Mitsuhiro Nishizawa, Diane Hu, Hassan Serhan, Bahram Saleh, Ralph S. Marcucio, Kazuhito Morioka
{"title":"具有抗菌特性的超细颗粒材料：一种减少脊柱植入物相关感染的新方法","authors":"Mitsuhiro Nishizawa, Diane Hu, Hassan Serhan, Bahram Saleh, Ralph S. Marcucio, Kazuhito Morioka","doi":"10.1002/jsp2.70091","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Background</h3>\n \n <p>Implant-associated infection remains a serious complication of instrumented spinal surgery. Since biofilm formation on the implant surface is a key factor in the pathogenesis of such infections, current preventive strategies include the use of implants with antibiotic coatings. However, these approaches raise concerns related to antibiotic resistance and cytotoxicity. Ultrafine-grained (UFG) stainless steel, characterized by nanoscale grain sizes, has demonstrated superior mechanical properties and potential antimicrobial effects. This study aimed to evaluate the antibacterial properties of UFG stainless steel implants against <i>Staphylococcus aureus</i> biofilm formation in both in vitro and in vivo models.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>UFG and conventional SUS316L stainless steel wires were incubated with bioluminescent <i>Staphylococcus aureus</i> Xen36 for up to 7 days in vitro. Biofilm formation was assessed using crystal violet (CV) staining, colony-forming unit (CFU) counting, and quantitative PCR (qPCR) for <i>16S rRNA</i> and <i>luxA</i> genes. In vivo antibacterial effects were evaluated using two mouse models: a subcutaneous pouch model and a postoperative spinal implant infection model. Wires were harvested at 1, 3, and 7 days post-infection and analyzed using the same assays.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>In vitro, UFG wires had significantly lower CFU counts than standard wires at 4 h (<i>p</i> = 0.0005), 1 day (<i>p</i> = 0.0001), and 3 days (<i>p</i> = 0.0314). In the subcutaneous pouch model, UFG wires showed significantly reduced bacterial load at Day 1 by CFU (<i>p</i> = 0.011). In the spinal implant model, CFU counts were significantly lower on UFG wires at Day 3 (<i>p</i> = 0.015).</p>\n </section>\n \n <section>\n \n <h3> Conclusions</h3>\n \n <p>UFG stainless steel implants demonstrated a significant reduction in early biofilm formation by <i>Staphylococcus aureus</i> in both in vitro and in vivo, suggesting a delay in the biofilm formation process. These findings support the potential of UFG materials as promising candidates for infection-resistant spinal implants.</p>\n </section>\n </div>","PeriodicalId":14876,"journal":{"name":"JOR Spine","volume":"8 3","pages":""},"PeriodicalIF":3.9000,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jsp2.70091","citationCount":"0","resultStr":"{\"title\":\"Ultrafine-Grained Materials With Antibacterial Properties: A Novel Approach to Reducing Spinal Implant-Associated Infections\",\"authors\":\"Mitsuhiro Nishizawa, Diane Hu, Hassan Serhan, Bahram Saleh, Ralph S. Marcucio, Kazuhito Morioka\",\"doi\":\"10.1002/jsp2.70091\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Background</h3>\\n \\n <p>Implant-associated infection remains a serious complication of instrumented spinal surgery. Since biofilm formation on the implant surface is a key factor in the pathogenesis of such infections, current preventive strategies include the use of implants with antibiotic coatings. However, these approaches raise concerns related to antibiotic resistance and cytotoxicity. Ultrafine-grained (UFG) stainless steel, characterized by nanoscale grain sizes, has demonstrated superior mechanical properties and potential antimicrobial effects. This study aimed to evaluate the antibacterial properties of UFG stainless steel implants against <i>Staphylococcus aureus</i> biofilm formation in both in vitro and in vivo models.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methods</h3>\\n \\n <p>UFG and conventional SUS316L stainless steel wires were incubated with bioluminescent <i>Staphylococcus aureus</i> Xen36 for up to 7 days in vitro. Biofilm formation was assessed using crystal violet (CV) staining, colony-forming unit (CFU) counting, and quantitative PCR (qPCR) for <i>16S rRNA</i> and <i>luxA</i> genes. In vivo antibacterial effects were evaluated using two mouse models: a subcutaneous pouch model and a postoperative spinal implant infection model. Wires were harvested at 1, 3, and 7 days post-infection and analyzed using the same assays.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>In vitro, UFG wires had significantly lower CFU counts than standard wires at 4 h (<i>p</i> = 0.0005), 1 day (<i>p</i> = 0.0001), and 3 days (<i>p</i> = 0.0314). In the subcutaneous pouch model, UFG wires showed significantly reduced bacterial load at Day 1 by CFU (<i>p</i> = 0.011). In the spinal implant model, CFU counts were significantly lower on UFG wires at Day 3 (<i>p</i> = 0.015).</p>\\n </section>\\n \\n <section>\\n \\n <h3> Conclusions</h3>\\n \\n <p>UFG stainless steel implants demonstrated a significant reduction in early biofilm formation by <i>Staphylococcus aureus</i> in both in vitro and in vivo, suggesting a delay in the biofilm formation process. 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引用次数: 0
摘要
背景：植入物相关感染仍然是脊柱固定手术的一个严重并发症。由于种植体表面的生物膜形成是此类感染发病机制的关键因素，目前的预防策略包括使用抗生素涂层的种植体。然而，这些方法引起了对抗生素耐药性和细胞毒性的担忧。超细晶（UFG）不锈钢具有纳米级的晶粒尺寸，具有优异的机械性能和潜在的抗菌效果。本研究旨在通过体外和体内模型评价UFG不锈钢植入物对金黄色葡萄球菌生物膜形成的抑菌性能。方法UFG与普通SUS316L不锈钢丝与金黄色葡萄球菌Xen36体外培养7 d。采用结晶紫（CV）染色、菌落形成单位（CFU）计数和16S rRNA和luxA基因的定量PCR （qPCR）评估生物膜的形成。采用两种小鼠模型：皮下袋模型和术后脊柱植入物感染模型来评估体内抗菌效果。在感染后1、3和7天采集金属丝，使用相同的方法进行分析。结果UFG丝在体外4 h （p = 0.0005）、1 d （p = 0.0001）和3 d （p = 0.0314）时CFU计数明显低于标准丝。在皮下袋模型中，CFU在第1天显著降低了UFG丝的细菌负荷（p = 0.011）。在脊柱植入物模型中，第3天UFG丝上的CFU计数明显降低（p = 0.015）。结论UFG不锈钢植入物在体外和体内均能显著减少金黄色葡萄球菌的早期生物膜形成，提示生物膜形成过程延迟。这些发现支持了UFG材料作为抗感染脊柱植入物的潜力。
Ultrafine-Grained Materials With Antibacterial Properties: A Novel Approach to Reducing Spinal Implant-Associated Infections
Background
Implant-associated infection remains a serious complication of instrumented spinal surgery. Since biofilm formation on the implant surface is a key factor in the pathogenesis of such infections, current preventive strategies include the use of implants with antibiotic coatings. However, these approaches raise concerns related to antibiotic resistance and cytotoxicity. Ultrafine-grained (UFG) stainless steel, characterized by nanoscale grain sizes, has demonstrated superior mechanical properties and potential antimicrobial effects. This study aimed to evaluate the antibacterial properties of UFG stainless steel implants against Staphylococcus aureus biofilm formation in both in vitro and in vivo models.
Methods
UFG and conventional SUS316L stainless steel wires were incubated with bioluminescent Staphylococcus aureus Xen36 for up to 7 days in vitro. Biofilm formation was assessed using crystal violet (CV) staining, colony-forming unit (CFU) counting, and quantitative PCR (qPCR) for 16S rRNA and luxA genes. In vivo antibacterial effects were evaluated using two mouse models: a subcutaneous pouch model and a postoperative spinal implant infection model. Wires were harvested at 1, 3, and 7 days post-infection and analyzed using the same assays.
Results
In vitro, UFG wires had significantly lower CFU counts than standard wires at 4 h (p = 0.0005), 1 day (p = 0.0001), and 3 days (p = 0.0314). In the subcutaneous pouch model, UFG wires showed significantly reduced bacterial load at Day 1 by CFU (p = 0.011). In the spinal implant model, CFU counts were significantly lower on UFG wires at Day 3 (p = 0.015).
Conclusions
UFG stainless steel implants demonstrated a significant reduction in early biofilm formation by Staphylococcus aureus in both in vitro and in vivo, suggesting a delay in the biofilm formation process. These findings support the potential of UFG materials as promising candidates for infection-resistant spinal implants.
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