Combined OVX and Concurrent Mechanical Disuse Induced Osteocytes Morphological Alteration, and Mitigation by Mechanobiology and Sclerostin Antibody

Osteoporosis and osteopenia are major health issues that mainly affect elderly people,women after menopause and immobilized patients.Our previous studies have proved that sclerostin antibody(Scl-Ab)can dramatically enhance bone formation and reduce bone resorption in a severe osteoporosis rat model with the combination of ovariectomy(OVX)and hindlimb immobilization(HLS).However,the mechanism in the cellular level is unclear.The objective of this study is to assess the effect of Scl-Ab on osteocytic morphology change in a combined OVX and HLS rat model via quantification of long-and short-axis and the ratio and osteocyte volume in midshaft cortical bone.Four-month-old virgin female SD rats were divided into 7 groups(n=11 per group):Sham+Veh,Sham+HLS+Veh,Sham+HLS+Scl-Ab,OVX+Veh,OVX+Scl-Ab,OVX+HLS+Veh,OVX+HLS+Scl-Ab.HLS was performed 2 weeks after sham or OVX surgery;and treatment was initiated at the time of HLS.Scl-Ab(25 mg/kg)or vehicle was subcutaneously injected twice per week for 5 weeks.Femurs were harvested at the end of study and embedded in PMMA and polished for SEM imaging.Cortical bone mid shaft osteocyte number per bone area was quantified under 1K magnification;the ratios between long axis and short axis of osteocytes were quantified under 2K magnification;osteocyte dendrite number and surface area were quantified under 5K magnification.Osteocyte dendrites width was quantified using 10K magnification.All the quantification was done by ImageJ.We have reported that multiple morphological and structural changes in osteocytes,including a decreased osteocyte density and reduced osteocyte dendrite number in HLS,OVX or the combination group and Scl-Ab’s ability to abolish these unfavorable alterations.We continued our SEM analysis on osteocytes and discovered that the oval shape of osteocyte under HLS,OVX or HLS+OVX has been distorted toward a spindle-like shape,with relatively longer long axis and shorter short axis,assuming osteocyte has a perfect spheroid shape.The ratio between long-and short-axis showed an increased trend in OVX and HLS condition,but Scl-Ab inhibited these increases(P<0.001,P<0.01,respectively).The volume decreased in HLS,OVX group,but Scl-Ab maintained osteocytes’volume in HLS condition(P<0.001).It indicates that cortical bone responds to HLS and/or OVX and Scl-Ab treatment via multiple cellular mechanisms,including density of osteocyte,dendrite number and osteocyte shape.The change of osteocyte shape may imply an altered cytoskeleton system within osteocyte and a subsequent disruption of mechanosensing ability for osteocyte,which lead to bone loss macroscopically.These data suggest Scl-Ab’s therapeutic potential could be related with its ability to maintain osteocyte’s morphologic and structural changes induced by OVX,HLS or both.

Numerical simulation on the adaptation of forms in trabecular bone to mechanical disuse and basic multi-cellular unit activation threshold at menopause

The objective of this paper is to identify the effects of mechanical disuse and basic multi-cellular unit （BMU） activation threshold on the form of trabecular bone during menopause. A bone adaptation model with mechanical- biological factors at BMU level was integrated with finite element analysis to simulate the changes of trabecular bone structure during menopause. Mechanical disuse and changes in the BMU activation threshold were applied to the model for the period from 4 years before to 4 years after menopause. The changes in bone volume fraction, trabecular thickness and fractal dimension of the trabecular structures were used to quantify the changes of trabecular bone in three different cases associated with mechanical disuse and BMU activation threshold. It was found that the changes in the simulated bone volume fraction were highly correlated and consistent with clinical data, and that the trabecular thickness reduced signi-ficantly during menopause and was highly linearly correlated with the bone volume fraction, and that the change trend of fractal dimension of the simulated trabecular structure was in correspondence with clinical observations. The numerical simulation in this paper may help to better understand the relationship between the bone morphology and the mecha-nical, as well as biological environment; and can provide a quantitative computational model and methodology for the numerical simulation of the bone structural morphological changes caused by the mechanical environment, and/or the biological environment.

Depletion of Glial Cell Line-Derived Neurotrophic Factor by Disuse Muscle Atrophy Exacerbates the Degeneration of Alpha Motor Neurons in Caudal Regions Remote from the Spinal Cord Injury

We have been previously reported that disuse muscle atrophy exacerbates both motor neuron (MN) degeneration in caudal regions remote from a spinal cord injury, and decrease in glial cell line-derived neurotrophic factor (GDNF) protein level in paralyzed muscle. In this study we found that disuse muscle atrophy exacerbated the decrease in GDNF protein level in the L4/5 spinal cord, which was not immunopositive for GDNF. Our results were consistent with the fact that in the lumbar spinal cord of rats with mid-thoracic contusion, GDNF expression was not detected, while expression of GDNF receptors (GFRα1 and RET) was. Our study showed that administration of exogenous recombinant GDNF into the atrophic muscle partially rescued α-MN degeneration in the L4/5 spinal cord. These results suggest that the depletion of GDNF protein by muscle atrophy exacerbates α-MN degeneration in caudal regions remote from the injury.

Lower limb suspension induces threshold-specific alterations of motor units properties that are reversed by active recovery

Purpose:This study aimed to non-invasively test the hypothesis that(a) short-term lower limb unloading would induce changes in the neural control of force production(based on motor units(MUs) properties) in the vastus lateralis muscle and(b) possible changes are reversed by active recovery(AR).Methods:Ten young males underwent 10 days of unilateral lower limb suspension(ULLS) followed by 21 days of AR.During ULLS,participants walked exclusively on crutches with the dominant leg suspended in a slightly flexed position(15°-20°) and with the contralateral foot raised by an elevated shoe.The AR was based on resistance exercise(leg press and leg extension) and executed at 70% of each participant’s 1repetition maximum,3 times/week.Maximal voluntary isometric contraction(MVC) of knee extensors and MUs properties of the vastus lateralis muscle were measured at baseline,after ULLS,and after AR.MUs were identified using high-density electromyography during trapezoidal isometric contractions at 10%,25%,and 50% of the current MVC,and individual MUs were tracked across the 3 data collection points.Results:We identified 1428 unique MUs,and 270 of them(18.9%) were accurately tracked.After ULLS,MVC decreased by 29.77%,MUs absolute recruitment/derecruitment thresholds were reduced at all contraction intensities(with changes between the 2 variables strongly correlated),while discharge rate was reduced at 10% and 25% but not at 50% MVC.Impaired MVC and MUs properties fully recovered to baseline levels after AR.Similar changes were observed in the pool of total as well as tracked MUs.Conclusion:Our novel results demonstrate,non-invasively,that 10 days of ULLS affected neural control predominantly by altering the discharge rate of lower-threshold but not of higher-threshold MUs,suggesting a preferential impact of disuse on motoneurons with a lower depolarization threshold.However,after 21 days of AR,the impaired MUs properties were fully restored to baseline levels,highlighting the plasticity of the components involved in neural control.

Effects of blood flow restriction without additional exercise on strength reductions and muscular atrophy following immobilization:A systematic review

Purpose:To investigate whether blood flow restriction(BFR) without concomitant exercise mitigated strength reduction and atrophy of thigh muscles in subjects under immobilization for lower limbs.Methods:The following databases were searched:PubMed,CINAHL,PEDro,Web of Science,Central,and Scopus.Results:The search identified 3 eligible studies,and the total sample in the identified studies consisted of 38 participants.Isokinetic and isometric torque of the knee flexors and extensors was examined in 2 studies.Cross-sectional area of thigh muscles was evaluated in 1 study,and thigh girth was measured in 2 studies.The BFR protocol was 5 sets of 5 min of occlusion and 3 min of free flow,twice daily for approximately 2 weeks.As a whole,the included studies indicate that BFR without exercise is able to minimize strength reduction and muscular atrophy after immobilization.It is crucial to emphasize,however,that the included studies showed a high risk of bias,especially regarding allocation concealment,blinding of outcome assessment,intention-to-treat analyses,and group similarity at baseline.Conclusion:Although potentially useful,the high risk of bias presented by original stodies limits the indication of BFR without concomitant exercise as an effective countermeasure against strength reduction and atrophy mediated by immobilization.

EFFECT OF ANGELICA SINENSIS ON AFFERENT DISCHARGE OF SINGLE MUSCLE SPINDLE IN TO ADS

Objective In drugs for i nvigorating blood circulation, to find a herb that can stimulate afferent discha rge of muscle spindle. Methods A single muscle spindle was isolated from sartorial mus cle of toad. Using air-gap technique, afferent discharge of the muscle spindle was recorded. Effects of Angelica Sinensis, Salvia Miltiorrhiza, and Safflower o n afferent discharge of the muscle spindle were observed. Results Angelica Sinensis could distinctly increase afferent di scharge frequency of the muscle spindle, and this increase was dose-dependent. But Salvia Miltiorrhiza and Safflower had no this excitatory effect. Conclusion It is known that Angelica Sinensis can invigorate bl ood circulation, and we have found its excitatory effect on muscle spindle which makes it possible to serve people with muscle atrophy if more evidences from cl inical experiments are available.

Hypo-activity induced skeletal muscle atrophy and potential nutritional interventions: A review

Periods of hypo-activity result in profound changes in skeletal muscle morphology and strength. This review primarily addresses the differential impact of de-training, bed-rest, limb immobilisation and unilateral lower limb suspension on muscle morphology, strength and fatigability. The degree of muscle atrophy differs depending on the hypo-activity model and the muscles in question, with the leg and postural muscles being the most susceptible to atrophy. Hypo-activity also results in the dramatic loss of strength that often surpasses the loss of muscle mass, and consequently, the nervous system and contractile properties adapt to adjust for this excessive loss of strength. In addition, the degree of muscle strength loss is different depending on the hypo-activity model, with immobilisation appearing to have a greater impact on strength than unloaded models. There is a step-wise difference in the magnitude of muscle loss so that, even after accounting for differential durations of interventions immobilisation ≥ unilateral lower limb suspension ≥ bed-rest ≥ de-training. Muscle fatigability varies between hypoactivity models but the results are equivocal and thismay be due to task-specific adaptations. This review also addresses potential nutritional interventions for attenuating hypo-activity induced muscle atrophy and strength declines, in the absence of exercise. Essential amino acid supplementation stands as a strong candidate but other supplements are good contenders for attenuating hypo-activity induced atrophy and strength losses. Several potential nutritional supplements are highlighted that could be used to combat muscle atrophy but extensive research is needed to determine the most effective.

The mechanisms and treatments of muscular pathological changes in immobilization-induced joint contracture: A literature review

The clinical treatment of joint contracture due to immobilization remains difficult.The pathological changes of muscle tissue caused by immobilization-induced joint contracture include disuse skeletal muscle atrophy and skeletal muscle tissue fibrosis.The proteolytic pathways involved in disuse muscle atrophy include the ubiquitin-proteasome-dependent pathway,caspase system pathway,matrix metalloproteinase pathway,Ca2+-dependent pathway and autophagy-lysosomal pathway.The important biological processes involved in skeletal muscle fibrosis include intermuscular connective tissue thickening caused by transforming growth factor-β1 and an anaerobic environment within the skeletal muscle leading to the induction of hypoxia-inducible factor-1α.This article reviews the progress made in understanding the pathological processes involved in immobilization-induced muscle contracture and the currently available treatments.Understanding the mechanisms involved in immobilization-induced contracture of muscle tissue should facilitate the development of more effective treatment measures for the different mechanisms in the future.

Alterations in renin-angiotensin receptors are not responsible for exercise preconditioning of skeletal muscle fibers

Endurance exercise training promotes a protective phenotype in skeletal muscle known as exercise pre-conditioning.Exercise preconditioning protects muscle fibers against a variety of threats including inactivity-induced muscle atrophy.The mechanism(s)responsible for exercise preconditioning remain unknown and are explored in these experiments.Specifically,we investigated the impact of endurance exercise training on key components of the renin-angiotensin system(RAS).The RAS was targeted because activation of the classical axis of the RAS pathway via angiotensinⅡtypeⅠreceptors(AT1Rs)promotes muscle atrophy whereas activation of the non-classical RAS axis via Mas receptors(MasRs)inhibits the atrophic signaling of the classical RAS pathway.Guided by prior studies,we hypothesized that an exercise-induced decrease in AT1Rs and/or increases in MasRs in skeletal muscle fibers is a potential mechanism responsible for exercise preconditioning.Following endurance exercise training in rats,we examined the abundance of AT1Rs and MasRs in both locomotor and respiratory muscles.Our results indicate that endurance exercise training does not alter the protein abundance of AT1Rs or MasRs in muscle fibers from the diaphragm,plantaris,and soleus muscles compared to sedentary controls(p>0.05).Furthermore,fluorescent angiotensinⅡ(AngⅡ)binding analyses confirm our results that exercise pre-conditioning does not alter the protein abundance of AT1Rs in the diaphragm,plantaris,and soleus(p>0.05).This study confirms that exercise-induced changes in RAS receptors are not a key mechanism that contributes to the beneficial effects of exercise preconditioning in skeletal muscle fibers.