| Objective: The effects of CGF on the proliferation and differentiation of dental pulp cells were observed in the process of human dental pulp cell culture;The root canal was treated with sodium hypochlorite,EDTA and chlorhexidine to observe the effects of different rinsing solutions on pulp cells adhering to the root canal wall;Observing the effect of erythropoietin(EPO)on the proliferation and differentiation of pulp cells under hypoxic conditions;Observing the clinical effect of CGF dental pulp revascularization and apical induction in the treatment of young permanent teeth;we hope to explore the factors and means that affect and protect pulp cells during pulp revascularization,To provide experimental reference for improving the prognosis of pulp revascularization through the proliferation and protection of pulp cells and CGF improves the success rate of pulp revascularization,so as to continuously improve the effect of pulp revascularization and better retain the young permanent teeth with pulp necrosis.Methods:1.Pulp cells were isolated and subcultured from healthy intact teeth under 18 years of age requiring pull out due to causes of orthodontics or impaction by modified tissue enzyme digestion.The patient's venous blood was extracted to prepare CGF gel,which was extruded into a film and cut into a size of 3×3mm for later use.This experiment was divided into four groups.In the experimental group,the medium were added to 1 CGF(low concentration),2 CGF(medium concentration)and 3 CGF(high concentration),respectively.In the control group,the medium added DMEM containing only 10%FBS.cck-8 was used to detect the effects of CGF on the proliferation of pulp cells,Alkaline phosphatase(ALP)was used to detect the effects of CGF on the differentiation of pulp cells at 3,5 and 7 days respectively.2.The mandibular single premolars extracted from the maxillofacial surgery clinic of Stomatology Hospital of Hebei Medical University were collected.High-speed corundum burs are used to open the dental pulp,extract the pulp,and cut off the crown with the carborundum piece at the enamel bone boundary.Root canals were treated with sodium hypochlorite(group A),EDTA(group B),and chlorhexidine(group C),dissected longitudinally to expose the root canal wall,cut into 1mm × 1.5mm × 1.5mm and placed in 75% ethanol.The remaining ethanol was rinsed with PBS after 30 min,and then placed in a 96-well plate to inoculate dental pulp cells.After 24 hours,observing the adherence of dental pulp cells and detect CCK-8.3.The experiment was divided into two groups under hypoxic conditions.The experimental group used 10% FBS DMEM medium containing 20 u / ml EPO to culture dental pulp cells.Control group only used DMEM with 10% FBS.The oxygen concentration of the incubator was set to 1%,and CCK-8 detected the effects of EPO on the proliferation of dental pulp cells,ALP detected the effects of EPO on the mineralization of dental pulp cells after 24 h,48h,72 h of culture.4.Collect 14 cases of young permanent teeth diagnosed with pulpitis or periapical periodontitis at the Stomatological Hospital of Hebei Medical University from March 2017 to December 2019.According to the patient's choice and the actual materials,the experimental group performed CGF pulp revascularization with i Root BP coverage,while the control group performed traditional apical induction.All the teeth met the inclusion criteria,and were reexamined regularly after the operation.The success rate of the treatment was evaluated by the clinical manifestations and X-ray films.5.Each group of data was statistically analyzed by SPSS21.0 software.Results:1.CGF group can promote the proliferation of dental pulp cells,and the proliferative capacity of dental pulp cells is also enhanced with the increase of time and CGF concentration.CCK-8 showed no significant difference in absorbance between the experimental group and the control group on the 3rd day(P> 0.05),but there were significant differences between the experimental group and the control group on the 5th and 7th day(P <0.05).The ALP kit showed that the alkaline phosphatase activity of the CGF group was higher than the control group on the 7th day,and there was a statistically significant difference in absorbance(P <0.05).2.the CCK-8 showed that the absorbance value of the Sodium hypochlorite group was higher than the other two groups after the root canal was treated with different root canal irrigants and dental pulp cells were cultured for 24 hours,and the difference was statistically significant(P <0.05).3.The results of CCK-8 showed that the absorbance of EPO group was higher than that of the control group under hypoxic conditions at 24 h,48h or 72h(P < 0.05).The results of ALP showed that there was no significant difference between the experimental group and the control group at 24 h and 48 h.At 72 h,the absorbance of the experimental group was higher than that of the control group,but the difference was not statistically significant(P > 0.05).4.Compared with the control group,the experimental group using CGF can better promote the development of roots through the clinical manifestations and X-ray films.Conclusions:1.High concentration of CGF can significantly promote the proliferation and differentiation of pulp cells,and improve the prognosis of pulp revascularization.2.Compared with sodium hypochlorite,EDTA and chlorhexidine,sodium hypochlorite can effectively improve the attachment of pulp cells to the root canal wall.3.EPO can protect dental pulp cells under hypoxia,promote their proliferation and anti-inflammatory effects.4.During pulp revascularization,CGF can promote the development of immature permanent teeth. |
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