Bioresponsive gingerol-loaded alginate-coated niosomal nanoparticles for targeting intracellular bacteria and cancer cells.

Targeting and treating intracellular pathogen infections has been long-standing challenge, particularly in light of the escalating prevalence of antimicrobial resistance. Herein, an optimum formulation of alginate (AL)-coated niosome-based carriers for delivery of herbal extract Gingerol (Gin) was developed to treat intracellular pathogen infections and cancer cells. We used Gin-Nio@AL as a model drug to assess its efficacy against Gram-negative/positive bacteria and breast cancer cell lines. Our investigation affirmed its heightened antibacterial and anticancer properties. The antibacterial activity of Gin-Nio@AL against intracellular Staphylococcus aureus (S. aureus) and pseudomonas aeruginosa (P. aeruginosa) was also tested. In the current study, the niosome nanoparticles containing herbal extract Gingerol were optimized regarding lipid content and Surfactant per Cholesterol molar ratio. The developed formulation provided potential advantages, such as smooth globular surface morphology, small diameter (240.68 nm), pH-sensitive sustained release, and high entrapment efficiency (94.85 %). The release rate of Gin from AL-coated niosomes (Gin-Nio@AL) in physiological and acidic pH is lower than uncoated nanoparticles (Gin-Nio). Besides, the release rate of Gin from niosomal formulations increased in acidic pH. The Gin-Nio@AL demonstrated good antimicrobial activity against S. aureus and P. aeruginosa, and compared to Gin-Nio, the MIC values decreased to 7.82 ± 0.00 and 1.95 ± 0.00 µg/mL, respectively. In addition, the time-kill assay results showed that the developed formulation significantly reduced the number of bacteria in both strains compared to other tested groups. The microtiter data and scanning electron microscope micrography showed that Gin-Nio@AL has a more significant inhibitory effect on biofilm formation than Gin-Nio and Gin. The cell cytotoxicity evaluation showed that Gin-Nio@AL reduced the survival rate of MDA-MB-231 cancer cells to 52.4 % and 45.2 % after 48 h and 72 h, respectively. The elimination of intracellular pathogens was investigated through a breast cancer cell infection in an in vitro model. Gin-Nio@AL exhibited an enhanced and sustained intracellular antibacterial activity against pathogens-infected breast cancer cells compared to other tested formulations. Overall, Gin-Nio@AL enables the triggered release and targeting of intra-extra cellular bacteria and cancer cells and provides a novel and promising candidate for treating intracellular pathogen infections and cancer cells.

In silico and in vitro studies of GENT-EDTA encapsulated niosomes: A novel approach to enhance the antibacterial activity and biofilm inhibition in drug-resistant Klebsiella pneumoniae.

Klebsiella pneumoniae (Kp) is a common pathogen inducing catheter-related biofilm infections. Developing effective therapy to overcome antimicrobial resistance (AMR) in Kp is a severe therapeutic challenge that must be solved. This study aimed to prepare niosome-encapsulated GENT (Gentamicin) and EDTA (Ethylenediaminetetraacetic acid) (GENT-EDTA/Nio) to evaluate its efficacy toward Kp strains. The thin-film hydration method was used to prepare various formulations of GENT-EDTA/Nio. Formulations were characterized for their physicochemical characteristics. GENT-EDTA/Nio properties were used for optimization with Design-Expert Software. Molecular docking was utilized to determine the antibacterial activity of GENT. The niosomes displayed a controlled drug release and storage stability of at least 60 days at 4 and 25 °C. GENT-EDTA/Nio performance as antimicrobial agents has been evaluated by employing agar well diffusion method, minimum bactericidal concentration (MBC), and minimum inhibitory concentration (MIC) against the Kp bacteria strains. Biofilm formation was investigated after GENT-EDTA/Nio administration through different detection methods, which showed that this formulation reduces biofilm formation. The effect of GENT-EDTA/Nio on the expression of biofilm-related genes (mrkA, ompA, and vzm) was estimated using QRT-PCR. MTT assay was used to evaluate the toxicity effect of niosomal formulations on HFF cells. The present study results indicate that GENT-EDTA/Nio decreases Kp's resistance to antibiotics and increases its antibiotic and anti-biofilm activity and could be helpful as a new approach for drug delivery.

pH-Responsive PEGylated Niosomal Nanoparticles as an Active-Targeting Cyclophosphamide Delivery System for Gastric Cancer Therapy.

A PEGylated niosomal formulation of cyclophosphamide (Nio-Cyclo-PEG) was prepared using a central composite design and characterized in terms of drug loading, size distribution, and average size. The stability of formulations was also studied at different conditions. In vitro cytotoxicity of drug delivery formulations was assessed on gastric cancer cells using MTT assay. The mechanism of cytotoxicity was studied at the transcriptional level by real-time PCR on Caspase3, Caspase9, CyclinD, CyclinE, MMP-2, and MMP-9 genes, while apoptosis was investigated with flow cytometry. The anti-metastatic property was evaluated using the scratch method. Propidium iodide staining was used to study the cell cycle. The results indicated that the as-designed nanocarrier exhibited a controlled drug release pattern with improved nanoparticle stability. It was found that the living cancer cells treated with Nio-Cyclo-PEG showed a significant decrease in number when compared with the niosomal carrier without PEG (Nio-Cyclo) and free drug (Cyclo). Moreover, the drug-loaded nanocarrier induced planned death (apoptosis) in the cancer cells through the regulation of Caspase3, Caspase9, CyclinD, CyclinE, MMP-9, and MMP-2 gene expression, indicating that the Nio-Cyclo-PEG formulation could significantly inhibit the cell cycle at the sub G1 phase as well as prevent the migration of cancer cells. In conclusion, Nio-Cyclo-PEG as developed in this study could serve as an active-targeting drug delivery nanocarriers for gastric cancer therapy with high efficacy and minimal side effects on healthy tissues/cells.