Inhibition of Biofilm Formation by Froriepia subpinnata Leaf Extract-Functionalized Magnetic Nanocomposites and Ciprofloxacin and Downregulation of pslA, algD, and pelA Genes in Clinical Isolates of Pseudomonas aeruginosa

Subject Areas : genetics

Mahsa Ahmadzadeh Kelangestani1 ¹ , Sajedeh Moazzami Mirak Mahaleh ² , Mohammad Nikpassand ³ , Mahdi Shahriarinour ⁴ , Najmeh Ranji ⁵

1 - Department of Biology, Ra.C., Islamic Azad University, Rasht, Iran.
2 - Department of Biology, Ra.C., Islamic Azad University, Rasht, Iran.
3 - Department of Chemistry, Ra.C., Islamic Azad University, Rasht, Iran.
4 - Department of Biology, Ra.C., Islamic Azad University, Rasht, Iran
5 - Department of Biology, Ra.C., Islamic Azad University, Rasht, Iran

Received: 2025-08-19 Accepted : 2025-09-07 Published : 2025-09-09

Keywords: Pseudomonas aeruginosa, Biofilm, Magnetic nanocomposites, Froriepia subpinnata, Ciprofloxacin,

Abstract :

introduction: Pseudomonas aeruginosa is an opportunistic hospital-acquired pathogen with multidrug resistance, in which biofilm formation plays a critical role in therapeutic failure. This study aimed to evaluate the effects of Froriepia subpinnata leaf extract-functionalized magnetic nanocomposites in combination with ciprofloxacin on the inhibition of biofilm formation and deregulation of biofilm-related gene in resistant clinical isolates.

Material and Methods: In this study, 26 clinical isolates of P. aeruginosa were collected from hospitals in Tehran, Iran, and their antibiotic susceptibility profiles were determined using the Kirby–Bauer method. Minimum inhibitory concentration (MIC) was evaluated by broth microdilution. The Froriepia subpinnata leaf extract-functionalized magnetic nanocomposites were synthesized and their anti-biofilm function were assessed in combination with ciprofloxacin. Synergistic activity was assessed by the checkerboard assay, biofilm formation was quantified using crystal violet staining, and the expression of pslA, algD, and pelA genes was analyzed by Q-RT-PCR.

Results: Physicochemical analysis confirmed the successful synthesis of Froriepia subpinnata leaf extract-functionalized magnetic nanoparticles with particle sizes ranging from 28 to 82 nm. All of isolates were resistant to ciprofloxacin, with MIC values ranging from 128 to 1024 µg/mL. Checker board analysis demonstrated a synergistic effect between the Froriepia subpinnata- functionalized nanocomposites and ciprofloxacin in the strain ATCC 9027 and several clinical isolates. Moreover, the combination treatment significantly led to decrease of biofilm formation and downregulation of pslA, algD, and pelA genes compared to treatment with ciprofloxacin alone (P<0.05).

Conclusion: The findings indicate that Froriepia subpinnata-functionalized magnetic nanocomposites can enhance the antibacterial efficacy of ciprofloxacin against resistant P. aeruginosa isolates by downregulating biofilm formation-related genes.

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