Universidade de São Paulo, Faculdade de Odontologia, Departamento de Dentística, São Paulo, São Paulo, Brasil.
Universidade de São Paulo, Faculdade de Odontologia, Departamento de Estomatologia, São Paulo, São Paulo, Brasil.
J Appl Oral Sci. 2019;27:e20180396. doi: 10.1590/1678-7757-2018-0396. Epub 2019 Apr 11.
Endodontic revascularization is based on cell recruitment into the necrotic root canal of immature teeth after chemical disinfection. The clinical outcome depends on the ability of surviving cells from the apical tissue to differentiate and promote hard tissue deposition inside the dentinal walls.
To investigate the effect of calcium hydroxide (CH) and modified triple antibiotic paste (mTAP - ciprofloxacin, metronidazole and cefaclor) on the viability and mineralization potential of apical papilla cells (APC) in vitro .
APC cultures were kept in contact with CH or mTAP (250-1000 µg/mL) for 5 days, after which cell viability was assessed using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Next, APCs were subjected to CH or mTAP at 250 µg/mL for 5 days before inducing the differentiation assay. After 14 and 21 days, calcium deposition was assessed by the Alizarin Red S staining method, followed by elution and quantification using spectrophotometry. Data were analyzed using ANOVA followed by Tukey post hoc test.
CH induced cell proliferation, whereas mTAP showed significant cytotoxicity at all concentrations tested. APC treated with CH demonstrated improved mineralization capacity at 14 days, while, for mTAP, significant reduction on the mineralization rate was observed for both experimental periods (14 and 21 days).
Our findings showed that CH induces cell proliferation and improves early mineralization, whereas mTAP was found cytotoxic and reduced the mineralization potential in vitro of APCs.
研究氢氧化钙(CH)和改良三联抗生素糊剂(mTAP-环丙沙星、甲硝唑和头孢克洛)对体外根尖乳头细胞(APC)活力和矿化潜能的影响。
将 APC 培养物与 CH 或 mTAP(250-1000μg/mL)接触 5 天,然后使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)测定法评估细胞活力。接下来,将 APC 在 250μg/mL 的 CH 或 mTAP 作用 5 天,然后进行诱导分化测定。14 和 21 天后,通过茜素红 S 染色法评估钙沉积,然后使用分光光度法洗脱和定量。使用方差分析(ANOVA)和 Tukey 事后检验对数据进行分析。
CH 诱导细胞增殖,而 mTAP 在所有测试浓度下均显示出显著的细胞毒性。用 CH 处理的 APC 在 14 天时显示出改善的矿化能力,而对于 mTAP,在两个实验期(14 天和 21 天)都观察到矿化率显著降低。
我们的研究结果表明,CH 诱导细胞增殖并改善早期矿化,而 mTAP 具有细胞毒性并降低体外 APC 的矿化潜能。