Cardiovascular Diseases
Conditions
Keywords
cilostazol, progenitor cells, cardiovascular disease, risk factors, angiogenesis, biological markers, endothelial function
Brief summary
1. The number and function of circulating endothelial progenitor cells (EPCs) are inversely associated with coronary risk factors and atherosclerotic diseases. 2. 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
1. titration of drugs 1. run-in period: eligible subjects are screened and baseline blood samples are obtained 2. 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 3. 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) 2. assays of human EPCs 1. colony formation by EPCs 2. quantification of EPCs and apoptotic endothelial cells 3. chemotactic motility, proliferation/viability and apoptosis assays 3. measurement of flow-mediated dilatation (FMD) of left brachial artery by sonography
Interventions
DRUGCilostazol
One tablet (100 mg) twice per day for 12 weeks
DRUGDummy Placebo
One tablet twice per day for 12 weeks
Sponsors
Department of Health, Executive Yuan, R.O.C. (Taiwan)
National Cheng-Kung University Hospital
Study design
Allocation
RANDOMIZED
Intervention model
PARALLEL
Primary purpose
TREATMENT
Masking
QUADRUPLE (Subject, Caregiver, Investigator, Outcomes Assessor)
Eligibility
Sex/Gender
ALL
Age
20 Years to No maximum
Healthy volunteers
No
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
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Circulating EPCs Number | 3 months | 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. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Viability (Proliferation) of EPCs | 3 months | 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. |
Other
| Measure | Time frame | Description |
|---|---|---|
| FMD of the Brachial Artery | 3 months | FMD in response to reactive hyperemia is measured in the left brachial artery in a quiet, temperature-controlled room after 10 min of bed rest. A high-resolution ultrasound machine equipped with a 7.5 mega Hertz linear array probe is used for the study. Arterial diameters are measured at the baseline and during reactive hyperemia. FMD is calculated as the percentage change in diameter compared with the baseline. |
Countries
Taiwan
Outcome results
None listed