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 important 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. Historically, the rat calvaria model has been employed to study the effects of various dental treatments on bone in vitro. However, there are morphological and functional differences which exist between bone cells derived from rat calvaria and human intraoral osseous tissue that impose certain limitations on the usefulness of the rat calvaria model for dental implant applications. Therefore, an in vitro culturing method for 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 osteoblasts is also vital. The specific objective of this study was to establish isolation and in vitro culturing methods utilizing human intraoral bone cells derived from dental implant preparation sites. This paper describes techniques for the harvesting of human bone cells from the intraoral derived osseous tissues and discuss the procedures for maintaining the primary intraoral bone cell culture. In addition, our studies utilize established protocols for the characterization of these cells as osteoblasts by means of alkaline phosphatase activity determination, identification of cellular osteonectin and osteocalcin antigens, establishing the presence of cells expressing type I collagen and determining the ability of cells to produce calcifications. The utilization of intraoral osseous tissue may prove useful for future dental implant research by providing an in vitro model system more closely related to conditions encountered clinically.
Mailhot
J M
JM
Department of Periodontics, Medical College of Georgia, Augusta 30912, USA. jmailhot.dentistr@mail.mcg.edu
Borke
J L
JL
D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D003757:Dental Implantation; D006801:Humans; D007150:Immunohistochemistry; D007568:Jaw / Q000166:cytology*; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D015676:Osteonectin / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods
D015676:Osteonectin; D015675:Osteocalcin; D003094:Collagen; D000469:Alkaline Phosphatase