The effects of shear stress on isolated receptor-ligand interactions of Staphylococcus epidermidis and human plasma fibrinogen using molecularly patterned microfluidics.
Staphylococcus epidermidis is an opportunistic pathogen that has been implicated in hospital-acquired infections, specifically related to implanted intravascular devices. S. epidermidis adhesion is a mechanism of colonization, leading to pathogenesis. Here we demonstrate an easily fabricated and robust parallel microfluidic platform to investigate the physiologically-relevant effects of fluid shear on S. epidermidis adhesion to human fibrinogen (hFg) with increased experimental throughput. In situ molecular patterning using fluid flow boundaries allows for isolation of the molecular interactions in highly defined shear stress environments, while keeping the device operation simple and reproducible. We characterize two modes of attachment of S. epidermidis to hFg coated surfaces. Single colonies adhere in high fractions at low shear stresses (~1 dyne cm(-2)) and adhesion decays with increasing shear. However, clusters of bacteria adhere the highest at median wall shear stress (up to 10 dyne cm(-2)), and adhesion subsequently decays above this critical shear stress. This initial characterization suggests a previously unobserved phenomenon of shear activated cell-cell adhesion in S. epidermidis, which acts to increase the overall attachment strength to hFg. Both of these modes of attachment are dependant upon the presence of intact hFg, indicating that adhesion is resultant from specific molecular recognition between the bacteria and human fibrinogen. This platform provides new insight into complex host-pathogen interactions, and will allow for further investigation of colonization and pathogenesis in more physiologically relevant conditions.
Department of Bioengineering, University of California, Los Angeles, CA, USA.
Westbrook M
WM
Weaver
Shivani
S
Dharmaraja
Vladana
V
Milisavljevic
Dino
D
Di Carlo
D001422:Bacterial Adhesion; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D005340:Fibrinogen / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008024:Ligands; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D055616:Microtechnology; D013212:Staphylococcus epidermidis / Q000502:physiology*; D013314:Stress, Mechanical; D013499:Surface Properties
D004129:Dimethylpolysiloxanes; D008024:Ligands; C013830:baysilon; D005340:Fibrinogen