Mechanism Of SLPI In The Differentiation Of Odontoblast

Cavities are a serious threat to human health and high rates of disease,which can lead to periodontitis and lead to cardiovascular disease,diabetes,sepsis.The main method for treating cavities now is to be filled with physics,but the method can be very painful to the patient,and it can’t be physically reversed.Therefore,human tooth regeneration is a very important research work with market value,social benefit and application prospect.The develop ment mechanism of teeth provides theoretical and experimental basis for the artificial cultivation of teeth.The development of teeth is a long and complex process,and it’s the result of the epithelial mass induction.These include the occurrence,formati on,differentiation of cells,matrix secretion and mineralization.Now,the shape of the tooth has been largely studied,but in terms of molecular mechanics,there’s still a lot of exploring.In the process of development of dentin,dentin matrix proteins play an important role,mainly including Col Ⅰ,BSP,OCN,ON,DSPP protein,etc.SLPI is a non-glycosylated single-stranded polypeptide protein secreted by mucosal epithelial cells,containing 107 amino acids,belonging to the family of whey acid protein.SLPI is composed of two highly homolog ous,homocysteine and C-terminal areas,the C-terminal area has the effect of inhibiting elastinase and the N-terminal area has antibacterial,anti-retroviral and anti-inflammatory activities.For the first time,we found that SLPI was associated with tooth devel opment.In addition,although there are reports about the differentiation of basal membrane and dentin,what role does SLPI play in the development of dentin? By what kind of molecular mechanism? It’s not clear.Therefore,it is necessary to probe into the mechanism of SLPI’s mechanism for the differentiation of dentin cell differentiation,basement membrane degradation and dentin,to reveal th e mechanism of molecular action and provide experimental and theoretical basis for the intervention treatment and t ooth repair and regeneration of incomplete tooth development and dentin development.In this paper,the model of MDPC-23 cells and mice were used,the cell and animal models of SLPI gene overexpression and silencing were established,and the role and mechanism of SLPI in tooth development were studied,which provided experimental basis and theoretical basis for the study of tooth regeneration.Experimental contents and results:1.The mouse source SLPI gene overexpression vector p LVX-SLPI-IRES-Zs Green1 was constructed with seamless cloning method,and the mouse source SLPI gene silencing carrier p LVX-SLPI-sh RNA2 was constructed with T4 connection method.2.The constructed plasmid was packed in slow virus and infected with MDPC-23 cells.Stable cell lines were obtained by flow separation.T he expression of SLPI m RNA in plasmid was established by RT-PCR experiment,and the expression of SLPI gene overexpression and silencing stable cell line was established successfully.3.By alizarin red staining and RT-PCR experiments study found that SLPI overexpression promote the differentiation and mineralization of MDPC-23 cells.Endogenous SLPI promote OCN,ON,Col Ⅰ,DSPP m RNA expression and inhibit BSP m RNA expression;Basement membrane promote differentiation and mineralization of odontoblasts.SLPI promote OCN,ON,Col Ⅰ,DSPP m RNA expression and inhibit BSP m RNA expression.4.Based on the development of wild and SLPI knock out mice tissues immunohistochemical and Masson experiments found that SLPI promote OCN,Col Ⅰ protein expression and the degradation of basement membrane,inhibit the expression of BSP and DSPP protein.The expression of ON is always low.When the expression of SLPI disappears,the basement membrane also disappears.It is necessary to further optimize that basement membrane board condition,and study the relationship between basement membrane and differentiation of dental pulp stem cell.