Contact information
Type
Scientific
Primary contact
Dr Ramon Estruch
ORCID ID
Contact details
Hospital Clínic de Barcelona
c/Villarroel nº170
Barcelona
08036
Spain
+34 932 27 93 65
restruch@clinic.ub.es
Additional identifiers
EudraCT number
ClinicalTrials.gov number
Protocol/serial number
AGL2004-08378-C02-02/ALI
Study information
Scientific title
Bioavailability of soluble phenols of cocoa. Scientific basis of the interaction of phenolic compounds and cellular and serum inflammatory biomarkers related to atherosclerosis: an open randomised cross-over controlled trial
Acronym
Study hypothesis
Soluble polyphenolic compounds of cocoa powder will reduce inflammatory biomarkers related to atherosclerosis. No adverse events will be observed.
Ethics approval
Institutional Review Board of the Hospital Clínic de Barcelona, approved on the 3rd June 2003.
Study design
Open randomised cross-over controlled trial
Primary study design
Interventional
Secondary study design
Randomised controlled trial
Trial setting
Other
Trial type
Other
Patient information sheet
Condition
Arteriosclerosis
Intervention
Initial wash-out period (15 days) followed by first intervention (28 days) and second intervention (28 days).
Intervention 1: 40 g/day of soluble cocoa powder dissolved in 250 mL of skim milk
Intervention 2: 250 mL of skim milk daily
There was no wash-out period between the two interventions. Since the period of each intervention was four weeks and the change of the variables studied occurred <15 days, we assumed that we can evaluate the effects of both interventions, comparing the results of the analysis performed at the end of each intervention.
Intervention type
Drug
Phase
Not Applicable
Drug names
Cocoa
Primary outcome measures
1. Leukocyte adhesion molecule expression
Lymphocyte and monocyte adhesion molecules on these cells will be marked with monoclonal antibodies (MAb) conjugated with fluorescein-isothiocyanate (FITC) and phycoerythrin (PE) by direct double immunofluorescence. The MAb of the adhesion molecules used will be: anti-CD11a (LFA-1), anti-CD40L, anti-CD11b (Mac-1) (Bender MedSystems Diagnostics, Austria), anti-Syalil Lewis (anti-CD15s) (Pharmingen, USA), anti-CD49d (VLA-4) (Cytogmos, Spain). The monoclonal antibodies used to mark the T-lymphocytes will be anti-CD2 and monocytes, anti-CD14 (Caltag Laboratories, USA).
2. Soluble adhesion molecules
The following serum soluble adhesion molecules will be determined by enzyme-linked immunosorbent assay (ELISA) kits: C-reactive protein (CRP), sICAM-1, sVCAM-1, sE-selectin, and sP-selectin, as well as sMCP-1, and IL-6 (Immunotech, Czech Republic).
3. Nuclear factor kppa B by western blot of peripheral blood mononuclear cells
4. Bioavailability of soluble phenolic compounds of cocoa powder by Liquid Chromatography/Mass Spectrometry/Mass Spectrometry (LC-MS/MS) analysis of plasma and urine metabolites
All outcomes will be measured at baseline and after each intervention period.
Secondary outcome measures
1. Medical record
A complete medical record will be obtained from all participants, which included data on cocoa intake, smoking and dietary habits. Blood pressure and heart rate will be measured with an electronic apparatus Omron HEM-705CP (Netherlands).
2. Nutrition assessment and general analyses
All participants will complete a validated nutritional questionnaire at baseline to determine the total quantity of calories ingested in the previous month as well as the proportion corresponding to carbohydrates, lipids and proteins. Overall nutrition will be determined by percentage of ideal weight, lean body mass and body mass index. Waist perimeter will be measured. The proteic nutrition will be determined on the basis of the following parameters: haemoglobin, total lymphocyte count, total proteins, albumin, prealbumin, transferrin and retinol-binding protein. Serum and intraerythrocytary folic acid concentrations will be measured, as well as serum vitamin A, B1, B12, C, E, B-carotenes, Zn, Mg and Se concentrations.
Moreover, the following measurements will also be obtained: red blood cell count, hematocrit, mean corpuscular volume, leukocyte count, glucose, creatinine, electrolytes, uric acid, transaminases, lactate dehydrogenase, alkaline phosphatase, gammaglutamyl transpeptidase and bilirrubin.
3. Coagulation tests
The following parameters will also be determined: platelet count, prothrombin time, and plasma fibrinogen.
4. Serum lipoproteins and others
Total cholesterol, triglycerides, cHDL, cLDL, Apo A1, Apo B, lipoprotein (a) and homocysteine will be determined.
5. Diet and exercise monitoring
All participants will follow an isocaloric diet prepared according to their personal preferences. Subjects will be asked to exclude all other cocoa-containing foods throughout the study and to limit the intake of foods containing high polyphenol content, such as virgin olive oil, red wine, tea, fruits, and vegetables. The diet will be strictly monitored during the study. Diet compliance will be assessed from 3-days (2 weekdays and 1 weekend day) diet records administered before each evaluation. This assessment will be administered by trained personnel. The foods ingested will be converted into nutritional values with the aid of the Professional Diet Balancer software (Cardinal Health Systems, Inc., USA). Physical activity will also be evaluated with the Minnesota Leisure Time Physical Activity questionnaire which has also been validated in Spain. Control of the diet and physical exercise will be carried out before and after each intervention, the same day on which the clinical examinations are performed and blood is withdrawn for immunologic studies.
All outcomes will be measured at baseline and after each intervention period.
Overall trial start date
01/12/2004
Overall trial end date
01/12/2007
Reason abandoned
Eligibility
Participant inclusion criteria
1. Males and females between 55 and 80 years old
2. Those without documented cardiovascular disease (ischemic heart disease, stroke, or peripheral vascular disease)
3. Those who have diabetes mellitus or two or more of the following factors:
3.1. Current smoking
3.2. Hypertension
3.3. Hypercholesterolemia (low-density Lipoprotein [LDL]-cholesterol >160 mg/dl)
3.4. High-density lipoprotein (HDL)-cholesterol <40 mg/dl
3.5. Obese (body mass index >30 kg/m^2)
3.6. Family history of premature coronary heart disease
4. Participant should give signed informed consent
Participant type
Patient
Age group
Senior
Gender
Both
Target number of participants
45
Participant exclusion criteria
1. Subjects with a previous history of cardiovascular disease (ischemic heart disease, stroke or peripheral vascular disease)
2. Any severe chronic disease
3. History of allergic reactions to any cocoa or milk components
Recruitment start date
01/12/2004
Recruitment end date
01/12/2007
Locations
Countries of recruitment
Spain
Trial participating centre
Hospital Clínic de Barcelona
Barcelona
08036
Spain
Sponsor information
Organisation
Ministry of Science and Innovation (Ministerio de Ciencia e Innovación) (Spain)
Sponsor details
C/Albacete
5
Madrid
28027
Spain
Sponsor type
Government
Website
Funders
Funder type
Government
Funder name
Ministry of Science and Innovation (Ministerio de Ciencia e Innovación) (Spain) (ref: AGL2004-08378-C02-02/ALI)
Alternative name(s)
Funding Body Type
Funding Body Subtype
Location
Results and Publications
Publication and dissemination plan
Not provided at time of registration
Intention to publish date
Participant level data
Not provided at time of registration
Results - basic reporting
Publication summary
1. 2009 results in http://www.ncbi.nlm.nih.gov/pubmed/19776136
2. 2012 results in http://www.ncbi.nlm.nih.gov/pubmed/21550218
Publication citations
-
Results
Monagas M, Khan N, Andres-Lacueva C, Casas R, Urpí-Sardà M, Llorach R, Lamuela-Raventós RM, Estruch R, Effect of cocoa powder on the modulation of inflammatory biomarkers in patients at high risk of cardiovascular disease., Am. J. Clin. Nutr., 2009, 90, 5, 1144-1150, doi: 10.3945/ajcn.2009.27716.
-
Results
Khan N, Monagas M, Andres-Lacueva C, Casas R, Urpí-Sardà M, Lamuela-Raventós RM, Estruch R, Regular consumption of cocoa powder with milk increases HDL cholesterol and reduces oxidized LDL levels in subjects at high-risk of cardiovascular disease., Nutr Metab Cardiovasc Dis, 2012, 22, 12, 1046-1053, doi: 10.1016/j.numecd.2011.02.001.