不同支持物及培养条件对幽门螺杆菌生物膜形成的影响

Effects of different supports and conditions on the biofilm formation of Helicobacter pylori

目的 研究在不同支持物和培养条件下幽门螺杆菌(Helicobacter pylori,H.pylori)生物膜造模方法。方法 选取H.pylori SS1作为实验菌株,通过液体培养、固体培养的方法,分别选取圆形玻片及硝酸纤维素膜(NC膜)为载体,将H.pylori SS1分别接种于无菌24孔板及固体培养基中NC膜上,37℃微需氧环境培养72 h,成膜后戊二醛固定,乙醇梯度脱水干燥,喷金,电镜下观察。结果 液体培养圆形玻片为载体条件下H.pylori成膜形态立体,可清晰看到其鞭毛结构,但其密度较差,生长状态一般,且成膜率较低;固体培养NC膜载体条件下H.pylori形态较扁平,但生长状态良好,同样可见其鞭毛结构,成膜率较高。结论 液体培养条件及固体培养条件下均可成功构建H.pylori生物膜模型,但电镜观察固体培养NC膜载体条件下生物膜生长状态更好,成膜率更高。体外实验更推荐以NC膜为载体的固体培养。

Objective To study the modeling method of Helicobacter pylori(H.pylori) biofilm under different supporting materials and conditions.Methods H.pylori SS1 as the experimental strains were selected.The circular glass slide and NC membrane were selected as the carrier.H.pylori SS1 was inoculated on NC membrane and 24-well sterile plate,and then cultured in liquid and solid,respectively,in 37 ℃ micro-aerobic environment for 72 hours.H.pylori SS1 was fixed with glutaraldehyde after biofilm formation,dehydrated and dried with ethanol gradient,sprayed with gold,and observed under electron microscope.Results The biofilm formed by H.pylori SS1 was solid under the condition of liquid culture on round glass.The flagellum structure can be clearly seen,while its density was relatively low,and the film formation rate was slow.Under the condition of solid culture on NC membrane carrier,H.pylori SS1 shape was relatively flat in shape,but in good condition.Its flagellum structure was also visible,and the film formation rate was high.Conclusion The model of H.pylori SS1 biofilm can be successfully constructed under both liquid and solid culture conditions.However,the growth state of biofilm is better on solid culture NC membrane carrier under electron microscope,also the film formation rate is higher.Solid culture on NC membrane is recommended for in vitro experiments.