The Impact of Bed Rest, Aging and NMES on Skeletal Muscle

Study Overview
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Brief Summary
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Loss of muscle mass is common phenotypic trait of muscular disuse and ageing. The loss of muscle mass affects, among others, the ability to maintain homeostasis of glucose metabolism and the energy reservoir in catabolic conditions, while also affecting mechanical muscle function which can cause detrimental impairments in general functional status and hence quality of life. However, a limited amount of research has attempted to elucidate molecular regulators of muscle mass loss following bed rest in older individuals and across genders. Consequently, the mechanistic drivers are unresolved and there are currently no effective therapeutic strategies to counteract muscle wasting and loss of function in individuals submitted to bed rest e.g. during hospitalization. Purpose The purpose is to examine the effects of 5 days of bed rest on muscle mass, including myofibrillar protein synthesis and breakdown, and muscle function, and elucidate molecular regulators of muscle mass loss and metabolic pathways, while also investigating if potential negative effects can be counteracted by daily NeuroMuscular Electrical Stimulation (NMES) across different age and genders. Methods The study is designed as a randomized controlled cross-over 5-day bed rest study including a group of healthy young (18-30 years) and healthy old (65-80 years) men and women. Participants will receive daily electrical stimulation (NMES) of the thigh muscles (30 min x 3/day) on one leg (ES), while the other leg serves as a control (CON). Participants will be tested at baseline (pre) and after (post) intervention for muscle strength, muscle power, balance, and muscle activation. Blood samples are collected at several time points and muscle biopsies are sampled pre- and post-intervention along with assessment of whole-body muscle mass and thigh muscle mass. Scientific exposition The results from the study can potentially provide insight into the adaptive mechanisms associated with NMES training and muscular disuse on both cellular- and whole-body level. The understanding of the underlying mechanisms is crucial for the application of NMES in a therapeutic context and will furthermore help us understand the basic mechanism regulating the skeletal muscle mass during both training and muscular disuse. Overall, the results can potentially help establishing treatments to counteract loss of muscle mass, muscle function and muscle health during periods of muscular disuse.

Official Title
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The Impact of Bed Rest and Aging on Muscle Mass and Muscle Function: Effects of Neuromuscular Electrical Stimulation

Conditions
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Myofibrillar Protein Synthesis, Disuse Atrophy (Muscle) of Lower Extremities, Muscle Function, Neuromuscular Electrical Stimulation

Intervention / Treatment
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* Other: Bed rest
* Other: Neuromuscular electrical stimulation


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Healthy Age between 18-30 or 65-80 years Injury free in the lower extremities (No previous or current knee injuries or knee pain) Normal weight Consumes normal diet Exclusion Criteria: Cognitive impairment affecting the ability to participate in the study. Health related contraindications to participating in the intervention (i.e., bed rest and/or NMES), such as eczema and rash on the lower extremities Smoker Obesity Not able to speak or understand Danish. Acute or chronic diseases such as diabetes, cancer, embolism, infection, cardio-vascular diseases Use of medication which affects myofibrillar protein synthesis or the skeletal muscle tissue Use of other medication (e.g. anticoagulants, adrenal cortex hormone [within the last 3 months] etc.) Previous or current use of anabolic steroids Previous participation in research trials involving deuterium oxide or another stable isotope tracer

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 80 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Crossover Assignment
Interventional Model Description: Participants are subjected to 5 days bed rest. One leg will receive 3/daily neuromuscular electrical stimulation. The contralateral leg will serve as control-leg undergo disuse only.
Masking: Single


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Bed rest- Control<br>One leg will be subjected to disuse by bed rest and will not receive further treatment during the bed rest period. | Other: Bed rest<br>* 5 days of strict bed rest<br>|
| Experimental: Bed rest + NMES<br>One leg will be subjected to disuse by bed rest and will in addition receive neuromuscular electrical stimulation of the quadriceps muscle 3 times/day. | Other: Bed rest<br>* 5 days of strict bed rest<br>Other: Neuromuscular electrical stimulation<br>* Unilateral neuromuscular electrical stimulation (m. Quadriceps)<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in Myofiber cross-sectional area | Histochemical analysis of type I and type II myofiber cross-sectional area | Change from baseline after bed rest intervention | 
| Assessment of myofibrillar protein synthesis | Quantification of myofibrillar protein synthesis using the stable-isotope amino acid tracer deuterium oxide (D2O) | Assessed during the period from pre-intervention biopsies (day 0, first day of bed rest) to post-intervention biopsies (day 5, last day and cessation of bed rest) | 
| Change in maximal isometric muscle strength and superimposed twitch | Maximal isometric voluntary quadriceps strength combined with the superimposed twitch technique to assess maximal strength and voluntary muscle activation | Change from baseline after bed rest intervention | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in total Akt protein assessed by Western blot | Assessment of Akt protein by Western Blot analysis using muscle tissue from vastus lateralis | Change from baseline after bed rest intervention | 
| Change in total mTOR protein assessed by Western blot | Assessment of mTOR protein by Western Blot analysis using muscle tissue from vastus lateralis | Change from baseline after bed rest intervention | 
| Change in total MuRF-1 protein assessed by Western blot | Assessment of MuRF-1 protein by Western Blot analysis using muscle tissue from vastus lateralis | Change from baseline after bed rest intervention | 
| Change in total Atrogin-1 protein assessed by Western blot | Assessment of Atrogin-1 protein by Western Blot analysis using muscle tissue from vastus lateralis | Change from baseline after bed rest intervention | 
| Change in total myostatin protein assessed by Western blot | Assessment of myostatin protein by Western Blot analysis using muscle tissue from vastus lateralis | Change from baseline after bed rest intervention | 
| Change in quadriceps muscle morphology and architecture by ultrasound scan | Ultrasound scan of rectus femoris and vastus lateralis muscle thickness and of vastus lateralis pennation angle | Change from baseline after bed rest intervention | 
| Change in body composition by DEXA scan | Assessment of whole body and regional lean mass and fat | Change from baseline after bed rest intervention | 
| Change in leg extensor power | Muscle power of the lower extremities assessed using the Nottingham power rig | Change from baseline after bed rest intervention | 
| Change in sway - postural balance | Measurement of displacement of center of pressure during unilateral and bilateral stance | Change from baseline after bed rest intervention | 
| Change in triglycerides | Fasting blood samples are collected for analysis of triglycerides | Day 0, day 2, day 4 and day 5 | 
| Change in cholesterol | Fasting blood samples are collected for later analysis of cholesterol | Day 0, day 2, day 4 and day 5 | 
| Change in C-reactive protein (CRP) | Fasting blood samples are collected for analysis of CRP values | Day 0, day 2, day 4 and day 5 |