Novel pulp treatment agent with stimulator for pulp regeneration

本研究的目的是分析促进血管化、牙髓伤口愈合和牙髓再生的蛋白质，以开发在牙髓损伤或牙髓炎时促进牙髓更早和更安全愈合的蛋白质疗法。虽然在研究之初就计划在大鼠牙髓创伤愈合模型中采用质谱分析的方法对牙髓刺激因子进行系统分析，但从大鼠切牙牙髓损伤后即刻、牙髓创伤愈合过程中以及牙髓愈合后的牙髓组织中获得的蛋白量不够，灵敏度不高。因此，二维电泳分析未检测到特殊斑点。推测该蛋白可能具有促进牙髓创伤愈合的高迁移作用。采用TAXIScan-FL体外水平趋化实验检测了各种迁移因子及其浓度梯度对人牙髓干细胞迁移的影响。BDNF的作用最强，SDF-1和bFGF的作用也相对较强，GDNF、VEGF、MMP 3和G-CSF的作用逐渐增强。终浓度为10 ng/μl时迁移细胞数最多。其中，G-CSF用海绵涂敷在离断牙髓上，以检查作为牙髓刺激因子的效果，因为它已经通过将造血干细胞从骨髓动员到外周血的效果而被批准作为药物。结果，在100 ng内几乎看不到效果。计划改变密度和支架，并在未来进行类似的检查。

The purpose of the present study was to analyze the protein that promotes vascularization, pulp wound healing and pulp regeneration to develop the protein therapy that promotes dental pulp healing earlier and safer at the time of pulp injury or the pulpitis. Though it was scheduled to analyze systematically the pulp stimulating factor by Mass spectrometry analyses in the rat dental pulp wound healing model at the beginning of the research, the amount of the protein obtained from the the dental pulp tissue immediately after rat incisor pulp injury, during pulp wound healing process and after pulp healing was not enough and the sensitivity was not high. Therefore, a special spot was not detected by the two-dimensional electrophoretic analyses. It was speculated that the protein to enhance pulp wound healing may have high migratory effect. The migratory effect of the variety of migration factors and their concentration gradient was examined in the human dental pulp stem cells in the horizontal chemotaxis assay by TAXIScan-FL in vitro. BDNF had the highest effect, SDF-1 and bFGF had also comparatively higher effect on migration, and GDNF, VEGF, MMP3, and G-CSF stimulated migration gradually. The final concentration, 10ng/μl was the highest in the number of migrating cells. Among these, G-CSF was applied on the amputated pulp with spongel to examine the effect as the pulp stimulating factor, because it had already been approved as a medicine by the effect of the mobilization of the hemopoietic stem cells from the bone marrow to the peripheral blood. As a result, the effect was hardly seen in 100ng. It is scheduled to change the density and scaffold and to examine it similarly in the future.