Restricted physical activity after volumetric muscle loss alters whole-body and local muscle metabolism.

Volumetric muscle loss (VML) is the traumatic loss of skeletal muscle, resulting in chronic functional deficits and pathological comorbidities, including altered whole-body metabolic rate and respiratory exchange ratio (RER), despite no change in physical activity in animal models. In other injury models, treatment with ß2 receptor agonists (e.g. formoterol) improves metabolic and skeletal muscle function. We aimed first to examine if restricting physical activity following injury affects metabolic and skeletal muscle function, and second, to enhance the metabolic and contractile function of the muscle remaining following VML injury through treatment with formoterol. Adult male C57Bl/6J mice (n = 32) underwent VML injury to the posterior hindlimb compartment and were randomly assigned to unrestricted or restricted activity and formoterol treatment or no treatment; age-matched injury naïve mice (n = 4) were controls for biochemical analyses. Longitudinal 24 h evaluations of physical activity and whole-body metabolism were conducted following VML. In vivo muscle function was assessed terminally, and muscles were biochemically evaluated for protein expression, mitochondrial enzyme activity and untargeted metabolomics. Restricting activity chronically after VML had the greatest effect on physical activity and RER, reflected in reduced lipid oxidation, although changes were attenuated by formoterol treatment. Formoterol enhanced injured muscle mass, while mitigating functional deficits. These novel findings indicate physical activity restriction may recapitulate following VML clinically, and adjunctive oxidative treatment may create a metabolically beneficial intramuscular environment while enhancing the injured muscle's mass and force-producing capacity. Further investigation is needed to evaluate adjunctive oxidative treatment with rehabilitation, which may augment the muscle's regenerative and functional capacity following VML. KEY POINTS: The natural ability of skeletal muscle to regenerate and recover function is lost following complex traumatic musculoskeletal injury, such as volumetric muscle loss (VML), and physical inactivity following VML may incur additional deleterious consequences for muscle and metabolic health. Modelling VML injury-induced physical activity restriction altered whole-body metabolism, primarily by decreasing lipid oxidation, while preserving local skeletal muscle metabolic activity. The ß2 adrenergic receptor agonist formoterol has shown promise in other severe injury models to improve regeneration, recover function and enhance metabolism. Treatment with formoterol enhanced mass of the injured muscle and whole-body metabolism while mitigating functional deficits resulting from injury. Understanding of chronic effects of the clinically available and FDA-approved pharmaceutical formoterol could be a translational option to support muscle function after VML injury.

Early initiation of electrical stimulation paired with range of motion after a volumetric muscle loss injury does not benefit muscle function.

NEW FINDINGS: What is the central question of this study? First, how does physical rehabilitation influence recovery from traumatic muscle injury? Second, how does physical activity impact the rehabilitation response for skeletal muscle function and whole-body metabolism? What is the main finding and its importance? The most salient findings were that rehabilitation impaired muscle function and range of motion, while restricting activity mitigated some negative effects but also impacted whole-body metabolism. These data suggest that first, work must continue to explore treatment parameters, including modality, time, type, duration and intensity, to find the best rehabilitation approaches for volumetric muscle loss injuries; and second, restricting activity acutely might enhance rehabilitation response, but whole-body co-morbidities should continue to be considered. ABSTRACT: Volumetric muscle loss (VML) injury occurs when a substantial volume of muscle is lost by surgical removal or trauma, resulting in an irrecoverable deficit in muscle function. Recently, it was suggested that VML impacts whole-body and muscle-specific metabolism, which might contribute to the inability of the muscle to respond to treatments such as physical rehabilitation. The aim of this work was to understand the complex relationship between physical activity and the response to rehabilitation after VML in an animal model, evaluating the rehabilitation response by measurement of muscle function and whole-body metabolism. Adult male mice (n = 24) underwent a multi-muscle, full-thickness VML injury to the gastrocnemius, soleus and plantaris muscles and were randomized into one of three groups: (1) untreated; (2) rehabilitation (i.e., combined electrical stimulation and range of motion, twice per week, beginning 72 h post-injury, for ∼8 weeks); or (3) rehabilitation and restriction of physical activity. There was a lack of positive adaption associated with electrical stimulation and range of motion intervention alone; however, maximal isometric torque of the posterior muscle group was greater in mice receiving treatment with activity restriction (P = 0.008). Physical activity and whole-body metabolism were measured ∼6 weeks post-injury; metabolic rate decreased (P = 0.001) and respiratory exchange ratio increased (P = 0.022) with activity restriction. Therefore, restricting physical activity might enhance an intervention delivered to the injured muscle group but impair whole-body metabolism. It is possible that restricting activity is important initially post-injury to protect the muscle from excess demand. A gradual increase in activity throughout the course of treatment might optimize muscle function and whole-body metabolism.

Resistance wheel running improves contractile strength, but not metabolic capacity, in a murine model of volumetric muscle loss injury.

The primary objective of this study was to determine if low- or high-resistance voluntary wheel running leads to functional improvements in muscle strength (i.e., isometric and isokinetic torque) and metabolic function (i.e., permeabilized fibre bundle mitochondrial respiration) after a volumetric muscle loss (VML) injury. C57BL/6J mice were randomized into one of four experimental groups at age 12 weeks: uninjured control, VML untreated (VML), low-resistance wheel running (VML-LR) and high-resistance wheel running (VML-HR). All mice, excluding the uninjured, were subject to a unilateral VML injury to the plantar flexor muscles and wheel running began 3 days post-VML. At 8 weeks post-VML, peak isometric torque was greater in uninjured compared to all VML-injured groups, but both VML-LR and VML-HR had greater (∼32%) peak isometric torque compared to VML. All VML-injured groups had less isokinetic torque compared to uninjured, and there was no statistical difference among VML, VML-LR and VML-HR. No differences in cumulative running distance were observed between VML-LR and VML-HR groups. Because adaptations in VML-HR peak isometric torque were attributed to greater gastrocnemius muscle mass, atrophy- and hypertrophy-related protein content and post-translational modifications were explored via immunoblot; however, results were inconclusive. Permeabilized fibre bundle mitochondrial oxygen consumption was 22% greater in uninjured compared to VML, but there was no statistical difference among VML, VML-LR and VML-HR. Furthermore, neither wheel running group demonstrated a change in the relative protein content of the mitochondrial biogenesis transcription factor, peroxisome proliferator-activated receptor γ coactivator 1-α (PGC-1α). These results indicate that resistance wheel running alone only has modest benefits in the VML-injured muscle. NEW FINDINGS: What is the central question of the study? Does initiation of a resistance wheel running regimen following volumetric muscle loss (VML) improve the functional capacity of skeletal muscle? What is the main finding and its importance? Resistance wheel running led to greater muscle mass and strength in mice with a VML injury but did not result in a full recovery. Neither low- nor high-resistance wheel running was associated with a change in permeabilized muscle fibre respiration despite runners having greater whole-body treadmill endurance capacity, suggesting resilience to metabolic adaptations in VML-injured muscle. Resistance wheel running may be a suitable adjuvant rehabilitation strategy, but alone does not fully mitigate VML pathology.

Compromised Muscle Properties in a Severe Hypophosphatasia Murine Model.

Hypophosphatasia (HPP) is a rare metabolic bone disorder characterized by low levels of tissue non-specific alkaline phosphatase (TNAP) that causes under-mineralization of the bone, leading to bone deformity and fractures. In addition, patients often present with chronic muscle pain, reduced muscle strength, and an altered gait. In this work, we explored dynamic muscle function in a homozygous TNAP knockout mouse model of severe juvenile onset HPP. We found a reduction in skeletal muscle size and impairment in a range of isolated muscle contractile properties. Using histological methods, we found that the structure of HPP muscles was similar to healthy muscles in fiber size, actin and myosin structures, as well as the α-tubulin and mitochondria networks. However, HPP mice had significantly fewer embryonic and type I fibers than wild type mice, and fewer metabolically active NADH+ muscle fibers. We then used oxygen respirometry to evaluate mitochondrial function and found that complex I and complex II leak respiration were reduced in HPP mice, but that there was no disruption in efficiency of electron transport in complex I or complex II. In summary, the severe HPP mouse model recapitulates the muscle strength impairment phenotypes observed in human patients. Further exploration of the role of alkaline phosphatase in skeletal muscle could provide insight into mechanisms of muscle weakness in HPP.

Temporal changes in the muscle extracellular matrix due to volumetric muscle loss injury.

PURPOSE/AIM: Volumetric muscle loss (VML) is a devastating orthopedic injury resulting in chronic persistent functional deficits, loss of joint range of motion, pathologic fibrotic deposition and lifelong disability. However, there is only limited mechanistic understanding of VML-induced fibrosis. Herein we examined the temporal changes in the fibrotic deposition at 3, 7, 14, 28, and 48 days post-VML injury. MATERIALS AND METHODS: Adult male Lewis rats (n = 39) underwent a full thickness ~20% (~85 mg) VML injury to the tibialis anterior (TA) muscle unilaterally, the contralateral TA muscle served as the control group. All TA muscles were harvested for biochemical and histologic evaluation. RESULTS: The ratio of collagen I/III was decreased at 3, 7, and 14 days post-VML, but significantly increased at 48 days. Decorin content followed an opposite trend, significantly increasing by day 3 before dropping to below control levels by 48 days. Histological evaluation of the defect area indicates a shift from loosely packed collagen at early time points post-VML, to a densely packed fibrotic scar by 48 days. CONCLUSIONS: The shift from early wound healing efforts to a fibrotic scar with densely packed collagen within the skeletal muscle occurs around 21 days after VML injury through dogmatic synchronous reduction of collagen III and increase in collagen I. Thus, there appears to be an early window for therapeutic intervention to prevent pathologic fibrous tissue formation, potentially by targeting CCN2/CTGF or using decorin as a therapeutic.

New Frontiers of Body Composition in Sport.

The body composition phenotype of an athlete displays the complex interaction among genotype, physiological and metabolic demands of a sport, diet, and physical training. Observational studies dominate the literature and describe the sport-specific physique characteristics (size, shape, and composition) of adult athletes by gender and levels of competition. Limited data reveal how body composition measurements can benefit an athlete. Thus, the objective is to identify purposeful measurements of body composition, notably fat and lean muscle masses, and determine their impact on the health and performance of athletes. Areas of interest include relationships among total and regional body composition measurements, muscle function, sport-specific performance, risk of injury, return to sport after injury, and identification of activity-induced fluid shifts. Discussion includes the application of specific uses of dual X-ray absorptiometry and bioelectrical impedance including an emphasis on the need to minimize measurement errors and standardize protocols, and highlights opportunities for future research. This focus on functional body composition can benefit the health and optimize the performance of an athlete.

Anterior Cruciate Ligament Reconstructed Female Athletes Exhibit Relative Muscle Dysfunction After Return to Sport.

We sought to examine the relationship between upper-leg compartmental lean mass, muscle-specific strength, and explosive strength following anterior cruciate ligament reconstruction. Twleve adolescent female athletes with prior anterior cruciate ligament reconstruction were individually-matched by age (16.4±0.9 vs. 16.4±1.0 yrs.), body mass index (23.2±2.1 vs. 23.2±2.7 kg/m2), and sport to 12 female athlete controls. One total-body and 2 lateral-leg dual X-ray absorptiometry scans measured total/segmental body composition. Isokinetic dynamometry measured knee extensor/flexor peak torque. Squat jumps on force platforms measured bilateral peak vertical ground reaction force. Paired t-tests assessed lean mass, peak torque, and force between previously-injured athletes' legs and between previously-injured and control athletes' legs. Previously-injured athletes' involved vs. non-involved leg demonstrated lower total (7.13±0.75 vs. 7.43±0.99 kg; p<0.01) and anterior (1.49±0.27 vs. 1.61±0.23 kg; p<0.01) and posterior (1.90±0.19 vs. 2.02±0.21 kg; p=0.04) upper-leg lean mass. Involved leg peak torque (1.36±0.31; 1.06±0.27; 0.97±0.19 Nm/kg) was lower vs. non-involved leg (1.71±0.36; 1.24±0.33; 1.04±0.15 Nm/kg; p<0.01-0.02) for extension at 60 and 120°/sec and flexion at 60°/sec and vs. controls' 'matched' leg (1.77±0.40 Nm/kg; p=0.01) for extension at 60°/sec. Involved leg force (296±45N) was lower vs. non-involved leg (375±55N; p<0.01) and vs. controls' 'matched' leg (372±88N; p=0.02). One-year post-anterior cruciate ligament reconstruction, adolescent female athletes' involved leg demonstrated relative muscle dysfunction.

Assessing Agreement of Lateral Leg Muscle and Bone Composition Using Dual X-ray Absorptiometry.

BACKGROUND: Recently, a lateral-view dual X-ray absorptiometry (DXA) scanning method for measuring leg total, lean, and fat masses demonstrated accuracy vs the standard whole-body frontal DXA scanning view on the GE Lunar iDXA. The current study examined the lateral scanning method's agreement using a Hologic Horizon A DXA scanner. METHODOLOGY: Thirty healthy college-age participants (16 female; X̅age = 21.5 ± 1.7 yr) received 3 DXA scans (1 whole-body, 2 lateral leg scans) to quantify leg composition in the frontal and lateral plane. To mark regions of interest for postscan analysis, metallic markers were placed at 60% of the length above and below each leg's lateral epicondyle. Using lateral subject positioning, leg composition was measured with participants lying on their right and left sides. Paired t tests examined the lateral DXA scanning method's agreement when quantifying total, fat, and lean masses, bone mineral content, and bone mineral density compared to measurements of equal area in the whole-body frontal scanning view. RESULTS: Comparisons of frontal and lateral view DXA scan measurements for right leg total mass (7.12 ± 0.91kg vs 6.39 ± 0.85kg), fat mass (1.70 ± 0.44kg vs 1.36 ± 0.33kg), lean mass (5.14 ± 1.05kg vs 4.77 ± 0.92kg), bone mineral content (0.28 ± 0.06kg vs 0.23 ± 0.05kg), and bone mineral density (1.39 ± 0.14g/cm2vs 1.36 ± 0.15g/cm2), respectively, were significantly different (p < 0.001-0.028). Similarly, comparisons of frontal and lateral left leg total mass (7.12 ± 0.97kg vs 6.38 ± 0.92kg), fat mass (1.70 ± 0.44kg vs 1.39 ± 0.36kg), lean mass (5.15 ± 1.12kg vs 4.76 ± 0.97kg), bone mineral content (0.28 ± 0.06kg vs 0.24 ± 0.06kg), and bone mineral density (1.39 ± 0.15g/cm2vs 1.36 ± 0.17g/cm2), respectively, were significantly different (p < 0.001-0.046). CONCLUSION: Unlike a previous study in which agreement of lateral vs frontal leg composition measurements of equal area was reported utilizing the GE Lunar iDXA, agreement was not observed using the Hologic Horizon A DXA scanner. Therefore, lateral view assessment may not be reliably performed on DXA scanner models produced by different manufacturers.

Body fat percent assessment between electrical impedance myography and dual X-ray absorptiometry.

OBJECTIVES: To compare total and regional body fat percent (BF%) measurements obtained using a handheld electrical impedance myography (EIM) device in comparison to BF% measured by dual X-ray absorptiometry (DXA). METHODS: Sixty-nine male and female (33 males/36 females; age = 21.9 ± 2.0 years, body mass index = 24.5 ± 3.6 kg/m2 ) college-age individuals participated in this study. Each participant's total and regional (ie, upper arms, upper legs, trunk) BF% was estimated using EIM and DXA. Metallic markers were used to delineate regional boundaries for analysis, including upper arms (biceps/triceps), upper legs (quadriceps/hamstrings), and trunk (abdominal region/low back region). Paired t-tests assessed the accuracy of BF% values estimated from EIM in comparison to BF% measured by DXA. RESULTS: Observations revealed EIM reported significantly lower BF% for upper left arm (P < .001), upper right arm (P < .001), upper right leg (P = .002), and trunk (P < .001) values. However, no significant differences were observed in total (P = .434) and upper left leg (P = .855) BF% between the two devices. CONCLUSIONS: This study's observations suggest that, EIM may be an accurate field method for measuring total BF%, but not regional BF%.

Total and Regional Body Composition of NCAA Division I Collegiate Basketball Athletes.

This study aimed to examine body composition using dual X-ray absorptiometry (DXA) in male and female NCAA Division I collegiate basketball athletes. Two-hundred ten (male [M]/female [F]=88/122) basketball athletes' total and regional fat mass, lean mass, bone mineral density, and visceral adipose tissue were measured. Athletes were classified as: point guards (M/F=27/34), shooting guards (M/F=18/27), small forwards (M/F=13/18), power forwards (M/F=21/27), and centers (M/F=9/16). ANOVA and Tukey's HSD assessed positional differences by sex. In males, centers and power forwards had greater total fat mass (p<0.025), lean mass (p≤0.001), and visceral adipose tissue (p<0.001) than other positions. Male centers had greater arm and leg fat mass and lean mass than point guards, shooting guards, and small forwards (p≤0.049), and greater arm bone mineral density than point guards (p=0.015). In females, centers had greater total fat mass (p<0.001) vs. other positions and greater total lean mass, arm fat and lean masses, arm and leg bone mineral density, and visceral adipose tissue vs. point guards and shooting guards (p≤0.005). Female point guards had lower total bone mineral density than power forwards (p=0.008). In conclusion, these sex- and position-specific total and regional body composition measurements in collegiate basketball players provide population-specific normative data.

Association of Compartmental Leg Lean Mass Measured by Dual X-Ray Absorptiometry With Force Production.

Raymond-Pope, CJ, Dengel, DR, Fitzgerald, JS, and Bosch, TA. Association of compartmental leg lean mass measured by dual X-ray absorptiometry with force production. J Strength Cond Res 34(6): 1690-1699, 2020-We recently reported a novel method for measuring upper leg anterior/posterior compartmental composition. The purpose of this study was to determine the association of this method with measures of muscle-specific and explosive strength and to compare this method with traditional dual energy X-ray absorptiometry (DXA) measurements of total and upper leg masses. We hypothesize this method will be related to muscle-specific strength measured by isokinetic dynamometry and explosive strength measured by jump mechanography. Nineteen NCAA Division I college athletes (10 women; age = 20.4 ± 1.4 years; height = 1.8 ± 0.1 m; body mass = 73.8 ± 17.0 kg) underwent 3 DXA scans (1 total body, 2 lateral) and knee extension/flexion strength assessment using isokinetic dynamometry at 3 velocities (60, 120, and 180°·s). A subset of 10 participants also completed a squat jump on a force platform on a different day. Pearson correlations compared 3 separate lean soft-tissue mass (LSTM) regions of interest (total leg, upper leg, and compartmental leg) with (a) isokinetic peak torque and (b) squat jump height, peak force, and peak and average rate of force development. Compartmental leg LSTM demonstrated similar correlations (r = 0.437-0.835) with peak torque in comparison with total leg (r = 0.463-0.803) and upper leg (r = 0.449-0.795) LSTM. Summed right and left total leg (r = 0.830-0.940), total upper leg (r = 0.824-0.953), and anterior (r = 0.582-0.798) and posterior (r = 0.750-0.951) compartmental leg LSTM demonstrated moderate-to-strong correlations with all squat jump variables, particularly jump height (p < 0.05). The lateral segmentation DXA scanning method demonstrated feasibility in assessing compartmental leg LSTM in relation with isokinetic and squat jump measurements-important outcomes when examining an athlete's response to training and rehabilitation.

Body Composition and Visceral Adipose Tissue in Female Collegiate Equestrian Athletes.

To examine measures of total and regional body composition using dual X-ray absorptiometry (DXA) in NCAA Division I collegiate equestrian athletes, 31 female collegiate equestrian athletes were matched to a population of normal controls by age and body mass index. Total and regional fat tissue mass (FM), lean tissue mass (LM), bone mineral density (BMD), and abdominal visceral adipose tissue (VAT) were measured by DXA. Equestrian athletes had a significantly (p=0.03) lower total body fat percentage (%fat) than controls. There were no significant differences in total LM and VAT between equestrian athletes and controls. However, equestrian athletes, when compared to the controls, had significantly lower leg %fat, leg FM and higher leg LM. The greater leg LM in equestrian riders resulted in a smaller upper to lower body LM ratio compared to controls. There was no difference in leg BMD between equestrian athletes and controls. There were no significant differences between the 2 styles of riding (i. e., hunt seat and western style) in regards to body composition. The lower total %fat in equestrian athletes seems to be influenced by differences in leg composition, with equestrian athletes having significantly more LM and less FM.

Total and Regional Body Composition of NCAA Division I Collegiate Baseball Athletes.

This study's purpose was to evaluate total, regional, and throwing versus non-throwing arm body composition measurements between various positions of NCAA Division I male baseball players using dual X-ray absorptiometry (DXA). Two hundred and one collegiate baseball athletes were measured using DXA. Visceral adipose tissue (VAT), total and regional fat mass (FM), lean mass (LM), and bone mineral density (BMD) were measured. Athletes were separated into: pitchers (n=92), catchers (n=25), outfielders (n=43), and infielders (n=41). ANOVA and Tukey's honest significant difference assessed total and regional differences between positions. Infielders had significantly (p<0.05) lower total LM than pitchers and outfielders. Additionally, outfielders had significantly lower total FM compared to pitchers and catchers. No significant differences between positions were observed for total BMD and VAT. Pitchers' and infielders' throwing arm demonstrated significantly greater total mass, FM, LM, and BMD compared to the non-throwing arm. Further, outfielders' throwing arm total mass, LM, and BMD were significantly higher vs. the non-throwing arm. Significant differences were observed in total and regional body composition measurements across position, in addition to differences in throwing arm vs. non-throwing arm composition. These measurement values are important to coaches and trainers as normative positional DXA data for collegiate baseball players.

Total and Regional Body Composition of NCAA Division I Collegiate Female Softball Athletes.

The purpose of this study was to evaluate total, regional, and throwing versus non-throwing arm body composition measures across the 4 major positions of NCAA Division I female softball players using dual X-ray absorptiometry (DXA) (n=128). Total and regional total mass (TM), fat mass (FM), lean mass (LM), bone mineral density (BMD), bone mineral content (BMC), and visceral adipose tissue were measured. Athletes were separated into: pitchers (n=32), catchers (n=13), outfielders (n=39), and infielders (n=44). ANOVA and Tukey's HSD assessed total and regional differences between positions. Although no significant total or regional LM differences were observed across positions, outfielders had significantly (p=0.006-0.047) lower total-body, arm, and trunk TM and FM, leg FM, and leg BMC in comparison to pitchers. The throwing arm had significantly (p<0.0001-0.018) greater LM, BMD, and BMC than the non-throwing arm for all positions. Notably, there were minimal body composition differences among softball positions, with the primary differences being that pitchers had larger total and regional fat values than outfielders. The throwing arm of all positions had greater LM, BMD, and BMC than the non-throwing arm. These values can be used by coaches and trainers as descriptive DXA data for collegiate softball players.

Exploring skeletal muscle tolerance and whole-body metabolic effects of FDA-approved drugs in a volumetric muscle loss model.

Volumetric muscle loss (VML) is associated with persistent functional impairment due to a lack of de novo muscle regeneration. As mechanisms driving the lack of regeneration continue to be established, adjunctive pharmaceuticals to address the pathophysiology of the remaining muscle may offer partial remediation. Studies were designed to evaluate the tolerance and efficacy of two FDA-approved pharmaceutical modalities to address the pathophysiology of the remaining muscle tissue after VML injury: (1) nintedanib (an anti-fibrotic) and (2) combined formoterol and leucine (myogenic promoters). Tolerance was first established by testing low- and high-dosage effects on uninjured skeletal muscle mass and myofiber cross-sectional area in adult male C57BL/6J mice. Next, tolerated doses of the two pharmaceutical modalities were tested in VML-injured adult male C57BL/6J mice after an 8-week treatment period for their ability to modulate muscle strength and whole-body metabolism. The most salient findings indicate that formoterol plus leucine mitigated the loss in muscle mass, myofiber number, whole-body lipid oxidation, and muscle strength, and resulted in a higher whole-body metabolic rate (p ≤ 0.016); nintedanib did not exacerbate or correct aspects of the muscle pathophysiology after VML. This supports ongoing optimization efforts, including scale-up evaluations of formoterol treatment in large animal models of VML.

Medial Gastrocnemius Muscle Properties of Children With Cerebral Palsy After Different Tone Treatments: A Pilot Study.

OBJECTIVE: Spasticity in children with cerebral palsy can be managed by a spectrum of approaches, from conservative therapy, to temporary botulinum toxin A injections, to permanent transection of sensory nerves with a selective dorsal rhizotomy. This pilot study investigated whether these three tone management approaches are associated with histological and biochemical properties of the medial gastrocnemius. DESIGN: A convenience sample of children with cerebral palsy undergoing gastrocnemius lengthening surgery was enrolled. Intraoperative biopsies were obtained from three individuals (one each: minimal tone treatment; frequent gastrocnemius botulinum toxin A injections; previous selective dorsal rhizotomy). All individuals had plantarflexor contractures, weakness, and impaired motor control before the biopsy. RESULTS: Differences between participants were observed for muscle fiber cross-sectional area, fiber type, lipid content, satellite cell density, and centrally located nuclei. The most pronounced difference was the abundance of centrally located nuclei in the botulinum toxin A participants (52%) compared with the others (3-5%). Capillary density, collagen area and content, and muscle protein content were similar across participants. CONCLUSIONS: Several muscle properties seemed to deviate from reported norms, although age- and muscle-specific references are sparse. Prospective studies are necessary to distinguish cause and effect and to refine the risks and benefits of these treatment options.

Response of terminal Schwann cells following volumetric muscle loss injury.

An often-overlooked component of traumatic skeletal muscle injuries is the impact on the nervous system and resultant innervation of the affected muscles. Recent work in a rodent model of volumetric muscle loss (VML) injury demonstrated a progressive, secondary loss of neuromuscular junction (NMJ) innervation, supporting a role of NMJ dysregulation in chronic functional deficits. Terminal Schwann cells (tSCs) are known to be vital for the maintenance of NMJ structure and function, in addition to guiding repair and regeneration after injury. However, the tSC response to a traumatic muscle injury such as VML is not known. Thus, a study was conducted to investigate the effect of VML on tSC morphological characteristics and neurotrophic signaling proteins in adult male Lewis rats that underwent VML injury to the tibialis anterior muscle using a temporal design with outcome assessments at 3, 7, 14, 21, and 48 days post-injury. The following salient observations were made; first, although there is a loss of innervation over time, the number of tSCs per NMJ increases, significantly so at 48 days post-injury compared to control. The degree of NMJ fragmentation was positively correlated with tSC number after injury. Moreover, neurotrophic factors such as NRG1 and BDNF are elevated after injury through at least 48 days. These results were unanticipated and in contrast to neurodegenerative disease models, in which there is a reduction in tSC number that precedes denervation. However, we found that while there are more tSCs per NMJ after injury, they cover a significantly smaller percent of the post-synaptic endplate area compared to control. These findings support a sustained increase in neurotrophic activity and tSC number after VML, which is a maladaptive response occurring in parallel to other aspects of the VML injury, such as over-accumulation of collagen and aberrant inflammatory signaling.

Independent of physical activity, volumetric muscle loss injury in a murine model impairs whole-body metabolism.

Volumetric muscle loss (VML) injuries result in a non-recoverable loss of muscle tissue and function due to trauma or surgery. Reductions in physical activity increase the risk of metabolic comorbidities over time, and it is likely that VML may reduce whole-body activity. However, these aspects remain uncharacterized following injury. Our goal was to characterize the impact of VML on whole-body physical activity and metabolism, and to further investigate possible muscle-specific metabolic changes. Adult male C57Bl/6J (n = 28) mice underwent a standardized VML injury to the posterior compartment of the hind limb, or served as injury naïve controls. Mice underwent longitudinal evaluation of whole-body physical activity and metabolism in specialized cages up to three times over the course of 8 weeks. At terminal time points of 4- and 8-weeks post-VML in vivo muscle function of the posterior compartment was evaluated. Additionally, the gastrocnemius muscle was collected to understand histological and biochemical changes in the muscle remaining after VML. The VML injury did not alter the physical activity of mice. However, there was a noted reduction in whole-body metabolism and diurnal fluctuations between lipid and carbohydrate oxidation were also reduced, largely driven by lower carbohydrate utilization during active hours. Following VML, muscle-specific changes indicate a decreased proportion of fast (i.e., type IIb and IIx) and a greater proportion of slow (i.e., type I and IIa) fibers. However, there were minimal changes in the capillarity and metabolic biochemical activity properties of the gastrocnemius muscle, suggesting a miss-match in capacity to support the physiologic needs of the fibers. These novel findings indicate that following VML, independent of changes in physical activity, there is whole-body diurnal metabolic inflexibility. Supporting future investigations into the chronic and overlooked co-morbidities of VML injury.