Effect of Cilostazol on Endothelial Progenitor Cells and Endothelial Function in High Risk for Cardiovascular Disease

Study Overview
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Brief Summary
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The number and function of circulating endothelial progenitor cells (EPCs) are inversely associated with coronary risk factors and atherosclerotic diseases. This double-blind, randomized, placebo-controlled trial to evaluate the effects of cilostazol on human early EPCs and endothelial function as well as the potential mechanisms of action in patients with high risk for cardiovascular disease.

Detailed Description
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titration of drugs run-in period: eligible subjects are screened and baseline blood samples are obtained study period: 12 weeks subjects with cilostazol and subjects with dummy placebo On the first day after the end of the study period, the follow-up data are obtained by the same procedure blood sampling and measurement of serum biomarkers obtained from peripheral veins in all study subjects at the run-in period and the end of the treatment period of the study sent for isolation, cell culture, and assays of human EPCs also stored for enzyme-linked immunosorbent assay (stromal cell derived factor-alfa1, adiponectin, soluble thrombomodulin, vascular endothelial growth factor) assays of human EPCs colony formation by EPCs quantification of EPCs and apoptotic endothelial cells chemotactic motility, proliferation/viability and apoptosis assays measurement of flow-mediated dilatation (FMD) of left brachial artery by sonography

Official Title
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Cilostazol Enhances the Number and Functions of Circulating Endothelial Progenitor Cells Mediated Through Multiple Mechanisms in Patients With High Risk for Cardiovascular Disease

Conditions
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Cardiovascular Diseases

Intervention / Treatment
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* Drug: Cilostazol
* Drug: Dummy Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: high-risk patients who have at least one of the following situations without pre-existing cardiovascular disease including peripheral artery disease or coronary artery disease: type 2 diabetes mellitus metabolic syndrome stage 3 (or more advanced) chronic kidney disease 2 or more coronary risk factors (male > 45 years or female > 55 years, hypertension, tobacco smoking, hyperlipidemia, family history of cardiovascular disease) Exclusion Criteria: ankle-brachial index less than 0.9 or more than 1.3 in one or both legs significant stenosis (more than 50% as compared to reference vessel) in peripheral artery on image study symptoms suggesting peripheral artery disease in at least one leg clinical or electrocardiographic evidence of coronary artery disease clinical evidence of cerebrovascular disease severe liver dysfunction (transaminases >10 times of upper normal limit, history of liver cirrhosis, or hepatoma) left ventricular ejection fraction (<50% by echocardiography) documented active malignancy chronic inflammatory disease known drug allergy history for cilostazol current use of cilostazol or any other cAMP-elevator premenopausal women

Ages Eligible for Study
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Minimum Age: 20 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Quadruple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Cilostazol<br>One tablet (100 mg) twice per day for 12 weeks | Drug: Cilostazol<br>* One tablet (100 mg) twice per day for 12 weeks<br>* Other names: Pletaal (brand name);|
| Placebo Comparator: Dummy Placebo<br>One tablet twice per day for 12 weeks | Drug: Dummy Placebo<br>* One tablet twice per day for 12 weeks<br>* Other names: Control;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Circulating EPCs Number | Peripheral blood mononuclear cells (one million cells in each) are suspended in 100 µL phosphate-buffered saline and incubated for 30 min with monoclonal antibodies against human peridinin chlorophyll protein-conjugated cluster of differentiation antigen-45, phycoerythrin-conjugated anti-human cluster of differentiation antigen-34 antibody and anti-human kinase insert domain receptor (KDR) antibody conjugated with Alexa Flour 647. Cells are washed and analyzed on a FACSCalibur flow cytometer with 100,000 events in the lymphocyte gate. EPCs, which are defined as negative for cluster of differentiation antigen-45 and positive for cluster of differentiation antigen-34 and KDR. Based on the peripheral blood mononuclear cell counts, the absolute number of circulating EPCs/µL is calculated. | 3 months | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Viability (Proliferation) of EPCs | 250,000 cells are seeded in each well of a 96-well plate and the cells are added with 200 μl of the culture medium and incubated at 37°C. Medium change is performed 3 days later. On 7th day, the plate is then re-incubated with 100 μl fresh medium and additional 50 μl of 2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2 Hydrogen-Tetrazolium-5-Carboxanilide reagent, and further incubated in dark at 37°C for 4 h. After incubation, the orange colored complex formed is read at 450 nm using a microplate reader with a 450 nm reference filter. | 3 months | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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progenitor cells, cardiovascular disease, risk factors, angiogenesis, biological markers, endothelial function, cilostazol