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

Sponsor

National Cheng-Kung University Hospital (Other)

Overall Status

Completed

CT.gov ID

NCT02194686

Collaborator

Department of Health, Executive Yuan, R.O.C. (Taiwan) (Other)

Enrollment

Location

Arms

Duration (Months)

3.7

Patients Per Site Per Month

Study Details

Study Description

Brief Summary

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.

Condition or Disease	Intervention/Treatment	Phase
Cardiovascular Diseases	Drug: Cilostazol Drug: Dummy Placebo	Phase 4

Detailed Description

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

Study Design

Study Type:

Interventional

Actual Enrollment :

71 participants

Allocation:

Randomized

Intervention Model:

Parallel Assignment

Masking:

Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)

Primary Purpose:

Treatment

Official Title:

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

Study Start Date :

Jan 1, 2013

Actual Primary Completion Date :

Mar 1, 2014

Actual Study Completion Date :

Aug 1, 2014

Arms and Interventions

Arm	Intervention/Treatment
Active Comparator: Cilostazol One tablet (100 mg) twice per day for 12 weeks	Drug: Cilostazol One tablet (100 mg) twice per day for 12 weeks Other Names: Pletaal (brand name)
Placebo Comparator: Dummy Placebo One tablet twice per day for 12 weeks	Drug: Dummy Placebo One tablet twice per day for 12 weeks Other Names: Placebo Control

Arm

Intervention/Treatment

Active Comparator: Cilostazol

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

Drug: Cilostazol

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

Other Names:

Pletaal (brand name)

Placebo Comparator: Dummy Placebo

One tablet twice per day for 12 weeks

Drug: Dummy Placebo

One tablet twice per day for 12 weeks

Other Names:

Placebo

Control

Outcome Measures

Primary Outcome Measures

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 Outcome Measures

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 Outcome Measures

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.

Eligibility Criteria

Criteria

Ages Eligible for Study:

20 Years and Older

Sexes Eligible for Study:

All

Accepts Healthy Volunteers:

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