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[对从生物材料中分离出的表皮葡萄球菌菌株表面特性的研究]

[Investigation of the surface properties of Staphylococcus epidermidis strains isolated from biomaterials].

作者信息

Sudağidan Mert, Erdem Ilker, Cavuşoğlu Cengiz, Ciftçloğlu Muhsin

机构信息

Izmir Yüksek Teknoloji Enstitüsü, Biyoteknoloji ve Biyomühendislik Merkezi Araştirma Laboratuvanri, Izmir.

出版信息

Mikrobiyol Bul. 2010 Jan;44(1):93-103.

Abstract

The surface properties of bacteria play an important role on adhesion to the biomaterial surface. In this study, the surface properties of Staphylococcus epidermidis strains isolated from clinically used polymeric biomaterial surfaces were investigated on the basis of zeta potential, hydrophobicity and surface topography. A total of 10 S.epidermidis strains isolated from intravenous catheters (n = 5), endotracheal tubes (n = 3) and central venous catheters (n = 2) which were used in the patients of pulmonary Intensive Care Unit, Ege University Medical Faculty Hospital, were included to the study. Seven of those isolates were biofilm producers, inhabiting biofilm genes, 2 were non-biofilm producers, however, inhabiting biofilm genes, and 1 was non-biofilm producer, inhabiting no biofilm genes. Zeta potential analysis have been performed in 3 different buffers (phosphate-buffered saline, 1 mM potassium chloride and 1 mM potassium phosphate buffer) and at different pH values (pH 4.1-8.2), in order to simulate in vivo environment of the biomaterials. Hydrophobicities of the strains were examined by bacterial adhesion to hydrocarbon (BATH) test and the surface topography of biofilms and slime layers were visualized by atomic force microscopy (AFM) and scanning electron microscopy (SEM) methods. It was found that all strains have negative zeta potential values (surface charge) in all buffers and pH values. In hydrophobicity analysis, the highest value (86%) was determined for non-biofilm forming S. epidermidis strain YT-169b (endotracheal tube isolate) and the lowest hydrophobicity (2.5%) was determined for biofilm forming S. epidermidis strain YT-212 (central venous catheter isolate). Biofilm and slime layers of the strains were imaginated by AFM and SEM analysis in microm scale. SEM analysis showed that bacteria highly adhered to rough surfaces on biomaterial surfaces and the produced slime layers covered the surface of bacteria. In conclusion, elucidating the surface properties of opportunistic pathogens in different physiologic buffers will give important clues for the production of non-adhesive materials and antibacterial surfaces for those bacteria. It was also estimated that designing the surface of the biomaterial to have negative surface charge in the body and to be as smooth as possible will hamper biofilm formation.

摘要

细菌的表面特性在其对生物材料表面的黏附中起着重要作用。在本研究中,基于zeta电位、疏水性和表面形貌,对从临床使用的聚合物生物材料表面分离出的表皮葡萄球菌菌株的表面特性进行了研究。本研究纳入了从伊兹密尔艾杰大学医学院医院重症监护病房患者使用的静脉导管(n = 5)、气管内导管(n = 3)和中心静脉导管(n = 2)中分离出的总共10株表皮葡萄球菌菌株。其中7株分离株是生物膜产生菌,携带生物膜相关基因;2株是非生物膜产生菌,但携带生物膜相关基因;1株是非生物膜产生菌,不携带生物膜相关基因。为了模拟生物材料的体内环境,在3种不同的缓冲液(磷酸盐缓冲盐水、1 mM氯化钾和1 mM磷酸钾缓冲液)以及不同的pH值(pH 4.1 - 8.2)下进行了zeta电位分析。通过细菌对碳氢化合物黏附试验(BATH)检测菌株的疏水性,并采用原子力显微镜(AFM)和扫描电子显微镜(SEM)方法观察生物膜和黏液层的表面形貌。结果发现,所有菌株在所有缓冲液和pH值下均具有负的zeta电位值(表面电荷)。在疏水性分析中,非生物膜形成的表皮葡萄球菌菌株YT - 169b(气管内导管分离株)的疏水性最高(86%),而生物膜形成的表皮葡萄球菌菌株YT - 212(中心静脉导管分离株)的疏水性最低(2.5%)。通过AFM和SEM分析在微米尺度上对菌株的生物膜和黏液层进行了成像。SEM分析表明,细菌高度黏附于生物材料表面的粗糙区域,并且所产生的黏液层覆盖了细菌表面。总之,阐明不同生理缓冲液中机会致病菌的表面特性将为生产针对这些细菌的非黏附材料和抗菌表面提供重要线索。据估计,设计生物材料表面使其在体内具有负表面电荷并尽可能光滑将阻碍生物膜的形成。

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