AN ISOLATION AND IN-VITRO CULTURING METHOD FOR HUMAN INTRAORAL BONE-CELLS DERIVED FROM DENTAL IMPLANT PREPARATION SITES

In dental implantology, the biocompatibility of the osseous tissue to the implant surface and to local environmental factors plays an import ant role in the process of healing. Bone cells derived from intraoral osseous tissue proves to be an important source of the osteoprogenitor cells required for healing of the periodontium around implants. Histo rically, the rat calvaria model has been employed to study the effects of various dental treatments on bone in vitro. However, there are mor phological and functional differences which exist between bone cells d erived from rat calvaria and human intraoral osseous tissue that impos e certain limitations on the usefulness of the rat calvaria model for dental implant applications. Therefore, an in vitro culturing method f or the isolation, growth and maintenance of human intraoral bone cell cultures derived from osseous tissues is truly warranted. In addition, a method for the accurate characterization of these bone cells as ost eoblasts is also vital. The specific objective of this study was to es tablish isolation and in vitro culturing methods utilizing human intra oral bone cells derived from dental implant preparation sites. This pa per describes techniques for the harvesting of human bone cells from t he intraoral derived osseous tissues and discuss the procedures for ma intaining the primary intraoral bone cell culture. In addition, our st udies utilize established protocols for the characterization of these cells as osteoblasts by means of alkaline phosphatase activity determi nation, identification of cellular osteonectin and osteocalcin antigen s, establishing the presence of cells expressing type I collagen and d etermining the ability of cells to produce calcifications. The utiliza tion of intraoral osseous tissue may prove useful for future dental im plant research by providing an in vitro model system more closely rela ted to conditions encountered clinically.