Salomão Priscila Maria Aranda, Oliveira Flávia Amadeu de, Rodrigues Paula Danielle, Al-Ahj Luana Polioni, Gasque Kellen Cristina da Silva, Jeggle Pia, Buzalaf Marilia Afonso Rabelo, Oliveira Rodrigo Cardoso de, Edwardson John Michael, Magalhães Ana Carolina
Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil.
Department of Pharmacology, University of Cambridge, Cambridge, United Kingdom.
PLoS One. 2017 Jun 14;12(6):e0179471. doi: 10.1371/journal.pone.0179471. eCollection 2017.
Titanium tetrafluoride (TiF4) has shown promising effect in preventing tooth lesions. Therefore, we compared the cytotoxicity of TiF4 with sodium fluoride (NaF) (already applied in Dentistry) considering different fluoride concentrations, pH values and experimental models.
Step 1) NIH/3T3 fibroblasts were exposed to mediums containing NaF or TiF4 (from 0.15 to 2.45% F), both at native and adjusted pH, for 6 h. Step 2) NIH/3T3 were exposed to NaF or TiF4 varnishes with 0.95, 1.95 or 2.45% F (native pH), for 6, 12 or 24 h. We applied MTT (1st and 2nd steps) and Hoescht/PI stain (2nd step) assays. Step 3) NIH/3T3 were exposed to NaF or TiF4 varnish (2.45% F), at native pH, for 6 or 12 h. The cell stiffness was measured by atomic force microscopy (AFM).
Step 1) All cells exposed to NaF or TiF4 mediums died, regardless of the F concentration and pH. Step 2) Both varnishes, at 1.90 and 2.45% F, reduced cell viability by similar extents (33-86% at 6 h, 35-93% at 12 h, and 87-98% at 24 h) compared with control, regardless of the type of fluoride. Varnishes with 0.95% F did not differ from control. Step 3) TiF4 and NaF reduced cell stiffness to a similar extent, but only TiF4 differed from control at 6 h.
Based on the results of the 3 experimental steps, we conclude that TiF4 and NaF have similar cytotoxicity. The cytotoxicity was dependent on F concentration and exposure time. This result gives support for testing the effect of TiF4 varnish in vivo.
四氟化钛(TiF4)在预防牙齿病变方面已显示出有前景的效果。因此,我们考虑不同的氟浓度、pH值和实验模型,比较了TiF4与已应用于牙科的氟化钠(NaF)的细胞毒性。
步骤1)将NIH/3T3成纤维细胞暴露于含有NaF或TiF4(氟含量为0.15%至2.45%)的培养基中,培养基的pH值为天然值和调节后的值,持续6小时。步骤2)将NIH/3T3细胞暴露于氟含量为0.95%、1.95%或2.45%(天然pH值)的NaF或TiF4清漆中,持续6、12或24小时。我们应用了MTT(第一步和第二步)和Hoescht/PI染色(第二步)检测。步骤3)将NIH/3T3细胞暴露于氟含量为2.45%的NaF或TiF4清漆中,天然pH值,持续6或12小时。通过原子力显微镜(AFM)测量细胞硬度。
步骤1)所有暴露于NaF或TiF4培养基的细胞均死亡,无论氟浓度和pH值如何。步骤2)与对照组相比,氟含量为1.90%和2.45%的两种清漆均使细胞活力降低了相似程度(6小时时为33% - 86%,12小时时为35% - 93%,24小时时为87% - 98%),无论氟的类型如何。氟含量为0.95%的清漆与对照组无差异。步骤3)TiF4和NaF使细胞硬度降低的程度相似,但仅在6小时时TiF4与对照组不同。
基于3个实验步骤的结果,我们得出结论,TiF4和NaF具有相似的细胞毒性。细胞毒性取决于氟浓度和暴露时间。这一结果为在体内测试TiF4清漆的效果提供了支持。