Jenway genova r0027

P. gingivalis strain ATCC 33277 was cultured in brain-heart-infusion and in broth-heart-brain extract (BHI; BD Bioxon, Milan, Italy) containing 5 µg/mL of hemin (Sigma-Aldrich, Munich, Germany) and 1 µg/mL of menadione (Sigma-Aldrich) under anaerobiosis using the anaerobic BBL-GasPak jar system (BD Biosciences) at 37 °C for 24 h.
After 24 h of culturing, bacteria were harvested by centrifugation for 10 min at 10,000 rpm and then washed and resuspended in Krebs-Ringer-Glucose (KRG) buffer (120 mM NaCl, 4.9 mM KCl, 1.2 mM MgSO₄, 1.7 mM KH₂PO₄, 8.3 mM Na₂HPO₄, 10 mM glucose, and 1.1 mM CaCl₂, pH 7.3). Bacterial growth was monitored spectrophotometrically (Jenway Genova R0027, Fischer Scientific, USA) at 675 nm. The bacterial density was visually adjusted to a turbidity of 0.5 McFarland (1 × 10⁸ colony-forming units; (CFU/mL) (Mc Farland, 1907 (link); Emani, Gunjiganur & Mehta, 2014 (link)). Ethical approval was given by the Ethics Committee of the School of Medicine (UNAM) with reference number C54-11.

Enterococcus faecalis ATCC 29212 and S. mutans strain ATCC 25175 were used in all the experiments. S. mutans was cultured under microaerophilic conditions with a candle jar at 37°C overnight in trypticase soy broth (TSB; Oxoid, Milan, Italy). E. faecalis was cultured under facultative anaerobiosis in brain heart infusion (BHI; BD Bioxon, Milan, Italy). Bacterial growth was monitored spectrophotometrically (Jenway Genova R0027, Fischer Scientific US) at 600 nm.
Ethical Approval was given by the Ethics committee of the Medicine Faculty UNAM with the reference number C54‐11.

P. gingivalis strain ATCC 33277 was cultured as previously described (Blancas-Luciano et al., 2022 (link)). Briefly P. gingivalis was cultured in brain-heart-infusion (BHI; BD Biox-on, Milan, Italy) containing 5 mg/mL of hemin (Sigma-Aldrich, Munich, Germany) and 1 mg/mL of menadione (Sigma-Aldrich) under anaerobiosis using the anaerobic BBL-GasPak jar system (BD Biosciences), at 37 °C for 24 h.
After 24 h of culturing, bacteria were harvested by centrifugation for 10 min at 8,200 x g and then washed and resuspended in Krebs-Ringer-Glucose (KRG) buffer (120 mM NaCl, 4.9 mM KCl, 1.2 mM MgSO₄, 1.7 mM KH₂PO₄, 8.3 mM Na₂HPO₄, 10 mM glucose, and 1.1 mM CaCl₂, pH 7.3). Bacterial growth was monitored spectrophotometrically (Jenway Genova R0027; Thermo Fischer Scientific, Waltham, MA, USA) at 675 nm. The bacterial density was visually adjusted to a turbidity of 0.5 McFarland (1 × 10⁸ colony-forming units); (CFU/mL) (Bollela, Sato & Fonseca, 1999 (link); Emani, Gunjiganur & Mehta, 2014 (link)).