Methods: Human gingival (HGF) and periodontal ligament fibroblasts (PDLF) established from tissue explants were used between their 5th and 8th passage in culture. Human whole blood from healthy subjects was collected in tubes containing acid-citrate-dextrose, centrifuged (1000 rpm for 20 min) to produce the PRP fraction. A second spin (3200 rpm for 10 minutes) produced the PPP fraction. Double-sided adhesive tape was used to fix 3 mm discs of each membrane to the bottom of a 24-well tissue culture plate. GoreTex ePTFE™, GoreTex Resolut™ and Inion™ membranes were studied. Each membrane was exposured to either PRP or PPP. Controls were treated with PBS alone. Membranes were seeded with HGF or PDLF (3x104cells/ml) and allowed to attach for 24 hours. Wells were then rinsed with PBS, fixed at room temperature for 24 hours using 10% buffered formalin and stained with hematoxylin. The number of attached cells per mm2 was counted using a graticule.
Results: All untreated membranes showed low attachment for both the HGF and PDLF (4-8 cells/mm2). PRP treated membranes significantly enhanced cell attachment (17-33 (HGF) cells/mm2, 18-45 (PDLF) cells/mm2. PPP treated membranes also showed enhanced cell attachment 14-38 (HGF) cells/mm2, 21-62 (PDLF) cells/mm2. ThePRP/PPP-treated Inion membranes showed the highest cell attachement (P<0.05) for both HGF and PDLF.
Conclusion: Pre-treatment of GTR membranes with freshly prepared PRP or PPP enhances the attachment of HGF and PDLF. This treatment may be used as a strategy for cell delivery to periodontal defects.
GTR membranes were provided by GoreTex and Inion Pty Ltd. Dr Qi Liu was supported by a grant from the Chinese Government.