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目的探讨金银花主要活性成分绿原酸对聚氯乙烯(PVC)生物材料表面金黄色葡萄球菌(S.a)生物被膜(BF)的体外抑制作用。方法采用肉汤-医用PVC薄片系统构建早期及成熟期的S.a BF体外模型,并用扫描电镜(SEM)鉴定;试管兑倍稀释法测定绿原酸的最低抑菌浓度;将实验对象分为空白对照组、绿原酸(0.125,0.25,0.5,1,2mg/ml)组;荧光正置相差显微镜观察BF形态学改变;结晶紫染色法定量载体表面生物总量;连续稀释法计数BF内活菌。结果成功在PVC生物材料表面建立不同时期的S.a BF;无论早期或成熟期,当绿原酸浓度达到0.5mg/ml及以上时,荧光显微镜下均可观察到PVC载体表面BF的形成较同期空白组明显减少,结晶紫染色法定量及BF内活菌计数均少于同期空白对照组(P<0.05)。结论金银花主要活性成分绿原酸可抑制PVC生物材料表面S.a BF的形成,且在一定浓度范围内随着绿原酸浓度增加,其对BF抑制作用增强。
Objective To investigate the inhibitory effect of chlorogenic acid, the main active component of honeysuckle, on the in vitro bioactivity of S.aureus biofilm on polyvinyl chloride (PVC) biomaterials. Methods The early and mature Sa BF in vitro models were constructed by using the broth-medical PVC sheet system and identified by scanning electron microscopy (SEM). The minimum inhibitory concentration of chlorogenic acid was determined by tube dilution test. The experimental subjects were divided into blank control Group, chlorogenic acid (0.125,0.25,0.5,1,2mg / ml) group; Fluorescence normal phase contrast microscope to observe the morphological changes of BF; Crystal violet staining method to quantify the total amount of surface bio-carrier; . Results Sa BF at different stages of PVC biomaterials was successfully established. The BF formation on the surface of PVC carrier was observed under the fluorescence microscope when the concentration of chlorogenic acid reached 0.5 mg / ml and above at the early or mature stage (P <0.05). The quantification by crystal violet staining and viable count in BF were less than those in the blank control group at the same period (P <0.05). Conclusion Chlorogenic acid, the main active component of honeysuckle, inhibits the formation of S.aBF on the surface of PVC biomaterials. With the increase of chlorogenic acid concentration in a certain concentration range, its inhibitory effect on BF is enhanced.