To determine the minimum inhibitory concentration (MIC), mTSB was inoculated with P. gingivalis and adjusted to OD600 0.08 on a spectrophotometer. Then, 100 µL of the suspension was distributed in a 96-well microplate (Thermo Scientific, Nunc MicroWell Plates, 163320, Roskilde, Zealand, Denmark). Following this, 8 mg/mL 1.5 MDa sodium hyaluronate (Lifecore Biomedical, HA15 M-5, Chaska, MN, USA) were two-fold serially diluted and incubated with the suspension. Additionally, 50 µg/mL azithromycin dihydrate (AK Scientific, G333, Union City, CA, USA) and 2 mg/mL Chlorhexidine Gluconate (Avalon Pharma, PS-2035, Riyadh, Saudi Arabia) were also two-fold serially diluted and included to serve as positive controls. Sodium chloride (9%) was used as a solvent for all three substances. The microwell plate was incubated as previously described for 24 h. Following incubation, 5 µL drops of each well were placed onto Brucella agar plates supplemented with hemin and vitamin K. The plates were incubated for 5 days to assess the growth visually from each concentration. The lowest concentration that had no or only minuscule growth was considered MIC. The procedures were conducted in triplicates.
Azithromycin dihydrate
Manufactured by AK Scientific
Sourced in United States
Azithromycin dihydrate is a chemical compound that belongs to the macrolide class of antibiotics. It is the dihydrate form of azithromycin, a widely used antibiotic medication. Azithromycin dihydrate is often used as a pharmaceutical ingredient in the production of various drug formulations.
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2 protocols using azithromycin dihydrate
1
Determining P. gingivalis MIC Using Hyaluronate
2
Microdilution Assay for P. gingivalis Susceptibility
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In a microdilution method (Standards, 2004 ), mTSB broth was inoculated with P. gingivalis from a previous culture and adjusted to an OD600 of 0.08. The suspension was then diluted and distributed in a microplate with 96 wells (Thermo Scientific, Nunc MicroWell Plates, 163320, Roskilde, Zealand, Denmark). Then, 1.5 MDa sodium hyaluronate (Lifecore Biomedical, HA15 M−5, Chaska, MN, United States), azithromycin dihydrate (AK Scientific, G333, Union City, CA, United States), and chlorhexidine gluconate (Avalon Pharma, PS-2035, Riyadh, Saudi Arabia) were serially diluted and added. The plate was incubated for 24 h as described previously. After incubation, 5 µL of each well was spread onto plates of Brucella agar. They were incubated for five days to visually evaluate growth at each concentration. MIC was defined as the lowest concentration at which no or minimal growth occurred.
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