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Completed

NCT Number: NCT01952756

Effect of Cilostazol Endothelial Progenitor Cells and Collateral Formation in Peripheral Occlusive Artery Disease (PAOD)

1. The number and function of circulating endothelial progenitor cells (EPCs) are inversely associated with coronary risk factors and atherosclerotic diseases such as PAOD. 2. This double-blind, randomized, placebo-controlled trial to evaluate the effects of cilostazol on human early EPCs and angiogenesis as well as the potential mechanisms of action in patients with mild-to-moderate PAOD.

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Key information

Conditions

Peripheral Arterial Diseases Arterial Occlusive Diseases Arteriosclerosis Atherosclerosis Cardiovascular Diseases Peripheral Arterial Disease Peripheral Vascular Diseases Vascular Diseases

Age range

20 year and older

Sex eligibility

All sexes

Study type

Interventional

Phase

Phase 4

Primary location

National Cheng Kung University Hospital

Tainan, 704, Taiwan

About this study

  • titration of drugs
  • run-in period: eligible subjects are screened and baseline blood samples are obtained
  • study period: 12 weeks
  • 24 subjects with cilostazol and 20 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
  • collateral vessels formation and distal run-off assessed by dual-energy multi-slice computed tomography angiography
  • echocardiographic examinations to evaluate left ventricular functions

Who can participate

Healthy volunteers accepted: No

Only the study team can determine whether someone qualifies for participation.

Inclusion criteria

  • ankle-brachial index (ABI) less than 0.9 in one or both legs but no obvious symptoms of intermittent claudication

Exclusion criteria

  • obvious symptoms of intermittent claudication
  • severe PAD (Fontaine grading > 3) or critical limb ischemia in at least one leg
  • severe liver dysfunction (transaminases >10 times of upper normal limit, history of liver cirrhosis, or hepatoma)
  • > stage 4 chronic kidney disease (end-stage renal disease with chronic dialysis not excluded)
  • left ventricular ejection fraction <50% by echocardiography
  • documented active malignancy
  • chronic inflammatory disease
  • planned coronary intervention or endovascular therapy or bypass surgery within 3 months
  • known drug allergy history for cilostazol
  • current use of cilostazol or any other cAMP-elevator
  • premenopausal women

Treatment and study plan

Cilostazol

Drug

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

Other names: Pletaal (brand name)

Dummy Placebo

Drug

One tablet twice per day for 12 weeks

Other names: Placebo, Control

Primary outcomes

  1. Circulating EPCs Number

    Time frame: 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 outcomes

  1. Colony Formation by EPCs

    Time frame: 3 months

    Peripheral blood mononuclear cells are isolated by density gradient centrifugation according to standard protocols. After centrifugation, cells are washed, resuspended in M199 medium supplemented with 20% (vol/vol) fetal bovine serum, 10 ng/ml vascular endothelial growth factor, 2 ng/ml basic-fibroblast growth factor, 10 ng/ml epidermal growth factor and 2 ng/ml insulin growth factor, and cultured in 24-well plates coated with human fibronectin for 7 days. EPCs cells are confirmed by uptake of acetyl-low density lipoprotein and lectin and by the expression of EPC markers. Cells are harvested after 7 days, fixed and stained with crystal violet reagent. The colony densities are quantified with an Olympus microscope at 100-fold magnification using an imaging measurement software.

Other outcomes

  1. Viability (Proliferation) of EPCs

    Time frame: 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.

Sponsors and collaborators

Lead sponsor

National Cheng-Kung University Hospital

Other

Collaborators

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

Registry information

Official study title

Cilostazol Enhances the Number and Functions of Circulating Endothelial Progenitor Cells and Collateral Formation Assessed by Dual-energy 128-row CT Angiography Mediated Through Multiple Mechanisms in Patients With Mild-to-moderate PAOD

Important dates

Study start
2012
Primary completion
2013
Study completion
2013
First posted
Sep 30, 2013
Registry last updated
Jul 18, 2014

OpenTrials presents study information sourced from ClinicalTrials.gov. The official registry record should be consulted for the latest information.

View the official ClinicalTrials.gov record (opens in a new tab)

This listing is for discovery and informational purposes only. It is not medical advice, does not guarantee that a study is recruiting, and does not determine eligibility. Contact the study team and a qualified healthcare professional when considering participation.

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