Browsing by Author "Braga, Gabriela Pacheco de Almeida"
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- Cytotoxic and antibiofilm properties of antibiotic-loaded thermoresponsive hydrogels for root canal therapyPublication . Duque, Cristiane; Braga, Gabriela Pacheco de Almeida; Carvalho, Juliana Machado de; Caiaffa, Karina Sampaio; Nunes, Gabriel Pereira; Rabelo, Rafaela Laruzo; Santos, Vanessa Rodrigues dos; Peres, Geórgia Rondó; Ribeiro, Lucas da Silva; Camargo, Emerson Rodrigues deDental infections can disrupt root development in immature permanent teeth, making traditional endodontic treatment challenging. Apexogenesis, a regenerative approach that promotes natural root development, offers a potential solution. However, issues related to disinfection and material biocompatibility still remain. The objective of this study was to evaluate the synergistic antimicrobial and antibiofilm properties of double and triple antibiotic combinations against common oral pathogens, and to incorporate the most effective combination into a thermosensitive hydrogel, to develop an alternative intracanal medication. Antibiotics were tested alone and in combination in planktonic and biofilm conditions of oral bacteria and Candida albicans. The antibiotic combinations with potential antimicrobial synergy were tested on Enterococcus faecalis biofilms in radicular dentin by confocal microscopy. Metronidazole (ME), ciprofloxacin (CI), and fosfomycin (FO) were incorporated into poly(N-vinylcaprolactam) (PNVCL) hydrogels, and their antibiofilm activity was compared to PNVCL hydrogels containing chlorhexidine (CHX) or calcium hydroxide (CH). The cytotoxicity of the hydrogels was assessed on MDPC-23 odontoblast-like cells using metiltetrazolium assays. A statistical analysis was performed using ANOVA followed by Tukey’s test (p < 0.05). The combination of ME + CI + FO showed superior antibiofilm effects in mono- and dual-species biofilms and on biofilms inside dentinal tubules, comparable to CHX. PNVCL hydrogels with ME + CI + FO significantly reduced E. faecalis biofilms in dentinal tubules, exhibiting a higher efficacy than PNVCL + CH. Cytotoxicity tests revealed minimal effects on cell viability for both PNVCL hydrogels with and without antibiotics. In conclusion, ME + CI + FO showed potent antimicrobial synergy and, when loaded in thermosensitive PNVCL hydrogel, demonstrated significant antibiofilm activity and low cytotoxicity. These findings emphasize the potential of this formulation as an effective and biocompatible endodontic medication, especially for the treatment of immature permanent teeth.
- EGCG and taxifolin modulate secretory activity and expression of dentinogenesis markers in odontoblast-like cellsPublication . Duque, Cristiane; Rabelo, Rafaela Laruzo; Braga, Gabriela Pacheco de Almeida; Soares, Igor Paulino Mendes; Souza, Maria Eduarda de; Chrisostomo, Daniela Alvim; Hebling, Josimeri; Costa, Carlos Alberto de Souza; Prakki, Anuradha; Regasini, Luís OctavioOdontoblasts are cells specialized in dentin matrix deposition and the first line of defense when the dentin–pulp complex is injured by pathological processes, such as dental caries and trauma. Natural compounds, such as flavonoids, could be useful to stimulate odontoblast activity and reparative dentinogenesis in vital pulp therapies, especially in immature permanent teeth. This study evaluated the effect of flavonoids on odontoblast secretory activity and the expression of dentinogenesis markers. The effect of flavonoids was evaluated on phenotypic mineralization markers (alkaline phosphatase (ALP) activity and mineralized nodule deposition) by colorimetric assays and on the expression of Alpl, Mmp2, Mmp9, Dmp1, and Dspp genes in odontoblast-like cells by quantitative polymerase chain reaction. Most of the flavonoids did not show toxicity between 100 and 25 μM. In distinct concentrations, epigallocatechin gallate (EGCG), taxifolin, myricetin, quercetin, and kaempferol stimulated the activity of ALP and increased mineralized nodule deposition. However, the highest effect on those phenotypic markers was observed after EGCG and taxifolin treatments. Then, they were selected for evaluation of gene expression. mRNA levels of Dmp1 and Dspp highly increased with taxifolin treatment, and Alpl expression was increased for both taxifolin and EGCG groups, without difference between them. Mmp2 and Mmp9 expression was not affected by these flavonoids. In conclusion, EGCG and taxifolin positively affect phenotypic mineralization markers; in particular, taxifolin highly stimulates early- and late-stage dentinogenesis genes.