Research On The Mechanism Of Gradient Nanostructured Titanium Promoting BMSCs Proliferation,Adhesion And Differentiation In Vitro Based On Genes Filtering And Analysis

Objective:Dental defects caused by various reasons affect the patient’s chewing and articulation function and are not conducive to aesthetics,seriously affecting the physical and mental health of patients.Within the range of indications,dental implants are an ideal way to address dentition defects.Titanium(Ti)and titanium-based alloy materials are ideal scaffolds for repairing bone defects,and the mechanism by which related genes induce bone marrow mesenchymal stem cells(BMSCs)osteogenic differentiation is currently a hot research topic.We use a technology called surface mechanical attrition treatment(SMAT)to prepare SMAT Ti with gradient nanostructure surfaces,and study the proliferation and differentiation of BMSCs inoculated on the surface and potential molecular mechanisms combined with bioinformatics analysis,providing theoretical and experimental basis for SMAT Ti implant application in oral clinics.Methods:1.The datasets GSE37237 and GSE84500 were obtained from the GEO database(Gene Expression Omnibus)and both datasets were based on osteogenesis induction group as experimental group and untreated group as control group.Differentially expressed genes(DEGs)and their functional abundances were studied.Search Tool for the Retrieval of Interacting Genes(STRING)was used to search for protein-protein interaction(PPI)and the expression of DEGs was visualized by Cytoscape software.In addition,differentially expressed genes were enriched by GO(Gene Ontology)and KEGG(Kyoto Encyclopedia of Genes and Genomes)and the results were visualized using R.2.SMAT Ti with gradient nanostructure surfaces was prepared using surface mechanical attrition treatment(SMAT)technology,and well-grown BMSCs were inoculated on the surface of the experimental group and the control group with medical pure titanium as the control group.Cell counting Kits-8,CCK-8),apoptosis,immunofluorescence techniques,alka-line phosphatase(ALP)staining,and alizarin red staining(ARS)staining were performed to study the proliferation,adhesion,and differentiation of BMSCs inoculated on SMAT Ti compared to medical pure titanium.3.Key genes were validated using real-time quantitative PCR(RT-q PCR).Results:1.A total of 279 DEGs were identified,which can be divided into 177 down-regulated genes and 102 up-regulated genes.In addition,DEGs’ enrichment pathways include regulation of actin cytoskeleton,regulation of inflammatory mediators of transient receptor potential(TRP)channels.The 10 upregulated key genes are concentrated in the AMP-activated protein kinase(AMPK)pathway.2.The results of cell counting Kits-8(CCK-8),apoptosis experiments,immunofluorescence techniques,alka-line phosphatase(ALP)staining and alizarin red staining(ARS)staining showed that the proliferation,adhesion and differentiation of BMSCs inoculated on SMAT Ti were significantly better than those of pure titanium.3.The RT-q PCR results showed that OMD,FOXO1 and MME were highly expressed during osteogenesis of BMSCs inoculated on SMAT Ti compared to the pure titanium.Conclusions:1.Regulation of actin cytoskeleton,regulation of inflammatory mediators of transient receptor potential(TRP)channels,peroxisome proliferation receptor(PPAR)pathway,and AMP-activated protein kinase(AMPK)signaling pathway may be involved in SMAT Ti to promote BMSCs osteogenic differentiation.2.The ability of SMAT Ti to promote the proliferation,adhesion and differentiation of BMSCs is significantly better than that of pure titanium.3.The key genes SMAT Ti that promote osteogenic differentiation of BMSCs may be OMD,FOXO1,and MME.