Upregulating Odonto/Osteogenic-differentiation of Dental Stem Cells by cAMP-loaded Layer-by-layer Self-assembly

Abstract

Aim: To investigate the effects of cAMP loaded by layer-by-layer (LBL) self-assembled delivery on odonto/osteogenic differentiation of stem-cells from apical-papilla (SCAPs). Methods: The treatment cells were SCAPs encapsulated with cAMP using layer-by-layer self-assembled gelatin and alginate polyelectrolytes (LBL-SCAPs). The expression of cAMP on the surface of LBL-SCAPs was detected by immunofluorescence after 24 hours. The cellular morphology of LBL-SCAPs and control cells was analyzed by scanning electron microscopy (SEM). Cell Counting Kit-8(CCK8) and live/dead staining were used to detected the proliferation and viability. The effect of cAMP-loaded treatment on odonto/osteogenic differentiation of SCAPs was assessed by Alizarin red staining. Results: LBL-SCAPs highly expressed cAMP which was detected by immunofluorescence after 24 hours. SEM assay showed that LBL-SCAPs exhibited a spheroid-like morphology, while control cells exhibited like a spindle cellular morphology. CCK8 and live/dead staining demonstrated that cAMP-loaded treatment had no effect on the proliferation and viability of SCAPs. Alizarin red staining revealed that LBL-SCAPs formed a greater number of mineralized nodules compared to the control cells. Conclusion: cAMP-loaded by layer-by-layer self-assembly encapsulated SCAPs enhances odonto/osteogenic differentiation of stem-cells from apical-papilla.