Navegando por Autor "Miranda, Diego Garcia"
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Item Exploring Antioxidant, Antimicrobial and Anti-Inflammatory Effects of Juglans regia and Pfaffia paniculata Extracts: implications for Intestinal Dysbiosis and Colorectal Cancer Risk Associated with Oral Pathogens(MDPI) Miranda, Diego Garcia; Ramos, Lucas de Paula; Attik, Nina; Silva, Nicole Van Der Heijde Fernandes; Camargo, Pyetra Claro; Araujo, Gabriela Ferraz de; Lopes, Nicole Fernanda dos Santos; Marcucci, Maria Cristina; Soares, Cristina Pacheco; Godoi, Bruno Henrique; Caires, Giovanna Arruda; Vigerelli, Hugo; Carrouel, FlorenceColorectal neoplasms rank as the third most prevalent cancer globally and stand as the second leading cause of cancer-related mortality. Its etiol- ogy is multifaceted, pointing to the role of microorganisms within the human microbiota in its development. Notably, the high prevalence of oral pathogens like Fusobacterium nucleatum and Parvimonas micra is implicated in inducing gut dysbiosis and stimulating the proliferation and metastasis of cancer cells. Therefore, this study aimed to evaluate in vitro the biological effects of extracts from Juglans regia and Pfaffia paniculata. Methods: Phytochemical analysis was carried out by HPLC, and the antioxidant effect was deter- mined by DPPH. Antimicrobial activity was investigated on F. nucleatum and P. micra planktonic and biofilms. Metabolic activity and genotoxicity were performed. Results: J. regia and P. paniculata expressed CE50 37.26 and 1367.57 mcg, respectively. The extracts exhibited a minimum bactericidal concentration of 1.73 and 0.48 mg/mL for J. regia and P. paniculata, respectively. Reduction superiorly 90% of P. micra biofilms. Metabolic activity was varied proportionally to the extract concentration, and no genotoxic effects were ob- served. Conclusions: The J. regia extract has great antioxidant activity and could be used as an alternative in combating pathogens associated with the onset of dysbiosis and tumor progression in colorectal neoplasms. Nevertheless, further studies are needed to validate their clinical applicability.Item Gymnema sylvestre as a Potential Anti-Inflammatory and Anti-Biofilm Agent Against Anaerobic Infections: An In Vitro Study(MDPI) Miranda, Diego Garcia; Tome, Fernanda Malagutti; Miguel, Manuela Maria Viana; Liberato, Sabrina Ferreira dos Santos; Marcucci, Maria Cristina; Vigerelli, Hugo; Rodrigues, Flavia Pires; Soares, Cristina Pacheco; Godoi, Bruno Henrique; Carrouel, Florence; Oliveira, Luciane Dias de; Ramos, Lucas de PaulaThis study evaluates the antimicrobial activity of the glycolic extract of G. sylvestre against anaerobic pathogens, along with its cytotoxicity, genotoxicity, anti-inflammatory activity, antioxidant effects, and phytochemical composition. Phytochemical analysis was conducted using high-performance liquid chromatography and liquid chromatography– mass spectrometry, while the antioxidant effect was assessed through a DPPH assay. An- timicrobial action was tested on planktonic cultures and biofilms of Porphyromonas gingivalis, Porphyromonas endodontalis, Parvimonas micra, and Fusobacterium nucleatum. Cytotoxicity was evaluated using mouse macrophages (RAW 264.7), rat fibroblasts (L929), and human keratinocytes (HaCaT). Anti-inflammatory effects were measured by an immunoenzymatic assay (ELISA) on RAW 264.7 cells. Statistical analysis was performed using a one-way ANOVA and Tukey’s test. Phytochemical analysis revealed the presence of phenolic com- pounds and flavonoids. The extract demonstrated a reduction of over 95% in biofilms of P. gingivalis, P. micra, and F. nucleatum within 5 min of treatment. Cell viability (HaCaT) remained above 80%. Antioxidant activity showed an EC50 of 353.43 μg/mL, achieving a 50% reduction in free radicals. A significant decrease in TNF-α (a pro-inflammatory cytokine) and an increase in IL-10 (an anti-inflammatory cytokine) were observed. In conclusion, the extract of G. sylvestre exhibits promising potential as a therapeutic agent for treating anaerobic infections, inflammation, and oxidative stress.Item Juglans regia and Pfaffia paniculata extracts: implications for periodontal disease treatment and correlation with Alzheimer's risk(Frontiers) Miranda, Diego Garcia; Carrouel, Florence; Attik, Nina; Araujo, Gabriela Ferraz; Lopes, Nicole Fernanda dos Santos; Marcucci, Maria Cristina; Rodrigues, Flavia Pires; Caires, Giovanna Arruda; Vigerelli, Hugo; Godoi, Bruno Henrique; Soares, Cristina Pacheco; Ramos, Lucas de PaulaPeriodontal disease (PD) is a significant global health concern, affecting approximately 19% of the world’s population. It is one of the most prevalent diseases today, causing substantial socio-economic impacts and diminished quality of life. Recent research has also revealed a potential link between PD and Alzheimer’s disease. This study investigated the antimicrobial effects of Juglans regia and Pfaffia paniculata extracts against P. endodontalis and P. gingivalis, bacteria that cause PD and are related to Alzheimer’s risk. The study also assessed the impact of these extracts on macrophage metabolic activity, pro- and anti-inflammatory cytokine expression, and genotoxicity. The phytochemical analysis of the extract was carried out first. Antimicrobial activity was performed using the M11-A7 protocol (CLSI) for planktonic cultures on monotypic biofilms matured for 168 hours in anaerobiosis. Cell viability analysis was carried out using MTT on mouse macrophages (RAW 264-7), as well as genotoxicity assessment using micronuclei. The anti-inflammatory activity was evaluated using ELISA method, checking the cytokines IL-6, IL-1B, TNF-alpha, IL-17 and IL-10. Phytochemical analysis revealed the presence of Miquelianin, Regiolone and Gallic Acid in J. regia extract. For the P. paniculata extract, we identified the glycosides Pfaffoside C, Pfaffoside A, 3-O-β-D-glycopyranosyl-oleanolic acid and Beta-ecdysone. Antimicrobial activity revealed a MBC of 1.73 for the extract of J. regia and 0.48 for P. paniculata against P. endodontalis and P. gingivalis. All biofilms were reduced by more than 89% after treatment with the extracts for 5 min. Cytotoxicity evaluations revealed that cell viability remained above 50% at concentrations up to 0.216 mg/ml for J. regia and 0.015 mg/ml for P. paniculata. Neither extract exhibited genotoxicity. Furthermore, both demonstrated anti-inflammatory activity by promoting the production of the cytokine IL-10. In conclusion, the antimicrobial and anti-inflammatory activities of J. regia and P. paniculata extracts suggest their potential as treatments for oral dysbiosis, which may contribute to a reduced risk of neurodegenerative diseases.Item Ketoprofen Associated with Hyaluronic Acid Hydrogel for Temporomandibular Disorder Treatment: An In Vitro Study(MDPI) Miranda, Diego Garcia; Ramos, Lucas de Paula; Lopes, Nicole Fernanda dos Santos; Silva, Nicole Van Der Heijde Fernandes; Soares, Cristina Pacheco; Rodrigues, Flavia Pires; Morais, Vinicius de Paula; Sani-Taiariol, Thalita; Baldan, Mauricio Ribeiro; Vasconcellos, Luana Marotta Reis de; Borges, Alexandre Luiz Souto; Grosgogeat, Brigitte; Gritsch, KerstinTemporomandibular disorders (TMD) are a public health problem that affects around 12% of the global population. The treatment is based on analgesics, non-steroidal anti-inflammatory, corticosteroids, anticonvulsants, or arthrocentesis associated with hyaluronic acid-based viscosup- plementation. However, the use of hyaluronic acid alone in viscosupplementation does not seem to be enough to regulate the intra-articular inflammatory process. So, we propose to develop and evaluate the physicochemical and biological properties in vitro of hyaluronic acid hydrogels (HA) associated with ketoprofen (KET) as a new therapeutic treatment for TMD. The hydrogels were synthesized with 3% HA and 0.125, 0.250, 0.500, or 1% KET. Physicochemical analyses of Attenu- ated Total reflectance-Fourier transform infrared spectroscopy (FTIR), Thermogravimetry (TGA), Rheology by Frequency, Amplitude sweeps, temperature ramp, and scanning electron microscopy (SEM) were performed with or without sterilization and cycled. Cytocompatibility and genotoxicity (micronucleus assay) were performed in mouse macrophages (RAW 264-7) for 24 h. Results: FTIR spectrum showed characteristic absorptions of HA and KET. In the TGA, two mass loss peaks were observed, the first representing the water evaporation at 30 and 100 ◦C, and the second peaks be- tween 200 and 300 ◦C, indicating the degradation of HA and KET. Rheology tests in the oscillatory regime classified the hydrogels as non-Newtonian fluids, time-dependent, and thixotropic. Mouse macrophages (RAW 264-7) presented viability of 83.6% for HA, 50.7% for KET, and 92.4%, 66.1%, 65.3%, and 87.7% for hydrogels, in addition to the absence of genotoxicity. Conclusions: Hyaluronic acid associated with ketoprofen shows satisfactory physicochemical and biological properties for use as viscosupplementation. As a limiting point of this study, further research is needed to evaluate the pharmacodynamic, toxicological, and pharmacokinetic characteristics of a complete organism.