Alleviation Of Metabolic Endotoxemia In Adults With Metabolic Syndrome With Milk Fat Globule Membrane

Metabolic syndrome (MetS) adults (n = 24; 18-65 y) will be enrolled to complete a 2-arm, double-blind, randomized controlled, crossover trial. They will be randomized in 4-unit blocks to receive, for 13 d, a controlled diet with dairy milk (3.5% fat; 3 servings/d) enriched with milk fat globule membrane (MFGM, MEB) or a matched dairy milk that instead contains soy lecithin/phospholipid (control, COMP). All foods during each study period will be provided to ensure weight maintenance and to increase homogeneity of gut and host responses. Anthropometrics and blood pressure will be assessed at days 0, 7, and 13. Prior to (day 0) and after each 2-wk arm (day 13), a fasting blood sample will be collected to assess serum endotoxin and metabolic chemistries (glucose, lipids, insulin), and Toll-like receptor 4 /nuclear factor kappaB (TLR4/NFκB)-dependent genes from whole blood. A breath sample will be collected to assess the correlation analysis of plasma metabolic biomarkers. After the 2-week intervention, from fecal samples collected on day 13, the investigators will assess microbiota composition and function, short chain fatty acids (SCFA), and intestinal inflammatory markers (calprotectin, myeloperoxidase). On d 13, participants in the fasted state will receive a high-fat/high-glucose meal challenge to induce gut-derived endotoxin translocation. At 30-minute intervals for 3-hour, the investigators will evaluate circulating endotoxin, glucose, and insulin; TLR4/NFκB-dependent genes will be assessed from whole blood at 0 hour and 3-hour. Gut permeability probes will be co-administered with the test meal challenge, and 24-hour urine will be collected to assess gut barrier integrity. Participants will then undergo a 2-week washout prior to receiving the alternative treatment and completing all procedures in an identical manner.

Study Overview

• Status: Completed
• Conditions: Metabolic Syndrome; Metabolic Endotoxemia
• Intervention / Treatment: Dietary supplement: MFGM-enriched full-fat dairy milk; Dietary supplement: Soy phospholipid/lecithin milk

Detailed Description

Background and hypothesis:

Our preclinical evidence shows that phospholipid-rich milk fat globule membrane (MFGM) attenuates lipopolysaccharide-induced increases in gut permeability and pro-inflammatory cytokines. MFGM also attenuates inflammation in association with a prebiotic and/or antimicrobial activity that modulates microbiota composition. Our central hypothesis is that MFGM-enriched dairy milk compared with a matched milk beverage containing soy lecithin (control) decreases metabolic endotoxemia and improves glucose tolerance in metabolic syndrome (MetS) adults by increasing gut barrier integrity in association with alleviating gut dysbiosis and inflammation.

Study Design:

The investigators will enroll male and female MetS adults (n = 24; 18-65 y) to complete a 2-arm, double-blind, randomized controlled, crossover trial. They will be randomized in 4-unit blocks to receive, for 14 days, a controlled diet with dairy milk (3.5% fat; 3 servings/d) enriched with MFGM-derived phospholipid or a matched dairy milk that instead contains coconut and palm oil (control). The investigators will provide all foods during each study period to ensure weight maintenance and to increase homogeneity of gut and host responses.

Subjects:

Participants will be recruited from Columbus, Ohio area. Participants having no history of liver or cardiovascular disease or cancer will be enrolled. They will have ≥3 established criteria for MetS: i) glucose (100-126 mg/dL), ii) waist circumference (>89 or >102 cm for F/M), iii) HDL-C (<50 or <40 mg/dL in F/M), iv) triglyceride >150 mg/dL, and iv) blood pressure >130/85 mmHg. Major exclusion criteria include: unstable body mass (±2 kg over prior 3-mo) vegetarian; food allergies or lactose intolerance; user of dietary supplements or probiotics (within past 1-mo); pregnancy, lactation, changes in birth control (within 6-month); any gastrointestinal disorders; chronic diarrhea; smoker; excess alcohol (>2 drinks/day); excess aerobic exercise (>7 h/week); recent antibiotic or anti-inflammatory agent use; blood pressure >140/90 mmHg.

Dietary Control:

The intervention will be performed in the Human Nutrition Metabolic Kitchen under the auspice of a registered dietitian (PI Bruno). In each 2-wk intervention, participants' diet will be rigorously controlled. All foods will be prepared, packaged, and provided every 3-4 days to supply a weight maintenance (i.e. eucaloric) diet. To assess compliance, participants will return MFGM/coconut/palm oil milk bottles for counting and any uneaten food portions for weighed measurement. Milk beverages will also be formulated to contain para-aminobenzoic acid (PABA; 80 mg/milk serving). Spot urine samples will be collected 4 times during each study arm coinciding when participants pick up test foods. Urinary PABA will be measured by spectrophotometry. Separate from this, participants will also keep food logs to document any dietary deviation. Diets will be standardized at 50-60% of energy from carbohydrate with low fiber intakes (~15 g/day) similar to Americans' diets to prevent potential masking of the benefits of MFGM, 15-20% from protein, and 25-30% from fat. Importantly, other than test beverages provided as part of the eucaloric diet, diets will be otherwise devoid of significant amounts of dairy foods, fermented products, and probiotics to prevent confounding effects.

Measurements:

Anthropometrics and blood pressure will be assessed at days 0, 7, and 13. Prior to (day 0), at day 7 and after each 2-wk arm (day 13), a fasting blood sample will be collected to assess serum endotoxin and metabolic chemistries (glucose, insulin, lipids (triglyceride, total and HDL cholesterol), and TLR4/NFκB-dependent genes from whole blood. A breath sample will be collected to assess the correlation analysis of plasma metabolic biomarkers. After the 2-week intervention, from fecal samples collected on day 13, the investigators will assess microbiota composition and function, SCFAs, and intestinal inflammatory markers (calprotectin, myeloperoxidase). During this period, participants will also record daily stool characteristics using a 7-point Bristol Stool scale. On day 13, participants in the fasted state will receive a high-fat/high-glucose meal challenge to induce gut-derived endotoxin translocation. At 30-minute intervals for 3 hours, the investigators will evaluate circulating endotoxin, glucose, and insulin; TLR4/NFκB-dependent genes will be assessed from whole blood at 0 hour and 3-hour. Gut permeability probes will be co-administered with the test meal challenge, and 24-hour urine will be collected to assess gut barrier integrity. Participants will then undergo a 2-week washout prior to receiving the alternative treatment and completing all procedures in an identical manner.

• Study Type: Interventional
• Enrollment (Actual): 30
• Phase: Not Applicable

Contacts and Locations

Study Locations

• United States
  • Ohio
    • Columbus, Ohio, United States, 43201
      • The Ohio State University

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 65 years (Adult, Older Adult)
• Accepts Healthy Volunteers: No

Description

Inclusion Criteria:

• Serum Glucose 100-126 mg/dl
• Waist C >89/102 cm; F/M
• Serum HDL-C: <50/40 mg/dl; F/M
• Serum triglyceride: >150 mg/dl
• Blood Pressure >130/85 mmHg

Exclusion Criteria:

• Unstable body mass (±2 kg over prior 3-mo)
• Vegetarian
• Food allergies or lactose intolerance
• User of dietary supplements or probiotics (within past 1-mo)
• Pregnancy, lactation, changes in birth control (within 6-mo)
• Any gastrointestinal disorders
• Chronic diarrhea
• Smoker
• Excess alcohol (>2 drinks/d)
• Excess aerobic exercise (>5 h/wk)
• Recent antibiotic or anti-inflammatory agent use
• Blood Pressure >140/90 mmHg

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Prevention
• Allocation: Randomized
• Interventional Model: Crossover Assignment
• Masking: Double

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: MFGM-enriched full-fat dairy milk; Participants in this arm will receive MFGM-enriched full-fat dairy milk (3 servings/d) that contains MFGM at 10% phospholipid (relative to total lipid content) delivering MFGM at ~10-times that in full-fat dairy milk.	Dietary supplement: MFGM-enriched full-fat dairy milk; MetS adults with metabolic endotoxemia will be enrolled to complete a 2-arm, double-blind, randomized controlled, crossover trial to test the independent benefits of MFGM. For each 2-wk arm, they will receive MFGM-enriched full-fat dairy milk (3 servings/d) or a matched dairy milk that instead contains soy phospholipid/lecithin (control). Milks have been formulated with soy lecithin or MFGM at 10% phospholipid (relative to total lipid content). This delivers MFGM at ~10-times that in full-fat dairy milk, which reflects that consumers obtain MFGM from dairy foods other than whole milk.
Placebo Comparator: Soy phospholipid/lecithin milk; Participants in this arm will receive a matched dairy milk that instead contains soy phospholipid/lecithin.	Dietary supplement: Soy phospholipid/lecithin milk; MetS adults with metabolic endotoxemia will be enrolled to complete a 2-arm, double-blind, randomized controlled, crossover trial to test the independent benefits of MFGM. For each 2-wk arm, they will receive MFGM-enriched full-fat dairy milk (3 servings/d) or a matched dairy milk that instead contains soy phospholipid/lecithin (control). Milks have been formulated with soy lecithin or MFGM at 10% phospholipid (relative to total lipid content).

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Serum Endotoxin	Data are biomarker fasting concentrations	Day 13
Serum Endotoxin	Data are biomarker area under the time-concentration curve (0-3 hours) on day 13	Day 13 (0, 30, 60, 90, 120, 150, 180 minutes post-meal challenge)

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Plasma Glucose	Data are biomarker area under the time-concentration curve (0-3 hours) on day 13	Day 13 (0, 30, 60, 90, 120, 150, 180 minutes post-meal challenge)
Plasma Glucose	Data are biomarker fasting concentrations	Day 13
Plasma Insulin	Data are biomarker fasting concentrations	Day 13
Plasma Insulin	Data are biomarker area under the time-concentration curve (0-3 hours) on day 13	day 13 (0, 30, 60, 90, 120, 150, 180 minutes post-meal challenge)
Plasma HDL-C	Data are biomarker fasting concentrations	Day 13
Plasma Triglyceride	Data are biomarker fasting concentrations	Day 13
Plasma Total Cholesterol	Data are biomarker fasting concentrations	Day 13
Plasma Glucagon-like Peptide-1	Data are biomarker fasting concentrations	Day 13
Urine Lactulose/Mannitol	Data are the 24-hour urinary excretion ratio of lactulose relative to mannitol post-meal challenge on day 13	Day 13
Urine Sucralose/Erythritol	Data are the 24-hour urinary excretion ratio of sucralose relative to erythritol post-meal challenge on day 13	5-24 hours
Fecal Acetate	Data are biomarker fecal concentrations	Day 13
Fecal Butyrate	Data are biomarker fecal concentrations	Day 13
Fecal Propionate	Data are biomarker fecal concentrations	Day 13
Plasma Gastric Inhibitory Polypeptide	Data are biomarker fasting concentrations	Day 13
Fecal Calprotectin	Data are biomarker fecal concentrations	Day 13
Fecal Myeloperoxidase	Data are biomarker fecal concentrations	Day 13
Whole-blood Messenger RNA Expression of Toll-like Receptor-4 at Fasting	Data are expression levels of whole blood target gene normalized to housekeeping gene (β-actin) expression levels.	Day 13
Whole-blood Messenger RNA Expression of Interleukin-6 at Fasting	Data are expression levels of whole blood target gene normalized to housekeeping gene (β-actin) expression levels.	Day 13
Whole Blood Messenger RNA Expression of Tumor Necrosis Factor-alpha at Fasting	Data are expression levels of whole blood target gene normalized to housekeeping gene (β-actin) expression levels.	Day 13
Whole Blood Messenger RNA Expression of Toll-like Receptor-4 at 3 h Post-meal Challenge	Data are expression levels of whole blood target gene normalized to housekeeping gene (β-actin) expression levels measure at 3-hours post-meal challenge	Day 13, 3 h post-meal
Whole Blood Messenger RNA Expression of Interleukin-6 at 3 h Post-meal Challenge	Data are expression levels of whole blood target gene normalized to housekeeping gene (β-actin) expression levels measure at 3-hours post-meal challenge	Day 13, 3 h post-meal
Whole Blood Messenger RNA Expression of Tumor Necrosis Factor-alpha at 3 h Post-meal Challenge	Data are expression levels of whole blood target gene normalized to housekeeping gene (β-actin) expression levels measure at 3-hours post-meal challenge	Day 13, 3 h post-meal

Collaborators and Investigators

• Sponsor: Ohio State University
• Investigators: Principal Investigator: Richard Bruno, PhD, Ohio State University

Publications and helpful links

General Publications

• Pokala A, Kraft J, Taormina VM, Michalski MC, Vors C, Torres-Gonzalez M, Bruno RS. Whole milk dairy foods and cardiometabolic health: dairy fat and beyond. Nutr Res. 2024 Jun;126:99-122. doi: 10.1016/j.nutres.2024.03.010. Epub 2024 Mar 21.
• Bruno RS, Pokala A, Torres-Gonzalez M, Blesso CN. Cardiometabolic health benefits of dairy-milk polar lipids. Nutr Rev. 2021 Dec 8;79(Suppl 2):16-35. doi: 10.1093/nutrit/nuab085.
• Quarles WR, Pokala A, Shaw EL, Ortega-Anaya J, Hillmann L, Jimenez-Flores R, Bruno RS. Alleviation of Metabolic Endotoxemia by Milk Fat Globule Membrane: Rationale, Design, and Methods of a Double-Blind, Randomized, Controlled, Crossover Dietary Intervention in Adults with Metabolic Syndrome. Curr Dev Nutr. 2020 Jul 25;4(9):nzaa130. doi: 10.1093/cdn/nzaa130. eCollection 2020 Sep.
• Pokala A, Quarles WR, Ortega-Anaya J, Jimenez-Flores R, Cao S, Zeng M, Hodges JK, Bruno RS. Milk-Fat-Globule-Membrane-Enriched Dairy Milk Compared with a Soy-Lecithin-Enriched Beverage Did Not Adversely Affect Endotoxemia or Biomarkers of Gut Barrier Function and Cardiometabolic Risk in Adults with Metabolic Syndrome: A Randomized Controlled Crossover Trial. Nutrients. 2023 Jul 23;15(14):3259. doi: 10.3390/nu15143259.

Study record dates

Study Major Dates

• Study Start (Actual): July 1, 2019
• Primary Completion (Actual): December 20, 2020
• Study Completion (Actual): December 20, 2020

Study Registration Dates

• First Submitted: February 28, 2019
• First Submitted That Met QC Criteria: February 28, 2019
• First Posted (Actual): March 4, 2019

Study Record Updates

• Last Update Posted (Estimated): November 18, 2025
• Last Update Submitted That Met QC Criteria: November 4, 2025
• Last Verified: November 1, 2025

More Information

Terms related to this study

• Keywords: obesity; inflammation; endotoxin; gut health; metabolic endotoxemia; milk fat globule membrane
• Additional Relevant MeSH Terms: Pathologic Processes; Nutrition Disorders; Metabolic Diseases; Overnutrition; Body Weight; Glucose Metabolism Disorders; Insulin Resistance; Hyperinsulinism; Overweight; Pathological Conditions, Signs and Symptoms; Nutritional and Metabolic Diseases; Signs and Symptoms; Obesity; Metabolic Syndrome; Inflammation
• Other Study ID Numbers: 2018H0564

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No