Collagen-coated glass beads containing glycerine for the measurement of platelet adhesion and inhibition of platelet adhesion by dilazep

• Published in: Blood & Vessel Vol. 18; no. 2; pp. 142 - 144
• Main Authors: HAMAZAKI, Tomohito, URAKAZE, Masaharu, KANBARA, Katsumi, SAWAZAKI, Shigeki, YANO, Saburo
• Format: Journal Article
• Language: English
• Published: The Japanese Society on Thrombosis and Hemostasis 1987
• Subjects: collagen; dilazep; glass bead column; glycerine; platelet adhesion
• ISSN: 0386-9717; 1884-2372
• DOI: 10.2491/jjsth1970.18.142

Using glycerine as a preservative, a collagen-coated glass bead column was made for the measurement of platelet adhesion (PA). Bovine achilles tendon collagen (250mg) was homogenized with 1mM HCl and centrifuged at 3000rpm for 10min. To the suspension was added glycerine (1/10), and 0.6ml of this collagen suspension was passed through a 10cm long glass bead column (Igakushoin-Kikai, Tokyo). Then the column was half-dried by passing 200ml of air through the column, which was stored at -20°C. To 9ml of human blood was added 1ml of 37mM Tris-HCl buffer, pH7.4, containing 25mM EDTA. PRP and PPP were obtained by centrifugation. RRP was adjusted for platelet count (150, 000/μl) with PPP. One-third ml of adjusted PRP was passed through the column, which was warmed to room temperature beforehand, platelet count in the eluate was counted, and the percentage of PA was calculated. Platelet adhesing activity of collagen in the column was the stablest when the collagen suspension contained 10% glycerine, followed by 20, 3 and 0% in decreasing order of stability. During storage of collagen-coated glass bead column using 10% glycerine at -20°C for 40 days, PA measured with PRP from the same volunteer decreased only 5 points (from 49% to 44%). The coefficiency of varience was less than 5 points (10%). PA measurement was stable for the first 2h afer blood sampling. Sixteen old patients suffering from cerebral arteriosclerosis without any anti-platelet treatment were divided into 2 groups. One group was treated with 150mg of Dilazep every day for 14 days. The other served as controls. PA was measured before and after 14 days of the treatment using the same lot of columns. PA was significantly decreased in the treated group but not in the control group. In conclusion, the addition of glycerine to a collagen-coated glass bead column increased the stability of collagen; and Dilazep decreased PA and appeared to be effective for the prevention of thrombosis.