelling of skeletal structures using structural optimization

Although various structural optimization techniques have a sound mathematical basis, the structural robustness and practical constructability of optimal designs pose a great challenge in the manufacturing stage. This paper presents an automated novel approach stemming from structural optimization and engineering principles, where discrete members of the structurally optimized designs are driven towards optimal utilization. The developed workflow unifies topology, layout and size optimization in a single parametric platform, which subsequently outputs a ready-to-manufacture CAD skeletal model which can be manufactured either additively or by assembly. All such outputs are checked and validated for structural requirements; strength, stiffness and stability in accordance with standard codes of practice. In the implementations, first, a topology-optimal model is generated and converted to a one-pixel-wide chain model using skeletonization. Herein, this paper uses a novel efficient method to extract the skeleton by using pixel-padding near the domain borders. Secondly, a spatial frame is extracted from the skeleton for its member size and layout optimization. Finally, the CAD model is generated using constructive solid geometry trees and the structural integrity of each member is assessed to ensure structural robustness prior to manufacturing. Various examples presented in the paper showcase the validity of the presented workflow across various structural engineering applications.

Improving the Recognition and Analysis of the Surendra Algorithm in Athletes’ Motion Capture

With the development of computer technology, its application in athletes’ motion capture is more and more extensive, which can be used to design suitable sensors by the Surendra algorithm. In recent years, more and more scholars have begun to use motion capture technology to study human motion posture, analyze and study human motion posture data and apply it to people’s work, study and life. Motion capture technology has also become a key technology in the field of human motion posture research and is playing an increasingly important role. In this paper, a three-dimensional skeletal model of the athlete’s body is first established based on dynamics, and the athlete’s movement characteristics are simulated by this model. Then, the athlete’s movement posture is judged to determine the appropriate form of movement expression. Then, the improved Surendra algorithm is used to detect the movement movements.

IL-17 signaling pathway: A potential therapeutic target for reducing skeletal muscle inflammation

BackgroundThe interleukin-17 (IL-17) signaling pathway is intricately linked with immunity and inflammation; however, the association between the IL-17 signaling pathway and skeletal muscle inflammation remains poorly understood. The study aims to investigate the role of the IL-17 signaling pathway in skeletal muscle inflammation and to evaluate the therapeutic potential of anti-IL-17 antibodies in reducing muscle inflammation. MethodsA skeletal muscle inflammation model was induced by cardiotoxin (CTX) injection in C57BL6/J mice. Following treatment with an anti-IL-17 antibody, we conducted a comprehensive analysis integrating single-cell RNA sequencing (scRNA-seq), bioinformatics, enzyme-linked immunosorbent assay (ELISA), immunofluorescence, and Western blot techniques to elucidate underlying mechanisms. ResultsscRNA-seq analysis revealed a significant increase in neutrophil numbers and activity in inflamed skeletal muscle compared to other cell types, including macrophages, T cells, B cells, endothelial cells, fast muscle cells, fibroblasts, and skeletal muscle satellite cells. The top 30 differentially expressed genes within neutrophils, along with 55 chemokines, were predominantly enriched in the IL-17 signaling pathway. Moreover, the IL-17 signaling pathway exhibited heightened expression in inflamed skeletal muscle, particularly within neutrophils. Treatment with anti-IL-17 antibody resulted in the suppression of IL-17 signaling pathway expression, accompanied by reduced levels of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α, as well as decreased numbers and activity of Ly6g+/Mpo+ neutrophils compared to CTX-induced skeletal muscle inflammation. ConclusionOur findings suggest that the IL-17 signaling pathway plays a crucial role in promoting inflammation within skeletal muscle. Targeting this pathway may hold promise as a therapeutic strategy for ameliorating the inflammatory micro-environment and reducing cytokine production.

KLF13 restrains Dll4-muscular Notch2 axis to improve the muscle atrophy.

Muscle atrophy can cause muscle dysfunction and weakness. Krüppel-like factor 13 (KLF13), a central regulator of cellular energy metabolism, is highly expressed in skeletal muscles and implicated in the pathogenesis of several diseases. This study investigated the role of KLF13 in muscle atrophy, which could be a novel therapeutic target. The effects of gene knockdown and pharmacological targeting of KLF13 on skeletal muscle atrophy were investigated using cell-based and animal models. Clofoctol, an antibiotic and KLF13 agonist, was also investigated as a candidate for repurposing. The mechanisms related to skeletal muscle atrophy were assessed by measuring the expression levels and activation statuses of key regulatory pathways and validated using gene knockdown and RNA sequencing. In a dexamethasone-induced muscle atrophy mouse model, the KLF13 knockout group had decreased muscle strength (N) (1.77±0.10 vs. 1.48±0.16, P<0.01), muscle weight (%) [gastrocnemius (Gas): 76.0±5.69 vs. 60.7±7.23, P<0.001; tibialis anterior (TA): 75.8±6.21 vs. 67.5±5.01, P<0.05], and exhaustive running distance (m) (495.5±64.8 vs. 315.5±60.9, P<0.05) compared with the control group. KLF13 overexpression preserved muscle mass (Gas: 100±6.38 vs. 120±14.4, P<0.01) and the exhaustive running distance (423.8±59.04 vs. 530.2±77.45, P<0.05) in an in vivo diabetes-induced skeletal muscle atrophy model. Clofoctol treatment protected against dexamethasone-induced muscle atrophy. Myotubes treated with dexamethasone, an atrophy-inducing glucocorticoid, were aggravated by KLF13 knockout, but anti-atrophic effects were achieved by inducing KLF13 overexpression. We performed a transcriptome analysis and luciferase reporter assays to further explore this mechanism, finding that delta-like 4 (Dll4) was a novel target gene of KLF13. The KLF13 transcript repressed Dll4, inhibiting the Dll4-Notch2 axis and preventing muscle atrophy. Dexamethasone inhibited KLF13 expression by inhibiting myogenic differentiation 1 (i.e., MYOD1)-mediated KLF13 transcriptional activation and promoting F-Box and WD repeat domain containing 7 (i.e., FBXW7)-mediated KLF13 ubiquitination. This study sheds new light on the mechanisms underlying skeletal muscle atrophy and potential drug targets. KLF13 regulates muscle atrophy and is a potential therapeutic target. Clofoctol is an attractive compound for repurposing studies to treat skeletal muscle atrophy.

Unraveling radiation-induced skeletal muscle damage: Insights from a 3D human skeletal muscle organoid model

BackgroundThree-dimensional (3D) organoids derived from human pluripotent stem cells (hPSCs) have revolutionized in vitro tissue modeling, offering a unique opportunity to replicate physiological tissue organization and functionality. This study investigates the impact of radiation on skeletal muscle response using an innovative in vitro human 3D skeletal muscle organoids (hSMOs) model derived from hPSCs. MethodsThe hSMOs model was established through a differentiation protocol faithfully recapitulating embryonic myogenesis and maturation via paraxial mesodermal differentiation of hPSCs. Key skeletal muscle characteristics were confirmed using immunofluorescent staining and RT-qPCR. Subsequently, the hSMOs were exposed to a clinically relevant dose of 2 Gy of radiation, and their response was analyzed using immunofluorescent staining and RNA-seq. ResultsThe hSMO model faithfully recapitulated embryonic myogenesis and maturation, maintaining key skeletal muscle characteristics. Following exposure to 2 Gy of radiation, histopathological analysis revealed deficits in hSMOs expansion, differentiation, and repair response across various cell types at early (30 min) and intermediate (18 h) time points post-radiation. Immunofluorescent staining targeting γH2AX and 53BP1 demonstrated elevated levels of foci per cell, particularly in PAX7+ cells, during early and intermediate time points, with a distinct kinetic pattern showing a decrease at 72 h. RNA-seq data provided comprehensive insights into the DNA damage response within the hSMOs. ConclusionsOur findings highlight deficits in expansion, differentiation, and repair response in hSMOs following radiation exposure, enhancing our understanding of radiation effects on skeletal muscle and contributing to strategies for mitigating radiation-induced damage in this context.

In vitro characterization of Trypanosoma cruzi infection dynamics in skeletal and cardiac myotubes models suggests a potential cell-to-cell transmission in mediating cardiac pathology.

Chagas disease predominantly affects the heart, esophagus, and colon in its chronic phase. However, the precise infection mechanisms of the causal agent Trypanosoma cruzi in these tissue types remain incompletely understood. This study investigated T. cruzi infection dynamics in skeletal (SM) and cardiac myotubes (CM) differentiated from H9c2(2-1) myoblasts (control). SM and CM were generated using 1% fetal bovine serum (FBS) without or with retinoic acid, respectively. Initial invasion efficiencies and numbers of released parasites were equivalent between undifferentiated and differentiated cells (~0.3-0.6%). Concomitantly, parasite motility patterns were similar across cell lines. However, CM demonstrated significantly higher infection kinetics over time, reaching 13.26% infected cells versus 3.12% for SM and 3.70% for myoblasts at later stages. Cellular automata modeling suggested an enhanced role for cell-to-cell transmission in driving the heightened parasitism observed in CM. The increased late-stage susceptibility of CM, potentially mediated by cell-to-cell transfer mechanisms of the parasite, aligns with reported clinical tropism patterns. The myotube infection models provide novel insights into Chagas disease pathogenesis that are not fully attainable through in vivo examination alone. Expanding knowledge in this area could aid therapeutic development for this neglected illness.