Initial attachment of osteoblasts to various guided bone regeneration membranes: an in vitro study

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Abstract
Guided bone regeneration (GBR) has proved to be a suitable and somehow predictable technique for promoting bone regeneration. A variety of synthetic and naturally derived GBR barriers have been used in clinics to facilitate bone regeneration. These barriers may differ in composition and structure and these may affect the outcomes of GBR. Therefore, the present study was undertaken to evaluate the in vitro ability of osteoblasts (MC3T3-E1) to attach to various GBR membranes. Six GBR/GTR (guided tissue regeneration) membranes [BioMend (BM), Resolut (RL), Guidor (GD), EpiGuide (EG), Gore-Tex (GT) and Millipore filter (MP)] were tested. For controls, cells were directly plated on culture dishes (CD). Each test membrane was secured to the bottom of a culture dish with a double-sided adhesive tape. All samples were triplicate. At 1.5 and 24 h after plating of 2 ml (5 x 10(4) cells/ml) of MC3T3-E1 (passage 7) cells, the specimens were rinsed with phosphate-buffered saline to wash out any unattached cells and then fixed with a 10% buffered formalin solution for 1 d. After washing with distilled water, the cells were stained with hematoxylin. The number of attached cells was counted under a light microscope equipped with an ocular-micrometer in a unit area of 0.25 mm(2) (five areas on each membrane). In addition, cell morphology attached to the membranes was evaluated under scanning electron microscope. Data were presented as mean +/- standard error and analyzed for statistical difference using a generalized Wilcoxon's test. Cell attachment at 1.5 h was as follows: MP (27.5 +/- 2.1) > RL (17.0 +/- 1.4) approximately equals BM (14.5 +/- 1.4) approximately equals EG (11.4 +/- 1.0) > GD (5.2 +/- 0.8) approximately equals GT (3.1 +/- 0.6); and at 24 h was: MP (67.6 +/- 3.6) > RL (35.8 +/- 1.8) > BM (15.4 +/- 0.9) approximately equals EG (13.3 +/- 1.3) > GD (5.9 +/- 0.7) approximately equals GT (5.6 +/- 1.3). At 24 h, the scanning electron microscope finding revealed that cells attached on MP, RL, BM and EG were flatter in shape, like cells on CD, than cells on GD and GT, where cells were rather round. Results from this study suggested that MP, BM, RL and EG enhanced the early osteoblast attachment. However, the true benefit of this observation in clinic remains to be determined.