Probable Limitations of Osteoblasts Isolation from the Periosteum and Alveolar Bone in a Dog Model

Bone defects in jaws create major problems for oral and maxillofacial surgery. To overcome the limitations of Autografts tissue engineering uses autogenus cells and synthetic scaffolds. Type of cells or cell sources have an important effect on the construction which is produced. The aim of this study was to evaluate the feasibility and probable limitations of osteoblast isolation from the periosteum and alveolar bone in an animal model and to compare their probable results quantitatively. Bone and periosteal samples were harvested from interdental septum and retromolar area of 4 dogs. Because no cell was grown new samples were harvested from edentulous ridge and buccal mandibular periosteum. Since no cell was isolated in next step samples were harvested from buccal area of maxillary premolars and explants in cell culture medium. After 2 weeks adherent cells reaching 80% confluent; cells were counted and passaged in cell culture flasks. Despite first good proliferation of periosteal cells of all dogs in P0, their growth was stopped and they become senescence after one week. The key problems in culture techniques are cell senescence and de-differentiation leading to lose the ability of proliferation. It seems there are probable reasons for isolating of osteoblasts including thermal damage of cells, contamination of culture with fibroblasts, inconsistency of medium and cell requirements, enzymatic damage from enzymes used for cell passage and high donor ages.