Biofilm Forming Bacteria and the Role of Human Ligands in this Process
University of Wales Institute Cardiff
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Wound healing is a complex process with many probable factors that can delay healing; wound infection, especially biofilm infection, is one of these delaying factors. The wound is colonized by bacteria and the bacteria begin to gain use from the wound and damage the wound healing process. Streptococci, Staphylococci and Pseudomonas are the most common bacteria associated with wound infection. Thus, in this study, the ability of three organisms, Streptococcus pyogenes, Pseudomonas aeruginosa and Staphylococcus aureus, to form biofilm and bind human ligands were examined by coating microtitre wells with fibrinogen or collagen. The capacity of these bacteria to form biofilm and bind human proteins was measured and finally the results were compared. Solid-phase protein binding assays were done in this research using 96 well microtitre plates. The coated wells were inoculated with mono-species bacteriaand incubated for one hour. The biofilms were fixed and stained in the microtitre wells and the results were obtained using a well microtitre plate reader. Results show that fibrinogen can bind to Streptococcus pyogenes biofilm strongly at all concentrations with the exception of 100ng concentration that appears weakly binding. Also fibrinogen can bind to Staphylococcus aureus biofilm at ≥ 400ng, whereas collagen can bind strongly at all concentration with S. pyogenes biofilm and at concentration ≥700ng collagen Staphylococus aureus biofilm. None of the two ligands bind to Pseudomonas aeruginosa at any concentration. This finding indicates that neither fibrinogen nor collagen is suitable to grow all types of biofilm in vitro. It is very important for further studies to provide information about biofilm-hosts by finding the appropriate ligand to grow most biofilm in vitro which will help in more understanding of bacterial biofilms and their role in chronic wound pathogens.
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