In Sequence Anti-Fibrotic Treatment and Rehabilitation following Volumetric Muscle Loss Injury.

Objective: Mitigation of local pathologic fibrotic tissue deposition is a target area of interest for volumetric muscle loss (VML); nintedanib has shown promise for reduction of fibrosis following VML. Herein studies investigate how in sequence anti-fibrotic treatment administered immediately following VML and delayed rehabilitation could improve functional recovery after VML. Approach: Adult male C57BL/6 mice (n=36) were VML injured or Naïve, and randomly assigned to nintedanib (6 mg/kg/day) for 2 weeks or were left untreated; additionally, mice were given access to a running wheel beginning at 2 weeks until 8 weeks. Terminally, mice underwent maximal in vivo functional testing in addition to quantification of muscle collagen content and fibrotic and myogenic markers. Results: Daily running distances (p=0.17) were similar across groups, but weekly averages were greatest in the VML anti-fibrotic group (p<0.01). As expected, 2 weeks post-VML, all VML-injured mice had lower maximal torque normalized to body and muscle mass than Naïve. By 8 weeks, running alone after VML did not recover function, but mice that received the anti-fibrotic treatment prior to running, had greater torque than those untreated (p<0.01), with functional measurements similar to naïve muscle thar ran, indicating improved functional recovery. Innovation: The ability to translate current FDA-approved pharmaceuticals, in a repurposing approach, is critical to mitigate the pathophysiologic consequences of VML in support of functional recovery. However, foundational and translational studies are still needed to understand feasibility and efficacy. Conclusions: Early prevention of fibrotic tissue deposition supports improvements in muscle quality and force chronically after VML injury.

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.

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.

The Effect of Direct and Remote Postexercise Ischemic Conditioning on Muscle Soreness and Strength 24 Hours After Eccentric Drop Jumps.

ABSTRACT: Lillquist, T, Mahoney, SJ, Kotarsky, C, McGrath, R, Jarajapu, Y, Scholten, SD, and Hackney, KJ. The effect of direct and remote postexercise ischemic conditioning on muscle soreness and strength 24 hours after eccentric drop jumps. J Strength Cond Res 37(9): 1870-1876, 2023-Strategic limb occlusion applied after exercise may facilitate recovery, not only in directly targeted tissue but also in remote areas of the body. The purpose of this study was to determine if postexercise ischemic conditioning (PEIC) applied directly to one leg facilitated recovery in the targeted leg and the contralateral leg that did not receive direct PEIC. Twenty active men participated in a single-blind, randomized, crossover design. Subjects completed 2 paired testing sessions (PEIC and control-SHAM) that included pre-assessments and 24-hour postassessments. Each paired testing session included an eccentric drop jump task, which has been shown to increase lower-body muscle soreness and decrease strength. After each drop jump task, occlusion cuffs were immediately applied. In the PEIC session, ∼198 mm Hg was applied directly to one leg (PEIC-Direct), whereas the contralateral leg received a nonphysiological stimuli of 20 mm Hg (PEIC-Remote). In the control-SHAM session, both legs directly and remotely received the 20 mm Hg pressure. Unilateral pre-assessments and 24-hour postassessments included muscle soreness using a visual analog scale and strength via peak torque assessment across the force-velocity spectrum (flexion/extension 60/60, 120/120, 180/180, 240/240, 300/300 °·s -1 ), and a maximal eccentric extension (30/30 °·s -1 ). Muscle soreness was significantly increased ( p < 0.05) at 24 hours compared with pretreatment except for PEIC-Direct (1.19 ± 0.78 vs. 2.32 ± 1.48, p = 0.096). Across the force-velocity spectrum, there were no significant differences observed between any associated pretest and posttest ( p > 0.05). PEIC applied directly to target leg after eccentric drop jumps attenuated perceived quadriceps muscle soreness 24 hours post; however, there was no effect on muscle strength.

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.