Browsing by Author "Duque, Cristiane"
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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.
- Exploring salivary biomarkers in pediatric obesity: a scoping reviewPublication . Sabella, Fernanda Maria; Katzenelson, Renata Thomaz; Carvalho, Fabíola Galbiatti de; Duque, Cristiane; Darrieux, Michelle; Marson, Fernando Augusto Lima; Parisotto, Thaís ManzanoChildhood obesity and overweight are linked to subclinical inflammatory conditions. The present manuscript aimed to undertake a scoping review exploring the relationship between childhood obesity and salivary biomarkers to answer the following question: “Are salivary biomarkers trustful factors/indicators for childhood obesity?” The main search terms used were: “obesity and salivary biomarkers and children” (Pubmed, Scielo, Scopus, Embase databases: 1999–2025). Assessed articles were carefully classified according to a predetermined criterion (Newcastle–Ottawa Scale), and the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) were considered. Papers involving children >13 years, duplicates/triplicates, literature reviews, and non-related to the question addressed were excluded. More than 30 salivary biomarkers were assessed in the thirteen studies appraised. Three studies were rated as having a high level of evidence, two as moderate, and eight as having a low level. Fourteen biomarkers were found to be significantly increased in childhood obesity/overweight (p < 0.05): leptin, insulin, α-amylase, tumor necrosis factor α, interleukin 6, vascular endothelial growth factor-A, C-reactive protein, monocyte chemotactic protein-1, resistin, phosphate, nitric oxide, interleukin 1β, uric acid and fetuin-A; and three were found to be significantly decreased (p < 0.05): adiponectin, secretory immunoglobulin A, and interleukin-12p70. In conclusion, the present review supported the idea that saliva might be a promising diagnostic tool in early life and that it is a significant source of obesity biomarkers in children.
- Methacrylated epigallocatechin gallate functionalized dental adhesives: antiproteolytic activity and dentin bonding studiesPublication . Duque, Cristiane; Chrisostomo, Daniela Alvim; Scaffa, Polliana Mendes Candia; Gouveia, Zach; Nascimento, Fabio Dupart; Plotnikov, Sergey V.; Prakki, AnuradhaObjectives: To assess the antiproteolytic effect of EGCG-methacrylate monomers and its inhibitory effect on gelatinolytic activity in the hybrid layer. Also, to investigate the effect of an adhesive material functionalized with EGCG-methacrylate monomers on immediate and long-term dentin-resin bond strength. Methods: Neat EGCG (E0) was reacted with three different ratios of methacryloyl ester and dissolved in ethyl acetate to obtain EGCG-methacrylates with hydroxyl functionalization at 33 % (M-E33), 67 % (M-E67) and 100 % (M-E100) levels. Resin composite blocks were built on human dentin surfaces using self-etching adhesive containing E0, M-E33, M-E67, and M-E100 at 1 wt%. Demineralized human dentin disks were immersed in deionized water (DW) or lactic acid (LA) and subsequently treated with DW, acetone (as controls), E0, M-E33, M-E67 and M-E100 diluted in acetone. Concentrations of solubilized type I collagen C-terminal (CTX and ICTP) and N-terminal (NTX) telopeptides were determined from 7-day extracts of dentin matrix specimens by ELISA assays. In situ zymography of adhesive-dentin interface slices from restored teeth was performed by confocal microscope after 24 h dentin treatment. Microtensile bond strength (µTBS) and failure pattern were evaluated after 24 h and 6 months. Data were analyzed using two-way ANOVA and Tukey post hoc test (p < 0.05). Results: All experimental groups statistically reduced the release of solubilized telopeptides from dentin samples in DW and LA. E0 and M-E100 incorporated into the adhesive system reduced the gelatinolytic activity within the hybrid layer. The lowest µTBS values for restored teeth were observed for E0 and M-E100 groups, after 24 h and 6 months, respectively. The most prevalent failure observed was classified as type 4, except for M-E100. Significance: EGCG-methacrylate monomers effectively protected collagen from degradation. When incorporated into adhesive systems, EGCG-methacrylates reduced gelatinolytic activity within the hybrid layer, and did not affect immediate and long-term bond strength values of restorations.
- Morin-loaded chitosan-poloxamer hydrogel as an osteoinductive delivery system for endodontic applicationsPublication . Pereira, Jesse Augusto; Martin, Victor; Araújo, Rita; Grenho, Liliana; Gomes, Pedro; Marto, Joana; Fernandes, Maria Helena; Santos, Catarina; Duque, CristianeConsidering the search for new biocompatible intracanal medicaments that can preserve remaining cells and stimulate bone tissue repair in the periapical region, this study aimed to synthesize and characterize the physicochemical properties of morin-loaded chitosan-poloxamer hydrogel (MCP) as well as to evaluate its osteogenic potential. Morin hydrate (M) was loaded into chitosan-poloxamer (CP) hydrogel and the resulting particles were characterized by infrared spectroscopy (FTIR), UV–vis spectrophotometer and scanning electron microscopy. Biological assays evaluated the metabolic activity, cell morphology and alkaline phosphatase (ALP) activity of human bone marrow stem cells (HBMSC) in three different settings, such as the exposure to dissolved morin, hydrogel's leachates and assembled particles by indirect contact. Cells cultured in standard culture conditions were used as control. The effect of CP and MCP particles on the formation of collagenous and mineralized tissues was also assessed within the organotypic model of segmented embryonic chick femora. Datasets were assessed for one-way analysis of variance (ANOVA), followed by Tukey's post hoc test (p < 0.05). Morin at 50 μg/mL was cytocompatible and increased ALP activity. CP and MCP particles showed stability, and morin was entrapped in the hydrogel matrix without changing its chemical structure. Cultures treated with 30-min CP and MCP hydrogel leachates presented significantly higher metabolic activity compared to control. By indirect contact, CP particles increased metabolic activity, but only MCP particles induced an upregulation of ALP activity in comparison to control. The amount of collagenous tissue and mineralized area on the fractured embryonic chick femora was greater in MCP particles compared to CP counterparts. Chitosan-poloxamer platforms are suitable systems to delivery morin, enhancing cell proliferation and bone mineralization, which upholds its application as intracanal medication for endodontic purposes.
- Root canal instrumentation of artificial primary teeth with rotary and reciprocating files: a micro-CT analysisPublication . Chrisostomo, Daniela Alvim; Danelon, Marcelle; Furlan, Renan Diego; Duarte, Marco Antônio Hungaro; Mello-Moura, Anna Carolina Volpi; Duque, CristianeThis study compared the outcomes of two endodontic instrumentation protocols (rotary or reciprocating files), using artificial primary teeth and micro-computed tomography. Twenty-four artificial primary molars were equally distributed into two groups of 12, according to the type of instrumentation – rotary files (Sequence Baby NiTi Files©) or reciprocating files (X1-Blue File NiTi files©). The following parameters were evaluated: root canal and dentin volumes, canal transportation and centering ability, risk of root perforation, and time of instrumentation. Statistically significant differences between the two instrumentation protocols considering root canal transportation and centering ability were identified. There was no difference in dentin thickness, fractures, and cracks comparing preoperative and postoperative time points for both endodontic files. Although the time of instrumentation was shorter for reciprocating files, rotary files promoted smaller root canal enlargement. Instrumentation with reciprocating and rotary files proved generally safe for pulpectomy in primary molars, promoting a negligible reduction in dentin volume, canal transportation, and centering ability, thereby preserving dentin thickness and lowering the risk of fractures.