Dark Chocolate and Platelet Function in Humans

NCT01099150 | Completed DMC

• 2 other identifiers: 60009/002
• Study Type: interventional
• Target: 42
• Locations: 1 country
• Sites: 1

Brief Summary

Cardiovascular disease is a major cause of mortality worldwide and responsible for one out of three global deaths. A main characteristic of cardiovascular disease is impaired blood flow and formation of blood clots. Platelets are clot-forming cells responsible for the prevention of bleeding. However, in disease conditions they may be overly activated, promoting blood clots and blockage of blood vessels. Consumption of diets rich in fruits and vegetables decreases mortality from cardiovascular disease through a number of mechanisms, including the prevention of platelet clotting and aggregation. There is some evidence suggesting that platelet aggregation may be modulated through a group of compounds known as flavan-3-ols, which are found in various foods, and especially in cocoa. However, the mechanisms by which those compounds affect platelet function are not yet fully understood. We designed a human study assessing the mechanisms by which flavan-3-ols from cocoa beneficially affect platelet function and the platelet proteome.

Conditions

Cardiovascular Disease

Keywords

Platelets; Platelet aggregation; Platelet activation; Platelet function; Light transmission aggregometry; Platelet Function Analyzer-100 (PFA-100); P-selectin; Fibrinogen receptor; Glycoprotein IIb-IIIa; Chocolate; Cocoa; Flavan-3-ols; Procyanidins

Outcome Measures

Primary Outcomes (1)

• Change in light transmission aggregometry of platelet-rich plasma

  * Using a Helena Platelet Aggregation Chromogenic Kinetics System-4 (PACKS-4) light transmission aggregometer * Induced by adenosine diphosphate (ADP) and thrombin receptor-activating peptide (TRAP)

  Post-prandial, up to 6 hours after chocolate consumption

Secondary Outcomes (14)

• Change in ex vivo bleeding time using the Platelet Function Analyzer-100 (PFA-100)

  Post-prandial, up to 6 hours after chocolate consumption

• Change in P-selectin expression and activation of the fibrinogen receptor by flow cytometry

  Post-prandial, up to 6 hours after chocolate consumption

• Levels of flavan-3-ols and their metabolites in plasma and urine

  Post-prandial, up to 6 hours after chocolate consumption

• Changes in the platelet proteome

  Post-prandial, 2 hours after chocolate ingestion

• Changes in thromboxane A2 production induced by ADP and TRAP

  Post-prandial, up to 6 hours after chocolate consumption

Study Arms (1)

42 healthy volunteers - crossover

EXPERIMENTAL

Acute consumption of three interventions (60 g dark chocolate enriched in flavan-3-ols, 60 g standard dark chocolate, or 60 g white chocolate) on three separate days (at least 2 weeks apart) in random order. Post-prandial measurements at t = 0 h, t = 2 h and t = 6 h.

Dietary Supplement: Dark chocolate enriched in flavan-3-ols and procyanidins; Dietary Supplement: Standard dark chocolate; Dietary Supplement: White chocolate

Interventions

Dark chocolate enriched in flavan-3-ols and procyanidins DIETARY_SUPPLEMENT

Acute consumption (within 15 minutes) of 60 g of chocolate containing \~900 mg of total flavan-3-ols and procyanidins.

Also known as: CocoanOX12%-containing chocolate

42 healthy volunteers - crossover

Standard dark chocolate DIETARY_SUPPLEMENT

Acute consumption (within 15 minutes) of 60 g of chocolate containing \~400 mg total flavan-3-ols and procyanidins.

42 healthy volunteers - crossover

White chocolate DIETARY_SUPPLEMENT

Acute consumption (within 15 minutes) of 60 g of white chocolate containing no flavan-3-ols and procyanidins.

42 healthy volunteers - crossover

Eligibility Criteria

• Age: 18 Years - 70 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• Healthy male and/or female volunteers, aged between 18 and 70 years

You may not qualify if:

• Subjects are excluded if:
• they are taking aspirin or aspirin-containing drugs, other anti-inflammatory drugs, or any drugs or herbal medicines known to alter platelet function or the haemostatic system in general (without a minimum washout period of one month)
• they are taking fish oils or evening primrose oil, or fat soluble vitamin supplements within the last 4 weeks
• they are taking any medicine known to affect lipid and/or glucose metabolism
• they are taking hormone replacement therapy
• they have any known clinical signs of diabetes, hypertension, renal, hepatic, hematological disease, gastrointestinal disorders, endocrine disorders, coronary heart disease, infection or cancer
• they are suffering from alcohol or any other substance abuse or are having eating disorders
• they are usually consuming a vegetarian diet
• they have a BMI below 18 or above 35 kg/ sqm
• they are undertaking more than 6 hours of vigorous exercise per week
• they are having an abnormal menstrual cycle
• they are pregnant
• they suffer from an allergy to cocoa or any of the ingredients contained within either of the chocolate bars
• they have been giving a pint of blood for transfusion purposes within the last month
• they have a low platelet count (\< 170 x 10E09/ L)

Sponsors & Collaborators

• University of Aberdeen: lead
• Rural and Environment Research and Analysis Directorate (RERAD, UK): collaborator
• Biotechnology and Biological Sciences Research Council: collaborator
• Natraceutical Industrial S.L.U., Valencia, Spain: collaborator

Study Sites (1)

University of Aberdeen Rowett Institute of Nutrition and Health

Aberdeen, Aberdeenshire, AB21 9SB, United Kingdom

Location

Related Publications (23)

• Baba S, Osakabe N, Yasuda A, Natsume M, Takizawa T, Nakamura T, Terao J. Bioavailability of (-)-epicatechin upon intake of chocolate and cocoa in human volunteers. Free Radic Res. 2000 Nov;33(5):635-41. doi: 10.1080/10715760000301151.

  PMID: 11200094 BACKGROUND

• Bordeaux B, Yanek LR, Moy TF, White LW, Becker LC, Faraday N, Becker DM. Casual chocolate consumption and inhibition of platelet function. Prev Cardiol. 2007 Fall;10(4):175-80. doi: 10.1111/j.1520-037x.2007.06693.x.

  PMID: 17917513 BACKGROUND

• de Roos B, Duthie SJ, Polley AC, Mulholland F, Bouwman FG, Heim C, Rucklidge GJ, Johnson IT, Mariman EC, Daniel H, Elliott RM. Proteomic methodological recommendations for studies involving human plasma, platelets, and peripheral blood mononuclear cells. J Proteome Res. 2008 Jun;7(6):2280-90. doi: 10.1021/pr700714x. Epub 2008 May 20.

  PMID: 18489134 BACKGROUND

• Flammer AJ, Hermann F, Sudano I, Spieker L, Hermann M, Cooper KA, Serafini M, Luscher TF, Ruschitzka F, Noll G, Corti R. Dark chocolate improves coronary vasomotion and reduces platelet reactivity. Circulation. 2007 Nov 20;116(21):2376-82. doi: 10.1161/CIRCULATIONAHA.107.713867. Epub 2007 Nov 5.

  PMID: 17984375 BACKGROUND

• Guerrero JA, Navarro-Nunez L, Lozano ML, Martinez C, Vicente V, Gibbins JM, Rivera J. Flavonoids inhibit the platelet TxA(2) signalling pathway and antagonize TxA(2) receptors (TP) in platelets and smooth muscle cells. Br J Clin Pharmacol. 2007 Aug;64(2):133-44. doi: 10.1111/j.1365-2125.2007.02881.x. Epub 2007 Apr 10.

  PMID: 17425630 BACKGROUND

• Hamed MS, Gambert S, Bliden KP, Bailon O, Singla A, Antonino MJ, Hamed F, Tantry US, Gurbel PA. Dark chocolate effect on platelet activity, C-reactive protein and lipid profile: a pilot study. South Med J. 2008 Dec;101(12):1203-8. doi: 10.1097/SMJ.0b013e31818859eb.

  PMID: 19005437 BACKGROUND

• Heptinstall S, May J, Fox S, Kwik-Uribe C, Zhao L. Cocoa flavanols and platelet and leukocyte function: recent in vitro and ex vivo studies in healthy adults. J Cardiovasc Pharmacol. 2006;47 Suppl 2:S197-205; discussion S206-9. doi: 10.1097/00005344-200606001-00015.

  PMID: 16794458 BACKGROUND

• Hermann F, Spieker LE, Ruschitzka F, Sudano I, Hermann M, Binggeli C, Luscher TF, Riesen W, Noll G, Corti R. Dark chocolate improves endothelial and platelet function. Heart. 2006 Jan;92(1):119-20. doi: 10.1136/hrt.2005.063362. No abstract available.

  PMID: 16365364 BACKGROUND

• Hubbard GP, Wolffram S, de Vos R, Bovy A, Gibbins JM, Lovegrove JA. Ingestion of onion soup high in quercetin inhibits platelet aggregation and essential components of the collagen-stimulated platelet activation pathway in man: a pilot study. Br J Nutr. 2006 Sep;96(3):482-8.

  PMID: 16925853 BACKGROUND

• Hubbard GP, Wolffram S, Lovegrove JA, Gibbins JM. Ingestion of quercetin inhibits platelet aggregation and essential components of the collagen-stimulated platelet activation pathway in humans. J Thromb Haemost. 2004 Dec;2(12):2138-45. doi: 10.1111/j.1538-7836.2004.01067.x.

  PMID: 15613018 BACKGROUND

• Innes AJ, Kennedy G, McLaren M, Bancroft AJ, Belch JJ. Dark chocolate inhibits platelet aggregation in healthy volunteers. Platelets. 2003 Aug;14(5):325-7. doi: 10.1080/0953710031000123681.

  PMID: 12944249 BACKGROUND

• Mavrommatis Y, O'Kennedy N, de Roos B et al. Effect of eicosapentaenoic acid or docosahexaenoic acid intervention on platelet aggregation: does soluble epoxide hydrolase play a role? unpublished, 2010

  BACKGROUND

• Murphy KJ, Chronopoulos AK, Singh I, Francis MA, Moriarty H, Pike MJ, Turner AH, Mann NJ, Sinclair AJ. Dietary flavanols and procyanidin oligomers from cocoa (Theobroma cacao) inhibit platelet function. Am J Clin Nutr. 2003 Jun;77(6):1466-73. doi: 10.1093/ajcn/77.6.1466.

  PMID: 12791625 BACKGROUND

• Nardini M, Natella F, Scaccini C. Role of dietary polyphenols in platelet aggregation. A review of the supplementation studies. Platelets. 2007 May;18(3):224-43. doi: 10.1080/09537100601078083.

  PMID: 17497435 BACKGROUND

• Natella F, Nardini M, Belelli F, Pignatelli P, Di Santo S, Ghiselli A, Violi F, Scaccini C. Effect of coffee drinking on platelets: inhibition of aggregation and phenols incorporation. Br J Nutr. 2008 Dec;100(6):1276-82. doi: 10.1017/S0007114508981459. Epub 2008 Apr 28.

  PMID: 18439332 BACKGROUND

• Natella F, Nardini M, Virgili F, Scaccini C. Role of dietary polyphenols in the platelet aggregation network - A review of the in vitro studies. Curr Top Nutraceut Res 4(1): 1-22, 2006.

  BACKGROUND

• Ostertag LM, O'Kennedy N, Kroon PA, Duthie GG, de Roos B. Impact of dietary polyphenols on human platelet function--a critical review of controlled dietary intervention studies. Mol Nutr Food Res. 2010 Jan;54(1):60-81. doi: 10.1002/mnfr.200900172.

  PMID: 20058256 BACKGROUND

• Pearson DA, Paglieroni TG, Rein D, Wun T, Schramm DD, Wang JF, Holt RR, Gosselin R, Schmitz HH, Keen CL. The effects of flavanol-rich cocoa and aspirin on ex vivo platelet function. Thromb Res. 2002 May 15;106(4-5):191-7. doi: 10.1016/s0049-3848(02)00128-7.

  PMID: 12297125 BACKGROUND

• Rechner AR, Kroner C. Anthocyanins and colonic metabolites of dietary polyphenols inhibit platelet function. Thromb Res. 2005;116(4):327-34. doi: 10.1016/j.thromres.2005.01.002. Epub 2005 Feb 8.

  PMID: 16038718 BACKGROUND

• Rein D, Paglieroni TG, Wun T, Pearson DA, Schmitz HH, Gosselin R, Keen CL. Cocoa inhibits platelet activation and function. Am J Clin Nutr. 2000 Jul;72(1):30-5. doi: 10.1093/ajcn/72.1.30.

  PMID: 10871557 BACKGROUND

• Schewe T, Sadik C, Klotz LO, Yoshimoto T, Kuhn H, Sies H. Polyphenols of cocoa: inhibition of mammalian 15-lipoxygenase. Biol Chem. 2001 Dec;382(12):1687-96. doi: 10.1515/BC.2001.204.

  PMID: 11843182 BACKGROUND

• Schramm DD, Wang JF, Holt RR, Ensunsa JL, Gonsalves JL, Lazarus SA, Schmitz HH, German JB, Keen CL. Chocolate procyanidins decrease the leukotriene-prostacyclin ratio in humans and human aortic endothelial cells. Am J Clin Nutr. 2001 Jan;73(1):36-40. doi: 10.1093/ajcn/73.1.36.

  PMID: 11124747 BACKGROUND

• Wang-Polagruto JF, Villablanca AC, Polagruto JA, Lee L, Holt RR, Schrader HR, Ensunsa JL, Steinberg FM, Schmitz HH, Keen CL. Chronic consumption of flavanol-rich cocoa improves endothelial function and decreases vascular cell adhesion molecule in hypercholesterolemic postmenopausal women. J Cardiovasc Pharmacol. 2006;47 Suppl 2:S177-86; discussion S206-9. doi: 10.1097/00005344-200606001-00013.

  PMID: 16794456 BACKGROUND

MeSH Terms

Conditions

Cardiovascular Diseases

Interventions

Proanthocyanidins

Intervention Hierarchy (Ancestors)

Flavonoids; Chromones; Benzopyrans; Pyrans; Heterocyclic Compounds, 1-Ring; Heterocyclic Compounds; Heterocyclic Compounds, 2-Ring; Heterocyclic Compounds, Fused-Ring; Tannins; Biopolymers; Polymers; Macromolecular Substances

Study Officials

• Baukje de Roos, MSc PhD

  University of Aberdeen Rowett Institute of Nutrition and Health

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: SINGLE
• Who Masked: INVESTIGATOR
• Purpose: PREVENTION
• Intervention Model: CROSSOVER
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: March 25, 2010
• First Posted: April 6, 2010
• Study Start: March 1, 2009
• Primary Completion: November 1, 2009
• Study Completion: May 1, 2011
• Last Updated: April 17, 2012

Record last verified: 2012-04

Locations

GB (1)