Adaptive expression of biofilm regulators and adhesion factors of Staphylococcus aureus during acute wound infection under the treatment of negative pressure wound therapy in vivo

Li,Tongtong; Wang,Guoqi; Yin,Peng; Li,Zhirui; Zhang,Lihai; Tang,Peifu

doi:10.3892/etm.2020.8679

Adaptive expression of biofilm regulators and adhesion factors of Staphylococcus aureus during acute wound infection under the treatment of negative pressure wound therapy in vivo

Authors:
- Tongtong Li
- Guoqi Wang
- Peng Yin
- Zhirui Li
- Lihai Zhang
- Peifu Tang
View Affiliations

Affiliations: Department of Orthopedics, Tianjin Hospital, Tianjin 300211, P.R. China, Department of Pediatrics, Chinese People's Liberation Army (PLA) General Hospital, Beijing 100853, P.R. China, Department of Orthopedics, Beijing Chao‑Yang Hospital, Beijing 100043, P.R. China, Department of Orthopedics, Chinese People's Liberation Army (PLA) General Hospital, Beijing 100853, P.R. China
Published online on: April 23, 2020 https://doi.org/10.3892/etm.2020.8679
Pages: 512-520
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Negative pressure wound therapy (NPWT) is gaining acceptance as a physical therapy for a wide variety of infected wounds. To gain insight into the response of bacteria to NPWT in vivo, the adaptive expression of biofilm regulators and adhesion factors of Staphylococcus aureus (S. aureus), the most frequently isolated pathogen in the clinic, during acute wound infection was investigated. A 3 cm full‑thickness dermal wound was created on each side of a rabbit back and inoculated with green fluorescent protein‑labeled S. aureus. NPWT was initiated at 6 h post inoculation, with the wound on the contralateral side as the untreated self‑control. The wounds were subjected to a 28 day observation period. Histological analysis, laser scanning confocal microscopy and scanning electron microscopy revealed a transition of S. aureus to a free‑living phenotype in tissues treated with NPWT, compared with microcolonies in untreated wounds. Viable bacteria counts showed a modest reduction in the bioburden of NPWT group on day 8 (P<0.001), with ~1x106 colony‑forming units/g tissue. Transcript analysis of biofilm‑ and colonization‑related genes were investigated using reverse transcription‑quantitative PCR on postoperative days 1, 2, 4 and 8. The poly‑beta‑1,6‑N‑acetyl‑D‑glucosamine synthase locus and holin‑like protein CidA/antiholin‑like protein LrgA network were less active in the NPWT group compared with the untreated control group. Accordingly, the expression profile switched to an elevated expression of the adhesive factors UDP‑phosphate N‑acetylglucosaminyl 1‑phosphate transferase (at days 0‑4) and fibronectin‑binding protein A and iron‑regulated surface determinant protein A at >4 days during both stages of colonization. Meanwhile, low expression levels of the effector molecule (RNAIII) of the accessory gene regulator type I (agr) system was detected in NPWT group, suggesting that the bacterial density in NPWT‑treated wounds was under the threshold for agr activation, thus not leading to an active and invasive infection. The wounds treated by NPWT healed completely on day 28, compared with an average of an 8.11% defect area in the control group (P<0.001). The results of the current study indicated that S. aureus responds to NPWT by regulating gene expression, manifesting a decrease in biofilm formation and an increase in bacterial colonization in vivo, which potentially benefits the wound repair and healing process.

View Figures

View References

1	Marimuthu K, Eisenring MC, Harbarth S and Troillet N: Epidemiology of Staphylococcus aureus surgical site infections. Surg Infect (Larchmt). 17:229–235. 2016.PubMed/NCBI View Article : Google Scholar
2	Heilmann C: Adhesion mechanisms of staphylococci. Adv Exp Med Biol. 715:105–123. 2011.PubMed/NCBI View Article : Google Scholar
3	Speziale P, Pietrocola G, Rindi S, Provenzano M, Provenza G, Di Poto A, Visai L and Arciola CR: Structural and functional role of Staphylococcus aureus surface components recognizing adhesive matrix molecules of the host. Future Microbiol. 4:1337–1352. 2009.PubMed/NCBI View Article : Google Scholar
4	Weidenmaier C, Kokai-Kun JF, Kulauzovic E, Kohler T, Thumm G, Stoll H, Götz F and Peschel A: Differential roles of sortase-anchored surface proteins and wall teichoic acid in Staphylococcus aureus nasal colonization. Int J Med Microbiol. 298:505–513. 2008.PubMed/NCBI View Article : Google Scholar
5	Burian M, Rautenberg M, Kohler T, Fritz M, Krismer B, Unger C, Hoffmann WH, Peschel A, Wolz C and Goerke C: Temporal expression of adhesion factors and activity of global regulators during establishment of Staphylococcus aureus nasal colonization. J Infect Dis. 201:1414–1421. 2010.PubMed/NCBI View Article : Google Scholar
6	Edwards R and Harding KG: Bacteria and wound healing. Curr Opin Infectious Dis. 17:91–96. 2004.PubMed/NCBI View Article : Google Scholar
7	Poutahidis T, Kearney SM, Levkovich T, Qi P, Varian BJ, Lakritz JR, Ibrahim YM, Chatzigiagkos A, Alm EJ and Erdman SE: Microbial symbionts accelerate wound healing via the neuropeptide hormone oxytocin. PLoS One. 8(e78898)2013.PubMed/NCBI View Article : Google Scholar
8	Lee K, Lee JH, Ryu SY, Cho MH and Lee J: Stilbenes reduce Staphylococcus aureus hemolysis, biofilm formation, and virulence. Foodborne Pathog Dis. 11:710–717. 2014.PubMed/NCBI View Article : Google Scholar
9	Bayles KW: The biological role of death and lysis in biofilm development. Nat Rev Microbiol. 5:721–726. 2007.PubMed/NCBI View Article : Google Scholar
10	Otto M: Staphylococcal infections: Mechanisms of biofilm maturation and detachment as critical determinants of pathogenicity. Annu Rev Med. 64:175–188. 2013.PubMed/NCBI View Article : Google Scholar
11	Thoendel M, Kavanaugh JS, Flack CE and Horswill AR: Peptide signaling in the staphylococci. Chem Rev. 111:117–151. 2011.PubMed/NCBI View Article : Google Scholar
12	Schierle CF, De la Garza M, Mustoe TA and Galiano RD: Staphylococcal biofilms impair wound healing by delaying reepithelialization in a murine cutaneous wound model. Wound Repair Regen. 17:354–359. 2009.PubMed/NCBI View Article : Google Scholar
13	Roche ED, Renick PJ, Tetens SP, Ramsay SJ, Daniels EQ and Carson DL: Increasing the presence of biofilm and healing delay in a porcine model of MRSA-infected wounds. Wound Repair Regen. 20:537–543. 2012.PubMed/NCBI View Article : Google Scholar
14	Anagnostakos K and Mosser P: Negative pressure wound therapy in the management of postoperative infections after musculoskeletal tumour surgery. J Wound Care. 23:191–194, 196-197. 2014.PubMed/NCBI View Article : Google Scholar
15	Hahn HM, Lee IJ, Woo KJ and Park BY: Silver-impregnated negative-pressure wound therapy for the treatment of lower-extremity open wounds: A prospective randomized clinical study. Adv Skin Wound Care. 32:370–377. 2019.PubMed/NCBI View Article : Google Scholar
16	Singh DP, Gowda AU, Chopra K, Tholen M, Chang S, Mavrophilipos V, Semsarzadeh N, Rasko Y and Holton Iii L: The effect of negative pressure wound therapy with antiseptic instillation on biofilm formation in a porcine model of infected spinal instrumentation. Wounds. 28:175–180. 2017.PubMed/NCBI
17	Glass GE, Murphy GRF and Nanchahal J: Does negative-pressure wound therapy influence subjacent bacterial growth? A systematic review. J Plast Reconstr Aesthet Surg. 70:1028–1037. 2017.PubMed/NCBI View Article : Google Scholar
18	Ćirković I, Jocić D, Božić DD, Djukić S, Konstantinović N and Radak D: The effect of vacuum-assisted closure therapy on methicillin-resistant Staphylococcus aureus wound biofilms. Adv Skin Wound Care. 31:361–364. 2018.PubMed/NCBI View Article : Google Scholar
19	Islam N, Kim Y, Ross JM and Marten MR: Proteomic analysis of Staphylococcus aureus biofilm cells grown under physiologically relevant fluid shear stress conditions. Proteome Sci. 12(21)2014.PubMed/NCBI View Article : Google Scholar
20	Castro SL, Nelman-Gonzalez M, Nickerson CA and Ott CM: Induction of attachment-independent biofilm formation and repression of Hfq expression by low-fluid-shear culture of Staphylococcus aureus. Appl Environ Microbiol. 77:6368–6378. 2011.PubMed/NCBI View Article : Google Scholar
21	Rosado H, Doyle M, Hinds J and Taylor PW: Low-shear modelled microgravity alters expression of virulence determinants of Staphylococcus aureus. Acta Astronautica. 66:408–413. 2010.
22	Li T, Wang G, Yin P, Li Z, Zhang L, Liu J, Li M, Zhang L, Han L and Tang P: Effect of negative pressure on growth, secretion and biofilm formation of Staphylococcus aureus. Antonie van Leeuwenhoek. 108:907–917. 2015.PubMed/NCBI View Article : Google Scholar
23	Institute of Laboratory Animal Resources (US). Committee on Care, Use of Laboratory Animals, and National Institutes of Health (US). Division of Research Resources: Guide for the care and use of laboratory animals. 8th edition. National Academies Press, Washington, DC, 2011.
24	Seth AK, Geringer MR, Nguyen KT, Agnew SP, Dumanian Z, Galiano RD, Leung KP, Mustoe TA and Hong SJ: Bacteriophage therapy for Staphylococcus aureus biofilm-infected wounds: A new approach to chronic wound care. Plast Reconstr Surg. 131:225–234. 2013.PubMed/NCBI View Article : Google Scholar
25	Liu D, Zhang L, Li T, Wang G, Du H, Hou H, Han L and Tang P: Negative-pressure wound therapy enhances local inflammatory responses in acute infected soft-tissue wound. Cell Biochem Biophys. 70:539–547. 2014.PubMed/NCBI View Article : Google Scholar
26	Liu D, Li Z, Wang G, Li T, Zhang L and Tang P: Virulence analysis of Staphylococcus aureus in a rabbit model of infected full-thickness wound under negative pressure wound therapy. Antonie van Leeuwenhoek. 111:161–170. 2018.PubMed/NCBI View Article : Google Scholar
27	Lalliss SJ, Stinner DJ, Waterman SM, Branstetter JG, Masini BD and Wenke JC: Negative pressure wound therapy reduces pseudomonas wound contamination more than Staphylococcus aureus. J Orthop Trauma. 24:598–602. 2010.PubMed/NCBI View Article : Google Scholar
28	Kamamoto F, Lima ALM, Rezende MR, Mattar-Junior R, Leonhardt MC, Kojima KE and Santos CCD: A new low-cost negative-pressure wound therapy versus a commercially available therapy device widely used to treat complex traumatic injuries: A prospective, randomized, non-inferiority trial. Clinics (Sao Paulo). 72:737–742. 2017.PubMed/NCBI View Article : Google Scholar
29	Morykwas MJ, Argenta LC, Shelton-Brown EI and McGuirt W: Vacuum-assisted closure: A new method for wound control and treatment: Animal studies and basic foundation. Ann Plast Surg. 38:553–562. 1997.PubMed/NCBI View Article : Google Scholar
30	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
31	Omar A, Wright JB, Schultz G, Burrell R and Nadworny P: Microbial Biofilms and Chronic Wounds. Microorganisms. 5(pii:E9)2017.PubMed/NCBI View Article : Google Scholar
32	Swanson EA, Freeman LJ, Seleem MN and Snyder PW: Biofilm-infected wounds in a dog. J Am Vet Med Assoc. 244:699–707. 2014.PubMed/NCBI View Article : Google Scholar
33	Boles BR and Horswill AR: Staphylococcal biofilm disassembly. Trends Microbiol. 19:449–455. 2011.PubMed/NCBI View Article : Google Scholar
34	Licker M, Moldovan R, Hogea E, Muntean D, Horhat F, Bădiţoiu L, Rogobete A, Tirziu E and Zambori C: Microbial biofilm in human health-An updated theoretical and practical insight. Romanian J Laboratory Med. 25:9–26. 2017.
35	Li T, Zhang L, Han LI, Wang G, Yin P, Li Z, Zhang L, Guo Q, Liu D and Tang P: Early application of negative pressure wound therapy to acute wounds contaminated with Staphylococcus aureus: An effective approach to preventing biofilm formation. Exp Ther Med. 11:769–776. 2016.PubMed/NCBI View Article : Google Scholar
36	Cornforth DM, Dees JL, Ibberson CB, Huse HK, Mathiesen IH, Kirketerp-Møller K, Wolcott RD, Rumbaugh KP, Bjarnsholt T and Whiteley M: Pseudomonas aeruginosa transcriptome during human infection 115: E5125-E5134, 2018.
37	Copeland H, Newcombe J, Yamin F, Bhajri K, Mille VA, Hasaniya N, Bailey L and Razzouk AJ: Role of negative pressure wound care and hyperbaric oxygen therapy for sternal wound infections after pediatric cardiac surgery. World J Pediatr Congenit Heart Surg. 9:440–445. 2018.PubMed/NCBI View Article : Google Scholar
38	Tahir S, Malone M, Hu H, Deva A and Vickery K: The Effect of negative pressure wound therapy with and without instillation on mature biofilms in vitro. Materials (Basel). 11(pii: E811)2018.PubMed/NCBI View Article : Google Scholar
39	Song J, Lays C, Vandenesch F, Benito Y, Bes M, Chu Y, Lina G, Romby P, Geissmann T and Boisset S: The expression of small regulatory RNAs in clinical samples reflects the different life styles of Staphylococcus aureus in colonization vs. infection. PLoS One. 7(e37294)2012.PubMed/NCBI View Article : Google Scholar
40	Huang C, Leavitt T, Bayer LR and Orgill DP: Effect of negative pressure wound therapy on wound healing. Curr Probl Surg. 51:301–331. 2014.PubMed/NCBI View Article : Google Scholar
41	Nie B and Yue B: Biological effects and clinical application of negative pressure wound therapy: A review. J Wound Care. 25:617–626. 2016.PubMed/NCBI View Article : Google Scholar
42	Patil PS, Evancho-Chapman MM, Li H, Huang H, George RL, Shriver LP and Leipzig ND: Fluorinated methacrylamide chitosan hydrogel dressings enhance healing in an acute porcine wound model. PLoS One. 13:e0203371. 2018.PubMed/NCBI View Article : Google Scholar