In vitro Antibiofilm Activity of Pomegranate (Punica granatum) Juice on Oral Pathogens

Abstract


 Introduction: Pomegranate (Punica granatum) fruit contains valuable ingredients, such as ellagitannins and flavonoids, that have many potential effects, including antibacterial, antifungal, and anti-inflammatory functions.
 
 Objectives: The aim of this study was to investigate the effects of pomegranate fruit juice on F. nucleatum and S. sanguinis monospecies and multispecies biofilm formation in vitro.
 
 Methods: Pomegranate juice was obtained using a juicer and diluted using a brain heart infusion (BHI) broth into five different concentrations. The biofilm assay was performed as follows: F. nucleatum and S. sanguinis were cultured separately in the BHI broth for 48 hours at 37°C in an anaerobic atmosphere. A 200 mL bacterial suspension (107 CFU/mL) was distributed into a 96-well plate and incubated for 24 hours to form a biofilm. Subsequently, pomegranate juice was added to the biofilm well and observed after 1 hours, 3 hours, 6 hours, and 24 hours. The biofilm mass was measured using a microplate reader (490 nm) after crystal violet staining. Chlorhexidine (0.2%) and the biofilms without treatment were used as the positive and negative controls, respectively. The data were statistically analyzed using one-way analysis of variance, with p<0.05 as the level of significance.
 
 Result: There was a significant biofilm reduction after treatment with pomegranate juice for all the concentrations and incubation times (p<0.05). The effective concentrations to inhibit the biofilm monospecies F. nucleatum and S. sanguinis and the multispecies were 6.25% (OD 0.148±0.019), 50% (OD 0.211±0.026), and 6.25% (OD 0.024±0.209), respectively.
 
 Conclusion: Pomegranate juice inhibits F. nucleatum and S. sanguinis biofilm formation as a monospecies and a multispecies. Future studies are needed to observe the mechanism of this active substance.

Anti-Biofilm Forming Activity of Natural Products Extract Punica Granatum L. and Magnifera Indica L

“…The dry extract obtained from the solvent was stored at -20°C for further use in various tests. 16 96 well microtiter plate method; Isolates from newly cultivated plates were then subcultured in Tryptone soy broth (TSB) containing 1% glucose and incubated for 24 hours at 37 C before diluting (1:100) with bacterial suspension. Individual wells of sterilized, polystyrene, flat-bottom tissue cultivation plates were subsequently loaded with 0.2 ml aliquots of the diluted cultures, with broth in well no 12, serving as a control to assure sterility and non-specic medium adherence.…”

“…13 Antimicrobial activity of Punica granatum L, peel, seeds and juice has been established against Escherichia coli, H.pylori, Klebsiella, Listeria monocytogenes, Pseudomonas aeruginous, and other microorganisms. 14 So our study is conducted to evaluate the biolm formation by Escherichia coli and the antibiofilm forming activity of pomegranate peels Punica granatum L. and Mango leave Mangifera Indica L. extracts against Escherichia coli. There combined synergistic eect using 96 well microtiter plate.…”