Controlled biosynthesis of AgCl nanoparticles by a thermotolerant <Emphasis Type="Italic">Aspergillus terreus</Emphasis> in the L-Tryptophan supplemented media: Characterization and antimicrobial activity

A simple and green method was developed for the extracellular biosynthesis of silver chloride nanoparticles, free from silver nanoparticles, using cell-free filtrate of a thermotolerant fungal strain Aspergillus terreus 8. The synthesized silver chloride nanoparticles exhibited characteristic absorption maximum at 275 nm. As-fabricated AgCl-NPs were characterized by UV-vis spectroscopy, XRD, SEM-EDX, and FT-IR. The biosynthesized silver chloride nanoparticles exhibited strong antimicrobial activity towards pathogenic microorganisms such as Fusarium oxysporum f. sp. vasinfectum and Verticillium dahliae. The synthesized silver chloride nanoparticles can be exploited as a promising new biocide bionanocomposite against pathogenic microorganisms.     

Biosynthesis of silver nanoparticles with the participation of extracellular Mn-dependent peroxidase from <Emphasis Type="Italic">Azospirillum</Emphasis>

The accumulation of nanoparticles of colloidal silver with spherical shape in culture liquid of Azospirillum brasilense has been shown by transmission electron microscopy. Bacterial extracellular Mn-peroxidases were found to participate in silver reduction from silver nitrate with the formation of nanoparticles. A mechanism of extracellular biosynthesis of silver nanoparticles by A. brasilense bacteria was proposed.     

Antimicrobial Activity of Silver Nanoparticles in a Carboxymethyl Chitin Matrix Obtained by the Microwave Hydrothermal Method

Silver nanoparticles have been obtained in a matrix of 6-O-carboxymethyl chitin in the presence of D-glucose as a reducing agent by microwave hydrothermal synthesis. The TEM results show that the silver nanoparticles have a spherical shape; the particle size range is 3–20 nm. The resulting colloidal solution of silver nanoparticles had a strong bacterial effect on gram-positive bacteria Staphylococcus aureus ATCC 21027 (=209 P), Bacillus subtilis ATCC 6633, and, to a lesser extent, on gram-negative bacteria Escherichia coli АТСС 25922. The synthesized silver nanoparticles showed pronounced fungistatic activity against A. niger INA 00760.     

Galactose-Containing Polymer-DOX Conjugates for Targeting Drug Delivery

A novel multifunctional drug delivery system was fabricated by conjugating galactose-based polymer, methoxy-poly(ethylene glycol)-block-poly(6-O-methacryloyl-D-galactopyranose) (mPEG-b-PMAGP) with doxorubicin (DOX) via an acid-labile carbamate linkage. The mPEG-b-PMAGP-co-DOX nanoparticles were spherical in shape, and the diameter determined by dynamic light scattering (DLS) was 54.84?±?0.58 nm, larger than that characterized by transmission electron microscopy (TEM). The in vitro drug release profiles were studied, and the release of DOX from the nanoparticles was pH-responsive. The cellular uptake behavior of free-DOX and mPEG-b-PMAGP-co-DOX nanoparticles by asialoglycoprotein (ASGP) receptor-positive cancer cell line (HepG2) and ASGP receptor-negative cancer cell lines (MCF-7 and A549 cells) was evaluated by confocal laser scanning microscopy (CLSM) and flow cytometry (FCM), respectively. The mPEG-b-PMAGP-co-DOX nanoparticles which contain galactose functional groups exhibited higher cellular uptake behavior via ASGP receptor-mediated endocytosis in HepG2 cells than in other two cancer cells. The in vitro cytotoxicity assay manifested that the mPEG-b-PMAGP-co-DOX nanoparticles exhibited higher anticancer efficacy against HepG2 cells than MCF-7 cells. These results indicated that the multifunctional mPEG-b-PMAGP-co-DOX nanoparticles possessing pH-responsible and hepatoma-targeting function have great potential to be used as a targeting drug delivery system for hepatoma therapy.     

Polysaccharide-based silver nanoparticles synthesized by <Emphasis Type="Italic">Klebsiella oxytoca</Emphasis> DSM 29614 cause DNA fragmentation in <Emphasis Type="Italic">E. coli</Emphasis> cells

Silver nanoparticles (AgNPs), embedded into a specific exopolysaccharide (EPS), were produced by Klebsiella oxytoca DSM 29614 by adding AgNO₃ to the cultures during exponential growth phase. In particular, under aerobic or anaerobic conditions, two types of silver nanoparticles, named AgNPs-EPS^aer and the AgNPs-EPS^anaer, were produced respectively. The effects on bacterial cells was demonstrated by using Escherichia coli K12 and Kocuria rhizophila ATCC 9341 (ex Micrococcus luteus) as Gram-negative and Gram-positive tester strains, respectively. The best antimicrobial activity was observed for AgNPs-EPS^aer, in terms of minimum inhibitory concentrations and minimum bactericidal concentrations. Observations by transmission electron microscopy showed that the cell morphology of both tester strains changed during the exposition to AgNPs-EPS^aer. In particular, an electron-dense wrapped filament was observed in E. coli cytoplasm after 3 h of AgNPs-EPS^aer exposition, apparently due to silver accumulation in DNA, and both E. coli and K. rhizophila cells were lysed after 18 h of exposure to AgNPs-EPS^aer. The DNA breakage in E. coli cells was confirmed by the comparison of 3-D fluorescence spectra fingerprints of DNA. Finally the accumulation of silver on DNA of E. coli was confirmed directly by a significant Ag⁺ release from DNA, using the scanning electrochemical microscopy and the voltammetric determinations.     

Forage Crop <Emphasis Type="Italic">Lolium multiflorum</Emphasis> Assisted Synthesis of AgNPs and Their Bioactivities Against Poultry Pathogenic Bacteria in In Vitro

Italian ryegrass is one of main feed for livestock animals/birds. It has potential antioxidant metabolites that can improve their health and protect them against various infectious diseases. In this work, we studied synthesis of silver nanoparticles assisted by forage crop Lolium multiflorum as a green synthesis way. Potential antibacterial efficacy of these synthesized nanosized silver nanoparticles against poultry pathogenic bacteria was then studied. Aqueous extract of IRG was used as reducing agent for bio-reduction of silver salt to convert Ag⁺ to Ag⁰ metallic nano-silver. Size, shape, metallic composition, functional group, and crystalline nature of these synthesized silver nanoparticles were then characterized using UV–Vis spectrophotometer, FESEM, EDX, FT-IT, and XRD, respectively. In addition, antibacterial effects of these synthesized AgNPs against poultry pathogenic bacteria were evaluated by agar well diffusion method. UV–Vis spectra showed strong absorption peak of 440–450 nm with differ reaction time ranging from 30 min to 24 h. FESEM measurements revealed particles sizes of around 20–100 nm, majority of which were spherical in shape while a few were irregular. These biosynthesized silver nanoparticles using IRG extract exhibited strong antibacterial activities against poultry pathogenic microorganisms, including Pseudomonas aeruginosa, Salmonella typhi, Escherichia coli, and Bacillus subtilis. Overall results confirmed that IRG plant extract possessed potential bioactive compounds for converting silver ions into nanosized silver at room temperature without needing any external chemical for redox reaction. In addition, such synthesized AgNPs showed strong antibacterial activities against pathogenic bacteria responsible for infectious diseases in poultry.     

Silver Nanoparticles in Epoxy Resin Deposited on Silicon Substrates for Light Trapping

Metallic silver nanoparticles were prepared in epoxy resin using N,N-dimethylformamide (DMF) as reducing agent of silver ions at room temperature. They were characterized by UV–Vis spectroscopy, atomic force microscopy (AFM), transmission electronic microscopy (TEM), and high-resolution transmission electronic microscopy (HRTEM). The silver nanoparticles showed broadband absorption spectra attributed to high-order plasmonic resonances. The morphology of the metallic particles corresponds to elongated particles and their aggregates with a size above 30 nm. These silver nanoparticles were deposited by the spin-coating process on crystalline silicon at room temperature. Then, the antireflective properties of these samples were measured. According to the observed results, it is inferred that the films of coalesced silver nanoparticles decrease the reflectance of crystalline silicon better than particles separated by large distances. These results are discussed in terms of forward scattering of large metallic nanoparticles where higher order multipolar modes are dominant and the retardation effects are very important.     

Antibacterial evaluation of silver nanoparticles synthesized from lychee peel: individual versus antibiotic conjugated effects

This paper describes the extracellular synthesis of silver nanoparticles from waste part of lychee fruit (peel) and their conjugation with selected antibiotics (amoxicillin, cefixim, and streptomycin). FTIR studies revealed the reduction of metallic silver and stabilization of silver nanoparticles and their conjugates due to the presence of CO (carboxyl), OH (hydroxyl) and CH (alkanes) groups. The size of conjugated nanoparticles varied ranging from 3 to 10 nm as shown by XRD. TEM image revealed the spherical shape of biosynthesized silver nanoparticles. Conjugates of amoxicillin and cefixim showed highest antibacterial activity (147.43 and 107.95%, respectively) against Gram-negative bacteria i.e. Alcaligenes faecalis in comparison with their control counterparts. The highest reduction in MIC was noted against Gram-positive strains i.e. Enterococcus faecium (75%) and Microbacterium oxydans (75%) for amoxicillin conjugates. Anova two factor followed by two-tailed t test showed non-significant results both in case of cell leakage and protein estimation between nanoparticles and conjugates of amoxicillin, cefixime and streptomycin. In case of MDA release, non-significant difference among the test samples against the selected strains. Our study found green-synthesized silver nanoparticles as effective antibacterial bullet against both Gram positive and Gram negative bacteria, but they showed a more promising effect on conjugation with selected antibiotics against Gram negative type.     

Elucidating the interactions and phytotoxicity of zinc oxide nanoparticles with agriculturally beneficial bacteria and selected crop plants

There is a growing interest in the use of bioinoculants to assist mineral fertilizers in improving crop production and yield. Azotobacter and Pseudomonas are two agriculturally relevant strains of bacteria which have been established as efficient bioinoculants. An experiment involving addition of graded concentrations of zinc oxide (ZnO) nanoparticles was undertaken using log phase cultures of Azotobacter and Pseudomonas. Growth kinetics revealed a clear trend of gradual decrease with Pseudomonas; however, Azotobacter exhibited a twofold enhancement in growth with increase in the concentration of ZnO concentration. Scanning electron microscopy (SEM), supported by energy-dispersive X-ray (EDX) analyses, illustrated the significant effect of ZnO nanoparticles on Azotobacter by the enhancement in the abundance of globular biofilm-like structures and the intracellular presence of ZnO, with the increase in its concentration. It can be surmised that extracellular mucilage production in Azotobacter may be providing a barrier to the nanoparticles. Further experiments with Azotobacter by inoculation of wheat and tomato seeds with ZnO nanoparticles alone or bacteria grown on ZnO-infused growth medium revealed interesting results. Vigour index of wheat seeds reduced by 40–50% in the presence of different concentrations of ZnO nanoparticles alone, which was alleviated by 15–20%, when ZnO and Azotobacter were present together. However, a drastic 50–60% decrease in vigour indices of tomato seeds was recorded, irrespective of Azotobacter inoculation.     

Association Between Chronic Exposure to Tobacco Smoke and Accumulation of Toxic Metals in Hair Among Pregnant Women

Tobacco smoke contains various toxic heavy metals that individuals are exposed to when they smoke. Despite the presence of heavy metals in tobacco smoke, the relationship between smoking and the accumulation of toxic metals in pregnant women after long-term exposure remains under discussion. We examined the association between long-term exposure to tobacco smoke and the accumulation of toxic metals in the hair of female participants. Our study recruited 252 women from the Shanxi and Hebei provinces of Northern China; these participants were self-reported non-active smokers, and had previously delivered healthy babies without birth defects. Scalp hair was collected and analyzed for nicotine and cotinine and five potentially toxic metals (specifically, silver, chromium, cadmium, mercury, and lead). Our results showed significant positive correlations between cotinine and four metals, including silver (r?=?0.369, p?<?0.001), cadmium (r?=?0.185, p?<?0.01), mercury (r?=?0.161, p?<?0.05), and lead (r?=?0.243, p?<?0.001). Significant positive correlations were also found between nicotine and three metals—specifically silver (r?=?0.331, p?<?0.001), cadmium (r?=?0.176, p?<?0.01), and lead (r?=?0.316, p?<?0.001). A logistic regression model showed significant associations between cotinine and potentially toxic metals including mercury, silver, and lead (with or without adjusting for potential confounders). We thus conclude that long-term passive smoking could potentially increase the exposure level of toxic metals including lead, silver, and mercury in our study, which are especially harmful for pregnant women and their unborn fetus.     

Growth of Ag-nanoparticles in an aqueous solution and their antimicrobial activities against Gram positive,Gram negative bacterial strains and Candida fungus

Silver nanoparticles (AgNPs) were synthesized using Ocimum sanctum (Tulsi) leaves aqueous extract as reducing as well as a capping agent in absence and presence of cetyltrimethylammonium bromide (CTAB). The resulting nanomaterials were characterized by UV–visible spectrophotometer, and transmission electron microscope. The UV–Vis spectroscopy revealed the formation of AgNPs at 400–450 nm. TEM photographs indicate that the truncated triangular silver nanoplates and/or spherical morphology of the AgNPs with an average diameter of 25 nm have been distorted markedly in presence of CTAB. The AgNPs were almost mono disperse in nature. Antimicrobial activities of AgNPs were determined by using two bacteria (Gram positive Staphylococcus aureus MTCC-3160), Gram negative Escherichia coli MTCC-450) and one species of Candida fungus (Candida albicans ATCC 90030) with Kirby-Bauer or disc diffusion method. The zone of inhibition seems extremely good showing a relatively large zone of inhibition in both Staphylococcus aureus, Escherichia coli, and Candida albicans strains.     

Chromosomal Sil system contributes to silver resistance in <Emphasis Type="Italic">E. coli</Emphasis> ATCC 8739

The rise of antibiotic resistance in pathogenic bacteria is endangering the efficacy of antibiotics, which consequently results in greater use of silver as a biocide. Chromosomal mapping of the Cus system or plasmid encoded Sil system and their relationship with silver resistance was studied for several gram-negative bacteria. However, only few reports investigated silver detoxification mediated by the Sil system integrated in Escherichia coli chromosome. Accordingly, this work aimed to study the Sil system in E. coli ATCC 8739 and to produce evidence for its role in silver resistance development. Silver resistance was induced in E. coli ATCC 8739 by stepwise passage in culture media containing increasing concentrations of AgNO₃. The published genome of E. coli ATCC 8739 contains a region showing strong homology to the Sil system genes. The role of this region in E. coli ATCC 8739 was assessed by monitoring the expression of silC upon silver stress, which resulted in a 350-fold increased expression. De novo sequencing of the whole genome of a silver resistant strain derived from E. coli ATCC 8739 revealed mutations in ORFs putative for SilR and CusR. The silver resistant strain (E. coli AgNO₃R) showed constitutive expression of silC which posed a cost of fitness resulting in retarded growth. Furthermore, E. coli AgNO₃R exhibited cross-resistance to ciprofloxacin and a slightly increased tolerance to ampicillin. This study demonstrates that E. coli is able to develop resistance to silver, which may pose a threat towards an effective use of silver compounds as antiseptics.     

Magnetic characterization of superparamagnetic nanoparticles pulled through model membranes

Background

To quantitatively compare in-vitro and in vivo membrane transport studies of targeted delivery, one needs characterization of the magnetically-induced mobility of superparamagnetic iron oxide nanoparticles (SPION). Flux densities, gradients, and nanoparticle properties were measured in order to quantify the magnetic force on the SPION in both an artificial cochlear round window membrane (RWM) model and the guinea pig RWM.

Methods

Three-dimensional maps were created for flux density and magnetic gradient produced by a 24-well casing of 4.1 kilo-Gauss neodymium-iron-boron (NdFeB) disc magnets. The casing was used to pull SPION through a three-layer cell culture RWM model. Similar maps were created for a 4 inch (10.16 cm) cube 48 MGOe NdFeB magnet used to pull polymeric-nanoparticles through the RWM of anesthetized guinea pigs. Other parameters needed to compute magnetic force were nanoparticle and polymer properties, including average radius, density, magnetic susceptibility, and volume fraction of magnetite.

Results

A minimum force of 5.04 × 10^-16 N was determined to adequately pull nanoparticles through the in-vitro model. For the guinea pig RWM, the magnetic force on the polymeric nanoparticles was 9.69 × 10^-20 N. Electron microscopy confirmed the movement of the particles through both RWM models.

Conclusion

As prospective carriers of therapeutic substances, polymers containing superparamagnetic iron oxide nanoparticles were succesfully pulled through the live RWM. The force required to achieve in vivo transport was significantly lower than that required to pull nanoparticles through the in-vitro RWM model. Indeed very little force was required to accomplish measurable delivery of polymeric-SPION composite nanoparticles across the RWM, suggesting that therapeutic delivery to the inner ear by SPION is feasible.

     

Overexpression of <Emphasis Type="Italic">AtGRDP2</Emphasis> gene in common bean hairy roots generates vigorous plants with enhanced salt tolerance

Proteins with glycine-rich repeats have been identified in plants, mammalians, fungi, and bacteria. Plant glycine-rich proteins have been associated to stress response. Previously, we reported that the Arabidopsis thaliana AtGRDP2 gene, which encodes a protein with a glycine-rich domain, plays a role in growth and development of A. thaliana and Lactuca sativa. In this study, we generated composite Phaseolus vulgaris plants that overexpress the AtGRDP2 gene in hairy roots generated by Agrobacterium rhizogenes. We observed that hairy roots harboring the AtGRDP2 gene developed more abundant and faster-growing roots than control hairy roots generated with the wild type A. rhizogenes. In addition, composite common bean plants overexpressing the AtGRDP2 gene in roots were more tolerant to salt stress showing increments in their fresh and dry weight. Our data further support the role of plant GRDP genes in development and stress response.     

Formation of Ag/AgCl nanoparticles in the matrix of the exopolysaccharide of a diazotrophic strain <Emphasis Type="Italic">Azotobacter chroococcum</Emphasis> XU1

A complex of Ag/AgCl nanoparticles was synthesized on the basis of the extracellular polysaccharide of Azotobacter chroococcum XU1 and 10 mM AgNO₃ solution. The complex was characterized by UV spectroscopy, X-ray diffraction, and scanning electron microscopy. Colloidal solutions of the complex had absorption peaks at 260 and 420 nm, indicating the formation Ag/AgCl nanoparticles. The size of the nanoparticles varied from 6 to 50 nm. The nanobiocomposite consisting of the exopolysaccharide matrix and Ag/AgCl nanoparticles exhibited a fungicidal effect against such plant pathogens as Fusarium oxysporum f. sp. vasinfectum and Verticillium dahliae.     

Fabrication of silver ion exchanged zeolite using scoria and its antibacterial activity

In this study, we examined the antibacterial activity of silver ion exchanged zeolite synthesized from Cheju Scoria. We synthesized zeolite in various NaOH concentrations, but zeolite synthesized in 4 M NaOH was most similar to type A zeolite. Using the synthesized zeolite, we prepared a silver ion exchanged zeolite for studies of antibacterial activity. Antibacterial tests using agar cultures of Escherichia coli (E. coli), with silver ion exchanged zeolite showed a zone of inhibition colonies bacteria did not grow near silver ion exchanged zeolite. Furthermore, spectrophotometry demonstrated a significantly low absorbance of E. coli culture mediums when silver ion exchanged zeolite was included indicating that E. coli propagation was prevented. Through results of these experiments, we conclude that synthesized products with sodalite crystal can be synthesized from Scoria, and these are suitable to produce silver ion exchanged zeolite with antibacterial activity.     

Comparative physiological and immunological characteristics of carps (cyprinidae) grown in aquaculture with different types of food

On the basis of the hematological, cytochemical, and biochemical blood indices, it has been established that the adaptive capabilities in common carp Cyprinus carpio and grass carp Ctenopharyngodon idella are higher than in silver carp hybrid Hypophthalmichthys molitrix × H. nobilis, while the potential for growth is, by contrast, higher in silver carp hybrid.     

Antimicrobial properties of ternary eutectic aluminum alloys

Several Escherichia coli deletion mutants of the Keio collection were selected for analysis to better understand which genes may play a key role in copper or silver homeostasis. Each of the selected E. coli mutants had a deletion of a single gene predicted to encode proteins for homologous recombination or contained functions directly linked to copper or silver transport or transformation. The survival of these strains on pure copper surfaces, stainless steel, and alloys of aluminum, copper and/or silver was investigated. When exposed to pure copper surfaces, E. coli ΔcueO was the most sensitive, whereas E. coli ΔcopA was the most resistant amongst the different strains tested. However, we observed a different trend in sensitivities in E. coli strains upon exposure to alloys of the system Al–Ag–Cu. While minor antimicrobial effects were detected after exposure of E. coli ΔcopA and E. coli ΔrecA to Al–Ag alloys, no effect was detected after exposure to Al–Cu alloys. The release of copper ions and cell-associated copper ion concentrations were determined for E. coli ΔcopA and the wild-type E. coli after exposure to pure copper surfaces. Altogether, compared to binary alloys, ternary eutectic alloys (Al–Ag–Cu) had the highest antimicrobial effect and thus, warrant further investigation.     

Adsorptive removal of harmful algal species <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> directly from aqueous solution using polyethylenimine coated polysulfone-biomass composite fiber

In recent times, the treatment of harmful algal blooms (HABs) became an important environmental issue to preserve and remediate water resources globally. In the present study, the adsorptive removal of harmful algal species Microcystis aeruginosa directly from an aqueous medium was attempted. Waste biomass (Escherichia coli) was immobilized using polysulfone and coated using the cationic polymer polyethylenimine (PEI) to generate PEI-coated polysulfone-biomass composite fiber (PEI-PSBF). The density of M. aeruginosa in an aqueous medium (BG11) was significantly decreased by treatment with PEI-PSBF. additionally, analysis using FE-SEM, confirmed that the removal of M. aeruginosa algal cells by PEI-PSBF was caused by the adsorption mechanism. According to the profiles of phosphorus for the algal cell growth in M. aeruginosa cultivating samples, we found that the adsorbed M. aeruginosa onto the PEI-PSBF lost their biological activity compared to the non-treated M. aeruginosa cells.     

Antifungal Activity of Chitosan-Coated Poly(lactic-co-glycolic) Acid Nanoparticles Containing Amphotericin B

Amphotericin B (AmB) is one of the most used drugs for the treatment of systemic fungal infections; however, the treatment causes several toxic manifestations, including nephrotoxicity and hemolytic anemia. Chitosan-coated poly(lactide-co-glycolide) (PLGA) nanoparticles containing AmB were developed with the aim to decrease AmB toxicity and propose the oral route for AmB delivery. In this work, the antifungal efficacy of chitosan-coated PLGA nanoparticles containing AmB was evaluated in 20 strains of fungus isolates from patients with vulvovaginal candidiasis (01 Candida glabrata and 03 Candida albicans), bloodstream infections (04 C. albicans and 01 C. tropicalis) and patients with urinary tract infection (04 Candida albicans, 02 Trichosporon asahii, 01 C. guilhermondii, 03 C. glabrata) and 01 Candida albicans ATCC 90028. Moreover, the cytotoxicity over erythrocytes was evaluated. The single-emulsion solvent evaporation method was suitable for obtaining chitosan-coated PGLA nanoparticles containing AmB. Nanoparticles were spherical in shape, presented mean particle size about 460 nm, positive zeta potential and encapsulation efficiency of 42%. Moreover, nanoparticles prolonged the AmB release. All the strains were susceptible to plain AmB and nanostructured AmB, according to EUCAST breakpoint version 8.1 (resistant > 1 μg/mL), using broth microdilution method. In C. albicans (urine, blood, and vulvovaginal secretion isolates, and 1 ATCC), the MIC value of AmB-loaded nanoparticles varied from 0.25 to 0.5 μg/mL and EUCAST varied from 0.03 to 0.5 μg/mL. In urine and vulvovaginal secretion isolates of C. glabrata, the MIC value of AmB-loaded nanoparticles varied from 0.25 to 0.5 μg/mL and EUCAST varied from 0.03 to 0.015 μg/mL. In urine isolates of C. guilhermondii, the MIC value of AmB-loaded nanoparticles was 0.12 μg/mL and EUCAST was 0.06 μg/mL. In blood isolates of C. tropicalis, the MIC value of AmB-loaded nanoparticles was 0.5 μg/mL and EUCAST was 0.25 μg/mL. Finally, in urine isolates of T asahii, the MIC value of AmB-loaded nanoparticles was 1 μg/mL and EUCAST varied from 0.5 to 1 μg/mL. In the cytotoxicity assay, plain AmB was highly hemolytic (100% in 24 h) while AmB-loaded chitosan/PLGA nanoparticles presented negligible hemolysis.