Virgin olive oil and high-density lipoprotein functionality: a model for tailoring functional food

ISRCTN	ISRCTN77500181
DOI	https://doi.org/10.1186/ISRCTN77500181
Protocol serial number	AGL2009-13517-C03-01
Sponsor	Spanish Ministry of Economy and Competitiveness (Spain)
Funder	Spanish Ministry Science and Technology (Spain) ref: AGL2009-13517-C03-01

Submission date: 06/03/2012
Registration date: 09/05/2012
Last edited: 25/04/2016

Recruitment status: No longer recruiting
Overall study status: Completed
Condition category: Nutritional, Metabolic, Endocrine

Prospectively registered

Protocol

Statistical analysis plan

Results

Individual participant data

Plain English summary of protocol

Background and study aims
The aim of this study is to find out whether consuming olive oil enriched with olive oil phenolic compounds (OOPC) and olive oil enriched with OOPC plus flavonoids from thyme increases quality and quantity of high density lipoprotein (HDL) compared with consuming normal virgin olive oil.

Who can participate?
Male and female hypercholesterolemic (>200mg/dL) patients aged 20-80 years.

What does the study involve?
Participants will receive 25 ml per day of one of the three types of olive oil in three separate periods of 3 weeks, preceded each one by a 2-week wash-out period.

What are the possible benefits and risks of participating?
The benefit for the participants will be the possibility of reducing their cardiovascular disease risk. No harmful effects have been found to be associated with olive oil consumption.

Where is the study run from?
Institut Hospital del Mar dInvestigacions Mèdiques (IMIM) , Barcelona, Spain

When is the study starting and how long is it expected to run for?
April to September 2012

Who is funding the study?
Spanish Ministry of Economy and Competitiveness

Who is the main contact?
Dr María-Isabel Covas
mcovas@imim.es
mfito@imim.es

Contact information

Dr Maria Isabel Covas
Scientific

Doctor Aiguader, 88
Barcelona
08003
Spain

Phone	+34 (0)933 160 720
Email	mcovas@imim.es

Study information

Primary study design	Interventional
Study design	Randomized controlled double-blind cross-over single-centre clinical supplementation trial
Secondary study design	Randomised cross over trial
Study type	Participant information sheet
Scientific title	Virgin Olive oil and HDL Functionality (VOHF): a model for tailoring functional food: the VOHF project
Study acronym	VOHF
Study objectives	A flavoured functional olive oil enriched not only with its own phenolics but also with complementary ones (regarding structure/function) from thyme could provide additional health benefits and consumers acceptation.
Ethics approval(s)	Clinical Research Ethical Committee of the "Institut Municipal dAssistència Sanitària (IMAS)", 09/09/2009, ref: 2009/3347/I
Health condition(s) or problem(s) studied	Hypercholesterolemic patients
Intervention	Current interventions as of 23/04/2014: Randomized, controlled, double-blind, cross-over, single-centre clinical supplementation trial, in which 33 subjects (19 men and 14 women) will be randomised to one of 3 orders of administration of 25 ml/day of raw: 1. Virgin olive oil (Control) 2. Functional Olive Oil 1 enriched with its own phenolic compounds (FOO1) 3. Functional Olive OIl 2 enriched with its olive oil phenolic compounds plus those of thyme (FOO2) This will be in 3 periods of 3 weeks Previous interventions: Randomized, double-blind, cross-over, single-centre clinical supplementation trial, in which 30 subjects (15 men and 15 women) will be randomised to one of 3 orders of administration of: 1. 25 mL/day of raw refined virgin olive oil (Control) 2. Functional Olive Oil 1 enriched with its own phenolic compounds (FOO1) 3. Functional Olive OIl 2 enriched with its olive oil phenolic compounds plus those of thyme (FOO2) This will be in 3 periods of 3 weeks
Intervention type	Supplement
Primary outcome measure(s)	1. HDL functionality: The following parameters will be measured at baseline and before and after each intervention: 1.1. Markers of compliance: Measurement of tyrosol and hydroxytyrosol in spot morning urine will be performed by GC/mass spectrometry 1.2. Parameters of HDL functionality: Isolation of HDL will be done by sequential centrifugation. 2. The following parameters will be measured in isolated HDL: 2.1. Fatty acid content, vitamin E, and total phenolic content in HDL by HPLC-DAD 2.2. Apolipoproteins (Apo)A1, ApoA2, and ApoA4 by ELISA 2.3. Cholesterol esther transfer protein (CETP) and lecitine cholesterol acyl transferase (LCAT) by fluorometry 2.4. Paraoxonase (Organophosphatase activity) PON1) and PAF-AH activities by spectrometry with automated mode 2.5. 3 chlorotyrosine and 3-nitro-tyrosine by GC/MS 2.6. Bilayer fluidity of HDL will be measured by confocal microscopy. Cholesterol efflux from cells will be measured in culture of macrophages. 3. OMICS Gene expression assay will be performed as previously described. Several candidate genes implicated in the increase of the HDL cholesterol and in the improvement of the HDL functionality in humans will be tested. Candidate genes: ABCA1: ATP-binding cassette, sub-family A (ABC1), member 1; ABCG1: ATP-binding cassette, sub-family G (WHITE), member 1; and ABCG4: ATP-binding cassette, sub-family G (WHITE), member 4 ApoA1: apolipoprotein A- I; and ApoE: apolipoprotein E; LPL: lipoprotein lipase; PAF-AH1B3: platelet-activating factor acetylhydrolase, isoform Ib, gamma subunit 29kDa; PPARalpha: peroxisome proliferator -activated receptor alpha; PPARgamma: peroxisome proliferator-activated receptor gamma; and PBPPAR: PPAR binding protein; RARA: retinoic acid receptor, alpha. Proteomic analyses, by separation with 2-DE (two-dimensional gel electrophoresis) and PMF (protein mass fingerprinting) with MALDI-TOF-MS (matrix -assisted laser desorption/ionization time of flight mass spectrometry ,will be performed in plasma samples.
Key secondary outcome measure(s)	Oxidative damage The following parameters will be measured at baseline and before and after each intervention: 1. Plasma lipids will be performed by enzymatic methods in a automated mode. In vivo circulating oxidized LDL and 3 chlorotyrosine derivates in urine will be determined by ELISA 2. Protein carbonyl content (PCC) by spectrometry 3. 8-isoprostane (8-epi PGF2á) 4. 8-hydroxydeoxyguanosine (8-OHdG) by enzyme-linked immunosorbent assay (ELISA) Added 23/04/2014: 5. Endothelial function 6. Intestinal microorganism study
Completion date	07/09/2012

Eligibility

Participant type(s)	Patient
Age group	Adult
Sex	All
Target sample size at registration	33
Key inclusion criteria	Current inclusion criteria as of 23/04/2014: 1. Hypercholesterolemic patients (>200 mg/dL) 2. Aged 20-80 years Previous inclusion criteria: 1. Hyperlipidemic patients 2. Patients with low (<40 mg/dL) HDL cholesterol 3. Aged 20-60 years
Key exclusion criteria	Current exclusion criteria as of 25/04/2014: 1. Smokers 2. Athletes with PA >3000 kcal/day 3. BMI >35 kg/m2 4. Diabetes, multiple allergies, intestinal diseases 5. Any condition that limits the mobility of the subject making study visits impossible 6. Life threatening illness such as cancer or severe disease with a lowered expected 3 year survival 7. Any other disease or condition that would worsen the adherence to the measurements or treatment Previous exclusion criteria: 1. Intake of antioxidant supplement or acetylsalicylic acid or any other drug with established antioxidative properties 2. Athletes with PA >3000 kcal/week in leisure-time 3. Obesity (BMI > 30 kg/m2) 4. Diabetes, multiple allergies, intestinal diseases 5. Any condition that limits the mobility of the subject making study visits impossible 6. Life threatening illness such as cancer or severe disease with a lowered expected 3 year survival 7. Any other disease or condition that would worsen the adherence to the measurements or treatment
Date of first enrolment	02/04/2012
Date of final enrolment	07/09/2012

Locations

Countries of recruitment

Spain

Study participating centre

Doctor Aiguader, 88

Barcelona
08003
Spain

Results and Publications

Individual participant data (IPD) Intention to share	No
IPD sharing plan summary	Not provided at time of registration
IPD sharing plan

Study outputs

Output type	Details	Date created	Date added	Peer reviewed?	Patient-facing?
Results article	results	10/06/2015		Yes	No
Participant information sheet	Participant information sheet	11/11/2025	11/11/2025	No	Yes

Editorial Notes

25/04/2016: Publication reference added.

On 23/04/2014 the following changes were made to the trial record:
1. The disease/condition/study domain field was changed from 'Hyperlipidemic patients with low HDL cholesterol' to 'Hypercholesterolemic patients'
2. The target number of participants was changed from 30 to 33.