In vitro evaluation of platelet/biomaterial interactions in an epifluorescent video microscopy combined with a parallel plate flow cell

Artif Organs. 1994 Aug;18(8):588-95. doi: 10.1111/j.1525-1594.1994.tb03383.x.

Abstract

Suitable evaluation systems are critical for ranking various biomaterials in order to develop a method to design and synthesize nonthrombogenic biomaterials. We have recently developed an in vitro test system to evaluate platelet/biomaterial interactions in whole blood. The system consists of a parallel plate flow cell and epifluorescent video microscopy (EVM). A glass coverslip coated with a polymer was incorporated into the flow cell, and blood was perfused using a syringe pump via a polymer-coated PVC tubing connected to the flow cell. Whole human blood was anticoagulated with heparin (2 U/ml), and the platelets were labeled with the fluorescent dye mepacrine (5 microM). This system permitted real-time and dynamic observations of platelet/biomaterial interactions in whole blood under a defined flow condition. In order to evaluate the feasibility of this system, two different segmented polyether-polyurethanes (SPEUs), PU-PTMG(650) and PU-PTMG(2000), were chosen as test polymers. Surface characteristics verified with electron spectroscopy for chemical analysis (ESCA) and contact angle measurements showed similar results in both SPEUs. Blood was perfused at a wall shear rate of 200 s-1 for 20 min. Excitation light was applied for 2 s at 1 min intervals. The real-time image was then analyzed at each time point for the percentage of surface area of platelet coverage. Plasma beta-thromboglobulin (beta-TG) levels were also measured before and after each run. PU-PTMG(650) showed a significantly higher number of adhered platelets than PU-PTMG(2000) at each time point.(ABSTRACT TRUNCATED AT 250 WORDS)

MeSH terms

  • Biocompatible Materials*
  • Blood Platelets / drug effects
  • Blood Platelets / physiology*
  • Fluorescent Dyes
  • Humans
  • Materials Testing / methods*
  • Microscopy, Fluorescence
  • Platelet Activation / drug effects
  • Platelet Adhesiveness / drug effects
  • Platelet Aggregation / drug effects
  • Polyurethanes
  • Quinacrine
  • Rheology / instrumentation
  • Rheology / methods
  • Rhodamines
  • Surface Properties
  • Video Recording
  • beta-Thromboglobulin / metabolism

Substances

  • Biocompatible Materials
  • Fluorescent Dyes
  • Polyurethanes
  • Rhodamines
  • beta-Thromboglobulin
  • polyetherurethane
  • acridine red
  • Quinacrine