Sipert Carla Renata, Oliveira Aline Pereira, Caldeira Celso Luiz
Universidade de São Paulo, Faculdade de Odontologia, Departamento de Dentística, São Paulo, Brasil.
J Appl Oral Sci. 2019 Feb 21;27:e20180291. doi: 10.1590/1678-7757-2018-0291.
The aim of this study was to investigate the cytotoxic effects of modified triple antibiotic paste and an experimental composition using calcium hydroxide on lipoteichoic acid (LTA)-primed apical papilla cells (APC).
Human APC were tested for in vitro cytotoxicity of modified Triple Antibiotic Paste (mTAP - Ciprofloxacin, Metronidazole and Cefaclor at 1:1:1) and of a paste of Ciprofloxacin, Metronidazole and Calcium hydroxide (CMC - 1:1:2) and modified CMC (mCMC - 2:2:1) by using MTT assay. The substances were reconstituted in DMEM at 1,000 µg/mL and ¼ serially diluted before being kept in contact with cells for 1, 3, 5 and 7 days. Further, cells were primed with 1 µg/mL of Enterococcus faecalis LTA for 7 days prior to the viability test with 1,000 µg/mL of each substance. Statistical analysis was performed using one-way analysis of variance (ANOVA) and two-way ANOVA respectively followed by Tukey's post-test. Significance levels were set at p<0.05.
In the first assay, the higher cytotoxic rates were reached by mTAP for all experimental periods. CMC was found toxic for APC at 5 and 7 days, whereas mCMC did not affect the cell viability. Only CMC and mCMC were able to induce some cellular proliferation. In the second assay, when considering the condition with medium only, LTA-primed cells significantly proliferated in comparison to LTA-untreated ones. At this context, mTAP and CMC showed similar cytotoxicity than the observed for LTA-untreated cells, while mCMC was shown cytotoxic at 7 days only for LTA-primed APC. Comparing the medications, mTAP was more cytotoxic than CMC and mCMC.
mTAP showed higher cytotoxicity than CMC and mCMC and the effect of topic antimicrobials might differ when tested against apical papilla cells under physiological or activated conditions.
本研究旨在探讨改良三联抗生素糊剂及一种含氢氧化钙的实验性组合物对脂磷壁酸(LTA)致敏的根尖乳头细胞(APC)的细胞毒性作用。
采用MTT法检测人APC对改良三联抗生素糊剂(mTAP - 环丙沙星、甲硝唑和头孢克洛,比例为1:1:1)、环丙沙星、甲硝唑和氢氧化钙糊剂(CMC - 1:1:2)及改良CMC(mCMC - 2:2:1)的体外细胞毒性。将这些物质在DMEM中以1000μg/mL重新配制,并进行1/4系列稀释,然后与细胞接触1、3、5和7天。此外,在使用每种物质1000μg/mL进行活力测试前7天,用1μg/mL粪肠球菌LTA对细胞进行致敏。分别采用单因素方差分析(ANOVA)和双因素方差分析,随后进行Tukey事后检验进行统计分析。显著性水平设定为p<0.05。
在第一次试验中,mTAP在所有实验期均达到较高的细胞毒性率。发现CMC在第5天和第7天对APC有毒性,而mCMC不影响细胞活力。只有CMC和mCMC能够诱导一些细胞增殖。在第二次试验中,仅考虑使用培养基的情况时,与未用LTA处理的细胞相比,用LTA致敏的细胞显著增殖。在此情况下,mTAP和CMC显示出与未用LTA处理的细胞相似的细胞毒性,而mCMC仅在第7天对用LTA致敏的APC显示出细胞毒性。比较这些药物,mTAP比CMC和mCMC具有更高的细胞毒性。
mTAP比CMC和mCMC显示出更高的细胞毒性,并且在生理或活化条件下针对根尖乳头细胞进行测试时,局部抗菌药物的效果可能不同。