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Effect of isochlorogenic acid on Aspergillus Fumigatus biofilm in vitro

Objective To investigate the effect of isochlorogenic acid on the biofilm of Aspergillus Fumigatus in early and mature stages in vitro. Methods The minimum inhibitory concentration(MIC) and the minimum fungicidal concentration(MFC) for isochlorogenic acid were measured by broth microdilution method according to CLSI M38-A2 guideline. The biofilm models in early and mature stages established under static condition were affected by different concentration gradients(64, 128, 256, 512, 1024 μg/m L) of isochlorogenic acid for 48 h in vitro, then biofilm quantitation was performed by crystal violet staining assay. Scanning electron microscope(SEM) was used to observe the morphological change of biofilm structure. The viability of Aspergillus Fumigatus hyphae in biofilm was detected by confocal laser scanning microscope(CLSM). The MIC and MFC of different groups were compared. Results Both the MIC and the MFC of isochlorogenic acid on Aspergillus Fumigatus isolate were above 1024 μg/m L. The results of CLSM demonstrated that isochlorogenic acid showed hardly any fungicidal effect to the mycelia in early or mature stage of biofilm. Under SEM, the structure of Aspergillus Famigatustus biofilm treated with isochlorogenic acid was destructed and the outline of mycelia appeared to be more clear than that without isochlorogenic acid due to the reduction of extracellular matrix.The in vitro action was performed while the concentration of isochlorogenic acid achieved the level of 256 μg/m L, within the concentrations of 256, 512, 1024 μg/m L, results of biofilm quantities indicated significant differences between serially increasing isochlorogenic acid concentration groups and untreated control group, with statistical significance(P 0.05). Conclusion Isochlorogenic acid exhibits destructive effect on the early and mature stages of Aspergillus Famigatustus biofilm in vitro, which shows a concentration-dependent manner.