Lab Med Online.  2016 Oct;6(4):214-220. 10.3343/lmo.2016.6.4.214.

Monitoring the Antiplatelet Effect of Cilostazol with Light Transmission Aggregometer: Two Cases of Possible Cilostazol Resistance

• Shim H
• Jang S
• Park CJ
• Chi HS
• Lee SW
• Park SW

• KMID: 2353422
• DOI: http://doi.org/10.3343/lmo.2016.6.4.214

Abstract

BACKGROUND
Coronary artery disease is an important cause of death in adults and stent insertion is one of the treatment modalities. The most severe adverse effect of a stent insertion is the formation of a thrombus; therefore, antiplatelet agents are used. The addition of cilostazol to low-dose aspirin and clopidogrel results in a better antiplatelet effect. However, laboratory tests to monitor the effect of cilostazol are insufficient.
METHODS
We tested the inhibitory effect of cilostazol using maximal platelet aggregation in 20 healthy volunteers. Conditions for incubation and concentrations of cilostazol and prostaglandin E1 (PGE1) were established and aggregation was induced by 5'-adenosine diphosphate (ADP) and measured with light transmission aggregometry (LTA). Blood samples were incubated with 1 µM and 2 µM cilostazol for 10 minutes at room temperature, and 80 nM PGE1 was added and incubated for an additional 10 minutes. Aggregation was induced by ADP and reactivity was evaluated.
RESULTS
The average maximum aggregation (MA) was 58.1% at 1 µM cilostazol and 22.0% when PGE1 was added. The average MA was 42.8% when cilostazol concentration was increased to 2 µM and 21.2% when PGE1 was added. Average inhibition of aggregation at 1 µM cilostazol was not statistically significant (P=0.085), but was significant (P=0.004) at 2 µM cilostazol. Aggregation was not inhibited even with 2 µM cilostazol and PGE1 in 2 volunteers, which suggests possible resistance to cilostazol.
CONCLUSIONS
We designed a method to monitor the effect of cilostazol using in vitro incubation with PGE1.

Keyword

Antiplatelet; Cilostazol; Light transmission aggregometry

MeSH Terms

Adenosine Diphosphate
Adult
Alprostadil
Aspirin
Cause of Death
Coronary Artery Disease
Healthy Volunteers
Humans
In Vitro Techniques
Methods
Platelet Aggregation
Platelet Aggregation Inhibitors
Stents
Thrombosis
Volunteers
Adenosine Diphosphate
Alprostadil
Aspirin
Platelet Aggregation Inhibitors

Figure

• 

  Fig. 1 Example of experimental protocol with participants. Maximal aggregation % of A) cilostazol 1 µM, ADP 20 µM, B) cilostazol 2 µM, ADP 20 µM, C) cilostazol 1 µM, PGE1 80 nM, ADP 20 µM, and D) cilostazol 2 µM, PGE1 80 nM, ADP 20 µM.

• 

  Fig. 2 Effect of PGE1 on maximal aggregation (%, Y-axis). Maximal aggregation % of A) ADP 20 µM, B) PGE1 100 nM, ADP 20 µM.

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  Fig. 3 Effect of addition of PGE1 on maximal aggregation (%, Y-axis). Maximal aggregation % of A) cilostazol 1 µM, ADP 20 µM, B) cilostazol 1 µM, ADP 20 µM, PGE1 80 nM.

• 

  Fig. 4 Decreasing aggregation % according to the concentration of cilostazol. A) % aggregation with no cilostazol, 1 µM of cilostazol and 1 µM of cilostazol and 80 nM of PGE1, B) % aggregation with no cilostazol, 2 µM of cilostazol and 2 µM of cilostazol and 80 nM of PGE1.

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  Fig. 5 Result of participants showing resistance to cilostazol. Maximal aggregation % (Y-axis) did not decrease with 2 µM of cilostazol and 80 nM of PGE1.

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  Fig. 6 Box plot distribution of % aggregation under 2 µM of cilostazol and 80 nM of PGE1. Two outlying results, probably resistant to cilostazol, are indicated by the arrow. Minimum value is 2.3%, maximum 74.1%, mean 21.1%, with standard deviation of 22.2%.

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