Cytotoxicity of intracanal dressings on apical papilla cells differ upon activation with E. faecalis LTA

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 » 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.

Effect of irrigation technique for removal of triple antibiotic paste on bond strength of MTA to root dentin

Abstract This study evaluated the bond strength of mineral trioxide aggregate (MTA) to root canal dentin after the performance of various irrigation procedures to remove triple antibiotic paste (TAP). A total of 56 single-rooted human mandibular premolars were instrumented using a rotary system to size 40 and divided randomly into a control group (no intracanal dressing) and three experimental groups (TAP application for 28 days). TAP was then removed by rinsing with 10 mL 2.5% NaOCl using three irrigation systems (Vibringe sonic irrigation, CanalBrush, and syringe irrigation). The coronal and middle parts of root canals were then obturated with MTA. After storage for 1 week, each specimen was embedded in an acrylic block and sectioned horizontally (2-mm-thick slices) at two levels (coronal and middle). Bond strength of MTA to root canal dentin was assessed in 28 samples per group via push-out test using a universal testing machine. Data from the four groups were compared using one-way analysis of variance. Tukey's test was used for multiple comparisons. Push-out bond strength values were significantly higher in the control and Vibringe groups than in the CanalBrush and syringe irrigation groups (p < 0.001). TAP removal from root canals with the Vibringe irrigation system may increase the push-out bond strength of MTA compared with the use of the CanalBrush or syringe irrigation.

[Studying the accelerated photostability of ciprofloxacin and lomefloxacin in tablets and eye drops]

In this work the photostability of two fluoroquinolones: ciprofloxacin and lomefloxacin has been detected in tables and eye drops formulations using light-stability cabinet. Ciprofloxacin and lomefloxacin were subjected to stress conditions. The degradation products were well separated from the peak of the active substance. The stability of these compounds has been studied both in containers and under direct light in the light-stability cabinet. Samples were assayed immediately and at 1,3,6 months by stability-indicating high performance liquid chromatography methods with photodiode array detector. The determination was performed on C18 [250 x4.6mm, 6 micro m]. The first mobile phase consisted of 0.025M phosphoric acid and acetonitrile [87:13] pumped at a flow rate 2ml/min for ciprofloxacin, while the second consisted of water, acetonitrile, triethylamine [80:20:0.3] pumped at a flow rate 1ml/min for lomefloxacin. The UV detector was operated at 278 nm for ciprofloxacin and 288 nm for lomefloxacin. The methods was suitably validated for linearity, accuracy, precision, robustness and selectivity. All validation parameters were within the acceptance range. Data analysis revealed that plastic and amber containers can not protect either ciprofloxacin or lomefloxacin in eye drops formulation from photodegradation after one month in the cabinet. White blister protect the two agents in tablets formulation after six months in the cabinet

Interaction of nalidixic acid and ciprofloxacin with wild type and mutated quinolone-resistance-determining region of DNA gyrase A.

The quinolones exert their anti-bacterial activity by binding to DNA gyrase A (GyrA), an essential enzyme in maintenance of DNA topology within bacterial cell. The mutations conferring resistance to quinolones arise within the quinolone-resistance-determining region (QRDR) of GyrA. Therefore, quinolones interaction with wild and mutated GyrA can provide the molecular explanation for resistance. Resistant strains of Salmonella enterica of our hospital have shown mutations in the QRDR of GyrA of serine 83 (to phenylalanine or tyrosine) or aspartic acid 87 (to glycine or tyrosine). In order to understand the association between observed resistance and structural alterations of GyrA with respect to quinolone binding, we have studied the interaction of mutated QRDR of GyrA with nalidixic acid and ciprofloxacin by molecular modeling using GLIDE v4. Analysis of interaction parameters like G-score has revealed reduced interaction between nalidixic acid/ciprofloxacin with QRDR of GyrA in all four mutated cases of resistant strains. The mutation of Ser83 to Phe or Tyr shows least binding for nalidixic acid, while Asp87 to Gly or Tyr exhibits minimal binding for ciprofloxacin. The study also highlights the important role of arginines at 21, 91 and His at 45, which form strong hydrogen bonds (at < 3 Å) with quinolones. The hydrophilic OH group of Serine 83, which is in close proximity to the quinolone binding site is replaced by aromatic moieties of Tyr or Phe in mutated GyrA. This replacement leads to steric hindrance for quinolone binding. Therefore, quinolone resistance developed by Salmonella appears to be due to the decreased selectivity and affinity of nalidixic acid/ciprofloxacin to QRDR of GyrA.