Gingival fibroblasts (GF) are the most abundant cell type in periodontal connective tissues, andhave distinct functional activities in the repair of periodontal tissues and in inflammatory periodontal diseases. Human gingival fibroblasts (hGF) can be used for periodontal tissue engineering. This study examined whether the alkaline phosphatase of hGF is enhanced by recombinant human BMP-4 and/or Anti human BMP-4 antibody. hGF was obtained from the excised gingival tissue of an implant patient undergoing 2nd surgery. The tissue was incubated at 37℃ in 5% CO2 and 95% humidity, and the cultivating media was changed every 2 days. The 2nd passage hGF cells were cultured in a medium containing Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum and 1 X antibiotic antimycotic solution. The control hGF was cultured for 7 days without rhBMP-4/Anti human BMP-4 antibody. The experimental groups were cultured for 7 with BMP-4 (10 ng/ml) and/or Anti human BMP-4 antibody. This study evaluated the differentiation of hGF to osteoblasts using alkaline phosphatase assay. In the experimental groups, the hGF showed abundant positive ALP staining. Among the experimental groups, the experimental group 3 (mixture of rhBMP-4 (20ng/㎖) and Anti human BMP-4 antibody (50000ng/㎖) showed most abundant positive ALP staining. In the control group, the hGF showed weak positive ALP staining. Overall, these results suggest that the ALP expression of hGF can enhanced by rhBMP-4 or mixture of rhBMP-4/ anti human BMP-4 antibody.

Ⅰ. Introduction
 Ⅱ. Materials and Methods
  1. Cell Isolation and Culture
  2. Osteogenic induction
  3. ALP assay
 Ⅲ. Results
  1. Morphological assay
  2. ALP assay
 Ⅳ. Discussion
 Ⅴ. References

• 이일권(School of Pharmacy, Chosun University) | IL Kwon Lee
• 장현선(Department. of Oral Pathology, Oral Biology Research Institute, School of Dentistry, Chosun University) | Hyun Seon Jang correspondence