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-way anova and 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 µm, in which cell viability was significantly lower than the control (α-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 α-MEM control (α-MEM osteogenic medium - L-ascorbic acid and β-glycerophosphate) (P < 0.05). CONCLUSIONS CaneCPI-1 was cytocompatible and also induced the differentiation of hDPCs in osteogenic phenotype in vitro. CaneCPI-1 is a promising molecule to induce pulp repair.

中文翻译：

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甘蔗胱抑素CaneCPI-1促进人牙髓细胞的成骨分化：对半胱氨酸蛋白酶抑制剂的新见解。

目的研究CaneCPI-1在人牙髓细胞（hDPCs）中诱导的生物相容性，细胞死亡类型，成骨生物活性和成骨标记物的mRNA表达。方法学暴露于CaneCPI-1且未暴露（对照）的hDPC通过3-（4,5-二甲基噻唑）-2,5-二苯基四唑溴化物（MTT）分析评估细胞活力；流式细胞仪检测细胞凋亡 通过计算百里酚酞释放量来碱性磷酸酶（ALP）的活性；通过qPCR检测骨形态发生蛋白2（BMP-2），矮子相关转录因子2（RUNX2），ALP，骨钙蛋白（OC），骨唾液蛋白（BSP）的基因表达；和茜素红染色可生产矿化的结核。通过单因素方差分析（方差分析）和土耳其的事后检验，两因素方差分析和Bonferroni事后检验或t检验对数据进行分析（P <0.05）。结果除了浓度为33.20 µm的CaneCPI-1（其细胞活力显着低于对照（无血清的α-MEM非成骨培养基））（P <0.05）外，CaneCPI-1不会诱导细胞凋亡，也没有细胞毒性作用。与对照或成骨性α-MEM对照（α-MEM成骨性）相比，CaneCPI-1组的ALP活性明显更高，BMP-2，RUNX2，ALP，OC和BSP基因的表达更高，矿化的结节产生更多。中等-L-抗坏血酸和β-甘油磷酸酯（P <0.05）。结论CaneCPI-1具有细胞相容性，并且还可以诱导hDPCs在成骨表型中分化。CaneCPI-1是诱导果肉修复的有前途的分子。细胞活力显着低于对照（无血清的α-MEM非骨形成培养基）（P <0.05）。与对照或成骨性α-MEM对照（α-MEM成骨性）相比，CaneCPI-1组的ALP活性明显更高，BMP-2，RUNX2，ALP，OC和BSP基因的表达更高，矿化的结节产生更多。中等-L-抗坏血酸和β-甘油磷酸酯（P <0.05）。结论CaneCPI-1具有细胞相容性，并且还可以诱导hDPCs在成骨表型中分化。CaneCPI-1是诱导果肉修复的有前途的分子。细胞活力显着低于对照（无血清的α-MEM非骨形成培养基）（P <0.05）。与对照或成骨性α-MEM对照（α-MEM成骨性）相比，CaneCPI-1组的ALP活性明显更高，BMP-2，RUNX2，ALP，OC和BSP基因的表达更高，矿化的结节产生更多。中等-L-抗坏血酸和β-甘油磷酸酯（P <0.05）。结论CaneCPI-1具有细胞相容性，并且还可以诱导hDPCs在成骨表型中分化。CaneCPI-1是诱导果肉修复的有前途的分子。与对照组或成骨性α-MEM对照（α-MEM成骨性培养基-L-抗坏血酸和β-甘油磷酸酯）相比，CaneCPI-1组的OC和BSP基因以及矿化的结节产生更多（P <0.05）。结论CaneCPI-1具有细胞相容性，并且还可以诱导hDPCs在成骨表型中分化。CaneCPI-1是诱导果肉修复的有前途的分子。与对照组或成骨性α-MEM对照（α-MEM成骨性培养基-L-抗坏血酸和β-甘油磷酸酯）相比，CaneCPI-1组的OC和BSP基因以及矿化的结节产生更多（P <0.05）。结论CaneCPI-1具有细胞相容性，并且还可以诱导hDPCs在成骨表型中分化。CaneCPI-1是诱导果肉修复的有前途的分子。