Bbl gaspak jar system

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.

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)).