86 research outputs found
12th International Conference of the European Chitin Society and 13th International Conference on Chitin and Chitosan (EUCHIS/ICCC 2015)
Chitosan/Cyclodextrin/TPP Nanoparticles Loaded with Quercetin as Novel Bacterial Quorum Sensing Inhibitors
The widespread emergence of antibiotic-resistant bacteria has highlighted the urgent need of alternative therapeutic approaches for human and animal health. Targeting virulence factors that are controlled by bacterial quorum sensing (QS), seems a promising approach. The aims of this study were to generate novel nanoparticles (NPs) composed of chitosan (CS), sulfo-butyl-ether-Ī²-cyclodextrin (CaptisolĀ®) and/or pentasodium tripolyphosphate using ionotropic gelation technique, and to evaluate their potential capacity to arrest QS in bacteria. The resulting NPs were in the size range of 250ā400 nm with CS70/5 and 330ā600 nm with CS70/20, had low polydispersity index (<0.25) and highly positive zeta potential ranging from Ī¶ ~+31 to +40 mV. Quercetin, a hydrophobic model flavonoid, could be incorporated proportionally with increasing amounts of CaptisolĀ® in the NPs formualtion, without altering significantly its physicochemical properties. Elemental analysis and FTIR studies revealed that CaptisolĀ® and quercetin were effectively integrated into the NPs. These NPs were stable in M9 bacterial medium for 7 h at 37 Ā°C. Further, NPs containing CaptisolĀ® seem to prolong the release of associated drug. Bioassays against an E. coli Top 10 QS biosensor revealed that CS70/5 NPs could inhibit QS up to 61.12%, while CS70/20 NPs exhibited high antibacterial effects up to 88.32%. These results suggested that the interaction between NPs and the bacterial membrane could enhance either anti-QS or anti-bacterial activities
Assessment of the Quorum Sensing Inhibition Activity of a Non-Toxic Chitosan in an N-Acyl Homoserine Lactone (AHL)-Based Escherichia coli Biosensor
New approaches to deal with drug-resistant pathogenic bacteria are urgent. We studied the antibacterial effect of chitosans against an Escherichia coli quorum sensing biosensor reporter strain and selected a non-toxic chitosan to evaluate its quorum sensing (QS) inhibition activity and its effect on bacterial aggregation. To this end, chitosans of varying degree of acetylation (DA) (12 to 69%) and molecular weight (Mw) (29 to 288 kDa) were studied. Only chitosans of low DA (~12%) inhibited bacterial growth, regardless of their Mw. A chitosan with medium degree of polymerization (named MDP) DA30, with experimental DA 42% and Mw 115 kDa was selected for further QS inhibition and scanning electron microscopy (SEM) imaging studies. MDP DA30 chitosan exhibited QS inhibition activity in an inverse dose-dependent manner (ā¤12.5 Āµg/mL). SEM images revealed that this chitosan, when added at low concentration (ā¤30.6 Āµg/mL), induced substantial bacterial aggregation, whereas at high concentration (234.3 Āµg/mL), it did not. Aggregation explains the QS inhibition activity as the consequence of retardation of the diffusion of N-acylated homoserine lactones (AHLs)
On the Fractionation and Physicochemical Characterisation of Self-Assembled ChitosanāDNA Polyelectrolyte Complexes
Chitosan is extensively studied as a carrier for gene delivery and is an attractive non-viral gene vector owing to its polycationic, biodegradable, and biocompatible nature. Thus, it is essential to understand the chemistry of self-assembled chitosanāDNA complexation and their structural and functional properties, enabling the formation of an effective non-viral gene delivery system. In this study, two parent chitosans (samples NAS-032 and NAS-075; Mw range ~118ā164 kDa) and their depolymerised derivatives (deploy nas-032 and deploy nas-075; Mw range 6ā14 kDa) with degrees of acetylation 43.4 and 4.7%, respectively, were used to form polyelectrolyte complexes (PECs) with DNA at varying [āNH3+]/ [āPO4ā] (N/P) molar charge ratios. We investigated the formation of the PECs using Ī¶-potential, asymmetric flow field-flow fractionation (AF4) coupled with multiangle light scattering (MALS), refractive index (RI), ultraviolet (UV) and dynamic light scattering (DLS) detectors, and TEM imaging. PEC formation was confirmed by Ī¶-potential measurements that shifted from negative to positive values at N/P ratio ~2. The radius of gyration (Rg) was determined for the eluting fractions by AF4-MALS-RI-UV, while the corresponding hydrodynamic radius (Rh), by the DLS data. We studied the influence of different cross-flow rates on AF4 elution patterns for PECs obtained at N/P ratios 5, 10, and 20. The determined rho shape factor (Ļ = Rg/Rh) values for the various PECs corresponded with a sphere morphology (Ļ ~ 0.77ā0.85), which was consistent with TEM images. The results of this study represent a further step towards the characterisation of chitosanāDNA PECs by the use of multi-detection AF4 as an important tool to fractionate and infer aspects of their morphology
Characterisation of the Interaction among Oil-In-Water Nanocapsules and Mucin
Mucins are glycoproteins present in all mucosal surfaces and in secretions such as saliva. Mucins are involved in the mucoadhesion of nanodevices carrying bioactive molecules to their target sites in vivo. Oil-in-water nanocapsules (NCs) have been synthesised for carrying N,Nā²-(di-m-methylphenyl)urea (DMTU), a quorum-sensing inhibitor, to the oral cavity. DMTU-loaded NCs constitute an alternative for the treatment of plaque (bacterial biofilm). In this work, the stability of the NCs after their interaction with mucin is analysed. Mucin type III from Sigma-Aldrich has been used as the mucin model. Mucin and NCs were characterised by the multi-detection asymmetrical flow field-flow fractionation technique (AF4). Dynamic light scattering (DLS) and Ī¶-potential analyses were carried out to characterise the interaction between mucin and NCs. According to the results, loading DMTU changes the conformation of the NC. It was also found that the synergistic interaction between mucin and NCs was favoured within a specific range of the mucin:NC ratio within the first 24 h. Studies on the release of DMTU in vitro and the microbial activity of such NCs are ongoing in our lab
Interaction Between Chitosan and Mucin: Fundamentals and Applications
The term chitosan (CS) refers to a family of aminopolysaccharides derived from chitin. Among other properties, CS is nontoxic, mucoadhesive and can be used for load and transport drugs. Given these and other physicochemical and biological properties, CS is an optimal biopolymer for the development of transmucosal drug delivery systems, as well as for the treatment of pathologies related to mucosal dysfunctions. Mucins are glycoprotein macromolecules that are the major components of mucus overlaying epithelia. CS interacts with mucin and adsorbs on and changes the rheology of mucus. However, CS and mucins denote families of polymers/macromolecules with highly variable chemical structure, properties, and behavior. To date, their interactions at the molecular level have not been completely unraveled. Also, the properties of complexes composed of CS and mucin vary as a function of the sources and preparation of the polymers. As a consequence, the mucoadhesion and drug delivery properties of such complexes vary as well. The breadth of this review is on the molecular interactions between CS and mucin. In particular, in vitro and ex vivo characterization methods to investigate both the interactions at play during the formation of CS-mucin complexes, and the advances on the use of CS for transmucosal drug delivery are addressed
Physicochemical characterization of FRET-labelled chitosan nanocapsules and model degradation studies
Sub-micron o/w emulsions coated with chitosan have been used for drug delivery, quorum sensing inhibition, and vaccine development. To study interactions with biological systems, nanocapsules have been fluorescently labelled in previous works, but it is often difficult to distinguish the released label from intact nanocapsules. In this study, we present advanced-labelling strategies based on Fƶrster Resonance Energy Transfer (FRET) measurements for chitosan-coated nanocapsules and investigate their dissolution and degradation. We used FRET measurements of nanocapsules loaded with equimolar concentrations of two fluorescent dyes in their oily core and correlated them with dynamic light scattering (DLS) count rate measurement and absorbance measurements during their disintegration by dissolution. Using count rate measurements, we also investigated the enzymatic degradation of nanocapsules using pancreatin and how protein corona formation influences their degradation. Of note, nanocapsules dissolved in ethanol, while FRET decreased simultaneously with count rate, and absorbance was caused by nanocapsule turbidity, indicating increased distance between dye molecules after their release. Nanocapsules were degradable by pancreatin in a dose-dependent manner, and showed a delayed enzymatic degradation after protein corona formation. We present here novel labelling strategies for nanocapsules that allow us to judge their status and an in vitro method to study nanocapsule degradation and the influence of surface characteristics
Investigating on the toxicity and bio-magnification potential of synthetic glitters on Artemia salina
Our research aims to assess the toxic impacts of polyethylene terephthalate (PET) glitters on Artemia salina as a model zooplankton. The mortality rate was assessed using a Kaplan Maier plot as a function of various microplastic dosages. The ingestion of microplastics was confirmed by their presence in digestive tract and faecal matter. Gut wall damage was confirmed by dissolution of basal lamina walls and an increase in the secretory cells. A significant decrease in the activities of cholinesterase (ChE) and glutathione-S-transferase (GST) were noted. A decrease in catalase activity could be correlated to an increase in the generation of reactive oxygen species (ROS). Cysts incubated in presence of microplastics exhibited delay in their hatching into āumbrellaā and āinstarā stages. The data presented in the study would be useful for scientists working on discovering new sources of microplastics, related scientific evidences, image data and model of study
Chitosan nanoencapsulation of flavonoids enhances their quorum sensing and biofilm formation inhibitory activities against an E.coli Top 10 biosensor
Phytochemicals have been found to be promising alternatives to conventional antibiotic therapies for the control of bacterial infections, as they may entail less selective pressure and hence reduce the development of resistance. This study involved examining the inhibition of biofilm formation and of quorum sensing (QS), and the cytotoxicity on mammalian cells of two flavonoids, quercetin and baicalein, in freeform and associated into chitosan-based nanocapsules. This was done by use of a transformed E. coli Top10 biosensor strain, while the cytotoxicity was evaluated on MDCK-C7 cells. In free form, application both flavonoids exhibited slight inhibitory activity on the QS response and biofilm formation, a scenario that was improved positively upon encapsulation with chitosan (Mw ā¼115,000 g/mol and DA ā¼42%). The association efficiency of 99% (quercetin) and 87% (baicalein) was determined, and each formulation had an average diameter of 190 Ā± 4 and 187 Ā± 2 nm, and zeta ( ) potential of +48.1 Ā± 2.03 and +48.4 Ā± 3.46 mV, respectively. Both types of systems were stable against aggregation in M9 and MEM media. The in vitro release kinetics data of both flavonoids seemed to be similar with only ā¼20% released over the first 5 h, orā¼10% over the first 4 h, respectively, with subsequent sudden release increase up to ā¼40% in both cases.The free phytochemicals seemed to be cytotoxic to MDCK-C7 cells at higher doses, however, upon nanoencapsulation, a cytoprotective effect was evidenced. We have gained proof-of-principle of the advantages of encapsulation of two bioactive flavonoids
New insights into the nature of the Cibacron brilliant red 3B-A ā Chitosan interaction
Cibacron brilliant red 3B-A (CBR) has been introduced to determine chitosan (CS) concentrations in solution, and several studies applied it to measure chitosan content in pharmaceutical formulations. So far, studies have relied on the absorbance band shift to 570 nm to determine the extent of the CBR ā CS interaction. In this study, we show that CBR forms micro- to nanometer sized aggregates with CS, depending on their charge ratio and that other photophysical changes in CBR are induced by this interaction. We found that, besides the bathochromic band shift, aggregation induces emission at 600 nm and emission quenching at 360 nm. We compared changes CS induced in absorbance and fluorescence emission of CBR with the CS monomer glucosamine and poly(allylamine) hydrochloride, which both contain amino groups, and found that similar but less intense photophysical changes also occur. Furthermore, CS-induced circular dichroism in CBR suggests a twisted, chiral structure of these aggregates that should match with the previously published in silico simulations of the structure of CS in solution. The low linear charge density of CS and its chiral conformation are considered responsible for the enhanced photophysical response of CBR interacting with the polycation
- ā¦