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Browsing by Subject "Fusobacterium nucleatum"
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Item Effect of Curcumin-loaded Photoactivatable Polymeric Nanoparticle on peri-implantitis-related biofilm(2022) Tonon, Caroline Coradi; Panariello, Beatriz; Chorilli, Marlus; Spolidorio, Denise Madalena Palomari; Duarte, Simone; Cariology, Operative Dentistry and Dental Public Health, School of DentistryCurcumin has been used as a photosensitizer (PS) for antimicrobial photodynamic chemotherapy (PACT). However, its low solubility, instability and poor bioavailability are a challenge for its in vivo application. This study aimed to synthesize curcumin-loaded polymeric nanoparticles (curcumin-NP) and to determine their antimicrobial and cytotoxic effects. Nanoparticles (NP) were synthesized by the nanoprecipitation method using polyprolactone as a polymer. Curcumin-NP was characterized by particle size, polydispersity index and zeta potential, scanning electron microscopy and curcumin encapsulation efficiency (EE). Curcumin-NP was compared to free curcumin solubilized in 10% DMSO as photosensitizers for PACT in single and multi-species Porphyromonas gingivalis, Fusobacterium nucleatum and Streptococcus oralis biofilms. Chlorhexidine 0.12% (CHX) and ultrapure water were used as positive and negative controls, respectively. The cytotoxic effect of curcumin-NP was evaluated on human periodontal ligament fibroblast cells (HPLF). Data were analyzed by ANOVA (α=0.05). Curcumin-NP exhibited homogeneity and stability in solution, small particle size and 67.5% EE of curcumin. Curcumin-NP presented antibiofilm activity at 500 µg/ml when photoactivated. Curcumin-NP and curcumin with and without photoactivation were not cytotoxic to HPLF cells. Curcumin-NP has antimicrobial and antibiofilm properties, with better effects when associated with blue-light, being a promising therapy for preventing and treating peri-implant diseases.Item Effect of epigallocatechin gallate on nicotinetreated Fusobacterium nucleatum biofilm(Office of the Vice Chancellor for Research, 2016-04-08) Kaur, Mandeep; Dhami, Amarjeet; Gregory, Richard L.Abstract Tea polyphenols such as epigallocatechin gallate (EGCG) have exhibited antimicrobial properties. Fusobacterium nucleatum is an oral bacterium that is associated with periodontal diseases. Biofilm adheres to the enamel surfaces of our teeth as plaque. Biofilm formation in the oral cavity leads to many complications such as caries and periodontal diseases. Those who smoke tend to have increased risk of periodontal diseases and F. nucleatum biofilm formation. The objective of this research was to determine the effects of EGCG (0.25 mg/ml) and varying concentrations of nicotine (0-32 mg/ml) on F. nucleatum biofilm. The study was conducted by treating F. nucleatum biofilm with various concentrations of nicotine (0-32 mg/ml) and EGCG. Biofilm formation was measured using a crystal violet dye staining assay and a spectrophotometer. Biofilm formation of F. nucleatum with EGCG and nicotine exhibited a significant decrease in biofilm formation at low concentrations of nicotine (0-4 mg/ml). EGCG alone without nicotine significantly reduces F. nucleatum biofilm formation. EGCG can be added to dental treatments such as toothpaste and mouthwash for those who smoke. Periodontal diseases lead to many health problems in other parts of the body, therefore it is important to find ways to decrease biofilm formation of F. nucleatum.Item Nicotine-Treated Fusobacterium nucleatum Binding to Collagen, Fibrinogen, and Fibronectin(Office of the Vice Chancellor for Research, 2016-04-08) Beshay, Y.S.; Gregory, R.L.Fusobacterium nucleatum, a gram-negative anaerobic bacterium found in dental plaque, causes periodontal diseases. Smoking is one of the risk factors that can increase periodontal problems and atherosclerosis. Atherosclerosis is initiated by oral bacteria (i.e., F. nucleatum) binding to surface proteins of endothelial cells, such as collagen, fibrinogen, and fibronectin. The main objective for this study was to test the binding of F. nucleatum to collagen, fibrinogen, and fibronectin under the effect of different concentrations of nicotine. F. nucleatum was grown overnight in brain-heart infusion (BHI) supplemented with yeast extract and 5% vitamin-K/hemin. Biofilm was grown for 48 hours in 0, 0.25, 0.5, 1, and 2 mg/mL of nicotine. Then, the biofilm cells were labeled with biotin 3-sulfo-N-hydroxy-succinimide ester sodium salt and fixed with 10% formaldehyde. A binding assay was conducted by coating a high-binding 96-well microtiter plate with 1 μg/mL of collagen, fibrinogen, or fibronectin. The plate was incubated overnight and blocked with 1% Bovine Serum Albumin (BSA), followed by the biotinylated and nicotine-treated F. nucleatum cells. ExtrAvidin-Peroxidase and OPD Peroxidase Substrate was used to visualize the binding. Optical density (OD) was measured with a spectrophotometer at 490 nm. Collagen, fibrinogen, and fibronectin binding assays demonstrated significantly higher absorbance with 2 mg/mL nicotine-treated F. nucleatum cells compared to untreated cells. The results indicated that an increase in nicotine concentration leads to an increase in F. nucleatum binding to collagen, fibrinogen, and fibronectin. This means that smokers may have an increased risk for atherosclerosis. Supported by Life-Health and Sciences Internship (LHSI).