Comparing the Effects of Chitosan Scaffolds Containing Various Divalent Metal Phosphates on Osteogenic Differentiation of Stem Cells from Human Exfoliated Deciduous Teeth

Inducing the differentiation of stem cells from human exfoliated deciduous teeth (SHEDs) proceeds with low efficiency, which greatly limits clinical applications. Divalent metal elements play an important role in osteoinductivity for bone remodeling because they can simulate bone formation and decrease bone resorption. The purpose of this study was to investigate the effect of some divalent metal phosphates on osteogenic differentiation from human exfoliated deciduous teeth. These divalent metal ions can be gradually released from the scaffold into the culture medium and continually induce osteoblastic differentiation. Experimental results revealed that SHEDs cultured in chitosan scaffolds containing divalent metal phosphates had notably increased osteoblastic differentiation compared with cells cultured without divalent metal phosphates. This effect was due to the high activity of alkaline phosphatase, as well as the bone-related gene expression of collagen type I, Runx2, osteopontin, osteocalcin, VEGF, and Ang-1, shown through RT-PCR and bone-related protein immunocytochemistry stains. A calcium-content assay further revealed significant enhancement of deposited minerals on the scaffolds after 21 days of culture, particularly for magnesium phosphate and zinc phosphate. Thus, divalent metals, except for barium phosphate, effectively promoted SHED cell differentiation and osteoblastic cell maturation. This study demonstrated that the divalent metal elements magnesium, strontium, and zinc could effectively induce SHED osteoblastic differentiation for use in tissue engineering and bone repair.