SPECIFIC AND QUANTITATIVE METHOD FOR ESTIMATION OF PLATELET-ADHESION TO FIBRILLAR COLLAGEN

  • 1 January 1979
    • journal article
    • research article
    • Vol. 94  (3) , 438-446
Abstract
A quantitative method (Sepharose test) was devised to measure blood platelet adhesion to fibrillar collagen. [14C]5HT[5-hydroxytryptamine]-labeled platelets were isolated from plasma, resuspended in EDTA buffer and incubated with buffer (control) or fibrillar collagen for 150 s at 33.degree. C. The mixtures were then filtered through Sepharose 2B columns. In controls the platelets were rapidly eluted, and this was confirmed after 51Cr labeling. [14C]5HT was recovered in 2 stages: 60% with platelets and 40% retarded, as free 5HT. After incubation with fibrillar collagen (50 .mu.g), platelets were retained with the fibrills on the top of the column, and only free [14C]5HT (released from the platelets) was eluted. The adhesion percentage depended on platelet number, collagen amount, its degree of polymerization, and time of incubation at 33.degree. C. [14C]5HT release was markedly diminished when incubation and filtration were performed at low temperature. ASA [acetylsalicyclic acid] used in vitro or in vivo in rabbits, did not change percentage of adhesion but significantly diminished total [14C]5HT amount eluted. This method offers a quantitative and reproducible system for differentiation of adhesion and release, independent of platelet aggregation. [This method was discussed in conjunction with the investigation of bleeding disorders or thrombotic tendencies and drug therapy monitoring.].

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