Influence of Harvesting Technique and Donor Site Location on In Vitro Growth of Osteoblastlike Cells from Facial Bone

Purpose: Donor morbidity is minimized when tissue engineering is applied to produce osteogenic grafts by growing osteoblasts on biomaterials. However, limiting factors are the origin, proliferation, and differentiation of osteoblasts. Therefore, the aim of this study was to evaluate the efficacy of growing osteoblasts from different types of bone samples and to assess the influence of the donor site. Materials and Methods: From 28 patients 37 bone specimens were obtained during removal of third molars in the maxilla and mandible. Seventeen specimens were bone chips and 20 were bone sludge. After subculturing primary cultures, histochemical and immunhistochemical tests (EZ4U test, BrdU labeling, ALP histochemistry, type I collagen immunohistochemistry, osteocalcin ELISA) were performed to determine cell proliferation, viability, and differentiation. Results: Both bone chips and bone sludge from the mandible and maxilla are suitable for culturing human osteoblastlike cells. However, bone chips were superior to bone sludge with respect to ability to grow cells, and maxillary bone was superior to mandibular bone in this regard. Harvesting technique had only little influence on the expression of cell differentiation markers (ALP, type I collagen, osteocalcin). Discussion and Conclusion: Chips from human membrane bone, especially from the maxilla, are suitable for culturing high numbers of differentiated osteoblastlike cells. These cells may be used to tissue engineer bone grafts, which may be used to enhance the implant placement site. Parole chiave: alveolar ridge augmentation, cell culture, cell proliferation and differentiation, implant placement, tissue engineering