Abstract

Objective 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).Material and Methods 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.Results 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.Conclusion 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.

Highlights

  • Regenerative Endodontic Procedures are considered one of the main issues discussed at contemporary Endodontics

  • Considering the importance of survival of apex surrounding cells after root canal disinfection prior to revascularization procedures, this study aimed to investigate the cytotoxicity of a modified triple antibiotic paste and formulations including ciprofloxacin, metronidazole and calcium hydroxide (CMC and modified Calcium hydroxide at distinct proportions: 1:1:2 (CMC)) on human cultured apical papilla cells under lipoteichoic acid (LTA)-untreated or LTA-primed conditions

  • To understand the effect of innate immunity activation on cytotoxic effect of intracanal dressings on ACP; part of the cells were primed with 1 μg/mL LTA from E. faecalis

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Summary

Introduction

Regenerative Endodontic Procedures are considered one of the main issues discussed at contemporary Endodontics. Based on cytotoxic data regarding TAP paste, CH is being proposed for revascularization procedures due to its biocompatibility and antimicrobial activity.7 Some microorganisms, such as Enterococcus faecalis, are resistant to this substance due to intrinsic mechanisms that lead to bacterial survival instead of the high pH levels resulting from hydroxyl ions release.. Considering the importance of survival of apex surrounding cells (including eventually remaining apical papilla cells) after root canal disinfection prior to revascularization procedures, this study aimed to investigate the cytotoxicity of a modified triple antibiotic paste (mTAP) and formulations including ciprofloxacin, metronidazole and calcium hydroxide (CMC and modified CMC) on human cultured apical papilla cells under LTA-untreated or LTA-primed conditions. The null hypothesis is that neither medications (mTAP, CMC or mCMC) or cellular condition (LTA-untreated or LTA-primed) will affect the cellular viability

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