P-PRP and other platelet suspensions were serially diluted with equal volume of acellular plasma or the corresponding buffer solutions. Series of diluted platelet suspensions were measured using a pocketable spectrophotometer (PiCOSCOPE, Ushio Inc., Tokyo, Japan) [6 (link)]. The spectrophotometer can be operated by remote control through a specific application installed on smart devices, including the iPad Air (Apple, Cupertino, CA, USA). Platelet suspensions were transferred into 0.2 mL highly transparent PCR tubes (Nippon Genetics Co., Ltd., Tokyo, Japan) and treated with 5 μM ADP (Wako Pure Chemicals, Osaka, Japan).
To prepare dysfunctional platelet models, we pretreated P-PRP with 0.1 mg/mL aspirin (acetylsalicylic acid; Wako Pure Chemicals, Osaka, Japan) or 10 μM H2O2 (Wako) for 30 min at 22–24 °C. The absorbance was measured at an interval of one minute at 615 nm (range of wavelength: 570–660 nm). At the end of measurement, each blank was measured as the absorbance of 100% aggregation.
To prepare dysfunctional platelet models, we pretreated P-PRP with 0.1 mg/mL aspirin (acetylsalicylic acid; Wako Pure Chemicals, Osaka, Japan) or 10 μM H2O2 (Wako) for 30 min at 22–24 °C. The absorbance was measured at an interval of one minute at 615 nm (range of wavelength: 570–660 nm). At the end of measurement, each blank was measured as the absorbance of 100% aggregation.
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