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| Changes in time-dependent killing efficiency of naphthoquinone after addition of AgNPs with the example of 3ChPL tested toward S. aureus strain ATCC 25923. Curves were obtained for concentrations of agents used alone or in combinations equal to 1× MBC (8 µg/mL) for 3ChPL and 0.25× MBC (3.6 µg/mL) for AgNPs. Results are reported as mean values of 9 replicates ± SD. Values indicated with similar letters are significantly different from each other (p < α, α = 0.01).
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Staphylococcus aureus is a human pathogen responsible for many antibiotic-resistant infections, for instance burn wound infections, which pose a threat to human life. Exploring possible synergy between various antimicrobial agents, like nanoparticles and plant natural products, may provide new weapons to combat antibiotic resistant pathogens. The o...
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... on time-dependent killing of bacterial cells confirmed the synergistic interaction of AgNPs and NQs (Figure 3). First of all, the number of bacterial cells treated with AgNPs at concentration corresponding to 0.25× MBC (3.6 µg/mL) combined with 3ChPL at its bactericidal concentration (8 µg/mL) dropped significantly. ...
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... Bacterial cell membrane disruption is supposedly the mechanism underlying the synergistic effects of 3ChPL and AgNPs. Synergistic interactions with AgNPs increased the efficacy of reduced 3ChPL concentrations, at which they are non-toxic to human keratinocytes (Krychowiak et al., 2018). Further studies have shown that AgNPs overcome intrinsic microbial resistance to NQs (Krychowiak-Maśnicka et al., 2021). ...
Plant cell and organ cultures are potential sources of valuable secondary metabolites that can be used as food additives, nutraceuticals, cosmeceuticals, and pharmaceuticals. Phytochemical biosynthesis in various in vitro plant cultures, in contrast to that in planta, is independent of environmental conditions and free from quality fluctuations.
Pharmaceutical application of plant biotechnology is of interest to almost all departments of the Faculty of Pharmacy and Institute of Pharmacology in Poland with a botanical profile (Pharmaceutical Botany, Pharmacognosy, and Pharmacology).
This study discusses the advances in plant biotechnology for the production of known metabolites and/or biosynthesis of novel compounds in plant cell and organ in vitro cultures in several scientific centers in Poland.
... To investigate synergistic activity of MF and AMP B with NPs, broth micro dilution assay was performed [31]. Briefly, JCuS NPs (0.1-1 lM/mL) and MF (1-10 lg/mL) were incubated in a 96 well plate, along with 100 lL of media and 50 lL of cells. ...
Vulvovaginal candidiasis (VVC) is a most common infection caused by Candida species especially, Candida albicans. Herein, we report that phytoprotein based nano scaffolds mitigates the standard antifungal drug (fluconazole) resistant C. albicans. Copper sulfide nanoparticles (CuS NPs) and the phytolectin, jacalin functionalized copper sulfide nanoparticles (JCuS NPs) were evaluated against C. albicans cells. The minimum inhibitory concentration of JCuS NPs (0.1 µM) was tenfold lower than CuS NPs (1 µM). Kill curve assay showed that the NPs exhibit anticandidal activity in a time dependent manner. Standard antifungal drugs like Amphotericin B (AMP B) and Micafungin (MF) were evaluated individually against VVC causing Candida sp. and also combined with NPs to determine their synergistic effect. Interestingly, MF which belongs to echinocandins group of antifungals exhibhited synergistic activity and AMP B which belongs to polyenes group displayed an additive effect. Molecular level studies revealed that JCuS NPs in combination with micafungin inhibits the drug resistant efflux pump of C. albicans by down-regulating the efflux genes. The results revealed that JCuS NPs and its combination with standard drugs have a better anti-candidal effect, suggesting that the lectins has the potential to serve as a drug carrier by targeting the cell surface glycans of Candida.
... Several studies have depicted the effective application of silver nanoparticles in conjugation with phytochemicals. Conjugation of silver nanoparticles with 3-chloroplumbagin showed a high synergistic effect with fractional bactericidal concentration index of 0.375 against the S. aureus strains (Krychowiak et al., 2018). Similarly, silver nanoparticles synthesized by using the aqueous extract of Swertia paniculata were more effective than the conventional application of the medicinal herbs against Pseudomonas aeruginosa and Klebsiella pneumoneae (Ahluwalia et al., 2018). ...
Nowadays, the pharma and food industries have been gearing up to meet the urgent need for anti-infective and anti-inflammatory nutritional formulations. In this way, several nutraceutical compounds are being re–evaluated due to their established bioactivities. Few compounds have been or may be efficiently targeted against infections, inflammatory conditions and for immune modulation. However, for successful management of these metabolic conditions, the nutraceuticals need to be designed into effective nutritional formulations. Over recent years there have been tremendous progress in the re-engineering of structurally delivery vehicles which provide stability, enhance bioaccessibility and bioavailability of these compounds. In this perspective, this review focuses on the structural and functional aspects of several such bio-based delivery vehicles like the micro and nano particles, nano-emulsions and liposome-based models. The aim is to bring forth recent information on the efficacious nutraceuticals and the suitable delivery vehicles which would be useful against infections and inflammatory conditions.
... INPs inhibit bacterial growth through different mechanisms [91,92]. However, combinatorial treatments of INPs with antibiotics, polymers, and antimicrobial peptides, can produce a synergistic effect and reduce the therapeutic doses [93,94]. ...
Since the discovery of antibiotics, humanity has been able to cope with the battle against bacterial infections. However, the inappropriate use of antibiotics, the lack of innovation in therapeutic agents, and other factors have allowed the emergence of new bacterial strains resistant to multiple antibiotic treatments, causing a crisis in the health sector. Furthermore, the World Health Organization has listed a series of pathogens (ESKAPE group) that have acquired new and varied resistance to different antibiotics families. Therefore, the scientific community has prioritized designing and developing novel treatments to combat these ESKAPE pathogens and other emergent multidrug-resistant bacteria. One of the solutions is the use of combinatorial therapies. Combinatorial therapies seek to enhance the effects of individual treatments at lower doses, bringing the advantage of being, in most cases, much less harmful to patients. Among the new developments in combinatorial therapies, nanomaterials have gained significant interest. Some of the most promising nanotherapeutics include polymers, inorganic nanoparticles, and antimicrobial peptides due to their bactericidal and nanocarrier properties. Therefore, this review focuses on discussing the state-of-the-art of the most significant advances and concludes with a perspective on the future developments of nanotherapeutic combinatorial treatments that target bacterial infections.
... These compounds show synergy with AgNPs and have an increased cytotoxic effect against Staphylococcus aureus (Li et al., 2011). Combination agents of AgNPs and naphthoquinones suggest a potential strategy to control antibiotic-resistant bacteria such as S.aureus (Krychowiak et al., 2018). ...
Plant derived drugs or formulations have always been explored because of their lower side effects and toxicities compared to synthetic drugs and they have been widely used as traditional and complementary medicines for the management of many diseases including cancer. The major challenges faced were the absorption of the plant-derived drugs, their stability, bioavailability, and transport to the intended sites inside the body. Recent progress in nanotechnology has helped to minimize these limitations and hence phyto-nanoformulations are slowly growing in preclinical trials as well as clinical use. The use of various nanostructures such as nano-micelles, lipid nanoparticles, carbon nanotubes, polymer nanoparticles, and nanoliposomes and various types of drug delivery vehicles such as polybutylcyanoacrylate, polylactic-co-glycolic acid, and lactoferrin has immensely helped in increasing the effectiveness of phytochemical drugs by increasing their stability, better pharmacokinetics and reducing the toxicity and side effects. Phyto-nanoformulations having natural product components such as curcumin, piperine, quercetin, berberine, scutellarin, baicalin, stevioside, silybin, gymnemic acid, naringenin, capsicum oleoresin, emodin, and resveratrol have been shown to improve the condition of patients diagnosed with diseases such as neurodegenerative disorders, diabetes, infection, and cancer. Phyto nanoformulations can also be used to treat disorders of the brain where the blood-brain barrier is impervious to the drugs. These phyto- nanoformulations have been shown to target several molecular cell-signaling and metabolic pathways. This chapter covers the compositions of phyto-nanoformulations and how they have been used to control several diseases.
... Both AMPs and AgNPs have demonstrated their potential in combating resistant microbes (Lara et al., 2010;Rai et al., 2012;Roque-Borda et al., 2021), and both have also been found to act synergistically with conventional antibiotics (Cassone and Otvos, 2010;Wan et al., 2016;Krychowiak et al., 2018;Ruden et al., 2019;Zharkova et al., 2019;Duong et al., 2021). Moreover, it has previously been reported by us as well as by others (Ruden et al., 2009;Salouti et al., 2016), that AMPs and AgNPs can enhance each other's antimicrobial activity, when used in combination. ...
Silver nanoparticles (AgNPs) and antimicrobial peptides or proteins (AMPs/APs) are both considered as promising platforms for the development of novel therapeutic agents effective against the growing number of drug-resistant pathogens. The observed synergy of their antibacterial activity suggested the prospect of introducing antimicrobial peptides or small antimicrobial proteins into the gelatinized coating of AgNPs. Conjugates with protegrin-1, indolicidin, protamine, histones, and lysozyme were comparatively tested for their antibacterial properties and compared with unconjugated nanoparticles and antimicrobial polypeptides alone. Their toxic effects were similarly tested against both normal eukaryotic cells (human erythrocytes, peripheral blood mononuclear cells, neutrophils, and dermal fibroblasts) and tumor cells (human erythromyeloid leukemia K562 and human histiocytic lymphoma U937 cell lines). The AMPs/APs retained their ability to enhance the antibacterial activity of AgNPs against both Gram-positive and Gram-negative bacteria, including drug-resistant strains, when conjugated to the AgNP surface. The small, membranolytic protegrin-1 was the most efficient, suggesting that a short, rigid structure is not a limiting factor despite the constraints imposed by binding to the nanoparticle. Some of the conjugated AMPs/APs clearly affected the ability of nanoparticle to permeabilize the outer membrane of Escherichia coli , but none of the conjugated AgNPs acquired the capacity to permeabilize its cytoplasmic membrane, regardless of the membranolytic potency of the bound polypeptide. Low hemolytic activity was also found for all AgNP-AMP/AP conjugates, regardless of the hemolytic activity of the free polypeptides, making conjugation a promising strategy not only to enhance their antimicrobial potential but also to effectively reduce the toxicity of membranolytic AMPs. The observation that metabolic processes and O 2 consumption in bacteria were efficiently inhibited by all forms of AgNPs is the most likely explanation for their rapid and bactericidal action. AMP-dependent properties in the activity pattern of various conjugates toward eukaryotic cells suggest that immunomodulatory, wound-healing, and other effects of the polypeptides are at least partially transferred to the nanoparticles, so that functionalization of AgNPs may have effects beyond just modulation of direct antibacterial activity. In addition, some conjugated nanoparticles are selectively toxic to tumor cells. However, caution is required as not all modulatory effects are necessarily beneficial to normal host cells.
... AgNPs combined with gentamicin and chloramphenicol indicate the synergistic action against Enterococcus faecalis and MRD pathogenic bacterium associated with hospital-acquired infections (Katva et al. 2018). Krychowiak et al. (2018) studied synergistic actions of AgNPs using four tested naphthoquinones (NQs): ramentaceone (RAM), plumbagin (PL), droserone (DR), and 3-chloroplumbagin (3ChPL) (FBC indexes ≤ 0.375), where the results showed an increase in bactericidal effect toward MDR S. aureus. ...
In more than two-thirds of the diabetic foot ulcer (DFUs) cases, lower limb amputation of foot ulceration is caused by the infection. The role of transition a metal complex as a therapeutic compound is becoming increasingly important. In vitro, four groups of antibiotics and one sulfa drug were tested against diabetic foot resistant bacteria. Using three concentrations of two different prepared metal complexes: copper (Cu) and silver (Ag) — isoniazid (Iso) and nicotinamide (Nicot) were tested against diabetic foot isolates. Results revealed that β-lactam drugs (cephradine and piperacillin) showed the minimum averages of MIC 265 μg/ml against Gram-positive and Gram-negative isolates. Silver isoniazid (Iso-Ag-1) metal complex was selected depending on the maximum averages of MIC against both types of clinical isolates. The combination between β-lactams and Iso-Ag-1 showed maximum FICI averages of 0.24 for Gram-positive and 0.28 for Gram-negative. In addition, a combination between Iso-Ag-1 with squilla chitosan nanoparticles (CSSq-nAg) showed averages of synergistic index by 0.23 against Staphylococcus aureus and 0.13, 0.30, and 0.27 against E. coli, K. pneumoniae, and Ps. aeruginosa, respectively. Final formula of Iso-Ag-1+CSSq-nAg + β-lactams (cephradine and piperacillin) showed a synergistic effect at FICI = 0.044 and 0.047, against G+ve and –ve, respectively. These two combinations showed a slight toxicity against the water flea Daphnia magna by 3.49 and 3.6 ppm, respectively. Results suggest the use of Iso-Ag-1-CSSq-nAg as enhancing agent in combination with β-lactams as a blind therapy in pharmaceutical preparations.
... This suggests that the phyto-fabricated AgNPs may suppress the chemical components in antibiotic discs. The combined activity of the AgNPs and antibiotics was in agreement with previous results for AgNPs using biogenic extracts [45][46][47]. Of note, antibacterial action for the antibiotic-associated AgNPs could be efficient for treating antibiotic-resistant bacteria in humans [21]. ...
... Of note, antibacterial action for the antibiotic-associated AgNPs could be efficient for treating antibiotic-resistant bacteria in humans [21]. [45][46][47]. Of note, antibacterial action for the antibiotic-associated AgNPs could be efficient for treating antibiotic-resistant bacteria in humans [21]. ...
The application of biological materials in synthesizing nanoparticles has become significant issue in nanotechnology. This research was designed to assess biogenic silver nanoparticles (AgNPs) fabricated using two aqueous extracts of Acacia arabica (Arabic Gum) (A-AgNPs) and Opophytum forsskalii (Samh) seed (O-AgNPs), which were used as reducing and capping agents in the NPs development, respectively. The current study is considered as the first report for AgNP preparation using Opophytum forsskalii extract. The dynamic light scattering, transmission electron microscopy, and scanning electron microscopy were employed to analyze the size and morphology of the biogenic AgNPs. Fourier transform infrared (FTIR) spectroscopy and chromatography/mass spectrometry (GC-MS) techniques were used to identify the possible phyto-components of plant extracts. The phyto-fabricated NPs were assessed for their antibacterial activity and also when combined with some antibiotics against Staphylococcus aureus (Gram-positive) and Pseudomonas aeruginosa and Escherichia coli (Gram-negative) and their anticandidal ability against Candida albicans using an agar well diffusion test. Furthermore, cytotoxicity against LoVo cancer cell lines was studied. The results demonstrated the capability of the investigated plant extracts to change Ag+ ions into spherical AgNPs with average size diameters of 91 nm for the prepared O-AgNPs and 75 nm for A-AgNPs. The phyto-fabricated AgNPs presented substantial antimicrobial capabilities with a zone diameter in the range of 10–29.3 mm. Synergistic effects against all tested strains were observed when the antibiotic and phyto-fabricated AgNPs were combined and assessed. The IC50 of the fabricated O-AgNPs against LoVo cancer cell lines was 28.32 μg/mL. Ten and four chemical components were identified in Acacia arabica (Arabic Gum) and Opophytum forsskalii seed extracts, respectively, by GC-MS that are expected as NPs reducing and capping agents. Current results could lead to options for further research, such as investigating the internal mechanism of AgNPs in bacteria, Candida spp., and LoVo cancer cell lines as well as identifying specific molecules with a substantial impact as metal-reducing agents and biological activities.
... The antibacterial activity of particular pure secondary metabolites found in D. muscipula extract was determined, as well as their interaction with AgNPs. Among the identified compounds (Table 3), we studied those which are commercially available, as well as those applied in our previous study [18,34], which represented naphthoquinones (plumbagin, 3-chloroplumbagin), flavonoids and their glycosides (quercetin, kaempferol, hyperoside), as well as phenolic acids (ellagic acid and 3,3′-di-O-methylellagic acid). None of the secondary metabolites studied were active against P. aeruginosa in the tested range of concentration, i.e., up to 512 µ g mL −1 . ...
... According to the study and our previous reports [18,34], the AgNP-naphthoquinone interaction is unique and highly specific. However, it is not clear whether its synergistic bactericidal potential against P. aeruginosa is based on an interaction with one molecular target or a particular cellular pathway, or comprises the result of multi-target action. ...
... The broth microdilution method was applied to determine bacterial susceptibility to tested agents, as described previously [34]. Experiments were performed according to CLSI guidelines for minimal bactericidal concentration (MBC) determination (CLSI 1996), as follows: a 2-fold serial dilutions of tested agents were performed on 96-well plates containing 100 µ L of solution per well. ...
Carnivorous plants are exemplary natural sources of secondary metabolites with biological activity. However, the therapeutic antimicrobial potential of these compounds is limited due to intrinsic resistance of selected bacterial pathogens, among which Pseudomonas aeruginosa represents an extreme example. The objective of the study was to overcome the intrinsic resistance of P. aeru-ginosa by combining silver nanoparticles (AgNPs) with secondary metabolites from selected carniv-orous plant species. We employed the broth microdilution method, the checkerboard titration technique and comprehensive phytochemical analyses to define interactions between nanoparticles and active compounds from carnivorous plants. It has been confirmed that P. aeruginosa is resistant to a broad range of secondary metabolites from carnivorous plants, i.e., naphthoquinones, flavonoids, phenolic acids (MBC = 512 µ g mL −1) and only weakly sensitive to their mixtures, i.e., extracts and extracts' fractions. However, it was shown that the antimicrobial activity of extracts and fractions with a significant level of naphthoquinone (plumbagin) was significantly enhanced by AgNPs. Our studies clearly demonstrated a crucial role of naphthoquinones in AgNPs and extract interaction, as well as depicted the potential of AgNPs to restore the bactericidal activity of naphthoquinones towards P. aeruginosa. Our findings indicate the significant potential of nanoparticles to modulate the activity of selected secondary metabolites and revisit their antimicrobial potential towards human pathogenic bacteria.
... Microbial resistance can be effectively overcome through strategies that are created on the basis of synergistic combinations of antimicrobial agents and also through drug designs developed on the basis of nanotechnology and phytomedicine (Krychowiak et al., 2018). Various studies have been carried out in the past few years that show that AgNPs may increase the antibacterial activities of antibiotics against susceptible as well as resistant bacteria, either in an additive manner or synergistically (Panáček et al., 2016). ...
For the first time, this study was carried out to investigate and evaluate the relative antibacterial activity of three different Nk-lysin peptides from human, chicken, and bovine activity compared to Gram-negative and Gram-positive bacteria as well as antiviral activity against rotavirus (strain SA-11) and finally mechanisms of action optionality. This report is the first of its kind that investigates the increased antimicrobial ability of (Nk-lysin + AgNPs) and (Nk-lysin + human IL-2) combinations against S. typhi activity by carrying out direct comparison under similar experimental settings. Our results showed that gram-negative and gram-positive microorganisms, including Streptococcus pyogenes , Streptococcus mutans , Escherichia coli , Pseudomonas aeruginosa , Klebsiella oxytoca , Shigella sonnei , Klebsiella pneumoniae and Salmonella typhimurium , are susceptible to NK-lysin treatment. It was shown in our findings that there was equal potentiality in mixture (Nk-lysin + AgNPs) and (Nk-lysin + human IL-2) for preventing the growth of S. typhi , however, when added together, there was minor increase in the level of action. In our study, the TOHO-1 gene was absent in treated bacteria. Following treatment with Nk-lysin peptides, the beta-lactamases genes (CTX-M-1, M-8, and M-9) were not found in any bacterial strains. The examination did not find any of the plasmid mediated quinolone resistance genes in the bacterial strains as a response to NK-lysin treatment. Nonetheless, no study has been carried out in the past that characterized the antiviral activity of bovine, human and chicken Nk-lysin peptides. Hence, this is the foremost study on the enhanced antimicrobial activity of human, bovine and chicken Nk-lysin peptides against Rotavirus (strain SA-11). The findings of the study demonstrated that the powerful antiviral activities were exhibited by Nk-lysin peptides against Rotavirus (strain SA-11). Based on the comparison between these peptides, it can be concluded that there is an evident potent antiviral activity of bovine Nk-lysin against Rotavirus (strain SA-11) as it restrains infection by up to 90%. However, growth was restricted by 80% by chicken Nk-lysin and by 50% by the human peptide.
