Good afternoon colleagues, I am testing the antimicrobial effect of a new Peptide (Lye Tx 1) against Periodontal Pathogenic Bacteria in Biofilms. To create biofilms, I am using the Calgary Device. Reading the paper of Harrison et al. 2010, the protocol suggest that pegs may need to be coated with some agent in order to promote bacterial adhesion.
Do you know whether is necessary coating the pegs when growing periodontal pathogenic biofilms in the Calgary Device?
Thank you very much.
Edison - not necessarily. I haven't worked with oral flora, but have experience with staphylococci and P. aeruginosa on this platform. In general, healthy well-suspended organisms will tend to adhere to the polystyrene or polycarbonate that the pin plate is typically made of. There is a difference between simple adhesion and actual biofilm formation, so adherent organisms will need some time to develop full biofilms. A couple of important points to consider are these:
1) The plankonic organisms not adhering to your pins will generally grow much faster than biofilm incorporate pins. This will tend to exhaust nutrients and oxygen in your wells. After a brief exposure to allow adherence, we replaced the media to allow the pin-adherent organisms to have a chance to compete. We 're-fed' plates that way every 12-24 hours to be sure that nonadherent organisms weren't swamping the system.
2) Unlike 'real world' biofilms, especially such as those related to dental enamel, there is no 'formal' adhesion between the biofilm and the pin. This means, at least for the first couple of days, that you'll need to be very gentle when removing and replacing the pins to avoid accidental sloughing of the biofilm matrix.
3) A method we considered, but never employed, was roughening the surface with a very fine sand paper to produce an irregular surface to allow easy adhesion and persistent growth on the pin.
4) Lastly, remember that in a typical 96-well type chamber with actively metabolizing cells, the partial pressure of oxygen falls sharply as one descends through the depth of the well. As a result, in your pin system the biofilm will probably be the thickest and most developed right at the fluid meniscus. Look for your effect there, not deep in the well at the tip of the pin.
Good luck! John
John, your answer is more than I expected, I really appreciate it. Can I get your email to be in contact with you? because this is my first time I´m working with this device and is a real challenge to me, and I am very exited with my research. Thanks again.
Edison - feel free to reach out at john 'at' akadeum.com
Have a great week. John
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