Development of in vitro Biofilm Model: Artificial Food Supplementation in Chemostat-type System

Characterisation of a sucrose‐independent in vitro biofilm model of supragingival plaque

“…To generate sucrose‐independent and sucrose‐dependent biofilms in 24‐well plates, we used the artificial saliva medium (basal medium mucin: BMM), modified from Monoi, Ohta, Morishima, and Ochiai (), containing 6.25 g/L mucin from porcine stomach (Sigma‐Aldrich Japan), 5 g/L proteose peptone (Becton Dickinson), 2.5 g/L tryptone (Becton Dickinson), 2.5 g/L yeast extract, 1.25 g/L potassium chloride (Wako Pure Chemical Industries, Ltd., Osaka, Japan), 0.25 g/L cysteine hydrochloride (Wako), 5 mg/L hemin, 1 mg/L menadione and 0.05% glucose (Wako) adjusted to pH 7.6, with or without 1% sucrose (Wako).…”

“…The harvested biofilms were disrupted by sonication (3 s, 200 μA, US‐150) using a probe sonicator (Nihon Seiki Kaisha Ltd, Tokyo, Japan) in 4 ml PBS. The biofilm suspension was then serially diluted and plated on blood agar for enumeration of A. naeslundii , on Mitis Salivarius agar (Becton Dickinson) for S. gordonii and S. mutans and on CVE agar (Monoi et al., ) for F. nucleatum and V. parvula . Blood agar plates were incubated aerobically at 37°C, while Mitis Salivarius agar and CVE agar plates were incubated anaerobically at 37°C.…”

Butyrate as a Potential Driver of a Dysbiotic Shift of the Tongue Microbiota

“…The collected tongue microbiota samples were inoculated into 2.5 mL BMM, which enables the cultivation of a wide variety of indigenous oral taxa ( 25 , 26 ). BMM (pH 7.5) contains 2.5 g/L porcine gastric mucin (type III; Sigma Chemical, St. Louis, MO), 2.0 g/L proteose peptone (BBL, Becton, Dickinson, Sparks, MD), 1.0 g/L Trypticase peptone (Becton, Dickinson), 1.0 g/L yeast extract (Becton, Dickinson), 0.5 g/L potassium chloride (Wako, Osaka, Japan), 0.1 g/L cysteine hydrochloride (Wako), 0.001 g/L hemin (Sigma Chemical), and 0.0002 g/L menadione (Sigma Chemical).…”