Sarcopenia in Elderly, Sarcopenia, Nutritional Intervention, Microbiome Analysis, Muscle Mass and Strength, Biomarkers / Blood, Quality of Life Outcomes
Conditions
Keywords
sarcopenia, elderly, pea proteins, vitamin D, snack
Brief summary
The ageing population makes it necessary to find effective strategies for the prevention of sarcopenia (the progressive loss of muscle mass and strength and a decline in physical performance) that can be counteracted with foods containing protein and adequate intake of vitamin D. This study will evaluate the effectiveness of consuming a food based on plant proteins and vitamin D supplementation. Intervention studies in humans conducted to date have mostly focused on the effect of animal proteins (mainly from whey) on disease progression. A study on the effect of pea proteins has not yet been conducted and will provide information on the effectiveness of these proteins in modulating markers linked to the disease. The effect on the gut microbiota will also be considered, as the existence of a gut-muscle axis has been suggested, in which microbial genera producing short-chain fatty acids have been linked to a positive effect on muscle mass through anabolic stimulation. Thus, the analysis of the modulation of the intestinal microbiota, through the dietary intervention proposed in this study, may represent a further step in research related to the prevention of this disease. Sarcopenic volunteers aged between 65 and 80 will be recruited to consume either a shortbread biscuit made with wheat flour enriched with hydrolysed pea protein and a vitamin D supplement in extra virgin olive oil, or a control biscuit and a placebo (extra virgin olive oil) for 12 weeks. The study will be randomised, parallel, single-blind. The effect of consuming the experimental biscuit and vitamin D supplementation compared to that of a traditional control biscuit and a placebo oil solution will be evaluated on certain markers related to sarcopenia. In particular, the following will be considered: muscle strength, measuring grip strength and leg strength (chair stand test); muscle mass through the measurement of appendicular muscle mass, and the calculation of the appendicular muscle mass index; physical performance using the Short Physical Performance Battery; the inflammatory response and other blood biomarkers related to sarcopenia. In addition, the following will be assessed: dietary habits through a food diary and quality of life through the SarQoL questionnaire. Finally, the effect of nutritional intervention on the modulation of the gut microbiota will be evaluated through 16S rRNA sequencing and bioinformatic analysis of the data.
Detailed description
Sarcopenia, characterized by the age-related loss of muscle mass, strength, and physical performance, is a growing public health concern given the global aging population. Physical activity-the standard treatment-is often not feasible for frail or bedridden elderly individuals, highlighting the need for alternative strategies. Nutritional interventions, particularly those involving protein and vitamin D supplementation, have shown promise in managing sarcopenia. This randomized, single-blind, parallel-design clinical trial aims to evaluate the effect of consuming a shortbread biscuit enriched with hydrolyzed pea protein, combined with a daily dose of vitamin D3 (20 μg/day, 800 IU/day), on muscle mass, muscle strength, physical performance, and gut microbiota composition in sarcopenic individuals aged 65-80 years. The study will enroll 74 participants meeting specific inclusion criteria. Participants will be randomly assigned to one of two groups: 1. Intervention Group: Receives a 50 g daily portion of a functional biscuit enriched with hydrolyzed pea protein, and 2 drops/day of a vitamin D3 supplement (providing 20 μg/day). 2. Control Group: Receives a similar biscuit without added protein and a placebo oil (extra virgin olive oil) in replace of vitamin D3. The intervention will last for 12 weeks. Measurements will be taken at baseline, mid-point (6 weeks), and study end (12 weeks), including handgrip strength, chair stand test, bioimpedance-based muscle mass, and SPPB. Blood samples will be analyzed for markers related to inflammation (e.g., CRP, IL-6, TNF-α), oxidative stress (oxLDL), anabolic signaling (IGF-1), and vitamin D status (25-hydroxycholecalciferol), among others. Gut microbiota composition and short-chain fatty acid (SCFA) production will be assessed via stool sample analysis and 16S rRNA sequencing. Participants will also complete quality-of-life (SarQoL) and food frequency questionnaires, along with 3-day dietary diaries at each time point. Compliance will be monitored via dietary diary entries and return of supplement containers. A sample size of 74 participants (32 per group + 15% dropout) was calculated to detect a 25% difference in treatment success (as defined by surpassing the sarcopenia threshold in ASMI). The study further investigates the potential of plant-based protein sources, particularly hydrolyzed pea protein, to serve as effective dietary interventions in the elderly. It also explores the gut-muscle axis, examining how nutritional modulation of the microbiota may impact sarcopenia-related outcomes. The trial is conducted at the Internal Medicine Unit 2 of the S. Maria della Misericordia Hospital (Udine, Italy) under the coordination of Prof. Alessandro Cavarape. All procedures follow ethical guidelines and include appropriate monitoring for adverse events.
Interventions
DIETARY_SUPPLEMENTVitamin D3 supplement
Two daily drops of vitamin D3 dissolved in extra virgin olive oil. Used in the Experimental Arm.
OTHERProtein-enriched biscuits
Daily portion (50 g) of biscuits made with wheat flour and pea protein hydrolysate. Used in the Experimental Arm.
OTHERControl biscuit
Daily portion (50 g) of control biscuits made with wheat flour, without pea protein hydrolysate.
OTHERPlacebo oil
Two daily drops of placebo oil (extra virgin olive oil without vitamin D3).
Sponsors
University of Udine
Study design
Allocation
RANDOMIZED
Intervention model
PARALLEL
Primary purpose
TREATMENT
Masking
TRIPLE (Subject, Caregiver, Outcomes Assessor)
Eligibility
Sex/Gender
ALL
Age
65 Years to 80 Years
Healthy volunteers
No
Inclusion criteria
Given the multifactorial, heterogeneous nature of sarcopenia, volunteers will be enrolled if they meet at least one of the following three diagnostic criteria : * Ishii equation values: patients with a score ≥105 for males and ≥120 for females will be included. The Ishii screening test is a widely used method that estimates the probability of sarcopenia using a score derived from an equation based on three variables: age, grip strength, and calf circumference. * Appendicular skeletal muscle mass index (ASMI): \<7.0 kg/m2 for males and \<5.5 kg/m2 for females. * Short physical performance battery score ≤8 In addition, the following inclusion criteria are considered: * Body mass index ≥19 \<30 kg/m2 * Ability to act
Exclusion criteria
* Age \<65 or \>80 years; * Body mass index \<19 or ≥30 kg/m²; * Diagnosis of cancer within the previous five years; * Regular intake of vitamin D supplements; * Dysphagia and difficulty chewing; * Allergy or other intolerances to gluten; * Allergy to eggs; * Intestinal disorders (Crohn's disease, ulcerative colitis, bacterial overgrowth syndrome, constipation, coeliac disease, irritable bowel syndrome) that could affect the gut microbiota; * Pacemaker recipients; * Acute inflammation (CRP \>10 mg/L); * Anaemia (haemoglobin \<13 g/dL in men; \<12 g/dL in women); * Renal failure (glomerular filtration rate (GFR) \>45 mL/min/1.73 m²); * Chronic liver disease (transaminases \< 40 IU in men, \< 35 IU/L in women); * Inability to act
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Change in Appendicular Skeletal Muscle Index (ASMI) | Screening, baseline, week 6 and week 12 (end of the treatment) | ASMI calculated as appendicular skeletal muscle mass in kilograms divided by height in meters squared (kg/m²). ASM and height will be measured separately and combined using this formula: "kg/m²". ASMI values \<7 kg/m2 for men, and \<5.5 kg/m2 for women indicate the condition of sarcopenia as indicated by the European Working Group on Sarcopenia in Older People. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Muscle strenght | Screening, baseline, week 6 and week 12 (end of the treatment) | The muscle strenght will be measured using two different analysis Handgrip Strength: Measured using Electronic Hand Dynamometer EH101, dominant hand, two attempts. The cut-off point for the diagnosis of sarcopenia are \<27 kg for men, and \<16 kg for women as indicated by the European Working Group on Sarcopenia in Older People. Chair Stand Test: Number of stands in 30 seconds without upper limb support. The cut of for the diagnosis of sarcopenia is \>15 s for five rises as indicated by the European Working Group on Sarcopenia in Older People. |
| Physical performance (SPPB score) | Screening, baseline, week 6 and week 12 (end of the treatment) | Short Physical Performance Battery (balance tests, 4-meter walk, 5 sit-to-stand repetitions). The scale for SPPB is between 0 and 12. The cut-off points for the diagnosis of sarcopenia is ≤8 point score as indicated by the European Working Group on Sarcopenia in Older People. |
| C-Reactive protein (CRP) | Baseline, week 6 and week 12 (end of the treatment) | CRP will be measured in serum using a multiplex ELISA assay and expressed in milligrams per liter (mg/L). |
| Gut micorbiome modulation and metabolites | Baseline at at week 12 (end of the treatment) | Quantification of SCFA production (via gas chromatography) and microbiota composition (16S sequencing, α-diversity, relative abundance of sarcopenia-associated genera: Roseburia, Eubacterium, Lachnospira, Ruminococcus). |
| Nutritional intake | Baseline, week 6 and week 12 (end of the treatment) | 3-day dietary record. This record will tracks all food and beverages consumed over 2 weekdays and 1 weekend day to analyze typical eating habits. |
| Nutritional habits | Enrollment | food frequency questionnaire (FFQ). This instrument allow monitoring the past dietary habits (previous year). |
| Quality of life of sarcopenic patients | Baseline, week 6 and week 12 (end of the treatment) | SarQoL questionnaire to assess health-related quality of life in sarcopenic subjects. The SarQoL scale is between 0 and 100. A cut-off ≤ 60 points is indicative of sarcopenia. However, these cutoffs are not diagnostic on their own but they have been proposed as screening indicators to flag individuals who may warrant further clinical evaluation for sarcopenia using established criteria. |
| Mini Nutritional Assessment (MNA®) | At the screening | The MNA® is a validated nutrition screening and assessment tool that can identify geriatric patients age 65 and above who are malnourished or at risk of malnutrition. The scale of MNA® is between 0 and 30. The cut-off points are 24-30 normal nutritional status; 17-23.5 risk of malnutrition; \<17 malnutrition. |
| Mini Mental State Examination (MMSE) | At the screening | It is an 11-question measure that tests five areas of cognitive function: orientation, registration, attention and calculation, recall, and language. The scale for MMSE goes from 0 to 30. Cut-off points are 24-30: No cognitive impairment; 18-23: Mild to moderate cognitive impairment; \< 18: Moderate to severe cognitive impairment. |
| Body mass index (BMI) | screening, baseline, week 6, week 12 (end of the treatment) | BMI will be calculated by dividing the body weigth in kg and the height in meters (m). |
| Interleukin-6 (IL-6) | baseline, week 6, week 12 (end of the treatment) | IL-6 will be measured in serum using a multiplex ELISA assay and expressed in picograms per milliliter (pg/mL). |
| Tumor Necrosis Factor-alpha (TNF-α) | baseline, week 6, week 12 (end of the treatment) | TNF-α will be measured in serum using a multiplex ELISA assay (Ella, ProteinSimple) and expressed in picograms per milliliter (pg/mL). |
| Leptin | baseline, week 6, week 12 (end of the treatment) | Leptin concentration in serum will be measured using multiplex assay kit (ELISA) and expressed as picograms per milliliter (pg/mL) |
| IGF-1 | Baseline, week 6, week 12 (end of the treatment) | Quantitative measurement of serum insulin-like growth factor-1 (IGF-1) concentration using the Thermo Fisher Human IGF-1 ELISA Kit and expressed as ng/mL. |
| Albumin | baseline, week 6, week 12 (end of the treatment) | Quantitative measurement of serum albumin concentration using the Thermo Fisher Human albumin ELISA Kit and expressed as ng/mL. |
| Oxidized LDL (oxLDL) | baseline, week 6 and week 12 (end of the treatment) | Quantitative measurement of fasting serum oxidized LDL (oxLDL) concentration using the Thermo Fisher Human Oxidized LDL ELISA Kit and expressed as pg/mL. |
| 25OH-D (Total 25-OH Vitamin D) | baseline, week 6, week 12 (end of the treatment) | Serum 25OH-D (Total 25-OH Vitamin D) will be measured using the FineTest® 25OH-D (Total 25-OH Vitamin D) ELISA Kit and expressed as ng/mL. |
| Human Agrin (C-terminal fragment) | baseline, week 6, week 12 (end of the treatment) | Serum Human Agrin (C-terminal fragment) will be quantified using the Human Agrin (C-terminal fragment) ELISA Kit and expressed as pg/mL. |
| Triglyceride (TG) | baseline, week 6, week 12 (end of the treatment) | Serum Triglyceride (TG) levels will be measured using Triglyceride (TG) Colorimetric Assay Kit and expressed as mmol/L. |
Countries
Italy
Contacts
PRINCIPAL_INVESTIGATORAlessandro Cavarape, M.D.
Azienda Sanitaria Universitaria del Friuli Centrale (ASU FC)
Outcome results
None listed