| Background:At present,regenerative endodontic treatment(RET)is a new method for the treatment of pulp diseases in young permanent teeth.The regenerative endodontic treatment is a treatment method that uses the principles of regenerative medicine and tissue engineering to restore and regenerate functional vital pulp.The three major components of tissue engineering are seed cells,biological scaffolds and growth factors,and the blood supply is also crucial.In recent years,a lot of studies have been done on pulp regeneration using dental stem cells.Pulp stem cells are derived from pulp and used as seed cells in pulp regeneration,which has a natural advantage.Depending on the time of development,human dental pulp stem cells can be divided into stem cells from human exfoliated deciduous teeth(SHED)and permanent teeth dental pulp stem cells(DPSC).SHED and DPSC originate from neural crest cells and have the characteristics of self-renewal,clonal formation and multidirectional differentiation of mesenchymal stem cells.At the same time,SHED and DPSC are located in the perivascular niche.On the one hand,they can parapocrine a variety of angiogenic regulatory proteins,and on the other hand,they can differentiate into vascular endothelial cells under specific microenvironment,thus promoting angiogenesis.Although there are a large number of studies on human pulp stem cells,most of the current studies focus on the general biological characteristics and mineralization ability of human pulp stem cells.There are relatively few reports on the angiogenic ability of SHED and DPSC,and the difference in angiogenic ability between the two has not been reported.Purpose:The purpose of this study was to study the differences between SHED and DPSC in general biological characteristics and angiogenic ability,and to provide experimental basis for clinical pulp regeneration therapy.Methods:1.The SHED and DPSC were isolated and cultured from pulp tissue of human deciduous teeth and young permanent teeth by modified tissue block enzymolysis.The surface markers of two kinds of pulp stem cells were detected by flow cytometry.After multidirectional induction,angiogenic,adipogenic and osteogenic differentiation abilities of the two types of cells were detected.CCK8 was used to detect the proliferation ability of two kinds of dental pulp stem cells.The migration ability of the two kinds of dental pulp stem cells was detected by scratch test.2.SHED and DPSC with good growth condition were induced in the direction of angiogenesis.The formation of vascular-like structures of the two kinds of cells after induction was detected by Matrigel tubule formation experiment,and the expression of angiogenic factors was detected by RT-q PCR and Western blot techniques.Results:1.Microscopically,both SHED and DPSC were found to be fibroblast-like.Flow cytometry showed that both cells had low expression of hematopoietic stem cell markers and high expression of mesenchymal stem cell markers.Both kinds of dental pulp stem cells can differentiate towards osteogenic and angiogenic directions,and are mesenchymal derived stem cells.CCK8 results showed that SHED had stronger proliferation ability than DPSC.The scratch experiment results show that SHED has better migration ability than DPSC.2.On Matrigel matrix glue,both SHED and DPSC could form vascular-like structure after angiogenic induction for 14 days,and the number of vascular mesh,nodes and segments formed by SHED was significantly higher than that of DPSC.RT-q PCR results showed that angiogenic genes were differentially expressed between the two types of cells,and SHED had a higher expression of angiogenic genes than DPSC.Western Blot results showed that on the 7th and 14 th day,the expression of vascular associated protein was higher in SHED than that in DPSC.Conclusion:Both SHED and DPSC isolated and cultured in this study were mesenchymal derived stem cells with multidirectional differentiation potential,and SHED has stronger proliferation and migration ability than DPSC.SHED has stronger angiogenic ability than DPSC in the vascular microenvironment and is an ideal source of seed cells for pulp regeneration. |
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