Tissue engineering of periosteal cell membranes in vitro

Abstract

Objectives: The cultivation of bone is a major focus in tissue engineering and oral implantology. Without a periosteal layer, instant or rapid development of a substantial cortical layer is unlikely for engineered bone grafts. The aim of this study was to test the ability of four collagen membranes to support and promote the proliferation of human periosteal cells. Materials and methods: Human periosteum cells were cultured using an osteogenic medium consisting of Dulbecco's modified Eagle's medium supplemented with fetal calf serum, penicillin, streptomycin and ascorbic acid at 37àwith 5% CO2. Four collagen membranes served as scaffolds: Bio-Gide, Chondro-Gide, Tutodent and Ossix Plus. Cell vitality was assessed by fluorescin diacetate (FDA) and propidium iodide (PI) staining, biocompatibility with LDH and BrdU, MTT, WST tests and scanning electron microscopy (SEM). Results: After 24 h, all probes showed viable periosteal cells. All biocompatibility tests revealed that proliferation on all membranes after treatment with eluate from membranes after a 24-h immersion in a serum-free cell culture medium was similar to the controls. Periosteal cells formed layers covering the surfaces of all four membranes 7 days after seeding in SEM. Conclusion: It can be concluded from our data that the collagen membranes can be used as scaffolds for the cultivation of periosteum layers with a view to creating cortical bone using tissue-engineering methods.