A novel model of chronic wounds: Importance of redox imbalance and biofilm-forming bacteria for establishment of chronicity

PLoS ONE

Volumn 9, Issue 10, 2014, Pages

  Dhall, Sandeep ^a,b   Do, Danh ^c   Garcia, Monika ^a   Wijesinghe, Dayanjan Shanaka ^d,e,f   Brandon, Angela ^g   Kim, Jane ^a   Sanchez, Antonio ^h   Lyubovitsky, Julia ^b   Gallagher, Sean ^h   Nothnagel, Eugene A ^a   Chalfant, Charles E ^d,e,f   Patel, Rakesh P ^g   Schiller, Neal ^c   Martins Green, Manuela ^a,b  

^a UNIVERSITY OF CALIFORNIA   (United States)

^b Bioengineering Interdepartmental Graduate Program University of California Riverside ^*  (United States)

^c UNIVERSITY OF CALIFORNIA   (United States)

^d VETERANS AFFAIRS MEDICAL CENTER   (United States)

^e VIRGINIA COMMONWEALTH UNIVERSITY   (United States)

^f VIRGINIA COMMONWEALTH UNIVERSITY   (United States)

^g University of Alabama at Birmingham   (United States)

^h Sepsis diagnostics expert group   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ANTIOXIDANT; REACTIVE NITROGEN SPECIES; REACTIVE OXYGEN METABOLITE; HERPESVIRUS ENTRY MEDIATOR LIGAND; TNFSF14 PROTEIN, MOUSE;

ADULT; ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; ANTIBIOTIC RESISTANCE; ANTIOXIDANT ASSAY; ARTICLE; BACTERIAL INFECTION; BACTERIAL LOAD; BACTERIUM CULTURE; BACTERIUM ISOLATION; BIOFILM; CHRONIC WOUND; COAGULASE NEGATIVE STAPHYLOCOCCUS; COLLAGEN FIBER; CONTROLLED STUDY; DISEASE MODEL; ENTEROBACTER CLOACAE; ENZYME ACTIVITY; EPITHELIZATION; GRAM NEGATIVE AEROBIC RODS AND COCCI; GRANULATION TISSUE; HISTOPATHOLOGY; INTERSTITIUM; MINIMUM INHIBITORY CONCENTRATION; MOUSE; NONHUMAN; OXIDATION REDUCTION STATE; OXIDATIVE STRESS; SKIN FLORA; STREPTOCOCCUS; VASCULARIZATION; WOUND HEALING IMPAIRMENT; ANIMAL; C57BL MOUSE; CHRONIC DISEASE; GENETICS; KNOCKOUT MOUSE; METABOLISM; MICROBIOLOGY; OXIDATION REDUCTION REACTION; PATHOLOGY; STAPHYLOCOCCAL SKIN INFECTION; STAPHYLOCOCCUS INFECTION; STREPTOCOCCUS INFECTION; WOUND HEALING;

ANIMALS; BIOFILMS; CHRONIC DISEASE; DISEASE MODELS, ANIMAL; DRUG RESISTANCE, BACTERIAL; MICE, INBRED C57BL; MICE, KNOCKOUT; OXIDATION-REDUCTION; OXIDATIVE STRESS; REACTIVE OXYGEN SPECIES; STAPHYLOCOCCAL INFECTIONS; STAPHYLOCOCCAL SKIN INFECTIONS; STREPTOCOCCAL INFECTIONS; TUMOR NECROSIS FACTOR LIGAND SUPERFAMILY MEMBER 14; WOUND HEALING;

EID: 84907970192     PISSN: None     EISSN: 19326203     Source Type: Journal    
DOI: 10.1371/journal.pone.0109848     Document Type: Article

References (93)

1. Sen CK, Gordillo GM, Roy S, Kirsner R, Lambert L, et al. (2009) Human skin wounds: a major and snowballing threat to public health and the economy. Wound Repair Regen 17: 763-771. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2810192&tool=pmcentrez&rendertype=abstract Accessed 2013 January 7.
2. Lazarus GS, Cooper DM, Knighton DR, Margolis DJ, Percoraro RE, et al. (1994) Definitions and guidelines for assessment of wounds and evaluation of healing. Wound Repair Regen: 165-170.
3. Wong VW, Gurtner GC (2012) Tissue engineering for the management of chronic wounds: current concepts and future perspectives. Exp Dermatol 21: 729-734. Available: http://www.ncbi.nlm.nih.gov/pubmed/22742728 Accessed 2013 April 15.
4. Roy S, Khanna S, Nallu K, Hunt TK, Sen CK (2006) Dermal wound healing is subject to redox control. Mol Ther 13: 211-220. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1389791&tool=pmcentrez&ren dertype=abstract Accessed 2013 January 5.
5. D'Autréaux B, Toledano MB (2007) ROS as signalling molecules: mechanisms that generate specificity in ROS homeostasis. Nat Rev Mol Cell Biol 8: 813-824. Available: http://www.ncbi.nlm.nih.gov/pubmed/17848967 Accessed 2013 February 27.
6. Sen CK, Roy S (2008) Redox signals in wound healing. Biochim Biophys Acta 1780: 1348-1361. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2574682&tool=pmcentrez&rendertype=abstract Accessed 2012 November 1.
7. Dröge W(2002) Free radicals in the physiological control of cell function. Physiol Rev 82: 47-95. Available: http://www.ncbi.nlm.nih.gov/pubmed/11773609
8. Yeoh-Ellerton S, Stacey MC (2003) Iron and 8-isoprostane levels in acute and chronic wounds. J Invest Dermatol 121: 918-925. Available: http://www.ncbi.nlm.nih.gov/pubmed/14632213
9. Wlaschek M, Scharffetter-Kochanek K (2005) Oxidative stress in chronic venous leg ulcers. Wound Repair Regen 13: 452-461. Available: http://www.ncbi.nlm.nih.gov/pubmed/16176453
10. Yang Q, Phillips PL, Sampson EM, Progulske-Fox A, Jin S, et al. (2013) Development of a novel ex vivo porcine skin explant model for the assessment of mature bacterial biofilms. Wound Repair Regen 21: 704-714. Available: http://www.ncbi.nlm.nih.gov/pubmed/23927831 Accessed 2013 December 2.
11. Hopf HW, Rollins MD (2007) Wounds: an overview of the role of oxygen. Antioxid Redox Signal 9: 1183-1192. Available: http://www.ncbi.nlm.nih.gov/pubmed/17536961 Accessed 2013 August 15.
12. Britland S, Ross-Smith O, Jamil H, Smith AG, Vowden K, et al. (2012) The lactate conundrum in wound healing: clinical and experimental findings indicate the requirement for a rapid point-of-care diagnostic. Biotechnol Prog 28: 917-924. Available: http://www.ncbi.nlm.nih.gov/pubmed/22581665 Accessed 2013 August 14.
13. Fazli M, Bjarnsholt T, Kirketerp-Møller K, Jørgensen A, Andersen CB, et al. (2011) Quantitative analysis of the cellular inflammatory response against biofilm bacteria in chronic wounds. Wound Repair Regen 19: 387-391. Available: http://www.ncbi.nlm.nih.gov/pubmed/21518086 Accessed 2013 August 15.
14. Warburg O (1956) On the Origin of Cancer Cells. Science (80-) 123: 309-314.
15. Hunt TK, Hopf H, Hussain Z (2000) Physiology of wound healing. Adv Skin Wound Care 13: 6-11. Available: http://www.ncbi.nlm.nih.gov/pubmed/11074996 Accessed 2013 August 14.
16. Dovi J V, Szpaderska AM, DiPietro L a (2004) Neutrophil function in the healing wound: adding insult to injury? Thromb Haemost: 275-280. Available: http://www.schattauer.de/index.php?id=1214&doi=10.1160/TH03-11-0720&no-cache=1 Accessed 2013 April 20.
17. McCarty SM, Cochrane C a, Clegg PD, Percival SL (2012) The role of endogenous and exogenous enzymes in chronic wounds: a focus on the implications of aberrant levels of both host and bacterial proteases in wound healing. Wound Repair Regen 20: 125-136. Available: http://www.ncbi.nlm.nih.gov/pubmed/22380687 Accessed 2013 May 19.
18. Schäfer M, Werner S (2008) Oxidative stress in normal and impaired wound repair. Pharmacol Res 58: 165-171. Available: http://www.ncbi.nlm.nih.gov/pubmed/18617006 Accessed 2012 November 1.
19. Luo J, Chen AF (2005) Nitric oxide: a newly discovered function on wound healing. Acta Pharmacol Sin 26: 259-264. Available: http://www.ncbi.nlm.nih.gov/pubmed/15715920 Accessed 2013 March 29.
20. Abd-El-Aleem S a, Ferguson MW, Appleton I, Kairsingh S, Jude EB, et al. (2000) Expression of nitric oxide synthase isoforms and arginase in normal human skin and chronic venous leg ulcers. J Pathol 191: 434-442. Available: http://www.ncbi.nlm.nih.gov/pubmed/10918219
21. Petreaca ML, Do D, Dhall S, McLelland D, Serafino A, et al. (2012) Deletion of a tumor necrosis superfamily gene in mice leads to impaired healing that mimics chronic wounds in humans. Wound Repair Regen 20: 353-366. Available: http://www.ncbi.nlm.nih.gov/pubmed/22564230 Accessed 2012 November 5.
22. Howell-Jones RS, Wilson MJ, Hill KE, Howard a J, Price PE, et al. (2005) A review of the microbiology, antibiotic usage and resistance in chronic skin wounds. J Antimicrob Chemother 55: 143-149. Available: http://www.ncbi.nlm.nih.gov/pubmed/15649989 Accessed 2012 November 9.
23. James G a, Swogger E, Wolcott R, Pulcini E . deLancey, Secor P, et al. (2007) Biofilms in chronic wounds. Wound Repair Regen 16: 37-44. Available: http://www.ncbi.nlm.nih.gov/pubmed/18086294 Accessed 2012 October 29.
24. Guo S, Dipietro L a (2010) Factors affecting wound healing. J Dent Res 89: 219-229. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2903966amp;tool=pmcentrez&rendertype=abstract Accessed 2014 April 28.
25. Martin JM, Zenilman JM, Lazarus GS (2010) Molecular microbiology: new dimensions for cutaneous biology and wound healing. J Invest Dermatol 130: 38-48. Available: http://www.ncbi.nlm.nih.gov/pubmed/19626034 Accessed 2014 May 8.
26. Petreaca ML, Yao M, Ware C, Martins-Green MM (2008) Vascular endothelial growth factor promotes macrophage apoptosis through stimulation of tumor necrosis factor superfamily member 14 (TNFSF14/LIGHT). Wound Repair Regen 16: 602-614. Available: http://www.ncbi.nlm.nih.gov/pubmed/19128255 Accessed 2014 February 18.
27. Gonzalez FJ (2005) Role of cytochromes P450 in chemical toxicity and oxidative stress: studies with CYP2E1. Mutat Res 569: 101-110. Available: http://www.ncbi.nlm.nih.gov/pubmed/15603755 Accessed 2014 May 8.
28. Song L, Wei X, Zhang B, Luo X, Liu J, et al. (2009) Role of Foxa1 in regulation of bcl2 expression during oxidative-stress-induced apoptosis in A549 type II pneumocytes. Cell Stress Chaperones 14: 417-425. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2728276&tool=pmcentrez&ren dertype=abstract Accessed 2014 May 8.
29. Kuhlicke J, Frick JS, Morote-Garcia JC, Rosenberger P, Eltzschig HK (2007) Hypoxia inducible factor (HIF)-1 coordinates induction of Toll-like receptors TLR2 and TLR6 during hypoxia. PLoS One 2: e1364. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2147045&tool=pmcentrez&ren dertype=abstract Accessed 2014 May 7.
30. Kundu TK, Velayutham M, Zweier JL (2012) Aldehyde Oxidase Functions as a Superoxide Generating NADH Oxidase: An Important Redox Regulated Pathway of Cellular Oxygen. Biochemistry 51: 2930-2939.
31. Shaw S, Jayatilleke E (1992) The Role of Cellular Oxidases and Catalytic iron in the pathogenesis of Ethanol-Induced Liver Injury. Life Sci 50: 2045-2052.
32. Moor AN, Tummel E, Prather JL, Jung M, Lopez JJ, et al. (2014) Consequences of age on ischemic wound healing in rats: altered antioxidant activity and delayed wound closure. Age (Dordr) 36: 733-748. Available: http://www.ncbi.nlm.nih.gov/pubmed/24443098 Accessed 2014 July 17.
33. Schneider LA, Korber A, Grabbe S, Dissemond J (2007) Influence of pH on wound-healing: a new perspective for wound-therapy? Arch Dermatol Res 298: 413-420. Available: http://www.ncbi.nlm.nih.gov/pubmed/17091276 Accessed 2012November 5.
34. Schreml S, Szeimies RM, Prantl L, Karrer S, Landthaler M, et al. (2010) Oxygen in acute and chronic wound healing. Br J Dermatol 163: 257-268. Available: http://www.ncbi.nlm.nih.gov/pubmed/20394633 Accessed 2012 December 4.
35. Stefaniak AB, Plessis J Du, John SM, Eloff F, Agner T, et al. (2013) International guidelines for the in vivo assessment of skin properties in non-clinical settings: part 1. pH. Skin Res Technol 19: 59-68. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3747458&tool=pmcentrez&rendertype=abstract Accessed 2014 July 22.
36. Lotito S, Blonder P, Francois AI, Rdmy C (1989) Correlation between intracellular pH and lactate levels in the rat brain during potassium cyanide induced metabolism blockade: a combined 31p-1H in vivo nuclear magnetic spectroscopy study. 97: 91-96.
37. Gethin G (2007) The significance of surface pH in chronic wounds. Wounds 3: 52-56. Available: http://www.woundsinternational.com/pdf/content-124.pdf Accessed 2014 January 29.
38. Bowler PG (2002) Wound pathophysiology, infection and therapeutic options. Ann Med 34: 419-427. Available: http://www.ncbi.nlm.nih.gov/pubmed/12523497
39. Dowd SE, Sun Y, Secor PR, Rhoads DD, Wolcott BM, et al. (2008) Survey of bacterial diversity in chronic wounds using pyrosequencing, DGGE, and full ribosome shotgun sequencing. BMC Microbiol 8: 43. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2289825&tool=pmcentrez&ren dertype=abstract Accessed 2014 June 2.
40. Christensen GD, Simpson W a, Younger JJ, Baddour LM, Barrett FF, et al. (1985) Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol 22: 996-1006. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=271866&tool=pmcentrez&rendertype=abstract
41. O'Toole G a, Kolter R (1998) Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Mol Microbiol 28: 449-461. Available: http://www.ncbi.nlm.nih.gov/pubmed/9632250
42. Stepanovic S, Vukovic D, Dakic I, Savic B, Svabic-Vlahovic M (2000) A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J Microbiol Methods 40: 175-179. Available: http://www.ncbi.nlm.nih.gov/pubmed/10699673
43. Kolodkin-Gal I, Cao S, Chai L, Böttcher T, Kolter R, et al. (2012) A self-produced trigger for biofilm disassembly that targets exopolysaccharide. Cell 149: 684-692. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3526955&tool=pmcentrez&rendertype=abstract Accessed 2014 January 21.
44. Kloos W, Schleifer K (1975) Isolation and Characterization of Staphylococci from Human Skin. Int J Syst Bacteriol 25: 62-79.
45. Qu Y, Daley AJ, Istivan TS, Garland SM, Deighton M a (2010) Antibiotic susceptibility of coagulase-negative staphylococci isolated from very low birth weight babies: comprehensive comparisons of bacteria at different stages of biofilm formation. Ann Clin Microbiol Antimicrob 9: 16. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2902406&tool=pmcentrez&render type=abstract
46. Cui B, Smooker PM, Rouch D a, Daley AJ, Deighton M a (2013) Differences between two clinical Staphylococcus capitis subspecies as revealed by biofilm, antibiotic resistance, and pulsed-field gel electrophoresis profiling. J Clin Microbiol 51: 9-14. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3536240&tool=pmcentrez&rendertype=abstract Accessed 2014 July 19.
47. Percival SL, Hill KE, Malic S, Thomas DW, Williams DW (2010) Antimicrobial tolerance and the significance of persister cells in recalcitrant chronic wound biofilms. Wound Repair Regen 19: 1-9. Available: http://www.ncbi.nlm.nih.gov/pubmed/21235682 Accessed 2014 January 29.
48. Parsek MR, Singh PK (2003) Bacterial biofilms: an emerging link to disease pathogenesis. Annu Rev Microbiol 57: 677-701. Available: http://www.ncbi.nlm.nih.gov/pubmed/14527295 Accessed 2014 January 22.
49. Siddiqui AR, Bernstein JM (2010) Chronic wound infection: facts and controversies. Clin Dermatol 28: 519-526. Available: http://www.ncbi.nlm.nih.gov/pubmed/20797512 Accessed 2012 December 17.
50. Cho I, Blaser MJ (2012) The human microbiome: at the interface of health and disease. Nat Rev Genet 13: 260-270. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3418802&tool=pmcentrez&rendertype=abstract Accessed 2014 January 22.
51. Stevens D, Lieberman M, McNitt T, Price J (1984) Demonstration of uronic acid capsular material in the cerebrospinal fluid of a patient with meningitis caused by mucoid Pseudomonas aeruginosa. J Clin Microbiol 19: 942-943. Available: http://jcm.asm.org/content/19/6/942.short Accessed 2014 February 8.
52. Bales PM, Renke EM, May SL, Shen Y, Nelson DC (2013) Purification and Characterization of Biofilm-Associated EPS Exopolysaccharides from ESKAPE Organisms and Other Pathogens. PLoS One 8: e67950. Available: http://dx.plos.org/10.1371/journal.pone.0067950 Accessed 2014 January 26.
53. Sindrilaru A, Peters T, Wieschalka S, Baican C, Baican A, et al. (2011) An unrestrained proinflammatory M1 macrophage population induced by iron impairs wound healing in humans and mice. 121: 985-997. doi:10.1172/JCI44490DS1
54. Bryan N, Ahswin H, Smart N, Bayon Y, Wohlert S, et al. (2012) Reactive oxygen species (ROS)-a family of fate deciding molecules pivotal in constructive inflammation and wound healing. Eur Cell Mater 24: 249-265. Available: http://www.ncbi.nlm.nih.gov/pubmed/23007910
55. Messonnier L, Kristensen M, Juel C, Denis C (2007) Importance of pH regulation and lactate/H+ transport capacity for work production during supramaximal exercise in humans. J Appl Physiol 102: 1936-1944. Available: http://www.ncbi.nlm.nih.gov/pubmed/17289910 Accessed 2013 August 15.
56. Shorrock SM, Kun S, Peura RA, Dum RM (2000) Determination of a Relationship between Bacteria Levels and Tissue pH in Wounds: Animal Studies. IEEE: 117-118.
57. Chen C-A, Druhan LJ, Varadharaj S, Chen Y-R, Zweier JL (2008) Phosphorylation of endothelial nitric-oxide synthase regulates superoxide generation from the enzyme. J Biol Chem 283: 27038-27047. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2556006&tool=pmcentrez&rendertype=abstract Accessed 2013 March 24.
58. Sullivan JC, Pollock JS (2006) Coupled and uncoupled NOS: separate but equal? Uncoupled NOS in endothelial cells is a critical pathway for intracellular signaling. Circ Res 98: 717-719. Available: http://www.ncbi.nlm.nih.gov/pubmed/16574911 Accessed 2014 May 8.
59. Erez A, Nagamani SCS, Shchelochkov O a, Premkumar MH, Campeau PM, et al. (2011) Requirement of argininosuccinate lyase for systemic nitric oxide production. Nat Med 17: 1619-1626. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3348956&tool=pmcentrez&rendertype=abstract Accessed 2014 May 8.
60. Miyazaki H, Matsuoka H, Cooke JP, Usui M, Ueda S, et al. (1999) Endogenous Nitric Oxide Synthase Inhibitor: A Novel Marker of Atherosclerosis. Circulation 99: 1141-1146. Available: http://circ.ahajournals.org/cgi/doi/10.1161/01.CIR.99.9.1141 Accessed 2014 May 8.
61. Moseley R, Hilton JR, Waddington RJ, Harding KG (2004) Original Research Articles - Basic Science Comparison of oxidative stress biomarker profiles between acute and chronic wound environments. Wound Repair Regen 12: 419-429.
62. Goel A, Spitz DR, Weiner GJ (2012) Manipulation of cellular redox parameters for improving therapeutic responses in B-cell lymphoma and multiple myeloma. J Cell Biochem 113: 419-425. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3374635&tool=pmcentrez&rendertype=abstract Accessed 2013 May 19.
63. Freeman T a, Parvizi J, Della Valle CJ, Steinbeck MJ (2009) Reactive oxygen and nitrogen species induce protein and DNA modifications driving arthrofibrosis following total knee arthroplasty. Fibrogenesis Tissue Repair 2: 5. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2785750&tool=pmcentrez&rendertype=abstract Accessed 2013 May 19.
64. Fraga CG, Shigenaga MK, Park JW, Degan P, Ames BN (1990) Oxidative damage to DNA during aging: 8-Hydroxy-2 ′-deoxyguanosine in rat organ DNA and urine. Proc Natl Acad Sci U S A 87: 4533-4537. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=54150&tool=pmcentrez&render type=abstract
65. Kikuchi S, Kobune M, Iyama S, Sato T, Murase K, et al. (2012) Improvement of iron-mediated oxidative DNA damage in patients with transfusion-dependent myelodysplastic syndrome by treatment with deferasirox. Free Radic Biol Med 53: 643-648. Available: http://www.ncbi.nlm.nih.gov/pubmed/22705364 Accessed 2013 May 19.
66. Beckman JS, Koppenol WH (1996) Nitric oxide, superoxide, and peroxynitrite: the good, the bad, and the ugly. AJP Cell Physiol 271: 1424-1437.
67. Berlett BS, Friguet B, Yim MB, Chock PB, Stadtman ER (1996) Peroxynitrite-mediated nitration of tyrosine residues in Escherichia coli glutamine synthetase mimics adenylylation: relevance to signal transduction. Proc Natl Acad Sci U S A 93: 1776-1780. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=39857&tool=pmcentrez&rendertype=abstract
68. Barraud N, Hassett DJ, Hwang S-H, Rice S a, Kjelleberg S, et al. (2006) Involvement of nitric oxide in biofilm dispersal of Pseudomonas aeruginosa. J Bacteriol 188: 7344-7353. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1636254&tool=pmcentrez&rendertype=abstract Accessed 2014 July 17.
69. Schreiber F, Beutler M, Enning D, Lamprecht-Grandio M, Zafra O, et al. (2011) The role of nitric-oxide-synthase-derived nitric oxide in multicellular traits of Bacillus subtilis 3610: biofilm formation, swarming, and dispersal. BMC Microbiol 11: 111. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3224222&tool=pmcentrez&rendertype=abstract Accessed 2014 July 22.
70. Kaplan JB (2010) Biofilm dispersal: mechanisms, clinical implications, and potential therapeutic uses. J Dent Res 89: 205-218. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3318030&tool=pmcentrez&ren dertype=abstract Accessed 2014 July 17.
71. Cathie K, Howlin R, Carroll M, Clarke S, Connett G, et al. (2014) Reducing Antibiotic Tolerance using Nitric Oxide in Cystic Fibrosis: report of a proof of concept clinical trial. Arch Dis Child 99: A159-A159. Available: http://adc.bmj.com/cgi/doi/10.1136/archdischild-2014-306237.367 Accessed 2014 July 22.
72. Rizk M, Witte MB, Barbul A (2004) Nitric oxide and wound healing. World J Surg 28: 301-306. Available: http://www.ncbi.nlm.nih.gov/pubmed/14961192 Accessed 2014 July 22.
73. Bedard K, Krause K (2007) The NOX Family of ROS-Generating NADPH Oxidases: Physiology and Pathophysiology. Physiol Rev 87: 245-313. doi:10.1152/physrev.00044.2005
74. Wink D a, Hines HB, Cheng RYS, Switzer CH, Flores-Santana W, et al. (2011) Nitric oxide and redox mechanisms in the immune response. J Leukoc Biol 89: 873-891. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3100761&tool=pmcentrez&rendertype=abstract Accessed 2014 July 22.
75. Clark RAF (2008) Oxidative stress and "senescent" fibroblasts in non-healing wounds as potential therapeutic targets. J Invest Dermatol 128: 2361-2364. Available: http://www.ncbi.nlm.nih.gov/pubmed/18787545 Accessed 2014 July 23.
76. Boles BR, Singh PK (2008) Endogenous oxidative stress produces diversity and adaptability in biofilm communities. Proc Natl Acad Sci U S A 105: 12503-12508. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2527941&tool=pmcentrez&rendertype=abstract
77. Liu X, Sun X, Wu Y, Xie C, Zhang W, et al. (2013) Oxidation-sensing regulator AbfR regulates oxidative stress responses, bacterial aggregation, and biofilm formation in Staphylococcus epidermidis. J Biol Chem 288: 3739-3752. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3567629&tool=pmcentrez&rendertype=abstract Accessed 2014 July 17.
78. Heck DE, Shakarjian M, Kim HD, Laskin JD, Vetrano AM (2010) Mechanisms of oxidant generation by catalase. Ann N Y Acad Sci 1203: 120-125. Available: http://www.ncbi.nlm.nih.gov/pubmed/20716293 Accessed 2013 May 19.
79. Chaudiere J, Wilhelmsen EC, Tappel a L (1984) Mechanism of selenium-glutathione peroxidase and its inhibition by mercaptocarboxylic acids and other mercaptans. J Biol Chem 259: 1043-1050. Available: http://www.ncbi.nlm.nih.gov/pubmed/6693375
80. Zhao G, Usui ML, Underwood RA, Singh PK, James GA, et al. (2012) Time course study of delayed wound healing in a biofilm-challenged diabetic mouse model. Wound Repair Regen 20: 342-352. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3349451&tool=pmcentrez&ren dertype=abstract Accessed 2013 November 6.
81. Costerton JW, Stewart PS, Greenberg EP (1999) Bacterial biofilms: a common cause of persistent infections. Science 284: 1318-1322. Available: http://www.ncbi.nlm.nih.gov/pubmed/10334980 Accessed 2013 September 30.
82. Scales BS, Huffnagle GB (2013) The microbiome in wound repair and tissue fibrosis. J Pathol 229: 323-331. Available: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3631561&tool=pmcentrez&rendertype=abstract Accessed 2014 January 22.
83. Youmans GP, Paterson PY, Sommers HM (1980) The Biologic and clinical basis of infectious diseases. J Am Med Assoc 244: 849. Available: http://books.google.com/books/about/The-Biologic-and-clinical-basis-of-infec.html?id=CaBrAAAAMAAJ&pgis=1 Accessed 2014 January 29.
84. Dalben M, Varkulja G, Basso M, Krebs VLJ, Gibelli M a, et al. (2008) Investigation of an outbreak of Enterobacter cloacae in a neonatal unit and review of the literature. J Hosp Infect 70: 7-14. Available: http://www.ncbi.nlm.nih.gov/pubmed/18632183 Accessed 2013 September 30.
85. Madsen SM, Westh H, Danielsen L, Rosdahl VT (1996) Bacterial colonization and healing of venous leg ulcers. APMIS 104: 895-899. Available: http://www.ncbi.nlm.nih.gov/pubmed/9048868
86. Gerding DN (1995) Foot infections in diabetic patients: the role of anaerobes. Clin Infect Dis 20 Suppl 2: S283-8. Available: http://www.ncbi.nlm.nih.gov/pubmed/7548576 Accessed 2013 September 30.
87. Sapico FL, Witte JL, Canawati HN, Montgomerie JZ, Bessman AN (n.d.) The infected foot of the diabetic patient: quantitative microbiology and analysis of clinical features. Rev Infect Dis 6 Suppl 1: S171-6. Available: http://www.ncbi.nlm.nih.gov/pubmed/6718934 Accessed 2013 September 30.
88. Sharp CS, Bessman AN, Wagner FW, Garland D (n.d.) Microbiology of deep tissue in diabetic gangrene. Diabetes Care 1: 289-292. Available: http://www.ncbi.nlm.nih.gov/pubmed/720182 Accessed 2013 September 30.
89. Hansson C, Hoborn J, Möller A, Swanbeck G (1995) The microbial flora in venous leg ulcers without clinical signs of infection. Repeated culture using a validated standardised microbiological technique. Acta Derm Venereol 75: 24-30. Available: http://www.ncbi.nlm.nih.gov/pubmed/7747531 Accessed 2013 September 30.
90. Gjødsbøl K, Christensen JJ, Karlsmark T, Jørgensen B, Klein BM, et al. (2006) Multiple bacterial species reside in chronic wounds: a longitudinal study. Int Wound J 3: 225-231. Available: http://www.ncbi.nlm.nih.gov/pubmed/16984578 Accessed 2012 November 9.
91. Brook I, Frazier EH (1998) Aerobic and anaerobic microbiology of chronic venous ulcers. Int J Dermatol 37: 426-428. Available: http://www.ncbi.nlm.nih.gov/pubmed/9646126
92. Stapley R, Owusu BY, Brandon A, Cusick M, Rodriguez C, et al. (2012) Erythrocyte storage increases rates of NO and nitrite scavenging: implications for transfusion-related toxicity. Biochem J 446: 499-508. Available: http://www.ncbi.nlm.nih.gov/pubmed/22720637 Accessed 2013 May 22.
93. DeLoney CR, Schiller NL (1999) Competition of Various beta -Lactam Antibiotics for the Major Penicillin-Binding Proteins of Helicobacter pylori: Antibacterial Activity and Effects on Bacterial Morphology. Antimicrob Agents Chemother 43: 2702-2709. Available: http://aac.asm.org/content/43/11/2702.long Accessed 2014 February 17.