Preparation of a pure autologous biodegradable fibrin matrix for tissue engineering

Parallel to the growing role of tissue engineering, the need for cell embed ding materials, which allow cells to stabilise in a three-dimensional distr ibution, has increased. Although several substances have been tested, fibri n is thus far the only one that permits the clinical application of culture d tissue. To date, autologous fibrinogen has usually been polymerised with bovine thrombin, which can cause severe immunological side effects. The obj ective of this study was to explore the practicability of obtaining autolog ous thrombin from a single patient in an adequate concentration and amount, Fibrinogen was cryoprecipitated from 200 mi of freshly-frozen plasma. Thro mbin was isolated from the supernatant through ion-exchange chromatography. The thrombin was first bound to Sephadex A-50 and then eluated using 2ml o f a salt buffer (2.0M NaCl in 0.015M trisodiumcitrate, pH 7.0). The activit y of the thrombin (51 NIH ml(-1) to 414 NIH ml(-1)) reached levels comparab le to those in commercially available fibrin glues (4-500 NIH ml(-1)). The study has shown that if is possible to obtain a sufficient amount of autolo gous thrombin from a single donor to create a fibrin matrix of high efficie ncy without the risk of immunological and infectious side effects.