Attachment, proliferation and differentiation of periodontal ligament cells on various guided tissue regeneration membranes

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Abstract
The purpose of this study was to evaluate the biological effects of guided tissue regeneration (GTR) membrane materials, per se, on the periodontal tissue regeneration. Rat periodontal ligament (PDL)‐derived cells were used to study the attachment, proliferation and differentiation, in vitro, on various GTR membranes. Five commercially available membranes–bovine type I collagen (BioMend®; BM), bovine type I atelocollagen (Tissue Guide®; TG), polylactic acid (Epi‐Guide®; EG), co‐polymer of polylactic acid and polyglycolic acid (Resolute®; RL) and expanded polytetrafluoroethylene: e‐PTFE (Gore Tex®; GT)–were examined. A 3×3 mm section of the membrane was fixed to the bottom of a 35×10 mm style culture dish and plated with 2 ml of cell suspension at an initial density of 5×104 cells/ml in culture medium with 10% fetal bovine serum. For cell growth analysis, the specimens were fixed with 10% buffered formalin and stained with hematoxylin at 1.5 hours and 1, 3 and 5 days after cell seeding. The number of cells included in a unit area of 0.25 mm2 were counted under light microscopy. As a comparative scaffold of cell proliferation, a plastic cover for cell culture slip (Celldesk®: CD) was used. For analysis of cell differentiation, activity of alkaline phosphatase (ALP) and calcification were histochemically revealed after 2‐week cultivation. The initial number of PDL cells attached to the membrane at 1.5 hours after cell seeding was different among membranes. RL, TG and EG had the same level of attached cell numbers as that on CD, while the cell numbers on GT and BM were significantly lower than that on CD (pp0.1). EG had a similar number of cell attachments to that at 1.5 hours throughout the experimental period. There was almost no cell proliferation on GT. Cell clusters of ALP positive cells and foci of calcification were seen on all membranes except for GT, where a scant number of cells were seen. Results from this study implied that GTR membrane materials, per se, may influence cell proliferation and differentiation in the process of periodontal tissue regeneration.

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