| BackgroundDental pulp stem cells have good proliferative capacity and multi-differentiation ability,so they are considered ideal seed cells for tooth regeneration.However,most of dental pulp stem cells in current studies were isolated from healthy dental pulp tissue.There exists stem cells in other pulp tissue from diseased teeth(teeth with dental caries,pulpitis,periodontitis,etc.).Studies have shown that dental pulp stem cells can be isolated from teeth with deep caries,which have stronger proliferation and mineralization capacity than dental pulp stem cells from normal teeth[1].Therefore,dental pulp stem cells isolated from unhealthy dental pulp tissue may also be used in tooth engineering.Vessels provide nutrients and oxygen to tissue,drain metabolic waste,thus angiogenesis is the basis for tissue repair and tissue regeneration.Due to the special anatomical structure of the dental pulp cavity,the pulp is surrounded by rigid dentin,and the main source of blood supply is only through the apical foramina,which lacks an effective collateral circulation.Therefore,angiogenesis is a key step in tissue repair and tissue regeneration.At present,there is no research report on the angiogenesis of dental pulp stem cells from teeth with deep caries.Therefore,our research intends to investigate the angiogenic and mineralized characteristics of dental pulp stem cells from teeth with deep caries and to explore the possibility in dental tissue engineering.Materials and Methods1.Research on biological characteristics of dental pulp stem cells from normal teeth and dental pulp stem cells from teeth with deep cariesThe dental pulp stem cells were isolated and cultured from normal pulp tissue and pulp tissue with deep caries by tissue enzyme digestion method.The dental pulp stem cells from normal teeth were named as nDPSCs and the dental pulp stem cells from teeth with deep caries were named as cDPSCs.Flow cytometry was used to detect cell surface markers of these cells;both were induced to differentiate into adipogenic,osteogenic,and chondrogenic directions to detect their multilineage differentiation potential;colony-formation assay was used to detect the clonality of both cells.The CCK8 assay detected the proliferation characteristics of both cells.qRT-PCR was used to detect the expression of the inflammatory mediators in both cells.2.Research on angiogenic and mineralized characteristics in dental pulp stem cells from normal teeth and dental pulp stem cells from teeth with deep caries in angiogenesis microenvironmentImmunohistochemical assay was used to detect the expression of angiogenesis related protein in normal dental pulp tissue and pulp tissue with deep caries,dental pulp stem cells from normal teeth and dental pulp stem cells from teeth with deep caries.The third passage of both cells in logarithmic growth phase were induced into angiogenesis.qRT-PCR was used to detect the expression of angiogenic factors and mineralization-related factors.The expression of VEGF and ALP in both cells was detected by Western Blot.Results1.Biological characteristics of dental pulp stem cells from normal teeth and dental pulp stem cells from teeth with deep cariesIn this experiment,dental pulp stem cells from normal teeth and dental pulp stem cells from teeth with deep caries were successfully isolated and cultured.Flow cytometry results showed that the surface marker molecules of mesenchymal stem cells were highly expressed on both cell surfaces,while the hematopoietic stem cell surface marker molecules were lowly expressed;both cells could be induced into osteogenic,adipogenic and chondrogenic differentiations and were mesenchymal derived stem cells;both cells had the colony-formation capability.Compared with dental pulp stem cells from normal teeth,dental pulp stem cells from teeth with deep caries had a stronger ability of colony-formation capability(P<0.01).CCK8 assay showed that the proliferation ability of dental pulp stem cells from teeth with deep caries was stronger than that of dental pulp stem cells from normal teeth(P<0.01).qRT-PCR results showed that the expression of inflammatory mediators in dental pulp stem cells from teeth with deep caries was more highly than that in dental pulp stem cells from normal teeth(P<0.01).2.Angiogenic and mineralized characteristics in dental pulp stem cells from normal teeth and dental pulp stem cells from teeth with deep caries in angiogenesis microenvironmentCompared with normal pulp tissue,pulp tissue with deep caries expressed higher angiogenesis related protein(P<0.01).In the angiogenesis microenvironment,the expression of angiogenic factors and mineralization-related factors was higher in dental pulp stem cells from teeth with deep caries than that in dental pulp stem cells from normal teeth(P<0.05);meanwhile,Western Blot results showed that the expression of angiogenic and mineralized related protein were higher in dental pulp stem cells from teeth with deep caries.Conclusion1.The dental pulp stem cells from normal teeth and dental pulp stem cells from teeth with deep caries isolated and cultured in this experiment were mesenchymal-derived stem cells and owned multi-directional differentiation potential.2.Dental pulp stem cells from teeth with deep caries had higher expression of inflammatory mediators,stronger colony-formation capacity and proliferation ability than dental pulp stem cells from normal teeth.3.In the angiogenesis microenvironment,dental pulp stem cells from teeth with deep caries had stronger angiogenesis and mineralization potential than dental pulp stem cells from normal teeth. |
PDF Full Text Request