Research On Dentin Matrix Components-controlled-released PCL/SF Ecletrospun As Periodontal Tissue Engineering Scaffold

Aim: Electrospun is the common method for preparing nanofibers.It could produce the large specific surface area,high porosity scaffold which can imitate extracellular matrix.When we add drug component into electrospinning solution,electrospun can be used as drug loading system.With the degradation of the electrospun composition,the drug will be released slowly and affect the surrounding cells and tissues.Method: First we fabricated treated dentin matrix and its extracted liquor,then purified the silk fibroin.Fabricated PCL/SF and PCL/SF/TDM nanofiber scaffold with electrospun.SEM was used to scan the scaffold surface micromorphology and detect the diameter of nanofibers.Explored the drug release of COL-1 and TGF-βwith Elisa kit.Human dental follicle cells were isolated and cultured from the human dental follicle tissue.FCM was used to detected cell surface antigen.Detect the osteogenic,adipogenic,osteogenic differentiation potential of DFCs by induced differentiation.After combined DFCs with the electrospun scaffold,immunofluorescence and CCK8 kit were used to reveal cells numbers and viability on the scaffold.SEM showed the morphology of cells on electrospun.Finally,real-time PCR and western blot were processed to ascertain the odontogenic and osteogenic differentiation degree of the DFCs cultured on the electrospun scaffold.Result:(1)PCL/SF and PCL/SF/TDM electrospun were constituted of successive and aligned nanofibers with average diameter was between 180nm-190nm;(2)COL-1 and TGF-β had similar drug release curve,most of them were release slowly within 10 days,there was no longer a significant increase in drug release after10 days;(3)Human dental follicle cells were isolated and cultured from human dental follicle tissue,and their surface antigens were positive for CD73,CD90,CD105,CD146 while negative for CD14,CD34,CD45.And they had the potential of osteogenic,adipogenic and neural differentiation.(4)The results showed that the fluorescence intensity of cells was similar to that of CCK8,and the cells cultured on the scaffolds were slower than those in the control group at the early stage,but there was no difference in the proliferation activity and the number of cells in the control group at seventh days(5)The cells were spindle-shaped,and the long axis of the cell was consistent with the direction of the nanofibers.(6)Odontogenic and osteogenic-related genes and proteins expression of electrospun groups were up-regulated compared with the control group,and the expression level increased with the passage of time,and what’s more PCL/SF/TDM group show higher expression of odontogenic markers than PCL/SF group.Conclusion: The PCL/SF/TDM electrospun scaffold prepared in this experiment has the ability of odontogenic differentiation and could induce dental follicle cells towards odontogenic differentiation so that it could be a promising scaffold for periodontal tissue engineering.