粪肠球菌
抗菌剂
化学
生物膜
大黄素
微生物学
适体
细菌
生物化学
生物
分子生物学
大肠杆菌
遗传学
基因
作者
Maryam Pourhajibagher,Abbas Bahador
标识
DOI:10.1016/j.pdpdt.2022.103020
摘要
• Flow cytometry assays revealed that Apt@EmoNp-DCD-1L had the highest selectivity to bind E. faecalis . • A significant reduction of Log 10 CFU/mL was observed in the groups treated with aPDT using Apt@EmoNp-DCD-1L. • aPDT using 4×MIC of Apt@EmoNp-DC-1 L showed a significant anti-biofilm activity over the other groups Despite the high success rate of root canal treatment, failures are observed in a broad range of cases. Therefore, the need for novel approaches with the development of new generations of antimicrobial agents and intracellular drug delivery systems as adjunctive therapy is undeniable. In this study, we investigated the antimicrobial effects of antimicrobial photodynamic therapy (aPDT) using dermcidin‑derived peptide DCD‑1L loaded onto aptamer-functionalized emodin nanoparticles (Apt@EmoNp-DCD-1L) against Enterococcus faecalis as one of the most common bacteria involved in recurrent root canal treatment failures. Following preparation of EmoNp-DCD-1L, the binding of selected labeled Apt to EmoNp-DCD-1L was performed, followed by the specificity of Apt@EmoNp-DCD-1L to E. faecalis was determined. The antimicrobial potential of aPDT was then assessed after the determination of the minimum inhibitory concentration (MIC) of Apt@EmoNp-DCD-1L. The molecular docking analysis was conducted to evaluate the potential binding modes of EmoNp to the proteins involved in E. faecalis pathogenesis. Eventually, the anti-virulence capacity of Apt@EmoNp-DCD-1L-mediated aPDT was investigated via quantitative real-time PCR (qRT-PCR) assay following measurement of intracellular reactive oxygen species (ROS) generation. The binding specificity of Apt@EmoNp-DCD-1L to E. faecalis was confirmed by flow cytometry. The results showed that the cell viability of E. faecalis exposed to aPDT groups employing the sub-MIC doses of Apt@EmoNp-DCD-1L (7.8 and 15.6 µM) was significantly reduced compared to the control group ( P < 0.05). Also, Apt@EmoNp-DCD-1L in combination with a blue laser light was capable of enhancing the anti-biofilm activity of aPDT against E. faecalis biofilm. Data obtained from the qRT-PCR analysis showed significant downregulation in the expression level of genes involved in bacterial biofilm formation after exposure to aPDT ( P < 0.05). : This in vitro study highlights that aPDT with the minimum concentration of Apt@EmoNp-DCD-1L can be considered as a targeted bio-theragnostic agent for the detection and elimination of E. faecalis in the dispersed and biofilm states.
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