Human osteoblasts attachment to guided tissue regeneration membranes which were coated either with platelet-rich plasma or platelet-poor plasma

Abstract

To determine whether human alveolar bone osteoblasts (HABO) attachment to commercial available guided tissue regeneration (GTR) membranes can be enhanced by coating with freshly prepared human platelet-rich plasma (PRP) and platelet poor plasma (PPP). Human osteoblasts established from tissue explants were used at 4 th passage in culture. Human whole blood from healthy subjects was collected and centrifuged twice to produce the PRP fraction and PPP fraction. Double-sided adhesive tape was used to fix 3 mm discs of each membrane and cover-slides to the bottom of a 24-well tissue culture plate. A (GoreTex-ePTFE), B (GoreTex-Resolut) and C (Inion-GTR) membranes were studied. Cover-slides were positive control. Membranes or cover-slides were exposed to PRP, PPP or PBS respectively for 2 hours. Membranes and cover-slides were seeded with osteoblasts (5 x 10(7) cells/L) and allowed to attach for 24 hours. After staining with hematoxylin, the number of attached cells per mm(2) was counted using a light microscope with graticule. The the ultrastructure of osteoblasts attachment to the membranes was observed by scanning electronic microscopy. PRP and PPP-treated membranes significantly enhanced osteoblasts attachment compared to PBS-treated membranes (P < 0.05). There was more osteoblasts attachment in the PRP-treated membranes than in the PPP-treated membranes (P < 0.05). Cover-slides showed more osteoblasts attachment than the three membranes (P < 0.05). B and C membranes showed higher cell attachment than A membranes (P < 0.05). SEM showed that osteoblasts attached to the membranes treated by PRP were spindle and stretched well, and there were platelets, fibrins in a interlaced mesh on the membranes, which appeared to grow in a multiplayer style. The osteoblasts attached to the membranes treated by PPP or PBS were round and partially attached. PRP and PPP could improve attachment of osteoblasts in the three membranes, and PRP altered and enhanced the way of the attachment to the membranes.