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

Journal 2019 · IF 3.331 · 53(1): 72-83 · 发表于 2019 年 8 月 16 日
作者: T. Ning,J. Shao,X. Zhang,X. Luo,X. Huang,H. Wu,S. Xu,B. Wu,D. Ma
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摘要

Aim To comparatively evaluate changes in the 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. Methodology 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. Results DPSCs were isolated from juvenile and adult rat dental pulp tissues. At low concentrations (0.1–1 μg mL −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 (0.5 μg mL −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. Conclusions 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.

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