Adhesion and proliferation of dental pulp stem cells on the chitosan-fibrin composite scaffold / 中国组织工程研究

ABSTRACT

BACKGROUND: With the rapid development of tissue engineering, a single biological scaffold material is hard to meet the needs of tissue engineering. Therefore, composite scaffolds with excellent performance will be obtained by combining two or more kinds of materials.OBJECTIVE: To detect the adherence and proliferation of dental pulp stem cells on the Chitosan-fibrin composite scaffold.METHODS: Dental pulp stem cells were isolated and extracted from C57 neonatal rats through modified enzyme-digestion method, and subcultured to the third generation, followed by adipogenic and osteogenic induction in vitro. Then, induced cells were identified. The chitosan-fibrinogen composite scaffold was prepared, and the pore size and porosity were determined. The chitosan-fibrin composite scaffold was co-cultured with passage 3 dental pulp stem cells to observe the cell proliferation by MTT assay, and the morphology of the composite scaffold, cell adhesion,proliferation and extracellular matrix secretion were observed under scanning electron microscope. In addition, the cells were inoculated directly on the bottom of culture plate as controls.RESULTS AND CONCLUSION: The dental pulp stem cells were successfully isolated and cultivated, and positive for osteogenic and adipogenic differentiation. The pore size and porosity of the composite scaffold was (105.32±22.10) μm and (87.714±1.276)%, respectively. The S-shaped proliferation curve in the experimental group was similar with that in the control group; the proliferation rate in the experimental group was significantly higher than that in the control group after 4-8 days of culture (P < 0.05). At the 2nd day after co-culture, the cells adhered tightly and grew well onto the composite scaffold; at the 4th day, enlarged cells began to proliferate obviously with abundant extracellular matrix; the surface and pores of the scaffold were full of cells at the 6th day. These results indicate that the chitosan-fibrin composite scaffold is suitable for the adhesion and proliferation of dental pulp stem cells.