Investagation of cbfa-1 in dental pulp tissue and stem cells

Objectives:The aim of this study was to investigate the expression of Cbfa1 in dental pulp tissue and dental pulp stem cells and its possible role in odontoblasts differentiation.. Methods: Dental pulp tissue from healthy teeth extracted for orthodontic reasons were digested by 3mg/ml Collagenase and 4mg/ml dispase. Cells were implanted in a six-well plate at a low density(1000-100000/well). Clonogenic cells were recorded and passaged when they reach 80% confluent. Cells were prepared for immunofluorescence and RT-PCR. Dental pulp tissues are fixed in 4% paraformaldehyde Envision immunohistochemistry technique were used to detect Cbfa1 in dental pulp . Gingiva tissue and cells were used as negative control. Antibody was oat-anti-human IgG(Santa-Cruz),Secondary antibody were PE labelled rabbit-anti-goat IgG and Biotin labelled rabbit-anti-goat IgG(DAKO). Results: The cells we get were negative for DSP,a marker of mature odotoblasts and positive for Stro-1,a relative specific marker of mesenchymal stem cell. Expression of Cbfa1was detected both in dental pulp tissue and stem cells, but not in gingival tissue and cells. Its mRNA was also found by RT-PCR . Conclusions:Cbfa-1 may be a transcriptional factor that is involved in odontoblast differentiation. Its exact function needs more research.