Sugarcane Cystatin Canecpi-1 Promotes Osteogenic Differentiation In Human Dental Pulp Cells: A New Insight Into Cysteine Proteases Inhibitors
INTERNATIONAL ENDODONTIC JOURNAL(2020)
摘要
Aim To investigate the biocompatibility, type of cell death, osteogenic bioactivity and mRNA expression of the osteogenic markers, induced by CaneCPI-1 in human dental pulp cells (hDPCs). Methodology hDPCs exposed to CaneCPI-1 and not exposed (control) were evaluated for cell viability by the 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay; apoptosis by flow cytometry; alkaline phosphatase (ALP) activity by calculation of thymolphthalein release; gene expression of bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (RUNX2), ALP, osteocalcin (OC), bone sialoprotein (BSP) by qPCR; and mineralized nodules production by using alizarin red staining. The data were analysed by one-way analysis of variance (anova) and Turkey's post-test, two-wayanovaand Bonferroni post-test or t-test (P < 0.05). Results CaneCPI-1 induced no apoptosis and had no cytotoxic effect, except in the concentration of 33.20 mu m, in which cell viability was significantly lower than the control (alpha-MEM nonosteogenic medium serum-free) (P < 0.05). There was significantly greater ALP activity, greater expression of the BMP-2, RUNX2, ALP, OC and BSP genes and greater mineralized nodules production in the CaneCPI-1 group in comparison with the control or osteogenic alpha-MEM control (alpha-MEM osteogenic medium - L-ascorbic acid and beta-glycerophosphate) (P < 0.05). Conclusions CaneCPI-1 was cytocompatible and also induced the differentiation of hDPCs in osteogenic phenotypein vitro. CaneCPI-1 is a promising molecule to induce pulp repair.
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关键词
canecystatin, cell differentiation, cysteine proteinase inhibitor, human dental pulp cells
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