A rabbit model of implant‑related osteomyelitis inoculated with biofilm after open femoral fracture

Zhang,Xiang; Ma,Yun‑Fei; Wang,Lei; Jiang,Nan; Qin,Cheng‑He; Hu,Yan‑Jun; Yu,Bin

doi:10.3892/etm.2017.5138

A rabbit model of implant‑related osteomyelitis inoculated with biofilm after open femoral fracture

Authors:
- Xiang Zhang
- Yun‑Fei Ma
- Lei Wang
- Nan Jiang
- Cheng‑He Qin
- Yan‑Jun Hu
- Bin Yu
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Affiliations: Department of Orthopaedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China
Published online on: September 19, 2017 https://doi.org/10.3892/etm.2017.5138
Pages: 4995-5001
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Currently, animal models used in research on implant‑associated osteomyelitis primarily use intramedullary fixation and initial inoculum of planktonic bacterial cells. However, these techniques have certain limitations, including lack of rotational stability and instable inoculation. To improve these models, the present study aimed to establish a novel rabbit model of implant‑associated osteomyelitis using biofilm as the initial inoculum following plate fixation of the femoral fracture. A total of 24 New Zealand White rabbits were randomly divided into two equal groups. Osteotomy was performed at the right femoral shaft using a wire saw following fixation with a 5‑hole stainless steel plate. The plates were not colonized with bacteria in group 1, but colonized with a biofilm of Staphylococcus aureus (American Type Culture Collection, 25923) in group 2. All the rabbits were sacrificed after 21 days for clinical, X‑ray, micro‑computed tomography and histological assessments of the severity of osteomyelitis. Scanning electron microscopy and confocal laser scanning microscopy were used for biofilm assessment. In group 2, pus formation, periosteal reaction, cortical destruction and absorption were observed in all the rabbits and biofilm formation was observed on all the plates. However, no pus formation was observed except for a slight inflammatory response and all the plates appeared clean without infection in group 1. The differences between the two groups were statistically significant regarding histologic scores and semi‑quantification of the bacteria on the plates (P<0.001). In the present study, a novel rabbit model of infection following internal plate fixation of open fracture was successfully established, providing a novel tool for the study of implant‑associated osteomyelitis.

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1	Arciola CR, Campoccia D, Ehrlich GD and Montanaro L: Biofilm-based implant infections in orthopaedics. Adv Exp Med Biol. 830:29–46. 2015. View Article : Google Scholar : PubMed/NCBI
2	Metsemakers WJ, Kuehl R, Moriarty TF, Richards RG, Verhofstad MH, Borens O, Kates S and Morgenstern M: Infection after fracture fixation: Current surgical and microbiological concepts. Injury. Sep 11–2016.(Epub ahead of print). View Article : Google Scholar
3	Worlock P, Slack R, Harvey L and Mawhinney R: An experimental model of post-traumatic osteomyelitis in rabbits. Br J Exp Pathol. 69:235–244. 1988.PubMed/NCBI
4	Reizner W, Hunter JG, O'Malley NT, Southgate RD, Schwarz EM and Kates SL: A systematic review of animal models for Staphylococcus aureus osteomyelitis. Eur Cell Mater. 27:196–212. 2014. View Article : Google Scholar : PubMed/NCBI
5	Donné J and Dewilde S: The challenging world of biofilm physiology. Adv Microb Physiol. 67:235–292. 2015. View Article : Google Scholar : PubMed/NCBI
6	Costerton JW: Cystic fibrosis pathogenesis and the role of biofilms in persistent infection. Trends Microbiol. 9:50–52. 2001. View Article : Google Scholar : PubMed/NCBI
7	Montanaro L, Speziale P, Campoccia D, Ravaioli S, Cangini I, Pietrocola G, Giannini S and Arciola CR: Scenery of Staphylococcus implant infections in orthopedics. Future Microbiol. 6:1329–1349. 2011. View Article : Google Scholar : PubMed/NCBI
8	Trampuz A and Zimmerli W: Diagnosis and treatment of infections associated with fracture-fixation devices. Injury. 37 Suppl 2:S59–S66. 2006. View Article : Google Scholar : PubMed/NCBI
9	Williams DL and Costerton JW: Using biofilms as initial inocula in animal models of biofilm-related infections. J Biomed Mater Res B Appl Biomater. 100:1163–1169. 2012. View Article : Google Scholar : PubMed/NCBI
10	Williams DL, Haymond BS, Woodbury KL, Beck JP, Moore DE, Epperson RT and Bloebaum RD: Experimental model of biofilm implant-related osteomyelitis to test combination biomaterials using biofilms as initial inocula. J Biomed Mater Res A. 100:1888–1900. 2012. View Article : Google Scholar : PubMed/NCBI
11	Trampuz A, Piper KE, Jacobson MJ, Hanssen AD, Unni KK, Osmon DR, Mandrekar JN, Cockerill FR, Steckelberg JM, Greenleaf JF and Patel R: Sonication of removed hip and knee prostheses for diagnosis of infection. N Engl J Med. 357:654–663. 2007. View Article : Google Scholar : PubMed/NCBI
12	Monsen T, Lövgren E, Widerström M and Wallinder L: In vitro effect of ultrasound on bacteria and suggested protocol for sonication and diagnosis of prosthetic infections. J Clin Microbiol. 47:2496–2501. 2009. View Article : Google Scholar : PubMed/NCBI
13	Rochford ET, Sabaté Brescó M, Zeiter S, Kluge K, Poulsson A, Ziegler M, Richards RG, O'Mahony L and Moriarty TF: Monitoring immune responses in a mouse model of fracture fixation with and without Staphylococcus aureus osteomyelitis. Bone. 83:82–92. 2016. View Article : Google Scholar : PubMed/NCBI
14	Glass GE, Chan JK, Freidin A, Feldmann M, Horwood NJ and Nanchahal J: TNF-alpha promotes fracture repair by augmenting the recruitment and differentiation of muscle-derived stromal cells. Proc Natl Acad Sci USA. 108:1585–1590. 2011. View Article : Google Scholar : PubMed/NCBI
15	Kishor C, Mishra RR, Saraf SK, Kumar M, Srivastav AK and Nath G: Phage therapy of staphylococcal chronic osteomyelitis in experimental animal model. Indian J Med Res. 143:87–94. 2016. View Article : Google Scholar : PubMed/NCBI
16	Odekerken JC, Arts JJ, Surtel DA, Walenkamp GH and Welting TJ: A rabbit osteomyelitis model for the longitudinal assessment of early post-operative implant infections. J Orthop Surg Res. 8:382013. View Article : Google Scholar : PubMed/NCBI
17	Moriarty TF, Campoccia D, Nees SK, Boure LP and Richards RG: In vivo evaluation of the effect of intramedullary nail microtopography on the development of local infection in rabbits. Int J Artif Organs. 33:667–675. 2010.PubMed/NCBI
18	Norden CW: Experimental osteomyelitis. I. A description of the model. J Infect Dis. 122:410–418. 1970. View Article : Google Scholar : PubMed/NCBI
19	Shiels SM, Bedigrew KM and Wenke JC: Development of a hematogenous implant-related infection in a rat model. BMC Musculoskelet Disord. 16:2552015. View Article : Google Scholar : PubMed/NCBI
20	Arens S, Kraft C, Schlegel U, Printzen G, Perren SM and Hansis M: Susceptibility to local infection in biological internal fixation. Experimental study of open vs minimally invasive plate osteosynthesis in rabbits. Arch Orthop Trauma Surg. 119:82–85. 1999. View Article : Google Scholar : PubMed/NCBI
21	Alt V, Lips KS, Henkenbehrens C, Muhrer D, Cavalcanti Oliveira MC, Sommer U, Thormann U, Szalay G, Heiss C, et al: A new animal model for implant-related infected non-unions after intramedullary fixation of the tibia in rats with fluorescent in situ hybridization of bacteria in bone infection. Bone. 48:1146–1153. 2011. View Article : Google Scholar : PubMed/NCBI
22	Svensson S, Trobos M, Hoffman M, Norlindh B, Petronis S, Lausmaa J, Suska F and Thomsen P: A novel soft tissue model for biomaterial-associated infection and inflammation-Bacteriological, morphological and molecular observations. Biomaterials. 41:106–121. 2015. View Article : Google Scholar : PubMed/NCBI
23	dos Reis JA Jr, de Carvalho FB, Trindade RF, de Assis PN, de Almeida PF and Pinheiro AL: A new preclinical approach for treating chronic osteomyelitis induced by Staphylococcus aureus: In vitro and in vivo study on photodynamic antimicrobial therapy (PAmT). Laser Med Sci. 29:789–795. 2014. View Article : Google Scholar
24	Gnanadhas DP, Elango M, Janardhanraj S, Srinandan CS, Datey A, Strugnell RA, Gopalan J and Chakravortty D: Successful treatment of biofilm infections using shock waves combined with antibiotic therapy. Sci Rep. 5:174402015. View Article : Google Scholar : PubMed/NCBI
25	Wimpenny J, Manz W and Szewzyk U: Heterogeneity in biofilms. FEMS Microbiol Rev. 24:661–671. 2000. View Article : Google Scholar : PubMed/NCBI
26	Christensen GD, Simpson WA, Younger JJ, Baddour LM, Barrett FF, Melton DM and Beachey EH: 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. 1985.PubMed/NCBI
27	Vassena C, Fenu S, Giuliani F, Fantetti L, Roncucci G, Simonutti G, Romanò CL, De Francesco R and Drago L: Photodynamic antibacterial and antibiofilm activity of RLP068/Cl against Staphylococcus aureus and Pseudomonas aeruginosa forming biofilms on prosthetic material. Int J Antimicrob Agents. 44:47–55. 2014. View Article : Google Scholar : PubMed/NCBI
28	Drago L, Agrappi S, Bortolin M, Toscano M, Romanò C and De Vecchi E: How to study biofilms after microbial colonization of materials used in orthopaedic implants. Int J Mol Sci. 17:2932016. View Article : Google Scholar : PubMed/NCBI