Quantitative Comparison and Analysis of Species-Specific Wound Biofilm Virulence Using an In Vivo, Rabbit-Ear Model
Abstract
BACKGROUND: Although bacterial biofilm is recognized as an important contributor to chronic wound pathogenesis, differences in biofilm virulence between species have never been studied in vivo. STUDY DESIGN: Dermal punch wounds in New Zealand white rabbit ears were inoculated with Klebsiella pneumoniae, Staphylococcus aureus, or Pseudomonas aeruginosa, or left uninfected as controls. In vivo biofilm was established and maintained using procedures from our previously published wound biofilm model. Virulence was assessed by measurement of histologic wound healing and host inflammatory mediators. Scanning electron microscopy (SEM) and bacterial counts verified biofilm viability. Extracellular polymeric substance (EPS)-deficient P aeruginosa was used for comparison. RESULTS: SEM confirmed the presence of wound biofilm for each species. P aeruginosa biofilm-infected wounds showed significantly more healing impairment than uninfected, K pneumoniae, and S aureus (p < 0.05), while also triggering the largest host inflammatory response (p < 0.05). Extracellular polymeric substance-deficient P aeruginosa demonstrated a reduced impact on the same quantitative endpoints relative to its wild-type strain (p < 0.05). CONCLUSIONS: Our novel analysis demonstrates that individual bacterial species possess distinct levels of biofilm virulence. Biofilm EPS may represent an integral part of their distinct pathogenicity. Rigorous examination of species-dependent differences in biofilm virulence is critical to developing specific therapeutics, while lending insight to the interactions within clinically relevant, polybacterial biofilms.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
A recent study demonstrated the ability of polymicrobial biofilms to impair healing in a rabbit ear model.
“a recent study demonstrated the ability of polymicrobial biofilms to impair healing in a rabbit ear model [39]”
From Interactions of Methicillin Resistant Staphylococcus aureus USA300 and Pseudomonas aeruginosa in Polymicrobial Wound Infection (2013) · cites this paper as [39]
Infection with either S. aureus or P. aeruginosa has been shown to delay wound closure in mouse and rabbit wound healing models.
“infection with either S. aureus or P. aeruginosa has been shown to delay wound closure in mouse and rabbit wound healing models [31], [37], [39], [40], [41].”
From Interactions of Methicillin Resistant Staphylococcus aureus USA300 and Pseudomonas aeruginosa in Polymicrobial Wound Infection (2013) · cites this paper as [39]
A recent study in a rabbit ear model found increased expression of pro-inflammatory cytokines TNFα and IL-1β with polymicrobial biofilm infection.
“Our results contrast those of a recent study in a rabbit ear model where increased expression of pro-inflammatory cytokines TNFα and IL-1β was seen with polymicrobial biofilm infection [39] and these differences may be explained by the variability between bacterial strains and animal models utilized.”
From Interactions of Methicillin Resistant Staphylococcus aureus USA300 and Pseudomonas aeruginosa in Polymicrobial Wound Infection (2013) · cites this paper as [39]
Few in vivo studies have focused on the synergistic effects of S. aureus and P. aeruginosa within the wound environment.
“However, despite the exceedingly high frequency of S. aureus and P. aeruginosa in chronic and acute wounds, few in vivo studies have focused on the synergistic effects of these prevalent bacteria within the wound environment [39], [42].”
From Interactions of Methicillin Resistant Staphylococcus aureus USA300 and Pseudomonas aeruginosa in Polymicrobial Wound Infection (2013) · cites this paper as [39]
Cites
The Involvement of Cell-to-Cell Signals in the Development of a Bacterial Biofilm
Davies, D. G., Parsek, M. R., Pearson, J. P., Iglewski, B. H., Costerton, J. W., & Greenberg, E. P. (1998) · Science · doi:10.1126/science.280.5361.295 · Cited by 3,396 · see citing works
Podschun, R., & Ullmann, U. (1998) · Clinical Microbiology Reviews · doi:10.1128/cmr.11.4.589 · Cited by 2,701 · see citing works
Lewis, K. (2001) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.45.4.999-1007.2001 · Cited by 1,988 · see citing works
Bacterial Biofilms: An Emerging Link to Disease Pathogenesis
Parsek, M. R., & Singh, P. K. (2003) · Annual Review of Microbiology · doi:10.1146/annurev.micro.57.030502.090720 · Cited by 1,563 · see citing works
James, G. A., Swogger, E., Wolcott, R., Pulcini, E. D., Secor, P., Sestrich, J., Costerton, J. W., & Stewart, P. S. (2008) · Wound Repair and Regeneration · doi:10.1111/j.1524-475x.2007.00321.x · Cited by 1,543 · see citing works
Edwards, R., & Harding, K. G. (2004) · Current Opinion in Infectious Diseases · doi:10.1097/00001432-200404000-00004 · Cited by 1,099 · see citing works
Incidence, outcomes, and cost of foot ulcers in patients with diabetes.
Ramsey, S. D., Newton, K. M., Blough, D. K., McCulloch, D. K., Sandhu, N., Reiber, G. E., & Wagner, E. H. (1999) · Diabetes Care · doi:10.2337/diacare.22.3.382 · Cited by 1,064 · see citing works
<scp>d</scp> -Amino Acids Trigger Biofilm Disassembly
Kolodkin-Gal, I., Romero, D., Cao, S., Clardy, J., Kolter, R., & Losick, R. (2010) · Science · doi:10.1126/science.1188628 · Cited by 874 · see citing works
Dowd, S. E., Sun, Y., Secor, P. R., Rhoads, D. D., Wolcott, B. M., James, G. A., & Wolcott, R. D. (2008) · BMC Microbiology · doi:10.1186/1471-2180-8-43 · Cited by 783 · see citing works
The Health Care Costs of Diabetic Peripheral Neuropathy in the U.S.
Gordois, A., Scuffham, P., Shearer, A. T., Oglesby, A. K., & Tobian, J. A. (2003) · Diabetes Care · doi:10.2337/diacare.26.6.1790 · Cited by 735 · see citing works
Rice, K. C., Mann, E. E., Endres, J. L., Weiss, E. C., Cassat, J. E., Smeltzer, M. S., & Bayles, K. W. (2007) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0610226104 · Cited by 674 · see citing works
Multiple bacterial species reside in chronic wounds: a longitudinal study
Gjødsbøl, K., Christensen, J. J., Karlsmark, T., Jørgensen, B., Klein, B. M., & Krogfelt, K. A. (2006) · International Wound Journal · doi:10.1111/j.1742-481x.2006.00159.x · Cited by 623 · see citing works
Vuong, C., Voyich, J. M., Fischer, E. R., Braughton, K. R., Whitney, A. R., DeLeo, F. R., & Otto, M. J. (2004) · Cellular Microbiology · doi:10.1046/j.1462-5822.2004.00367.x · Cited by 623 · see citing works
Quorum Sensing in Staphylococcus aureus Biofilms
Yarwood, J. M., Bartels, D. J., Volper, E. M., & Greenberg, E. P. (2004) · Journal of Bacteriology · doi:10.1128/jb.186.6.1838-1850.2004 · Cited by 597 · see citing works
Borlee, B. R., Goldman, A. D., Murakami, K., Samudrala, R., Wozniak, D. J., & Parsek, M. R. (2010) · Molecular Microbiology · doi:10.1111/j.1365-2958.2009.06991.x · Cited by 544 · see citing works
Health and Economic Impact of Surgical Site Infections Diagnosed after Hospital Discharge
Perencevich, E. N., Sands, K., Cosgrove, S. E., Guadagnoli, E., Meara, E. R., & Platt, R. G. (2003) · Emerging infectious diseases · doi:10.3201/eid0902.020232 · Cited by 512 · see citing works
Mutation of sarA in Staphylococcus aureus Limits Biofilm Formation
Beenken, K. E., Blevins, J. S., & Smeltzer, M. S. (2003) · Infection and Immunity · doi:10.1128/iai.71.7.4206-4211.2003 · Cited by 429 · see citing works
Burmølle, M., Thomsen, T. R., Fazli, M., Dige, I., Christensen, L. H., Homøe, P., Tvede, M., Nyvad, B., Tolker‐Nielsen, T., Givskov, M., et al. (2010) · FEMS Immunology & Medical Microbiology · doi:10.1111/j.1574-695x.2010.00714.x · Cited by 426 · see citing works
Jensen, P. Ø., Bjarnsholt, T., Phipps, R. K., Rasmussen, T. B., Calum, H., Christoffersen, L., Moser, C. E., Williams, P. M., Pressler, T., Givskov, M., et al. (2007) · Microbiology · doi:10.1099/mic.0.2006/003863-0 · Cited by 402 · see citing works
Microscopic and physiologic evidence for biofilm-associated wound colonization in vivo
Davis, S. C., Ricotti, C., Cazzaniga, A., Welsh, E., Eaglstein, W. H., & Mertz, P. M. (2008) · Wound Repair and Regeneration · doi:10.1111/j.1524-475x.2007.00303.x · Cited by 393 · see citing works
Staphylococcus quorum sensing in biofilm formation and infection
Kong, K., Vuong, C., & Otto, M. J. (2006) · International Journal of Medical Microbiology · doi:10.1016/j.ijmm.2006.01.042 · Cited by 385 · see citing works
Bacterial biofilms within the clinical setting: what healthcare professionals should know
Lindsay, D., & von Holy, A. (2006) · Journal of Hospital Infection · doi:10.1016/j.jhin.2006.06.028 · Cited by 380 · see citing works
Rashid, M. H., Rumbaugh, K. P., Passador, L., et al. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.170283397 · Cited by 375 · see citing works
Jesaitis, A. J., Franklin, M. J., Berglund, D. L., Sasaki, M., Lord, C. I., Bleazard, J. B., Duffy, J. E., Beyenal, H., & Lewandowski, Z. (2003) · The Journal of Immunology · doi:10.4049/jimmunol.171.8.4329 · Cited by 368 · see citing works
Schierle, C. F., De la Garza, M., Mustoe, T. A., & Galiano, R. D. (2009) · Wound Repair and Regeneration · doi:10.1111/j.1524-475x.2009.00489.x · Cited by 311 · see citing works
Self-citationQuorum Sensing and the Social Evolution of Bacterial Virulence
Rumbaugh, K. P., Diggle, S. P., Watters, C., Ross‐Gillespie, A., Griffin, A. S., & West, S. A. (2009) · Current Biology · doi:10.1016/j.cub.2009.01.050 · Cited by 302 · see citing works
Inhibitory Effects of d -Amino Acids on Staphylococcus aureus Biofilm Development
Hochbaum, A. I., Kolodkin‐Gal, I., Foulston, L., Kolter, R., Aizenberg, J., & Losick, R. M. (2011) · Journal of Bacteriology · doi:10.1128/jb.05534-11 · Cited by 293 · see citing works
Zhao, G., Hochwalt, P. C., Usui, M. L., Underwood, R. A., Singh, P. K., James, G. A., Stewart, P. S., Fleckman, P., & Olerud, J. E. (2010) · Wound Repair and Regeneration · doi:10.1111/j.1524-475x.2010.00608.x · Cited by 275 · see citing works
Interference with granulocyte function by Staphylococcus epidermidis slime
Johnson, G. M., Lee, D. A., Regelmann, W. E., Gray, E. D., Peters, G., & Quie, P. G. (1986) · Infection and Immunity · doi:10.1128/iai.54.1.13-20.1986 · Cited by 245 · see citing works
Multidrug-resistant Organisms in Military Wounds from Iraq and Afghanistan
Calhoun, J. H., Murray, C. K., & Manring, M. M. (2008) · Clinical Orthopaedics and Related Research · doi:10.1007/s11999-008-0212-9 · Cited by 188 · see citing works
Permeation of Antimicrobial Agents through Pseudomonas aeruginosa Biofilms: A Simple Method
Shigeta, M., Tanaka, G., Komatsuzawa, H., Sugai, M., Suginaka, H., & Usui, T. (2009) · Chemotherapy · doi:10.1159/000239587 · Cited by 187 · see citing works
Gurjala, A. N., Geringer, M. R., Seth, A. K., Hong, S. J., Smeltzer, M. S., Galiano, R. D., Leung, K. P., & Mustoe, T. A. (2011) · Wound Repair and Regeneration · doi:10.1111/j.1524-475x.2011.00690.x · Cited by 183 · see citing works
Self-citationEffect of Hyperbaric Oxygen and Growth Factors on Rabbit Ear Ischemic Ulcers
Zhao, L. L. (1994) · Archives of Surgery · doi:10.1001/archsurg.1994.01420340057010 · Cited by 166 · see citing works
Pseudomonas aeruginosa Forms Biofilms in Acute Infection Independent of Cell-to-Cell Signaling
Schaber, J. A., Triffo, W. J., Suh, S. J., Oliver, J. W., Hastert, M. C., Griswold, J., Auer, M., Hamood, A. N., & Rumbaugh, K. P. (2007) · Infection and Immunity · doi:10.1128/iai.00586-07 · Cited by 164 · see citing works
Hospital-acquired pressure ulcers: a comparison of costs in medical vs. surgical patients.
Beckrich, K., & Aronovitch, S. A. (2000) · PubMed · Cited by 153 · see citing works
Qin, Z., Yang, L., Qu, D., Molin, S., & Tolker‐Nielsen, T. (2009) · Microbiology · doi:10.1099/mic.0.028001-0 · Cited by 140 · see citing works
Patient-Level Estimates of the Cost of Complications in Diabetes in a Managed-Care Population
Ramsey, S. D., Newton, K. M., Blough, D. K., McCulloch, D. K., Sandhu, N., & Wagner, E. H. (1999) · PharmacoEconomics · doi:10.2165/00019053-199916030-00005 · Cited by 95 · see citing works
Shiau, A., & Wu, C. (1998) · Microbiology and Immunology · doi:10.1111/j.1348-0421.1998.tb01966.x · Cited by 89 · see citing works
Extracellular Polysaccharides Matrix — An Often Forgotten Virulence Factor in Oral Biofilm Research
Koo, H., Xiao, J., & Klein, M. I. (2009) · International Journal of Oral Science · doi:10.4248/ijos.09086 · Cited by 77 · see citing works
Treatment of Pseudomonas aeruginosa Biofilm–Infected Wounds with Clinical Wound Care Strategies
Seth, A. K., Geringer, M. R., Gurjala, A. N., et al. (2012) · Plastic & Reconstructive Surgery · doi:10.1097/prs.0b013e31823aeb3b · Cited by 76 · see citing works
Self-citationPainful venous ulcers: themes and stories about their impact on quality of life.
Krasner, D. L. (1998) · PubMed · Cited by 66 · see citing works
Weiss, E. C., Spencer, H. J., Daily, S. T., et al. (2009) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.01432-08 · Cited by 50 · see citing works
Virulence ofStaphylococcus epidermidisin a mouse model: significance of extracellular slime
Deighton, M., Borland, R., & Capstick, J. (1996) · Epidemiology and Infection · doi:10.1017/s0950268800001448 · Cited by 48 · see citing works
Nakagami, G., Sanada, H., Sugama, J., Morohoshi, T., Ikeda, T., & Ohta, Y. (2008) · Wound Repair and Regeneration · doi:10.1111/j.1524-475x.2007.00329.x · Cited by 39 · see citing works
Nakagami, G., Morohoshi, T., Ikeda, T., Ohta, Y., Sagara, H., Huang, L., Nagase, T., Sugama, J., & Sanada, H. (2011) · Wound Repair and Regeneration · doi:10.1111/j.1524-475x.2010.00653.x · Cited by 34 · see citing works
Effects of clinical isolates ofPseudomonas aeruginosaonStaphylococcus epidermidisbiofilm formation
Pihl, M., de Paz, L. E. C., Schmidtchen, A., et al. (2010) · FEMS Immunology & Medical Microbiology · doi:10.1111/j.1574-695x.2010.00707.x · Cited by 33 · see citing works
Drobni, M., Hallberg, K., Öhman, U., Birve, A., Persson, K., Johansson, I., & Strömberg, N. (2006) · BMC Microbiology · doi:10.1186/1471-2180-6-43 · Cited by 30 · see citing works
Evidence-Based Medicine in Wound Care
Carter, M. J., & Warriner, R. A. (2008) · Advances in Skin & Wound Care · doi:10.1097/01.asw.0000343719.38190.ad · Cited by 22 · see citing works
Fighting Infection in Chronic Wounds
Fleck, C. A. (2006) · Advances in Skin & Wound Care · doi:10.1097/00129334-200605000-00009 · Cited by 16 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Pastar, I., Nusbaum, A. G., Gil, J., Patel, S. B., Chen, J., Valdes, J., Stojadinovic, O., Plano, L. R., Tomic-Canic, M., & Davis, S. C. (2013) · PLoS ONE · doi:10.1371/journal.pone.0056846 · Cited by 391 · see citing works
- Authors
- Seth, A. K., Geringer, M. R., Galiano, R. D., Leung, K. P., Mustoe, T. A., & Hong, S. J.
- Affiliations
- Northwestern University (US)
- United States Army Institute of Surgical Research (US)
- Corresponding author
- Hong, Seok J.
- Venue
- Journal of the American College of Surgeons
- Published
- 2012-09
- Volume
- 215
- Issue
- 3
- Pages
- 388-399
- ISSN
- 1072-7515 (ISSN-L)
- OpenAlex
- W2087835752
- Cited by
- 65
- References
- 49
- Citations by year
- 2025: 1 · 2024: 7 · 2023: 6 · 2022: 6 · 2021: 2 · 2020: 5 · 2019: 2 · 2018: 3 · 2017: 6 · 2016: 9 · 2015: 4 · 2014: 8 · 2013: 6
- FWCI
- 2.31
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of the American College of Surgeons (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bacterial biofilms and quorum sensing
- Health Sciences › Medicine › Rehabilitation › Wound Healing and Treatments
- Health Sciences › Medicine › Surgery › Orthopedic Infections and Treatments
- Keywords
- wound biofilm, biofilm virulence, Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, extracellular polymeric substances, delayed wound healing, host inflammatory response, rabbit ear model
- MeSH terms
- Animals, Ear, External, Klebsiella pneumoniae, Microscopy, Electron, Scanning, Polysaccharides, Bacterial, Pseudomonas aeruginosa, Rabbits, RNA, Bacterial, RNA, Messenger, Staphylococcus aureus, Virulence, Wound Healing, Biomarkers, Soft Tissue Injuries, Biofilms, Inflammation Mediators, Reverse Transcriptase Polymerase Chain Reaction, Models, Animal, Bacterial Load
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
