Fluorescent Protein Mark Dental Tissue Engineering Study In Vivo

0bjective: We mixed the fluorescent protein labeled germ cells and gene transfecting the bone marrow stem cells and complexing with 3D printing polyvinyl alcohol/biphasic ceramic bone scaffold and gelatin sponge stand to construct heterogeneous tooth tissue engineering.In order to investigate their ability to differentiate into the teeth in vivo.Methods: 192 healthy SD rats were randomly divided into six groups,A:the cell pellet group(fluorescent protein labeled germ cells and gene transfecting the bone marrow stem cells);B:polyvinyl alcohol/biphasic ceramic bone scaffold group;C:cell clumps+polyvinyl alcohol/biphasic ceramic bone scaffold group;D:gelatin sponge group;E:cell clumps+gelatin sponge group;F:control group which are without any intervention.It is implanted into the renal subcapsular of rats under general anesthesia.We choose four time points after surgery to collect samples for testing,Masson staining;immunohistochemical staining and confocal slice observation.At the 5th,10 th,14th,28 th day each group collected8 samples.Results: Gross histological and Masson staining indicated:The 28 th day after morphological observation showed E group had dental tissue formation.Immunohistochemistry:Factorial analysis showed that C and E group in 5,10 days Dentin matrix protein1,bone morphogenetic protein 4 expression were highest and E group in 14 days expression Dentin sialoprotein were highest and were higher than the rest of the group the difference was statistically significant P < 0.05.Confocal microscopy showed:C and E group at each time point can observed in germ cell fluorescently marker.Conclusion: Fluorescently labeled germ cells and gene transfection bone marrow stem cells mixed culture,composite different scaffold successfully constructed of tooth tissue engineering tooth differentiation,its mechanism of into teeth needs further study.