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Applied and Environmental Microbiology, February 2007, p. 699-705, Vol. 73, No. 3
0099-2240/07/$08.00+0     doi:10.1128/AEM.02428-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Novel Quantitative Biosystem for Modeling Physiological Fluid Shear Stress on Cells{triangledown}

Eric A. Nauman,1,{dagger} C. Mark Ott,2,{dagger} Ed Sander,1 Don L. Tucker,3 Duane Pierson,2 James W. Wilson,4,{ddagger} and Cheryl A. Nickerson4,{ddagger}*

School of Mechanical Engineering, Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana 47907-2088,1 Habitability and Environmental Factors Division, NASA-Johnson Space Center, Houston, Texas 77058,2 University Space Research Association, Division of Space Life Science, Houston, Texas 77058,3 Department of Microbiology and Immunology, Program in Molecular Pathogenesis and Immunity, Tulane Center of Excellence in Bioengineering, Tulane University Health Sciences Center, New Orleans, Louisiana 701124

Received 16 October 2006/ Accepted 22 November 2006

The response of microbes to changes in the mechanical force of fluid shear has important implications for pathogens, which experience wide fluctuations in fluid shear in vivo during infection. However, the majority of studies have not cultured microbes under physiological fluid shear conditions within a range commonly encountered by microbes during host-pathogen interactions. Here we describe a convenient batch culture biosystem in which (i) the levels of fluid shear force can be varied within physiologically relevant ranges and quantified via mathematical models and (ii) large numbers of cells can be planktonically grown and harvested to examine the effect of fluid shear levels on microbial genomic and phenotypic responses. A quantitative model based on numerical simulations and in situ imaging analysis was developed to calculate the fluid shear imparted by spherical beads of different sizes on bacterial cell cultures grown in a rotating wall vessel (RWV) bioreactor. To demonstrate the application of this model, we subjected cultures of the bacterial pathogen Salmonella enterica serovar Typhimurium to three physiologically-relevant fluid shear ranges during growth in the RVW and demonstrated a progressive relationship between the applied fluid shear and the bacterial genetic and phenotypic responses. By applying this model to different cell types, including other bacterial pathogens, entire classes of genes and proteins involved in cellular interactions may be discovered that have not previously been identified during growth under conventional culture conditions, leading to new targets for vaccine and therapeutic development.

* Corresponding author. Mailing address: Center for Infectious Diseases and Vaccinology, The Biodesign Institute, Arizona State University, P.O. Box 875401, Tempe, AZ 85287-5401. Phone: (480) 727-7520. Fax: (470) 727-8943. E-mail: cheryl.nickerson{at}asu.edu.

{triangledown} Published ahead of print on 1 December 2006.

{dagger} E.A.N. and C.M.O. contributed equally to this work.

{ddagger} Present address: Center for Infectious Diseases and Vaccinology, The Biodesign Institute, Arizona State University, Tempe, AZ 85287-5401.

Applied and Environmental Microbiology, February 2007, p. 699-705, Vol. 73, No. 3
0099-2240/07/$08.00+0     doi:10.1128/AEM.02428-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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