Construction of a rabbit model of knee prosthesis infection: environmental factors in vivo affect Staphylococcus epidermidis and biofilms

doi:10.3969/j.issn.2095-4344.2015.39.002

Abstract

Abstract:

BACKGROUND: Bacterial biofilm is the main cause of the infection of the prosthesis. In vitro experiments confirmed that hypertonic sodium chloride and ethanol can apparently promote the formation of staphylococcal biofilms. There are no reports on the effects of ethanol and hypertonic environment surrounding the prosthesis on the formation of biofilms.
OBJECTIVE: To evaluate the effects of different environment factors surrounding the prosthesis on the growth of Staphylococcus epidermidis and bacterial biofilm formation after replacement.
METHODS: White rabbit models infected with Staphylococcus epidermidis on the prosthesis were established, and were randomly divided into hypertonic sodium chloride, ethanol and control groups (n=15). The bacteria were injected with 0.1 mL 4% sodium chloride and 4% ethanol into the knee of rabbits in the hypertonic sodium chloride
and ethanol groups. The rabbits were injected with 0.1 mL 0.9% sodium chloride in the control group. Three rabbits were sacrificed at 2, 4, 6, 8 and 16 days after inoculated with bacteria. Synovial fluid, prosthesis and tissue surrounding infection were obtained. Bacterium was cultured to extract total RNA. The ica operon transcription levels were detected in the gene levels. Adhesion of bacteria on the surface of the prosthesis was observed using a scanning electron microscope. Tissues surrounding the prosthesis were observed using hematoxylin-eosin staining.
RESULTS AND CONCLUSION: Histological examination revealed that inflammatory cell infiltration was observed in all the rabbits at 4 days after injection. Colony formation was found at 16 days after injection. At 6 days after injection, inflammatory cell infiltration was observed in the ethanol and control groups. Scanning electron microscope showed that compared with the control group, the bacteria adhered to the prosthetic surface became more in the hypertonic sodium chloride and ethanol groups at 6, 8 and 16 days (P < 0.05). At 6, 8 and 16 days, the expression of icaA mRNA was significantly higher in the hypertonic sodium chloride and ethanol groups than in the control group (P < 0.05). These data showed that the environment factors could affect the growth of Staphylococcus epidermidis and bacterial biofilm formation.

中国组织工程研究杂志出版内容重点：人工关节；骨植入物；脊柱；骨折；内固定；数字化骨科；组织工程

Key words: Prosthesis Implantation, Infection, Biofilms, Staphylococcus epidermidis

CLC Number:

R318

Shen You-liang, Zhu Tong-e, Zhang Jing-jing, Qi Chao, Yu Teng-bo. Construction of a rabbit model of knee prosthesis infection: environmental factors in vivo affect Staphylococcus epidermidis and biofilms[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(39): 6240-6245.

Figures/Tables 5

References

[1] Trampuz A,Widmer AF.Infection associated with orthopedic implants,Curr Opin Infect Dis. 2006;19(4):349-354.
[2] Shahi A, Parvizi J. Prevention of Periprosthetic Joint Infection. Arch Bone Jt Surg.2015;3(2):72-81.
[3] Bergin C,Speroni KG, Travis T,et al.Effect of preoperative incentive spirometry patient education on penitent outcomes in the knee and hip joint replacement population. J Perianesth Nurs. 2014;29(1):20-27.
[4] Deirmengian C, Kardos K, Kilmartin P, et al. Combined measurement of synovial fluidα-Defensin and C-reactive protein levels: highly accurate for diagnosing periprosthetic joint infection. J Bone Joint Surg Am. 2014; 96(17):1439- 1445.
[5] Vuong C,Otto M. Staphylococcus epidermidis infections. Microbes Infeet. 2002;4:481-489．
[6] Hartford O,O’Brien L,Sehofield K,et al. The Fbe(SdrG) protein oI Staphylococcus epidermidis HB promotes bacterial adherenee to fibrinogen. Microbiology. 2001;147: 2545-2552.
[7] 吴秋季,李强,张绍,等.手术室环境预防人工关节置换后感染的有效性[J].中国组织工程研究,2013,17(39):2095-4344.
[8] Zhuang YF, Wei MY. Risk factors of incision in fections after artificial total knee arthroplasty and counter measures. Chin J Nosocomiol. 2013;23(14):3390-3391.
[9] Knobloch JK, Bartscht K, Sabottke A, et al. Biofilm formation by Staphylococcus epidermidis depends on functional RsbU, an activator of the sigB operon: differential activation mechanisms due to ethanol and salt stress. J Bacteriol. 2001; 183(8):2624-2633.
[10] Milisavljevic V, Tran LP, Batmalle C, et al. Benzyl alcohol and ethanol can enhance the pathogenic potential of clinical Staphylococcus epidermidis strains. Am J Infect Control. 2008;36(8):552-558.
[11] 王志酬,曹力,龚时国,等.兔膝关节置换术后感染模型的建立[J].中华实验外科杂志,2010,27(2):258-260
[12] Gallo J, Kolár M, Florschütz AV, et al. In vitro testing of gentamicin-vancomycin loaded bone cement to prevent prosthetic joint infection. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2005;149(1):153-158.
[13] Thalhamer T, McGrath M, Harnett M. MAPKs and their relevance to arthritis and inflammation. Rheumatology. 2008; 47(4):409-414.
[14] Watanabe K, Minowa T, Takeda S, et al. Outcomes of knee arthrodesis following infected total knee arthroplasty: a retrospective analysis of 8 cases. Mod Rheumatol. 2014;24: 243-249.
[15] 毕衡.尺桡骨开放性骨折伤口感染的临床特征[J].中国骨与关节损伤杂志,2013,28(1);89-90.
[16] Knobloch JK, Jager S, Homtkotte MA, et al. RsbU-dependentegulation of Staphylococcus epidermldis biofilm formation is mediated via the alternative sigma factor sigmaB by repression of the negative regulator gene icaR. Infect Immun. 2004;72(7):3838-3848.
[17] Christensen GD, Simpson WA, Bisno AL, et al. Adherence of slim-produceing Staphylococcus epidermidis to smooth sufaces. Infect Immun. 1982;37(1):318-326.
[18] Del Pozo JL, Patel R. Clinical practice. Infection associated with prosthetic joints. N Engl J Med. 2009;361:787-794.
[19] Bémer P, Plouzeau C, Tande D, et al. Evaluation of 16S Rrna gene PCR sensitivity and specificity for diagnosis of prosthetic joint infection: a prospective multicenter cross-sectional study. J Clin Microbiol. 2014; 10:3583-3589.
[20] Christiane G, Angelika K. Characterization of the N-acetylglucosaminyl transferase activity involved in the biosynthesis of the Staphylococcus epidermidis polysaccharide intercellular adhesin. J Biol Chem. 1998; 279(29):18586-18593.
[21] 王子明,王爱民,唐桂阳.骨水泥型人工股骨头置换和术后感染模型的建立[J].中华实验外科杂志,2007,24(4):509.
[22] Conlon KM, Humphreys H, O'Gara JP. icaR encodes a transcriptional repressor involved in environmental regulation of ica operon expression and biofilm formation in Staphylococcus epidermidis. J Bacteriol. 2002;184(16): 4400-4408.
[23] Stewart PS, Costerton JW. Antibiotic resistance of bacteria in biofilms. Lancet. 2001;358:135-138
[24] Zmistowski B, Della Valle C, Bauer TW, et al. Diagnosis of periprosthetic joint infection. J Arthroplasty. 2014; 29(2 Suppl): 77-83.
[25] Bartok B, Firestein GS. Fibroblast-like synoviocytes: key effector cells in rheumatoid arthritis. Immunol Rev. 2010; 233(1):233-255.
[26] Broekhuizen CA, de Boer L, Schipper K, et al. Peri-implant tissue is an important niche for Staphylococcus epidermidis in experimental biomaterial-associated infection in mice. Infect Immue. 2007;75(3):1129-1136.
[27] Mack D, Davies AP, Harris LG, et al. Microbial interactions in Staphylococcus epidermidis biofilms. Anal Bioanal Chem. 2007;387(2):399-408.
[28] Peel TN, Dowsey MM, Buising KL, et al. Cost analysis of debridement and retention for management of prosthetic joint infection. Clin Microbiol Infect.2013;19: 181-186.
[29] Milisavljevic V, Tran LP, Batmalle C, et al. Benzyl alcohol and ethanol can enhance the pathogenic potential of clinical Staphylococcus epidermidis strains American journal of infection control. Am J Infect Control. 2008;36(8):552-558.
[30] Yang ZG, Chen WP, Wu LD. Effectiveness and safety of tranexamic acid in reducing blood loss in total knee arthroplasty :a meta-analysis. J Bone Joint Surg Am. 2012; 94(13):1153-1159.
[31] Greenwood-Quaintance KE, Uhl JR, Hanssen AD, et al. Diagnosis of prosthetic joint infection by use of PCR-electrospray ionization mass spectrometry. J Clin Microbiol. 2014;52:642-649.
[32] Ghanem E, Parvizi J, Burnett RS, et al. Cell count and differential of aspirated fluid in the diagnosis of infection at the site of total knee arthroplasty. J Bone Joint Surg Am. 2008; 90:1637-1643.