Oliveira AP. Influence of intracanal medications used in regenerative endodontic procedures on survival of apical papilla cells in vitro [thesis]. São Paulo: Universidade de São Paulo, Faculdade de Odontologia; 2014. Versão Corrigida.Regenerative Endodontic procedures have provided changes in treatment of patients with immature teeth and apical periodontitis enabling full root development and lower incidence of dental fracture. Intracanal dressings are used for disinfection; however, their effect on apical papilla cells is poorly elucidated. Additionally, the effect of these substances on cells previously subjected to proinflammatory condition is still unknown. The aim of this study was to evaluate the cytotoxicity of intracanal dressings used in Endodontics regenerative procedures on cultured human apical papilla cells at physiologic and activated condition. Cell culture was established from the apical papilla removed from an extracted immature third molar. The substances studied were triple antibiotic modified paste: ciprofloxacin, metronidazole and cephalosporin (1:1:1); CFC: ciprofloxacin, metronidazole and calcium hydroxide (1:1:2) and modified CFC: ciprofloxacin , metronidazole and calcium hydroxide (2:2:1). Part of the cells was stimulated previously with lipotheichoic acid (LTA) of Enterococcus faecalis por 7 days. Once plated, cells were exposed to increasing concentration of the medications for 1, 3, 5 and 7 days. Cell viability was evaluated by means of 3-bromide (4.5-dimetiliazol-2-yl) -2.5-difeniltetraze (MTT) and Nitric Oxide (NO) release was assessed by the Griess method. The statistical analysis was done through analysis of variance with 1 criteria (ANOVA) followed by Tukey test with 5% of significance level. Modified CFC was the medication that demonstrated the less cytotoxic effect on cell viabilityat the experimental periods studied while CFC promoted significant decrease on cell viability specially after 7 days of contact. The modified triple antibiotic paste resulted in important alteration of cell viability being considered the most citotoxic. Cellular activation by LTA resulted in increased levels of mitochondrial activity for all medications being more evident at the longer experimental periods. Cellular activation also contributed to higher levels of nitric oxide release. In conclusion, the cytotoxic effect of the tested medications is dependent on concentration, time of contact and cellular condition, being the triple antibiotic modified paste the most cytotoxic in high concentrations leading clinically in the decreased of the cells viability of the apical papilla, decreasing the success of regenerative procedures.
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.
RESUMOSuplementação com óleos vegetais tem sido usada frequentemente na dieta de equinos atletas. O objetivo deste trabalho foi avaliar o metabolismo lipídico em testes de longa duração em equinos suplementados com óleo de soja ou óleo de arroz. Doze equinos da raça Árabe, não treinados, foram distribuídos em dois grupos, e cada grupo foi suplementado com óleo de soja ou de arroz por um período de seis semanas. Antes e após esse período, esses animais foram submetidos a um teste de longa duração (TLD) em esteira, quando foram coletadas amostras de sangue antes, durante e após o exercício. No TLD houve aumento nos valores séricos de ácidos graxos livres (AGL) e redução nos valores de triglicérides nos animais após suplementação nos dois grupos (P<0,05). Em ambos os grupos não houve alterações significativas nos valores de HDL no TLD após a suplementação. Nos animais após suplementação com óleo houve aumento da disponibilidade de energia no exercício de longa duração por meio do aumento de AGL e da manutenção dos níveis de lactato. As alterações no metabolismo lipídico causadas pela suplementação com óleo vegetal podem favorecer o desempenho desses animais em testes de longa duração. sido observados em animais que realizam exercícios de resistência, para os quais a utilização de ácidos graxos para obtenção de ATP é muito importante, pois promove economia de glicogênio e gera uma menor quantidade de calor, melhorando, assim, o rendimento (González, 2007).
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