3D Porous Chitosan Scaffolds Suit Survival And Neural Differentiation Of Dental Pulp Stem Cells

Aim To observe 3 d a porous chitosan scaffold for the survival of the dental pulp stem cells (DPSCs) and their influence to differentiate into nerve cells.Methods The DPSCs were collected from the dental pulp of adult human third molars. We created small 3D porous chitosan scaffolds through freezedrying and determine the Scaffold Swelling Ratio. Cell viability was measured using the Cell Counting K.it-8 (CCK-8) assay. Scanning electron microscopy was used to analyze the surface of the chitosan scaffolds containing attached DPSCs. The successful neural differentiation of DPSCs was assayed by RT-PCR, western blotting, and immunofluorescence to detect the expression of Nestin、MAP-2、GFAP and CNP.Results At a swelling rate of ～84.33 ± 10.92%, the scaffold displayed high porosity and interconnectivity of pores, as revealed by SEM. Cell counting kit-8 assay established the biocompatibility of the chitosan scaffold, supporting the growth and survival of DPSCs. The successful neural differentiation of DPSCs was assayed by RT-PCR, western blotting, and immunofluorescence. We found that the scaffold-attached DPSCs showed high expression of Nestin that decreased sharply following induction of differentiation. Exposure to the differentiation media also increased the expression of neural molecular markers Microtubule-associated protein2, glial fibrillary acidic protein, and 20,30-cyclic nucleotide phosphodiesterase.Conclusion This study demonstrates that the granular 3D chitosan scaffolds are non-cytotoxic, biocompatible, and provide a conducive and favorable microenvironment for attachment, survival, and neural differentiation of DPSCs. These scaffolds have enormous potential to facilitate future advances in treatment of cerebral degenerative disease.