In vivo protein adsorption onto polymers: a transmission electron microscopic study

This paper reports in vivo protein adsorption onto polymers, including Biomer, PEO grafted Biomer (B-PEO-4K), heparin immobilized Biomer with PEO spacers (B-PEO-4K-HEP), and HEMA-Styrene block copolymer (H-S). Vascular grafts (6 mm ID, 7 cm in length) were fabricated with Biomer, coated on their luminal surfaces with test polymers, and implanted into the abdominal aorta of dogs. After 3 weeks-1 month, the grafts were retrieved and processed for TEM and SEM. TEM measured the thickness of adsorbed protein layers stained with a OsO4 solution, and the distribution pattern of adsorbed proteins (albumin, IgG and fibrinogen) using the immunoperoxidase technique. Retrieved grafts of Biomer and B-PEO-4K showed mural thrombi along the graft length, while thrombus formation on B-PEO-4K-HEP and H-S grafts was limited to the anastomotic sites. SEM pictures of B-PEO-4-HEP and H-S surfaces demonstrated clear morphology, with minimal platelet adhesion and activation, and microthrombi. Biomer and B-PEO-4K demonstrated a thick proteinaceous layer (1000-2000 A), whereas B-PEO-4K-HEP and H-S showed what can be described as a monolayer protein thickness (200-300 A). B-PEO-4K-HEP and H-S showed a monolayer-like adsorbed protein pattern, with high concentrations of albumin and IgG, and less fibrinogen, while Biomer and B-PEO-4K showed multilayered patterns with relatively high concentrations of fibrinogen, and less albumin. These results suggest that the surface properties of polymer may control protein adsorption pattern, and the composition of adsorbed protein is essential to in vivo long-term blood compatibility.