METHOD FOR IMMUNOLOCALIZATION OF EXTRACELLULAR PROTEINS IN ASSOCIATION WITH THE IMPLANT SOFT-TISSUE INTERFACE

A method allowing immunolocalization of macromolecules surrounding bio material implants has been developed. Plugs of pure titanium were inse rted into the abdominal wall of rats. One week after implantation plug s with adjacent tissue were gently fixed and cryoprotected before bein g rapidly frozen in LN(2)-cooled propane, followed by cryosubstitution , using methanol, and low temperature infiltration and UV curing of th e methacrylate LR-Gold. Before sectioning, bulk metal was removed elec trochemically, leaving the surface oxide intact, appearing in sections as a dense line in contact with the tissue. Postembedding immunolocal ization for rat albumin, IgG, C3c, fibrinogen and fibronectin was perf ormed at the light and electron microscopic levels. Our observations i ndicate that 1 wk after implant insertion, a network of fibrin and fib ronectin decorates the tissue surface that faces the implant, but with out any obvious concentration to the implant surface.