Structural Evolution of Staphylococcus Aureus Biofilm on the Surface of Orthopedic Titanium Plate in Acute Pha
1.Department of Orthopaedics，Affiliated Zhangzhou Hospital of Fujian Medical University
2.Fujian Medical University
3.Department of Orthopaedics，Second People’s Hospital of Nanyang
Abstract：Objective To observe the biofilm structure of Staphylococcus aureus on the surface of titanium plates at different time points during the acute phase (4 weeks) by laser confocal microscopy and scanning electron microscopy.Methods Staphylococcus aureus was cultured in vitro with orthopaedic titanium plates to form biofilms at different time points.After 1 week，2 weeks，3 weeks and 4 weeks of culture，the titanium plates were taken out，the biofilms points were stained with protein and propidium iodide dyes according to the different time points.After staining，the morphological structure of bacterial biofilms was observed by laser confocal microscope and scanning electron microscope.Results Laser confocal images and electron microscope images of biofilms at different time points were obtained.It was observed that a small amount of extracellular polysaccharide polymer was formed in bacteria at 1 week，and the internal spatial structure was disordered，which indicating the formation of early biofilms.With extension of the culture time，the amount of bacterial exopolysaccharide polymer gradually increased，the internal gaps contact with pores，and the spatial structure became complicated gradually.At 4 weeks，a large number of exopolysaccharide polymers were formed，and the inner membrane structure was wellstructured.It meant that a mature biofilm was formed.The number of viable bacteria in each time point had no difference(P＞0.05)，and the biofilm thickness between each time point had significant difference(P＜0.05). Conclusion Staphylococcus aureus bacterial biofilm has difference between early and late phase.It is a dynamic evolution processfrom from early biofilm to late mature biofilm，which is manifested in the maturity and stability of bacterial biofilm exopolysaccharide polymers and internal spatial structures.
［1］Pavoni GL，Giannella M，Falcone M，et al.Conservative medical therapy of prosthetic joint infections：Retrospective analysis of an 8-year experience［J］.Clin Microbiol Infect，2004，10(9)：831-837.
［2］Springer BD，Scuderi GR.Evaluation and management of the infected total knee arthroplasty［J］.Instr Course Lect，2013(62)：349-361.
［3］Inabathula A，Dilley JE，Ziemba-Davis M，et al.Extended oral antibiotic prophylaxis in high-risk patients substantially reduces primary total hip and knee arthroplasty 90-day infection rate［J］.J Bone Joint Surg(Am)，2018，100(24)：2103-2109.
［4］Montanaro L，Speziale P，Campoccia D，et al.Scenery of staphylococcus implant infections in orthopedics［J］.Future Microbiol，2011，6(11)：1329-1349.
［5］Komnos GA，Manrique J，Goswami K，et al.Periprosthetic joint infection in patients who have multiple prostheses in place：What should be done with the silent prosthetic joints［J］.J Bone Joint Surg(Am)，2020，102(13)：1160-1168.
［6］Boclé H，Lavigne JP，Cellier N，et al.〖JP2〗Effectiveness of early switching from intravenous to oral antibiotic therapy in Staphylococcus aureus prosthetic bone and joint or orthopedic metalware-associated infections.BMC Musculoskelet Disord，2021，22(1)：315.
［7］Zimmerli W，Trampuz A，Ochsner PE.Prostheticjoint infections［J］.N Engl J Med，2004，351(16)：1645-1654.
［8］Schwartz AM，Farley KX，Guild GN，et al.Projections and epidemiology of revision hip and knee arthroplasty in the united states to 2030［J］.J Arthroplasty，2020，35(6S)：S79-S85.
［9］H-iby N，Bjarnsholt T，Moser C，et al.ESCMID guideline for the diagnosis and treatment of biofilm infections 2014［J］.Clin Microbiol Infect，2015，21(Suppl 1)：S1-25.
［10］李舟，杨明坤，何坤林，等.医用钛板表面涂层2-MPC对细菌生物膜形成的影响［J］.实用骨科杂志，2020，26(6)：513-516.
［11］Pant N，Eisen DP.Non-antimicrobial adjuvant strategies to tackle biofilm-related staphylococcus aureus prosthetic joint infections［J］.Antibiotics (Basel)，2021，10(9)：1060.
［12］Zimmerli W.Clinical presentation and treatment of orthopaedic implant-associated infection［J］.J Intern Med，2014，276(2)：111-119.
［13］Scheper H，Gerritsen LM，Pijls BG，et al.Outcome of debridement，antibiotics，and implant retention for staphylococcal hip and knee prosthetic joint infections，focused on rifampicin use：A systematic review and meta-analysis［J］.Open Forum Infect Dis，2021，8(7)：298.
［14］Sousa A，Carvalho A，Pereira C，et al.Economic impact of prosthetic joint infection-an evaluation within the portuguese national health system［J］.J Bone Jt Infect，2018，3(4)：197-202.
［15］Unnanuntana A，Bonsignore L，Shirtliff ME，et al.The effects of farnesol on Staphylococcus aureus biofilms and osteoblasts.An in vitro study［J］.J Bone Joint Surg (Am)，2009，91(11)：2683-2692.
［16］Nostro A，Cellini L，Ginestra G，et al.Staphylococcal biofilm formation as affected by type acidulant［J］.APMIS，2014，122(7)：648-653.
［17］Nishitani K，Sutipornpalangkul W，de Mesy Bentley KL，et al.Quantifying the natural history of biofilm formation in vivo during the establishment of chronic implant-associated Staphylococcus aureus osteomyelitis in mice to identify critical pathogen and host factors.J Orthop Res，2015，33(9)：1311-1319.
［18］Takahashi C，Muto S，Yamamoto H.A microscopy method for scanning transmission electron microscopy imaging of the antibacterial activity of polymeric nanoparticles on a biofilm with an ionic liquid［J］.J Biomed Mater Res B Appl Biomater，2017，105(6)：1432-1437.
［19］罗延智，曾华，张志宏，等.聚维酮碘消毒剂对金黄色葡萄球菌生物膜的体外影响研究［J］.中华医院感染学杂志，2017，27(21)：5010-5013.
［20］陈明礼，罗延智，曾文容，等.碘伏对金黄色葡萄球菌生物膜的抗菌作用［J］.中华创伤杂志，2020，36(8)：736-742.
［21］Fridkin SK，Hageman JC，Morrison M，et al.Methicillinresistant Staphylococcus aureus disease in three communities［J］.N Engl J Med，2005，352(14)：1436-1444.
［22］Hota B，Ellenbogen C，Hayden MK，et al.Community-associated methicillin-resistant Staphylococcus aureus skin and soft tissue infections at a public hospital：Do public housing and incarceration amplify transmission?［J］.Arch Intern Med，2007，167(10)：1026-1033.
［23］Manasherob R，Mooney JA，Lowenberg DW，et al.Tolerant small-colony variants form prior to resistance within a staphylococcus aureus biofilm based on antibiotic selective pressure［J］.Clin Orthop Relat Res，2021(479)：1471-1481.
［24］Mukherjee S，Bassler BL.Bacterial quorum sensing in complex and dynamically changing environments［J］.Nat Rev Microbiol，2019，17(6)：371-382.
［25］Gupta TT，Gupta NK，Pestrak MJ，et al.Staphylococcus aureus aggregates on orthopedic materials under varying levels of shear stress［J］.Appl Environ Microbiol，2020，86(19)：e1234.
［26］Yu J，Jiang F，Zhang F，et al.Virtual screening for novel sara inhibitors to prevent biofilm formation of staphylococcus aureus in prosthetic joint infections［J］.Front Microbiol，2020 (11)：587175.
［27］Stoodley P，Nistico L，Johnson S，et al.Direct demonstration of viable Staphylococcus aureus biofilms in an infected total joint arthroplasty.A case report［J］.J Bone Joint Surg(Am)，2008，90(8)：1751-1758.
［28］Flemming HC，Neu TR，Wozniak DJ.The EPS matrix：The “house of biofilm cells”［J］.J Bacteriol，2007，189(22)：7945-7947.
［29］周浩，陈歌，尹一然，等.保留假体清创联合局部万古霉素治疗全膝关节置换术后早期感染［J］.实用骨科杂志，2020，26(5)：404-407；411.
［30］Grammatopoulos G，Kendrick B，McNally M，et al.Outcome following debridement，antibiotics，and implant retention in hip periprosthetic joint infection-an 18-year experience［J］.J Arthroplasty，2017，32(7)：2248-2255.
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