Bauer Sandra M, Santschi Elizabeth M, Fialkowski James, Clayton Murray K, Proctor Richard A
School of Veterinary Medicine, the Department of Statistics, and the College of Medicine, University of Wisconsin, Madison, WI 53706, USA.
Vet Surg. 2004 Jul-Aug;33(4):376-81. doi: 10.1111/j.1532-950X.2004.04054.x.
To quantify the adhesion of Staphylococcus aureus to 4 equine bone surfaces passivated in a balanced polyionic solution (Plasmalyte) or hyperimmune equine plasma (Polymune plasma).
In vitro comparative study.
Third metacarpal bone (MC3) surface explants from 9 equine cadavers.
Approximately 1 cm(2) sections of periosteum were removed from MC3 and stapled to sterile stainless steel screens. Three bone surface explants were cut using a surgical saw to present 1 cm(2) surfaces of subperiosteal bone, cut cortical bone, or endosteum. Duplicate explants of each surface were immersed for 1 hour in Plasmalyte or hyperimmune equine plasma. Each explant was then placed in a well of a 6-well sterile tissue culture plate with the surface of interest exposed. Each surface was inoculated with approximately 100 colony-forming units of S. aureus in 10 microL of Mueller Hinton broth and incubated for 6 hours at 37 degrees C. After gentle rinsing to remove non-adherent bacteria, samples were sonicated for 5 minutes at 60 kHz to loosen adhered bacteria. The number of adherent bacteria was determined by serial dilutions and incubation of the sonicate. Scanning electron microscopy (SEM) was performed on samples identically treated from an additional horse to confirm bacterial removal by sonication from all surfaces and support quantitative culture results.
Less S. aureus adhered to periosteum than to cortical bone, cut cortical bone, and endosteal surfaces, which were all similar. Exposure of all surfaces to hyperimmune plasma reduced S. aureus adherence compared with Plasmalyte exposure; SEM supported these conclusions.
Less bacteria adhere to periosteum than other bone surfaces. Hyperimmune plasma reduces bacterial adhesion to all bone tissue surfaces.
Understanding the factors that affect bacterial adhesion to bone will facilitate development of improved intraoperative lavage solutions to reduce the morbidity and mortality associated with postoperative infection.
量化金黄色葡萄球菌对4种在平衡多离子溶液（Plasmalyte）或超免疫马血浆（Polymune血浆）中钝化的马骨表面的黏附情况。
体外比较研究。
来自9具马尸体的第三掌骨（MC3）表面外植体。
从MC3上取下约1平方厘米的骨膜切片，用订书钉固定在无菌不锈钢筛网上。使用手术锯切割出3个骨表面外植体，以呈现1平方厘米的骨膜下骨、切割的皮质骨或骨内膜表面。每个表面的复制品外植体在Plasmalyte或超免疫马血浆中浸泡1小时。然后将每个外植体放入6孔无菌组织培养板的孔中，使感兴趣的表面暴露。每个表面接种约100个金黄色葡萄球菌菌落形成单位于10微升的穆勒-欣顿肉汤中，并在37℃下孵育6小时。轻轻冲洗以去除未黏附的细菌后，将样本在60千赫兹下超声处理5分钟以松动黏附的细菌。通过系列稀释和超声处理液的孵育来确定黏附细菌的数量。对来自另一匹马的经相同处理的样本进行扫描电子显微镜（SEM）检查，以确认通过超声处理从所有表面去除了细菌，并支持定量培养结果。
与皮质骨、切割的皮质骨和骨内膜表面相比，金黄色葡萄球菌在骨膜上的黏附较少，而这些表面的黏附情况相似。与暴露于Plasmalyte相比，所有表面暴露于超免疫血浆均降低了金黄色葡萄球菌的黏附；SEM支持了这些结论。
与其他骨表面相比，细菌在骨膜上的黏附较少。超免疫血浆可降低细菌对所有骨组织表面的黏附。
了解影响细菌对骨黏附的因素将有助于开发改进的术中灌洗溶液，以降低与术后感染相关的发病率和死亡率。
