Chen L, Hobden J A, Masinick S A, Hazlett L D
Department of Anatomy Cell Biology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
Curr Eye Res. 1998 Mar;17(3):231-7. doi: 10.1076/ceyr.17.3.231.5226.
To determine whether environmental factors or bacterial viability affect the binding of two strains of P. aeruginosa to mouse cornea.
Scarified corneas were placed in organ culture and inoculated with P. aeruginosa cell suspensions containing either ATCC 19660 or PAO1 bacterial strains classed as cytotoxic or invasive, respectively. Eyes were incubated in vitro for 1 h after bacterial application at different pH or temperature conditions or in PBS containing various divalent cations. The adhesion of heat-killed or formalin-fixed bacteria was tested similarly. Scanning electron microscopy (scanning EM) was used to quantitate adherent bacteria.
P. aeruginosa ATCC 19660 showed an increase in binding at pH 8.0, favored higher temperatures and required both calcium and magnesium for optimum binding. Adherence of PAO1 was enhanced at pH 6.5 and decreased at pH 8.0. This strain favored binding at lower temperatures and did not require either divalent cation for optimum binding. In addition, the presence of magnesium ions resulted in reduced binding for this strain. Both strains exhibited less binding ability after formalin fixation or heat killing.
Environmental factors and bacterial viability are important factors which influence the ability of both cytotoxic and invasive strains of P. aeruginosa to bind to the scarified cornea.
确定环境因素或细菌活力是否会影响两株铜绿假单胞菌与小鼠角膜的结合。
将划痕角膜置于器官培养中,接种分别归类为具有细胞毒性或侵袭性的含ATCC 19660或PAO1菌株的铜绿假单胞菌细胞悬液。在不同pH值或温度条件下或在含有各种二价阳离子的PBS中接种细菌后,将眼睛在体外孵育1小时。类似地测试热灭活或福尔马林固定细菌的粘附情况。使用扫描电子显微镜(扫描电镜)对粘附细菌进行定量。
铜绿假单胞菌ATCC 19660在pH 8.0时结合增加,在较高温度下更有利,并且需要钙和镁两者才能实现最佳结合。PAO1的粘附在pH 6.5时增强,在pH 8.0时降低。该菌株在较低温度下更有利于结合,并且不需要任何一种二价阳离子即可实现最佳结合。此外,镁离子的存在导致该菌株的结合减少。福尔马林固定或热灭活后,两株菌的结合能力均降低。
环境因素和细菌活力是影响细胞毒性和侵袭性铜绿假单胞菌菌株与划痕角膜结合能力的重要因素。