Department of Genetics and Evolution, São Carlos Federal University, São Carlos, São Paulo, Brazil.
Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil.
J Dent Res. 2021 Nov;100(12):1344-1350. doi: 10.1177/00220345211011590. Epub 2021 May 10.
The sugarcane cystatin (CaneCPI-5) was recently cloned and showed strong binding force to dental enamel and protection against initial erosion. However, evaluations on its safety and efficacy in a situation closer to the clinical condition are necessary. In the present study we analyzed 1) the cytotoxicity of CaneCPI-5 on human gingival fibroblasts (HGFs); 2) the ability of CaneCPI-5 to reduce enamel erosion and erosion+abrasion in situ. In part 1, HGFs were treated with CaneCPI-5 (0.025, 0.05, 0.1, 0.5 or 1.0 mg/mL) or no treatment (control). The cytotoxicity was assessed after 60 s and 24 h by mitochondrial activity (MTT), confocal microscopy, and hematoxylin/eosin staining. In part 2, 15 volunteers participated in a double-blind crossover protocol consisting of 3 phases, according to the following treatments: 1) 0.1 mg/mL CaneCPI-5; 2) SnCl/NaF/AmF (Elmex; positive control); 3) water (negative control). The volunteers wore an appliance containing 4 bovine enamel specimens for 5 d. Each day, the specimens were individually treated with 50 µL of the tested solutions per 60 s and then subjected to erosive challenges (0.1% citric acid, pH 2.5, for 90 s, 4 times per day). After the first and last erosive challenge each day, 2 samples were abraded (toothbrushing, 15 s). Enamel wear was measured by contact profilometry. One or two-way analysis of variance (ANOVA)/Tukey's or Sidak's tests ( < 0.05) were applied. Regardless of the concentration and the experimental time, CaneCPI-5 did not decrease the cell viability compared to the negative control ( < 0.05). Erosion+abrasion led to significantly greater wear compared to erosion only. For both conditions, the lowest wear was found for SnCl and CaneCPI-5, which did not differ significantly from each other, but showed significant protection when compared to the negative control. In conclusion, CaneCPI-5 is safe on HGFs and reduces enamel erosive wear to the same extent as a commercial solution used to control erosive tooth wear (ETW).
甘蔗半胱氨酸蛋白酶抑制剂(CaneCPI-5)最近被克隆,并表现出对牙釉质的强结合力和对初始侵蚀的保护作用。然而,在更接近临床情况的情况下,对其安全性和功效进行评估是必要的。在本研究中,我们分析了 1)CaneCPI-5 对人牙龈成纤维细胞(HGFs)的细胞毒性;2)CaneCPI-5 减少釉质侵蚀和侵蚀+磨损的能力。在第 1 部分中,用 CaneCPI-5(0.025、0.05、0.1、0.5 或 1.0mg/ml)或无处理(对照)处理 HGFs。用线粒体活性(MTT)、共聚焦显微镜和苏木精/伊红染色法在 60s 和 24h 后评估细胞毒性。在第 2 部分,15 名志愿者参与了一项双盲交叉试验,该试验包括 3 个阶段,根据以下治疗方法进行:1)0.1mg/ml CaneCPI-5;2)SnCl/NaF/AmF(Elmex;阳性对照);3)水(阴性对照)。志愿者佩戴一个含有 4 个牛牙釉质标本的装置 5 天。每天,将标本单独用 50µl 测试溶液处理 60s,然后进行侵蚀挑战(0.1%柠檬酸,pH2.5,90s,每天 4 次)。每天第一次和最后一次侵蚀挑战后,对 2 个样本进行磨损(刷牙,15s)。通过接触式轮廓测量法测量牙釉质磨损。应用单因素或双因素方差分析(ANOVA)/Tukey 或 Sidak 检验(<0.05)。无论浓度和实验时间如何,CaneCPI-5 与阴性对照相比均未降低细胞活力(<0.05)。侵蚀+磨损导致的磨损明显大于仅侵蚀。对于两种情况,磨损最低的是 SnCl 和 CaneCPI-5,它们彼此之间没有显著差异,但与阴性对照相比,具有显著的保护作用。总之,CaneCPI-5 对 HGFs 是安全的,并能减少牙釉质侵蚀性磨损,与用于控制侵蚀性牙磨损(ETW)的商业解决方案效果相当。