Biosynthesis Strategy of Gold Nanoparticles and Biofabrication of a Novel Amoxicillin Gold Nanodrug to Overcome the Resistance of Multidrug-Resistant Bacterial Pathogens MRSA and E. coli.

The prevalence of multidrug-resistant (MDR) bacteria has recently increased dramatically, seriously endangering human health. Herein, amoxicillin (Amoxi)-conjugated gold nanoparticles (AuNPs) were created as a novel drug delivery system to overcome MDR bacteria. MDR bacteria were isolated from a variety of infection sources. Phenotype, biotype, and 16S rRNA gene analyses were used for isolate identification. Additionally, Juniperus excelsa was used for the production of AuNPs. The conjugation of AuNPs with Amoxi using sodium tri-polyphosphate (TPP) as a linker to produce Amoxi-TPP-AuNPs was studied. The AuNP and Amoxi-TPP-AuNP diameters ranged from 15.99 to 24.71 nm, with spherical and hexagonal shapes. A total of 83% of amoxicillin was released from Amoxi-TPP-AuNPs after 12 h, and after 3 days, 90% of the medication was released. The Amoxi-TPP-AuNPs exhibited superior antibacterial effectiveness against MRSA and MDR E. coli strains. Amoxi-TPP-AuNPs had MICs of 3.6-8 µg mL-1 against the tested bacteria. This is 37.5-83 fold higher compared to values reported in the literature. Amoxi-TPP-AuNPs exhibit a remarkable ability against MRSA and E. coli strains. These results demonstrate the applicability of Amoxi-TPP-AuNPs as a drug delivery system to improve therapeutic action.

Biofabrication strategy of silver-nanodrug conjugated polyhydroxybutyrate degrading probiotic and its application as a novel wound dressing.

Wound infections with rising incidences of multi-drug resistant bacteria are among the public health problems worldwide. The current study describes wound dressing materials made from biodegradable polyhydroxybutyrate (PHB) combined with AgNPs and gelatin (AgNPs/Gelatin/PHB). Microbial PHB was mixed with gelatin (1:2) to form a polymer matrix which was loaded with different concentrations of AgNPs (8.3-133 µg/mL). The statistical results of AgNPs synthesizing based on Box-Behnken design revealed that 1.247 mM silver nitrate and 24.054 % of Corchorus olitorius leaf extract concentration at pH (8.07) were the optimum values for the biosynthesis. UV-Vis spectroscopy, FTIR study and XRD reflects that nanoparticles are formed. The UV-Vis spectroscopy of Gelatin/PHB/AgNPs exhibited two specific bands at 298 nm and 371 nm, which confirm the formation of the conjugate. AgNPs had MICs and MBCs of (24.9, 24.9, and 12.45 µg/mL) and (33.25, 33.25, and 16.6 µg/mL) against (Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus). The MIC and MBC of AgNPs/Gelatin/PHB against the same tested bacteria were 31.1 µg and 41.5 µg, respectively. AgNPs/Gelatin/PHB exhibit excellent antimicrobial efficacy against bacteria. Sterilized gauze loaded with 31.1 µg of AgNPs/Gelatin/PHB acted as an effective wound dressing. Thus, the study highlights the importance of wound dressings developed from degradable AgNPs/Gelatin/PHB in enhancing antimicrobial efficiency and facilitating a better wound healing process.

Prevalence and Molecular Characterization of Methicillin-Resistant Staphylococcus aureus from Nasal Specimens: Overcoming MRSA with Silver Nanoparticles and Their Applications.

Staphylococcus aureus is a cause of high mortality in humans and therefore it is necessary to prevent its transmission and reduce infections. Our goals in this research were to investigate the frequency of methicillin-resistant S. aureus (MRSA) in Taif, Saudi Arabia, and assess the relationship between the phenotypic antimicrobial sensitivity patterns and the genes responsible for resistance. In addition, we examined the antimicrobial efficiency and application of silver nanoparticles (AgNPs) against MRSA isolates. Seventy-two nasal swabs were taken from patients; MRSA was cultivated on Mannitol Salt Agar supplemented with methicillin, and 16S rRNA sequencing was conducted in addition to morphological and biochemical identification. Specific resistance genes such as ermAC, aacA-aphD, tetKM, vatABC and mecA were PCR-amplified and resistance plasmids were also investigated. The MRSA incidence was ~49 % among the 72 S. aureus isolates and all MRSA strains were resistant to oxacillin, penicillin, and cefoxitin. However, vancomycin, linezolid, teicoplanin, mupirocin, and rifampicin were effective against 100% of MRSA strains. About 61% of MRSA strains exhibited multidrug resistance and were resistant to 3-12 antimicrobial medications (MDR). Methicillin resistance gene mecA was presented in all MDR-MRSA strains. Most MDR-MRSA contained a plasmid of > 10 kb. To overcome bacterial resistance, AgNPs were applied and displayed high antimicrobial activity and synergistic effect with penicillin. Our findings may help establish programs to control bacterial spread in communities as AgNPs appeared to exert a synergistic effect with penicillin to control bacterial resistance.

Biosynthesis of silver nano-drug using Juniperus excelsa and its synergistic antibacterial activity against multidrug-resistant bacteria for wound dressing applications.

We report here the synthesis of silver nanoparticles (AgNPs) from an aqueous extract of Juniperus excelsa and their use as an antimicrobial agent on their own or in combination with antibiotics in inhibiting multidrug-resistant bacteria (MDR). One strategy of bacterial infection control in wound healing is AgNP biosynthesis. We collected bacterial strains of patient skin infections from Al-Adwani Hospital. Phenotyping, biotyping, and molecular characterizations were applied using 16S rRNA gene analysis of bacterial isolates. Our results identified tested MDR bacteria Staphylococcus aureus strains (methicillin-resistant and methicillin-susceptible) and Proteus mirabilis. Gas chromatography/mass spectrometry (GC/MS) analysis was used to identify the Juniperus excelsa biomolecules in the leaf extract acting as both reducing and capping agents in the biosynthesis of AgNPs. The AgNPs appeared hexagonal and spherical in shape upon transmission electron microscope (TEM) analysis. The AgNP sizes ranged from 16.08 to 24.42 nm. X-ray diffraction (XRD) analysis confirmed the crystalline nature of the particles. The minimum inhibitory concentrations (MICs) of the AgNPs against the tested MDR bacteria ranged from 48 to 56 µg/ml, while the minimum bactericidal concentrations (MBCs) of the AgNPs against the tested strains ranged from 72 to 96 µg/ml. The AgNPs showed a good synergistic efficacy with Cefaclor, Cefoxitin, and Erythromycin. Their efficiency showed a threefold increase in the inhibition of tested strains when used in wound dressing, due to the AgNPs potentially activating the antibiotics. Consequently, we can use AgNPs with Cefaclor, Cefoxitin, and Erythromycin antibiotics as alternative antimicrobial agents, and they could be utilized in wound dressing to prevent microbial infections.

Nanoformulation of Biogenic Cefotaxime-Conjugated-Silver Nanoparticles for Enhanced Antibacterial Efficacy Against Multidrug-Resistant Bacteria and Anticancer Studies.

OBJECTIVES: Due to the expanded bacterial genetic tolerance to antibiotics through different mechanisms, infectious diseases of MDR bacteria are difficult for treatment. Consequently, we synthesized drug conjugated nanoparticles to dissolve this problem. Moreover, the present study aims to display the cell death status treated with cefotaxime-CS-AgNPs and also, apoptosis pathways of human RPE-1 normal cells and human MCF-7 breast cancer cells. METHODS: Here, we demonstrate the possibility to synthesize AgNPs and conjugate them with cefotaxime to survey the probability of cefotaxime-CS-AgNPs as an antimicrobial agent against cefotaxime-resistant strains E. coli and MRSA. RESULTS: TEM showed the size of AgNPs, CS-AgNPs and cefotaxime-CS-AgNPs ranged from 7.42 to 18.3 nm, 8.05-23.89 nm and 8.48-25.3 nm, respectively, with a spherical shape. The cefotaxime-CS-AgNPs enhanced the high antimicrobial properties compared to AgNPs or pure antibiotic. The MIC of Cefotaxime-CS-AgNPs ranged from 3 µg/mL to 8 µg/mL against tested E. coli and MRSA bacteria. Consequently, the highest reduction in the MIC of cefotaxime-CS-AgNPs was noted against tested strains ranging from 22% to 96%. Comparing cefotaime-CS-AgNPs to AgNPs we showed that cefotaime-CS-AgNPs have no cytotoxic effect on normal cells at even 12 µg/mL for 24 hrs. The IC50 for the AgNPs and cefotaxime-CS-AgNPs was 12 µg/mL for human RPE-1 normal cells and human MCF-7 breast cancer cell lines. The pro-apoptotic genes p53, p21, and Bax of cancer cell lines significantly upregulated followed by downregulated by anti-apoptotic gene Bcl-2 after 48 hrs at 24 µg/mL, and this concentration represents the most effective dose. CONCLUSION: Results enhanced the conjugating utility in old unresponsive cefotaxime to AgNPs to restore its efficiency against previous strains and demonstrated potential therapeutic applications of cefotaxime-CS-AgNPs. Moreover, this research gives remarkable insights for designing nanoscale delivery and curative systems that have a pronounced cytotoxic activity on cancer cells and are safe to normal cells.

Formulation of Ceftriaxone Conjugated Gold Nanoparticles and Their Medical Applications against Extended-Spectrum ß-Lactamase Producing Bacteria and Breast Cancer.

Gold nanoparticles (AuNP) and their conjugates have been gaining a great deal of recognition in the medical field. Meanwhile, extended-spectrum ß-lactamases (ESBL)-producing bacteria are also demonstrating a challenging problem for health care. The aim of this study was the biosynthesis of AuNP using Rosa damascenes petal extract and conjugation of ceftriaxone antibiotic (Cef-AuNP) in inhibiting ESBL-producing bacteria and study of in vitro anticancer activity. Characterization of the synthesized AuNP and Cef-AuNP was studied. ESB-Lproducing strains, Acinetobacter baumannii ACI1 and Pseudomonas aeruginosa PSE4 were used for testing the efficacy of Cef-AuNP. The cells of MCF-7 breast cancer were treated with previous AuNP and Cef-AuNP at different time intervals. Cytotoxicity effects of apoptosis and its molecular mechanism were evaluated. Ultraviolet-visible spectroscopy and Fourier transform infrared spectroscopy established the formation of AuNP and Cef-AuNP. Transmission electron microscope demonstrated that the formed nanoparticles were of different shapes with sizes of 15~35 nm and conjugation was established by a slight increase in size. Minimum inhibitory concentration (MIC) values of Cef-AuNP against tested strains were obtained as 3.6 and 4 µg/ml, respectively. Cef-AuNP demonstrated a decrease in the MIC of ceftriaxone down to more than 27 folds on the studied strains. The biosynthesized AuNP displayed apoptotic and time-dependent cytotoxic effects in the cells of MCF-7 at a concentration of 0.1 µg/ml medium. The Cef-AuNP have low significant effects on MCF-7 cells. These results enhance the conjugating utility in old unresponsive ceftriaxone with AuNP to restore its efficiency against otherwise resistant bacterial pathogens. Additionally, AuNP may be used as an alternative chemotherapeutic treatment of MCF-7 cancer cells.