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具有长效抗菌活性的新型乳胶导尿管的细胞毒性评估。

The cytotoxicity assessment of the novel latex urinary catheter with prolonged antimicrobial activity.

机构信息

Chair and Department of Medicinal Chemistry, Medical University of Lublin, Jaczewskiego 4, Lublin 20-090, Poland.

出版信息

J Biomed Mater Res A. 2011 Aug;98(2):222-8. doi: 10.1002/jbm.a.33110. Epub 2011 May 4.

Abstract

The purpose of this study was to evaluate the in vitro biocompatibility of the novel Sparfoxacin (SPA)-treated latex catheter with previously performed prolonged antimicrobial activity. Rectangular-shaped test samples of silicone latex catheter were fabricated according to patented procedure permitting the immobilization of SPA on heparin (HP)-coated catheter by means of mixed, covalent and non-covalent bonds. Samples subjected to cytotoxicity assay were divided into four groups: (1) the untreated catheter, (2) HP-coated catheter, (3) HP-coated catheter with SPA immobilized in low SPA concentration solution (SPA-L treated sample), and (4) high SPA concentration solution (SPA-H treated sample). Then the samples were placed directly into green monkey kidney (GMK) cell monolayer for 24 h. After the incubation period, cytotoxicity was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The degree of cytotoxicity of each sample was evaluated according to the reference value represented by the control cells cultured without catheter sample. Statistical significance was determined by repeated t-test (P < 0.05). The cytotoxic effect of treated and untreated catheters was also estimated by microscopic observations of GMK cells morphological changes. SPA-treated catheters demonstrated high survival rates in MTT assay (>93%) on the contrary to the untreated catheters (6.13%) and HP-coated catheters (80.90%). Moreover, microscopic observation of GMK cells exposed to SPA-treated samples revealed no morphological changes and no cell growth reduction. We suggest that SPA-treated latex catheters are biofilm formation resistant (as we revealed in our previous work), considerably less toxic than untreated ones, and can be undoubtedly used in urological practice.

摘要

本研究旨在评估新型斯巴氟沙星(SPA)处理乳胶导尿管的体外生物相容性,此前已进行了延长抗菌活性的研究。根据专利程序,制备了硅酮乳胶导尿管的矩形测试样本,该程序允许 SPA 通过混合、共价和非共价键固定在肝素(HP)涂层导尿管上。进行细胞毒性测定的样品分为四组:(1)未处理的导尿管,(2)HP 涂层导尿管,(3)SPA 固定在低 SPA 浓度溶液中的 HP 涂层导尿管(SPA-L 处理样品),和(4)高 SPA 浓度溶液(SPA-H 处理样品)。然后将样品直接放入绿猴肾(GMK)单层细胞中孵育 24 小时。孵育期后,通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四唑溴盐(MTT)测定法测定细胞毒性。根据未培养导管样品的对照细胞的参考值评估每个样品的细胞毒性程度。通过重复 t 检验(P <0.05)确定统计学意义。通过 GMK 细胞形态变化的显微镜观察还评估了处理和未处理导管的细胞毒性。与未处理的导管(6.13%)和 HP 涂层的导管(80.90%)相比,SPA 处理的导管在 MTT 测定中显示出高存活率(>93%)。此外,暴露于 SPA 处理样品的 GMK 细胞的显微镜观察显示没有形态变化和细胞生长减少。我们建议 SPA 处理的乳胶导管具有抗生物膜形成的能力(如我们在之前的工作中所揭示的),毒性明显小于未处理的导管,并且可以在泌尿科实践中毫无疑问地使用。

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