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

Phase 4CompletedNCT02194686

National Cheng-Kung University Hospital71 enrolled

Overview

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.

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

Study Type

INTERVENTIONAL

Allocation

RANDOMIZED

Purpose

TREATMENT

Masking

QUADRUPLE

Enrollment

Conditions

Cardiovascular Diseases

Interventions

CilostazolDRUG

One tablet (100 mg) twice per day for 12 weeks

Dummy PlaceboDRUG

One tablet twice per day for 12 weeks

Eligibility

Sex: ALLMin age: 20 Years

Medical Language ↔ Plain English

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

Locations (1)

National Cheng Kung University Hospital

Tainan, Taiwan

Outcomes

Primary Outcomes

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.

Time frame: 3 months

Secondary Outcomes

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.

Time frame: 3 months

Data from ClinicalTrials.gov

This platform is for informational purposes only and does not constitute medical advice. Always consult a qualified healthcare professional.