Effect of Creatine+HMB Supplementation in Physically Active Older Adults

NCT05951439 | Completed DMC

• 1 other identifier: IR 24/2023
• Study Type: interventional
• Target: 30
• Locations: 1 country
• Sites: 1

Brief Summary

Sarcopenia (SAR) is the loss of muscle strength and mass caused by aging. It is accompanied by a progressive decline in physical and cognitive abilities, increasing the risk of falls. This loss of muscle mass leads to pathophysiological changes at the neuromuscular and tendon levels as a consequence, among others, of alterations in the balance between protein synthesis and degradation, inflammation (INF), or alterations in the anabolic/catabolic state (ACS). These alterations are caused by oxidative stress (OS), when reactive oxygen species-toxic metabolites produced by cells using oxygen-exceed the defensive capacity of the antioxidant system. Therapeutic strategies to modulate SAR are based on exercise and nutrition programs. Multicomponent physical exercise programs have shown improvements in parameters related to sarcopenia. Likewise, the use of nutritional supplements such as creatine (CRE) has demonstrated improvements in muscle function in older adults. CRE may reduce INF and OS in the general population. On the other hand, beta-hydroxy-beta-methylbutyrate (HMB) also appears to improve muscle function in older adults by promoting myogenesis. However, the effects of these supplements in older adults have only been observed when used in isolation. In this regard, our research team observed that a 10-week regimen combining 3 g/day of CRE + 3 g/day of HMB (CRE-HMB) improved muscle recovery (better ACS) and physical performance in athletes subjected to high muscle wear. However, to date, it has not been determined whether this combination improves muscle function, OS, INF, and ACS in women with SAR. Therefore, the working hypothesis is that the CRE-HMB combination could improve muscle function and physical performance, as well as OS, INF, and ACS in individuals with high muscle loss, such as those with SAR. Thus, a randomized double-blind crossover study is proposed to analyze the effect of 12 weeks-6 weeks of intervention + 3 weeks of washout + 3 weeks of intervention-of co-supplementation with 3 g/day of CRE + 3 g/day of HMB (CRE-HMB), combined with 4 sessions/week of multicomponent physical exercise, on muscle function, OS, INF, and ACS in 40 physically active men and women over 60 years of age. These 40 participants will be divided into two groups of 20 (20 in the placebo group and 20 in the CRE-HMB group). At the assessment points (baseline, at 6 weeks, after the 3-week washout period, and after another 6 weeks), body composition, nutritional intake, muscle strength, and performance tests will be evaluated. Blood samples will also be collected to determine biochemical markers of OS, INF, and ACS. It is expected that CRE-HMB co-supplementation for 6 + 6 weeks, together with multicomponent physical exercise, in physically active older adults will improve muscle strength, muscle mass, and performance. Additionally, improvements in OS, INF, and ACS levels are anticipated.

Conditions

Aging; Sarcopenia; Nutrition, Healthy

Keywords

aging; sarcopenia; nutrition; HMB; creatine; exercise; GAA

Outcome Measures

Primary Outcomes (52)

• Body composition (Day 0 and at 6 weeks of intervention).

  The evaluation of body composition was performed by a dietician-nutritionist of the research group at each control point. Prior to the other tests, participants was weighed and measured by seaside telescopic staghater MOD 220. A bioelectrical impedance analysis (BIA) of body composition was also performed to minimize the effect of hydration status on all measurements. Participants was instructed to refrain from caffeine and alcohol 24 h before, as well as, if performed 12 h before the test according to published guidelines for BIA. To predict the MMEA, a tanita TA-MC580- 1 PLUS impedance meter (Tanita Europe B.V., UK) will be used according to the manufacturer's guidelines. The MMEA was predicted through the Equation of Sergi et al, (80): MMEA (kg) = -3.964 + (0.227 RI) + (0.095 \* body mass) + (1.384 \* sex) + (0.064 \* XC), where RI is the standardized resistance for height and XC is the reactance.

  6 weeks

• Body composition (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  The evaluation of body composition was performed by a dietician-nutritionist of the research group at each control point. Prior to the other tests, participants was weighed and measured by seaside telescopic staghater MOD 220. A bioelectrical impedance analysis (BIA) of body composition was also performed to minimize the effect of hydration status on all measurements. Participants was instructed to refrain from caffeine and alcohol 24 h before, as well as, if performed 12 h before the test according to published guidelines for BIA. To predict the MMEA, a tanita TA-MC580- 1 PLUS impedance meter (Tanita Europe B.V., UK) will be used according to the manufacturer's guidelines. The MMEA was predicted through the Equation of Sergi et al, (80): MMEA (kg) = -3.964 + (0.227 RI) + (0.095 \* body mass) + (1.384 \* sex) + (0.064 \* XC), where RI is the standardized resistance for height and XC is the reactance.

  6 weeks

• Nutritional evaluation (Day 0 and at 6 weeks of intervention).

  The nutritional evaluation was performed by a dietitian-nutritionist of the research group at each control point. The nutritional analysis of the participants was performed using a multidimensional approach that included both food consumption frequency (T1 and T4) and adherence to the Mediterranean diet (T1, T2, T3, T4) and dietary habits (T1 and T4). The participants' dietary data were obtained through surveys and interviews, which provided information on the frequency and quantity of key food consumption. The content of macronutrients (carbohydrates, proteins and fats) and micronutrients (vitamins and minerals) was analysed, as well as the consumption patterns of foods rich in antioxidants, healthy fats and fibre.

  6 weeks

• Nutritional evaluation (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  The nutritional evaluation was performed by a dietitian-nutritionist of the research group at each control point. The nutritional analysis of the participants was performed using a multidimensional approach that included both food consumption frequency (T1 and T4) and adherence to the Mediterranean diet (T1, T2, T3, T4) and dietary habits (T1 and T4). The participants' dietary data were obtained through surveys and interviews, which provided information on the frequency and quantity of key food consumption. The content of macronutrients (carbohydrates, proteins and fats) and micronutrients (vitamins and minerals) was analysed, as well as the consumption patterns of foods rich in antioxidants, healthy fats and fibre.

  6 weeks

• Assessment of muscle strength and performance. Short Physical Performance Battery (Day 0 and at 6 weeks of intervention).

  Short Physical Performance Battery (SPPB): consists of 3 tests to be used to assess lower extremity function following standard protocols: * Balance by balance test. * Muscle strength test in the legs, sit to up test. * Mobility test using gait speed, gait speed test. It is important to follow the sequence of the tests, because if the patient starts to stand and sit, he/she may become fatigued and perform falsely poorly in the other two tests. The average test duration is between 6 and 10 min. Each test will be scored from 0 to 4. A score of 0 represent an inability to complete the test, and a score of 4 represent the highest level of performance. Therefore, total scores (SPPB scores) range from 0 to 12, and a higher SPPB score indicate a higher level of physical fitness.

  6 weeks

• Assessment of muscle strength and performance. Short Physical Performance Battery (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  Short Physical Performance Battery (SPPB): consists of 3 tests to be used to assess lower extremity function following standard protocols: * Balance by balance test. * Muscle strength test in the legs, sit to up test. * Mobility test using gait speed, gait speed test. It is important to follow the sequence of the tests, because if the patient starts to stand and sit, he/she may become fatigued and perform falsely poorly in the other two tests. The average test duration is between 6 and 10 min. Each test will be scored from 0 to 4. A score of 0 represent an inability to complete the test, and a score of 4 represent the highest level of performance. Therefore, total scores (SPPB scores) range from 0 to 12, and a higher SPPB score indicate a higher level of physical fitness.

  6 weeks

• Assessment of muscle strength and performance. Timed-up-and-go (Day 0 and at 6 weeks of intervention).

  The evaluation of muscular strength and physical performance was performed by a strength and conditioning coach graduated in physical activity and sport sciences, which was carried out at each control point. The tests to be performed to the participants was: Timed-up-and-go (TUG): Timed-up-and-go is used to assess mobility, balance and agility. Participants are instructed to get up from a chair without assistance (starting point), walk forward three meters (m), walk back to the chair and sit down again (end of test). They are asked to perform the test at a normal, everyday pace, using marks on the floor to indicate where to turn. The total time used from the beginning to the end of the test is recorded.

  6 weeks

• Assessment of muscle strength and performance. Timed-up-and-go (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  The evaluation of muscular strength and physical performance was performed by a strength and conditioning coach graduated in physical activity and sport sciences, which was carried out at each control point. The tests to be performed to the participants was: Timed-up-and-go (TUG): Timed-up-and-go is used to assess mobility, balance and agility. Participants are instructed to get up from a chair without assistance (starting point), walk forward three meters (m), walk back to the chair and sit down again (end of test). They are asked to perform the test at a normal, everyday pace, using marks on the floor to indicate where to turn. The total time used from the beginning to the end of the test is recorded.

  6 weeks

• Assessment of muscle strength and performance. 400-meter Walk Test Perfomance (Day 0 and at 6 weeks of intervention).

  The evaluation of muscular strength and physical performance was performed by a strength and conditioning coach graduated in physical activity and sport sciences, which will be carried out at each control point. The tests to be performed to the participants was: 400-meter Walk Test Perfomance: measures mobility and cardiovascular fitness through a 400-meter timed walk (min). Cones are placed 20 m apart, around which participants must move 10 times, back and forth, as fast as possible. Only one attempt was allowed.

  6 weeks

• Assessment of muscle strength and performance. 400-meter Walk Test Perfomance (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  The evaluation of muscular strength and physical performance was performed by a strength and conditioning coach graduated in physical activity and sport sciences, which will be carried out at each control point. The tests to be performed to the participants was: 400-meter Walk Test Perfomance: measures mobility and cardiovascular fitness through a 400-meter timed walk (min). Cones are placed 20 m apart, around which participants must move 10 times, back and forth, as fast as possible. Only one attempt was allowed.

  6 weeks

• Assessment of muscle strength and performance. Grip strength (Day 0 and at 6 weeks of intervention).

  The evaluation of muscular strength and physical performance was performed by a strength and conditioning coach graduated in physical activity and sport sciences, which will be carried out at each control point. The tests to be performed to the participants was: Grip strength: isometric hand strength is a commonly used measure to establish upper body skeletal muscle function and has been widely used as a general indicator of physical fitness. Grip strength was measured using a validated hydraulic hand-held dynamometer (Jamar plus +, if instruments, Adelaide, Australia). Values will be recorded in kilograms. Grip strength will be measured while seated, elbow in 90° flexion and wrist in neutral position. Participants was asked to apply maximum grip strength in 3 different attempts with the left and right hands. Between each attempt, a 30 s rest was allowed. The measurement in which the maximum grip strength is obtained from the 3 attempts will be used.

  6 weeks

• Assessment of muscle strength and performance. Grip strength (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  The evaluation of muscular strength and physical performance was performed by a strength and conditioning coach graduated in physical activity and sport sciences, which will be carried out at each control point. The tests to be performed to the participants was: Grip strength: isometric hand strength is a commonly used measure to establish upper body skeletal muscle function and has been widely used as a general indicator of physical fitness. Grip strength was measured using a validated hydraulic hand-held dynamometer (Jamar plus +, if instruments, Adelaide, Australia). Values will be recorded in kilograms. Grip strength will be measured while seated, elbow in 90° flexion and wrist in neutral position. Participants was asked to apply maximum grip strength in 3 different attempts with the left and right hands. Between each attempt, a 30 s rest was allowed. The measurement in which the maximum grip strength is obtained from the 3 attempts will be used.

  6 weeks

• Isometric strength of legs and back (Day 0 and at 6 weeks of intervention)

  A BASELINE traction dynamometer (back and leg dynamometer) with adjustable handle and base platform was used. The objective was to assess the maximum isometric strength of the back and leg extensor muscles (kg). The subject stood with their feet shoulder-width apart on the dynamometer platform. The legs were slightly bent and the back was in a neutral, inclined position. The dynamometer was adjusted so that the subject could grasp the handle with their arms fully extended downwards (near the knees). The subject was asked to pull the handle upwards, using their back and leg strength, without bending their arms or leaning their trunk backwards. The movement was isometric; force was applied without visible displacement. The contraction lasted about 3 to 5 seconds. Two attempts were made with at least 30 seconds of rest between them. The maximum value achieved was recorded.

  6 Weeks

• Isometric strength of legs and back (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  A BASELINE traction dynamometer (back and leg dynamometer) with adjustable handle and base platform was used. The objective was to assess the maximum isometric strength of the back and leg extensor muscles (kg). The subject stood with their feet shoulder-width apart on the dynamometer platform. The legs were slightly bent and the back was in a neutral, inclined position. The dynamometer was adjusted so that the subject could grasp the handle with their arms fully extended downwards (near the knees). The subject was asked to pull the handle upwards, using their back and leg strength, without bending their arms or leaning their trunk backwards. The movement was isometric; force was applied without visible displacement. The contraction lasted about 3 to 5 seconds. Two attempts were made with at least 30 seconds of rest between them. The maximum value achieved was recorded.

  6 Weeks

• Isometric arm flexion strength (Day 0 and at 6 weeks of intervention)

  A BASELINE traction dynamometer with adjustable handle and base platform was used. The objective was to assess the maximum isometric strength of the upper body (kg), specifically the arm flexor muscles (biceps). The subject stood with their feet shoulder-width apart on the dynamometer platform. Their legs were slightly bent and their back was straight and upright. The dynamometer was adjusted so that the subject could grasp the handle with their arms bent at 90°, elbows close to the body and wrists in a supine position. The subject was asked to pull the handle upwards, exerting force with their arms, without leaning the trunk backwards or using momentum. The movement was isometric; force was applied without visible displacement. The contraction lasted about 3 to 5 seconds. Two attempts were made with a 30-second rest between them. The maximum value achieved was recorded

  6 Weeks

• Isometric arm flexion strength (Day 0 after 3 weeks washout and at 6 weeks of intervention)

  A BASELINE traction dynamometer with adjustable handle and base platform was used. The objective was to assess the maximum isometric strength of the upper body (kg), specifically the arm flexor muscles (biceps). The subject stood with their feet shoulder-width apart on the dynamometer platform. Their legs were slightly bent and their back was straight and upright. The dynamometer was adjusted so that the subject could grasp the handle with their arms bent at 90°, elbows close to the body and wrists in a supine position. The subject was asked to pull the handle upwards, exerting force with their arms, without leaning the trunk backwards or using momentum. The movement was isometric; force was applied without visible displacement. The contraction lasted about 3 to 5 seconds. Two attempts were made with a 30-second rest between them. The maximum value achieved was recorded

  6 Weeks

• 30-second push-up test (Day 0 and at 6 weeks of intervention)

  This test was performed to assess the muscle strength and endurance (push) of the upper body (mainly pectorals, deltoids and triceps) and the stabilising strength of the lumbar-abdominal girdle. It consisted of performing as many push-ups as possible in 30 seconds from a plank position. The participant is placed in a high plank position with arms extended, hands shoulder-width apart, and body aligned from ankles to head. Each push-up must be performed with a full range of motion: lower the chest close to the floor (about 5-8 cm) and fully extend the elbows when rising. Only correctly executed repetitions were counted and recorded.

  6 Weeks

• 30-second push-up test (Day 0 after 3 weeks washout and at 6 weeks of intervention)

  This test was performed to assess the muscle strength and endurance (push) of the upper body (mainly pectorals, deltoids and triceps) and the stabilising strength of the lumbar-abdominal girdle. It consisted of performing as many push-ups as possible in 30 seconds from a plank position. The participant is placed in a high plank position with arms extended, hands shoulder-width apart, and body aligned from ankles to head. Each push-up must be performed with a full range of motion: lower the chest close to the floor (about 5-8 cm) and fully extend the elbows when rising. Only correctly executed repetitions were counted and recorded.

  6 Weeks

• 30-second supine trunk flexion test (Day 0 and at 6 weeks of intervention)

  Crunch Test: the objective was to assess abdominal muscle endurance. The subject lay supine with knees bent at 90° and feet flat on the floor and arms crossed over the chest. At the evaluator's signal, the subject performed as many trunk flexions as possible in 30 seconds. A repetition was counted as valid when the shoulders clearly lifted off the floor without lifting the lower back and returned to the starting position. The total number of correct repetitions was recorded.

  6 Weeks

• 30-second supine trunk flexion test (0 day after 3 weeks washout and at 6 weeks of intervention)

  Crunch Test: the objective was to assess abdominal muscle endurance. The subject lay supine with knees bent at 90° and feet flat on the floor and arms crossed over the chest. At the evaluator's signal, the subject performed as many trunk flexions as possible in 30 seconds. A repetition was counted as valid when the shoulders clearly lifted off the floor without lifting the lower back and returned to the starting position. The total number of correct repetitions was recorded.

  6 Weeks

• Isometric pull-up strength test (Day 0 and at 6 weeks of intervention)

  Flexed Arm Hang Test: used to assess the isometric strength and muscular endurance of the upper body, especially the arms, shoulders and back. Hanging from the bar, keep your head above the bar for as long as possible. The subject stands under a fixed horizontal bar, such as a pull-up bar. With the help of a bench or assistance, they position themselves with their elbows bent and their chin above the level of the bar. The palms may be pronated. The stopwatch starts when the person is suspended without assistance, with their feet completely off the ground. The test ends when the chin falls below the bar, the arms are extended, or the subject lets go.

  6 Weeks

• Isometric pull-up strength test (Day 0 after 3 weeks washout and at 6 weeks of intervention)

  Flexed Arm Hang Test: used to assess the isometric strength and muscular endurance of the upper body, especially the arms, shoulders and back. Hanging from the bar, keep your head above the bar for as long as possible. The subject stands under a fixed horizontal bar, such as a pull-up bar. With the help of a bench or assistance, they position themselves with their elbows bent and their chin above the level of the bar. The palms may be pronated. The stopwatch starts when the person is suspended without assistance, with their feet completely off the ground. The test ends when the chin falls below the bar, the arms are extended, or the subject lets go.

  6 Weeks

• Seated dumbbell arm curl test (Day 0 and at 6 weeks of intervention)

  The dynamic strength of the arm flexor muscles was measured. The participant sat on a chair without armrests, with their back straight and their feet flat on the floor. They held a dumbbell in their dominant hand with their palm facing upwards and their arm fully extended down at their side. In 30 seconds, the participant completely flexed their elbow, bringing the dumbbell towards their chest without moving their shoulder, and then extended it again. Complete repetitions were counted. A 2 kg dumbbell was used for women and a 4 kg dumbbell for men.

  6 Weeks

• Seated dumbbell arm curl test (Day 0 after 3 weeks washout and at 6 weeks of intervention)

  The dynamic strength of the arm flexor muscles was measured. The participant sat on a chair without armrests, with their back straight and their feet flat on the floor. They held a dumbbell in their dominant hand with their palm facing upwards and their arm fully extended down at their side. In 30 seconds, the participant completely flexed their elbow, bringing the dumbbell towards their chest without moving their shoulder, and then extended it again. Complete repetitions were counted. A 2 kg dumbbell was used for women and a 4 kg dumbbell for men.

  6 Weeks

• Quality of Life Assessment (Day 0 and at 6 weeks of intervention)

  The WHOQOL-BREF test was used in all controls. This questionnaire is a shortened version of the WHOQOL-100 questionnaire, developed by the World Health Organisation (WHO) to assess quality of life. It is one of the most widely used instruments internationally due to its psychometric robustness, cross-cultural applicability and brevity.There were 26 questions divided into 4 domains (Physical, Phychological, Social and Enviromental). Plus 2 questions on perception of quality of life and health that were not counted.

  6 weeks

• Quality of Life Assessment (Day 0 after 3 weeks washout and at 6 weeks of intervention)

  The WHOQOL-BREF test was used in all controls. This questionnaire is a shortened version of the WHOQOL-100 questionnaire, developed by the World Health Organisation (WHO) to assess quality of life. It is one of the most widely used instruments internationally due to its psychometric robustness, cross-cultural applicability and brevity.There were 26 questions divided into 4 domains (Physical, Phychological, Social and Enviromental). Plus 2 questions on perception of quality of life and health that were not counted.

  6 weeks

• Inflammation markers. CRP (Day 0 and at 6 weeks of intervention).

  CRP was assessed in duplicate from frozen stored serum. PCR levels using an enzyme-linked immunostorbent assay based on purified proteins and purified anti-CRP antibodies (Calbiochem, San Diego, CA).

  6 weeks

• Inflammation markers. CRP(Day 0 after 3 weeks washout and at 6 weeks of intervention).

  CRP was assessed in duplicate from frozen stored serum. PCR levels using an enzyme-linked immunostorbent assay based on purified proteins and purified anti-CRP antibodies (Calbiochem, San Diego, CA).

  6 weeks

• Inflammation markers. Cytokine levels and cytokine soluble receptors (Day 0 and at 6 weeks of intervention).

  IL-1 (soluble IL-1 receptor \[IL-1SR\]), IL-2 (soluble IL-2 receptor \[il-2SR\]), IL-6 (soluble IL-6 receptor \[IL-6SR\]), IL-7 (soluble IL-7 receptor \[IL-7SR\]) was measured using enzyme-linked immunostorbent assay kits (Research and Development Systems, Minneapolis, MN).

  6 weeks

• Inflammation markers. Cytokine levels and cytokine soluble receptors (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  IL-1 (soluble IL-1 receptor \[IL-1SR\]), IL-2 (soluble IL-2 receptor \[il-2SR\]), IL-6 (soluble IL-6 receptor \[IL-6SR\]), IL-7 (soluble IL-7 receptor \[IL-7SR\]) was measured using enzyme-linked immunostorbent assay kits (Research and Development Systems, Minneapolis, MN).

  6 weeks

• Inflammation markers. TNF-α (Day 0 and at 6 weeks of intervention).

  TNF-α (TNFSR1 and TNFSR2) levels was assessed in duplicate from frozen stored serum.

  6 weeks

• Inflammation markers. TNF-α (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  TNF-α (TNFSR1 and TNFSR2) levels was assessed in duplicate from frozen stored serum.

  6 weeks

• Oxidative stress markers. Reduced (GSH) and oxidized glutathione (GSSG) (Day 0 and at 6 weeks of intervention).

  Reduced (GSH) and oxidized glutathione (GSSG) was determined using colorimetric kits. Reducing power will be calculated by the equation GSH/(GSH + GSSG).

  6 weeks

• Oxidative stress markers. Reduced (GSH) and oxidized glutathione (GSSG) (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  Reduced (GSH) and oxidized glutathione (GSSG) was determined using colorimetric kits. Reducing power will be calculated by the equation GSH/(GSH + GSSG).

  6 weeks

• Oxidative stress markers. TBAR (Day 0 and at 6 weeks of intervention).

  Lipid peroxidation was analyzed in plasma as thiobarbituric acid reactive substances (TBAR). The TBAR was determined after treatment of the sample with 4% butylated hydroxytoluene and an additional reaction with 0.375% thiobarbituric acid in 0.25 M HCl and 1% Triton X-100 (15 min, at 100°C).

  6 weeks

• Oxidative stress markers. TBAR (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  Lipid peroxidation was analyzed in plasma as thiobarbituric acid reactive substances (TBAR). The TBAR was determined after treatment of the sample with 4% butylated hydroxytoluene and an additional reaction with 0.375% thiobarbituric acid in 0.25 M HCl and 1% Triton X-100 (15 min, at 100°C).

  6 weeks

• Oxidative stress markers. Malondialdehye equivalents (MDA) (Day 0 and at 6 weeks of intervention).

  Malondialdehye equivalents (MDA) was calculated according to the absorbance at 535 nm against a blank (lacking TBA) and using 1,1,2,2-tetroxyethylpropane as standard.

  6 weeks

• Oxidative stress markers. Malondialdehye equivalents (MDA) (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  Malondialdehye equivalents (MDA) was calculated according to the absorbance at 535 nm against a blank (lacking TBA) and using 1,1,2,2-tetroxyethylpropane as standard.

  6 weeks

• Oxidative stress markers. Superoxide dismutase (SOD) (Day 0 and at 6 weeks of intervention).

  Superoxide dismutase (SOD) was estimated by inhibition of adrenaline autooxidation, read at 480 nm in a spectrophotometer.

  6 weeks

• Oxidative stress markers. Superoxide dismutase (SOD) (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  Superoxide dismutase (SOD) was estimated by inhibition of adrenaline autooxidation, read at 480 nm in a spectrophotometer.

  6 weeks

• Oxidative stress markers. Nitric oxide (NO) (Day 0 and at 6 weeks of intervention).

  Nitric oxide (NO) was estimated spectrophotometrically as a function of nitrite generation.

  6 weeks

• Oxidative stress markers. Nitric oxide (NO) (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  Nitric oxide (NO) was estimated spectrophotometrically as a function of nitrite generation.

  12 weeks

• Oxidative stress markers. Total antioxidant capacity (TAC) (Day 0 and at 6 weeks of intervention).

  Total antioxidant capacity (TAC) was analyzed in duplicate samples, and absorbance will be measured using a microplate reader.

  6 weeks

• Oxidative stress markers. Total antioxidant capacity (TAC) (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  Total antioxidant capacity (TAC) was analyzed in duplicate samples, and absorbance will be measured using a microplate reader.

  6 weeks

• Hormones. Total serum testosterone (Day 0 and at 6 weeks of intervention).

  Total serum testosterone was measured using the enzyme from commercially available enzyme-linked immunosorbent assay (ELISA) kits (DRG Testosterone ELISA KIT, DRG Instruments GmbH, Marburg,® Germany).

  6 weeks

• Hormones. Total serum testosterone (Day 0 after 3 weeks washout and at 6 weeks of intervention)..

  Total serum testosterone was measured using the enzyme from commercially available enzyme-linked immunosorbent assay (ELISA) kits (DRG Testosterone ELISA KIT, DRG Instruments GmbH, Marburg,® Germany).

  6 weeks

• Hormones. Sex hormone binding globulin (SHBG) (Day 0 and at 6 weeks of intervention).

  Sex hormone binding globulin (SHBG) was measured by chemiluminescence based on the immunometric method (immunite, 2000), which will be from the same supplier as testosterone.

  6 weeks

• Hormones. Sex hormone binding globulin (SHBG) (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  Sex hormone binding globulin (SHBG) was measured by chemiluminescence based on the immunometric method (immunite, 2000), which will be from the same supplier as testosterone.

  6 weeks

• Hormones. Cortisol (Day 0 and at 6 weeks of intervention).

  Cortisol was measured by enzyme-linked fluorescent assay in a microplate reader (Cytation 5 Cell Imaging Multi-Mode Reader -Biotek Instruments, USA).

  6 weeks

• Hormones. Cortisol (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  Cortisol was measured by enzyme-linked fluorescent assay in a microplate reader (Cytation 5 Cell Imaging Multi-Mode Reader -Biotek Instruments, USA).

  6 weeks

• Hormones. Total testosterone/cortisol and total testosterone/SHBG ratios (Day 0 and at 6 weeks of intervention).

  Total testosterone/cortisol and total testosterone/SHBG ratios was calculated from serum testosterone, cortisol and SHBG concentrations.

  6 weeks

• Hormones. Total testosterone/cortisol and total testosterone/SHBG ratios (Day 0 after 3 weeks washout and at 6 weeks of intervention).

  Total testosterone/cortisol and total testosterone/SHBG ratios was calculated from serum testosterone, cortisol and SHBG concentrations.

  6 weeks

Study Arms (2)

Intervention Group 1

EXPERIMENTAL

CRE-HMB mixture

Dietary Supplement: CRE + HMB

Intervention Group 2

PLACEBO COMPARATOR

placebo

Dietary Supplement: Placebo

Interventions

CRE + HMB DIETARY_SUPPLEMENT

3 g/day of CRE and 3 g/day of HMB

Intervention Group 1

Placebo DIETARY_SUPPLEMENT

6 g/day of inulin

Intervention Group 2

Eligibility Criteria

• Age: 55 Years - 80 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• Participants will be age between 55 and 80 years.
• All individuals over the age of 55 who performed Integral Physical Conditioning training for a minimum of 120 minutes per week.
• They did not take the study supplement or any other supplement related to performance or muscle development.
• Voluntarily agreeing to participate in the study and having signed the informed consent.
• Not suffering any physical impediment to perform multicomponent physical exercise.

You may not qualify if:

• All individuals who were not in the age range studied.
• They did not meet the minimum weekly Comprehensive Physical Conditioning training requirement.
• All those who had suffered from or were being treated for heart, liver or kidney conditions.
• Those who were being treated with NSAIDs.
• Those who refused to stop taking supplements and/or follow the study protocol.

Sponsors & Collaborators

• Universidad de Burgos: lead

Study Sites (1)

Juan Mielgo Ayuso

Burgos, Burgos, 09001, Spain

Location

MeSH Terms

Conditions

Sarcopenia; Motor Activity

Condition Hierarchy (Ancestors)

Muscular Atrophy; Neuromuscular Manifestations; Neurologic Manifestations; Nervous System Diseases; Atrophy; Pathological Conditions, Anatomical; Pathological Conditions, Signs and Symptoms; Signs and Symptoms; Behavior

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: TRIPLE
• Who Masked: PARTICIPANT, INVESTIGATOR, OUTCOMES ASSESSOR
• Purpose: PREVENTION
• Intervention Model: CROSSOVER
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Assistan Professor

Model Details: Cross Randomized double-blind study

Study Record Dates

• First Submitted: June 18, 2023
• First Posted: July 19, 2023
• Study Start: November 8, 2024
• Primary Completion: February 22, 2025
• Study Completion: February 22, 2025
• Last Updated: August 29, 2025

Record last verified: 2024-12

Data Sharing

• IPD Sharing: Will not share

Locations

ES (1)