Abstract: Objective Mixed culture of RFP markers of tooth germ cells and bFGF/BMP2 transfecting the bone marrow stem cells(BMSCs ) was conducted and polyvinyl alcohol (PVA)/biphasic ceramic bone scaffold (DCCP) stand was painted threedimensionally to construct heterogeneous tooth tissue engineering(HTTE). In order to investigate their ability to differentiate in vivo. Methods 128 healthy SD rats were randomly divided into four groups: the cell pellet group; PVA/DCCP scaffold group; cell clumps + PVA/DCCP scaffold group; control group which was without any intervention. The HTTE was implanted into the renal subcapsular of rats under general anesthesia. Four time points after surgery were chosen to collect samples for testing, Immunohistochemical staining and confocal slice observation. At the 5th, 10th, 14th, 28th day, 8 samples were collected from each group. Results Immunohistochemistry: Factorial analysis showed that cell clumps + PVA/DCCP scaffold group in 5, 10 days DMP1, BMP4 expression were the highest and were higher than the rest groups, the expression level gradually decreased over time, and the difference was statistically significant P<0.05; DSP in each group at each time point was negative, the difference was not statistically significant P>0.05. Confocal microscopy showed: cell clumps +PVA/DCCP scaffolds group at each time point biopsy tissue could be observed in tooth germ cell fluorescently marker. Conclusion RFP labeled tooth germ cells and bFGF/BMP2 transfecting BMSCs mixed culture, composite PVA/DCCP scaffold successfully construct tooth tissue engineering and tooth differentiation, but its differentiation mechanism needs further study.

Key words: red fluorescent protein, the tooth germ cells, bone marrow stem cells, PVA/DCCP, tooth tissue engineering