The purpose of this study was performed to investigate the mineralization and differentiation of osteobalsts for bone regeneration in vitro and the effect of rate of the composition in periodontal cells on mineralization.
For this study, healthy gingival tissues were surgically obtained from the patients during 1st premolar extraction for the purposes of orthodontic treament. Gingival tissue was washed several time with Phosphate buffered saline contained high concentration of antibiotics and antifungal agent, and cultured in Dulbecco´s Modified Eagle´s Medium(DMEM, Gibco, U.S.A.) Every cell were cultured in state at 37 C, 100% of humidity, 5% of CO2 incubator. Bone marrow stromal cells were isolated from 5-day-old rat femur with using medium irrigation mathod by syringe. Cell suspension medium were centrifuged at 1500 rpm for 5 min and then cultured in the petri dish. Two kinds of cell were freezed and stocked in the liquid nitrogen tank until experiment.
Cell were incubated into the 24multi-well plate with 5 x 104 cell/well of medium at 37 ° C, 100% of humidity 5% CO2 incubator for 24 hours. After discarded of the supernatent of medium, 0.5m of medium were reapplied and incubated. And counted the number of cell using the hemocytometer and inverted light microscope.
We have measured the number of mineralized nodule with using Alizarin red S. staining in microscope; Furthermore every cell were observed the morphological change between every rate of co-culture of the two kinds of cell.
The results were as follows;
The rate of proliferation of co-culture cell revealed high rate tendency compared the bone marrow stromal cell only and low growth rate to compared with gingival fibroblast only.
The tendency of formation of the mineralized nodule were observed dose-depend pattern of bone marrow stromal
cell.
It is concluded that the gingival fibroblast may inhibit the formation of mineralized nodule in the culture of the bone marrow stromal cell.
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