Multivitamin and Mineral Supplementation and Healthy Aging (MiMIC)

As the global population of older adults continues to expand, it is imperative to identify accessible and evidence-based interventions to support healthy aging. Among the key biological hallmarks of aging, mitochondrial dysfunction is highly modifiable and especially responsive to lifestyle interventions such as diet and exercise. Importantly, our clinical studies demonstrate that mitochondrial bioenergetics plays a key role in the physical and cognitive abilities of older adults. Thus, identifying interventions that can improve mitochondrial bioenergetics is a promising avenue for the development of healthy aging interventions.

Multivitamin and mineral (MVM) dietary supplements have been used to support general health and nutritional status for years, particularly in older adults who are more susceptible to nutritional deficiencies due to factors such as reduced appetite, malabsorption, and polypharmacy. Observational studies consistently demonstrated that older adults exhibit higher rates of nutritional deficiencies, and that MVM supplementation may ameliorate these burdens. More recently, evidence suggests that MVM use may also confer functional health benefits; for example, the COSMOS-Mind trial demonstrated efficacy of MVM on global cognition, memory, and executive function in 2262 participants. Notably, most MVM studies are longitudinal and occur over the course of many years. A significant gap remains in understanding the acute effects of MVM supplementation on health and aging. Can rapid, measurable improvements in health be observed with MVM use? Moreover, what are the cellular mechanisms that can underlie the benefits of MVM use?

Description of Study Intervention: The study cohort will consist of 150 sedentary men and women aged 40-60-years of age. A double-blind, three-arm, placebo-controlled randomized clinical trial will be conducted. Participants will receive a daily oral tablet containing one of two possible formulations of vitamins and minerals or a placebo. The three groups will be MVM "GOLD" blend formula, the US Restage MVM "US CORE", and the placebo group. Specific formulations of the three tablets are detailed in the 2025 Stability Provisional Expiry for Centrum Gold Blend Tablets (FN-2492-0001), Centrum US Restage Blend Tablets (FN-2493-0001, and Placebo of Centrum Gold Blend Tablets (FN-0436-0231) -

Assessments:

The baseline study visit will occur at least 2 weeks after, but no more than 4 weeks after, the initial screening visit (actigraph will be deployed at screening and will be worn for 2 weeks).

At each study visit (V1, V2, V3, V4) the participant will undergo a fasted blood-draw followed by a snack break, assessment of vital signs, and physical assessments as listed below.

Sample Collection: A licensed phlebotomist will perform a venipuncture blood draw at the forearm, arm, or antecubital fossa of either arm and collect 20 ml of blood into three 10ml Acid Citrate Dextrose (ACD) tubes, one serum-separating tube (SST), and one ethylenediaminetetraacetic acid (EDTA) tube for each blood-draw visit (V1, V2, V3, V4). On visits V2 and V3, we will collect three additional ACD tubes for fresh cell assays. Our previous studies indicate that samples are stable for 8 hours at room temperature.

Within 1 hour of sampling, blood will be processed to separate blood components. Blood will be spun and components will be frozen, incuding platelets and PBMCs from all four blood-draw visits (V1-V4) for frozen respirometry. During V1 (baseline) and V3 (week 12) visits, live cells will be isolated for fresh respirometry. Fresh respirometry will include platelets, PBMCs, monocytes, and lymphocytes.

Clinical Laboratory Measurements: tubes of whole blood will be provided to the UCSD Center for Advanced Laboratory Medicine (CALM) Labs as well as the Associated Regional and University Pathologists, Inc (ARUP) Laboratories in Salt Lake City, UT to run various blood tests, including: compete blood count, comprehensive metabolic panel, c-reactive protein, sedimentation rate, lipid panel, and serum levels of cortisol, vitamins D, B12, B6, E, C, and beta-carotene.

Assessment of Vital Signs: Measurements of height, weight, heart rate, and blood pressure will be administered by trained study staff.

Wireless Activity Monitor: Objective assessments of physical activity will be measured using the ActiGraph GT3X+ accelerometer (ActiGraph, LLC; Pensacola, FL). The GT3X+ is a lightweight (19g) triaxial solid state accelerometer, with a dynamic range of +/- 6g and a user-specified sampling rate of 30-100hz (in 10hz increments) (19). The device is approximately 2''x2'x0.5'' in size and is worn on the hip attached to a belt around the waist. The device has 512MB of non-volatile flash memory and a 3.7V prismatic lithium-ion battery allowing for 40 days of raw acceleration data to be collected at 30Hz.

This method employs the seven-step algorithm identified as best-practice for the collection, processing, and summarization of PA accelerometer data collected with wearable systems (20). We will provide wear time instructions, and a brief self-report wear time log for participants to note periods that the device was taken off. Participants will be asked to wear the ActiGraph for seven days and will be prompted twice via telephone during the monitoring period (wear days 2 and 5) to assist with compliance. Upon return, we will immediately download and screen ActiGraph data by hour for completeness and possible irregularities/malfunction according to best practice recommendations (21).

Accelerometer outcome variables include (i) mean minutes per day of sedentary time, (ii) mean minutes per day of light intensity physical activity, and (iii) minutes per week of moderate-to-vigorous intensity physical activity (MVPA). Additional variables will also be computed based on patterns of how sedentary time and MVPA is accumulated (e.g., minutes of sedentary time per day occurring in bouts ≥30 minutes; minutes per week of MVPA occurring in bouts of ≥ 10 minutes).

Grip Strength: Hand grip strength will be measured in both hands using an adjustable grip strength dynamometer (BL5001 Hydraulic Hand Dynamometer). Participants will have a chance to familiarize themselves with the measurement by making a submaximal effort on both hands to feel how the instrument will react. For the assessment, the participant will stand and hold the dynamometer in their hand with their arm down at their side. The participant will be instructed to take a deep breath in and squeeze as hard as possible as they exhale. The measurement will be repeated twice on each hand, alternating between each side, and the highest score for each hand will be recorded to the nearest kilogram.

Submaximal Graded Exercise Test: Participants will undergo Submaximal Graded Exercise Test (GXT) on a treadmill at EPARC under the supervision of a trained exercise physiologist. Participants will complete a walking protocol at a self-selected speed, with the treadmill incline increasing every two minutes to progressively increase cardiovascular demand. The test will continue until the participant reaches 85% of their age-predicted maximal heart rate (calculated as 220 bpm - age) or experiences volitional fatigue, whichever occurs first.

The test itself is expected to last approximately 5-10 minutes, excluding warm-up and recovery periods, with the total session lasting about 20 minutes. For the first visit (V1), we will closely monitor blood pressure and heart rate before, during, and after the test, including at least five minutes into recovery and again before the participant leaves the lab. If there is a risk detected during blood pressure monitoring, (systolic >200, or an increase in diastolic during exercise), then BP monitoring will continue throughout all visits. For every visit, continuous heart rate monitoring will be performed via Polar chest strap, and ratings of perceived exertion (RPE) will be collected at regular intervals during the test.

In the unlikely event of a cardiac event or medical emergency, EPARC staff are trained in emergency procedures, and an automated external defibrillator (AED) and supplemental oxygen are available on-site. The protocol aligns with standardized exercise testing guidelines to provide reliable and meaningful physiological data while prioritizing participant safety.

Dual-energy X-ray Absorptiometry (DXA) Bone Mineral Density/Content (BMD/BMC) and Body Composition BMD/BMC and body composition will be assessed by dual energy x-ray absorptiometry (DXA) on a Prodigy Advance densitometer (GE/Lunar, Madison, WI). Densitometry will be performed by the same technician certified by the State of California Radiologic Health Branch. The scan sites include the spine (L1-L4), proximal femur (femoral neck and total hip), and whole body. The DXA whole body scans will be used to determine whole body BMD/BMC as well as body composition (fat and fat-free mass, and %fat). The total time required for subject positioning and scanning of the spine and hip is about 5 minutes, and ~6 minutes for the total body, in standard scan mode. Radiation exposure for all 3 scans is approximately 8-10 μSv (0.8-1.0 mRem), which is approximately equal to a day of background exposure. Since the long-term effects of exposure to a fetus are not known, pregnant women will not be scanned. Quality assurance tests are performed each morning of testing. The mean coefficient of variation (% CV) in our laboratory, based on precision studies of 30 individuals measured twice on the same day is: 0.64% for the total hip, 1.39 for femoral neck, 0.82% for the spine (L1-L4), 0.85% for total body BMD, 1.89% for total body fat mass, and 0.7% for total lean mass.

Subject Preparation Subjects are instructed, prior to the day of their scan, to dress in single-layer, loose clothing free of any metal, plastic, or similar materials, including all jewelry as well as leather or other dense fabrics. Women are encouraged to wear a halter-top or something similar. Subjects should be normally hydrated, and if they take a calcium supplement, not to take it on the day of their scan. Scans should not be done within 10-14 days following medical testing requiring any contrast agent.

Weekly Electronic Surveys: Weekly electronic surveys will be sent using REDCap. Electronic surveys will include the Diet Quality Questionnaire (DQQ), Warwick-Edinburgh Mental Well-Being Scale (WEMWBS), Short Form Survey (SF-36) quality of life survey, SATED sleep survey, and Pittsburgh Sleep Quality Index (PSQI) [sleep surveys are only administered monthly]. Participants will fill out an electronic adherence log each week to keep track of study compliance.