Ageing affects the proliferation and mineralization of rat dental pulp stem cells under inflammatory conditions.

Abstract

To comparatively evaluate changes inthe proliferation and mineralization abilities of dental pulp stem cells (DPSCs) from juvenile and adult rats in a lipopolysaccharide (LPS)-induced inflammatory microenvironment to provide a theoretical basis for the age-related differences observed in DPSCs during repair of inflammatory injuries. DPSCs were isolated from juvenile (JDPSCs) and adult rats (ADPSCs), and senescence-associated β-galactosidase staining was used to compare senescence between JDPSCs and ADPSCs. Effects of LPS on JDPSCs and ADPSCs proliferation were investigated by cell counting kit-8 assays and flow cytometry. Alizarin red staining, quantitative reverse transcription polymerase chain reaction and Western blot assay were used to examine the effects of LPS on mineralization-related genes and proteins in JDPSCs and ADPSCs. Immunohistochemistry was used to compare interleukin-1β (IL-1β) and osteocalcin (OCN) expression in the pulpitis model. Unpaired Student's t-tests and one-way anova were used for statistical analysis. DPSCs were isolated from juvenile and adult rat dental pulp tissues. At low concentrations (0.1-1μgmL-1 ), LPS significantly promoted the proliferation of JDPSCs (P<0.01) and ADPSCs (P<0.01 or P<0.05), with the effect being stronger in JDPSCs than in ADPSCs. In addition, mineralized nodules and the expression of mineralization-related genes (OCN, DSPP, ALP, BSP) increased significantly after stimulation with LPS (0.5μgmL-1 ) in JDPSCs and ADPSCs (P<0.01 or P<0.05), and JDPSCs displayed a more obvious increase than ADPSCs. Western blots revealed OCN and ALP expression levels in JDPSCs treated with LPS were significantly upregulated (P<0.05); meanwhile, ALP expression in ADPSCs increased slightly but significantly (P<0.05), and OCN expression was not affected. Finally, IL-1β expression was significantly higher (P<0.05) and OCN expression was significantly lower (P<0.05) in the inflamed dental pulp of adult rats than in juvenile rats. A certain degree of inflammatory stimulation promoted the proliferation and mineralization of DPSCs; however, this effect declined with age. The DPSCs of adult donors in an inflammatory microenvironment have a weaker repair ability than that of juvenile donors, who are better candidates for tissues damage repair.