Polydispersity
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This research paper presents a novel approach to the green synthesis of silver nanoparticles (AgNPs) using viticultural waste, allowing to obtain NP dispersions with distinct properties and morphologies (monodisperse and polydisperse AgNPs, referred to as mAgNPs and pAgNPs) and to compare their biological activities. Our synthesis method utilized the ethanolic extract of Vitis vinifera pruning residues, resulting in the production of mAgNPs and pAgNPs with average sizes of 12 ± 5 nm and 19 ± 14 nm, respectively. Both these AgNPs preparations demonstrated an exceptional stability in terms of size distribution, which was maintained for one year. Antimicrobial testing revealed that both types of AgNPs inhibited either the growth of planktonic cells or the metabolic activity of biofilm sessile cells in Gram-negative bacteria and yeasts. No comparable activity was found towards Gram-positives. Overall, pAgNPs exhibited a higher antimicrobial efficacy compared to their monodisperse counterparts, suggesting that their size and shape may provide a broader spectrum of interactions with target cells. Both AgNP preparations showed no cytotoxicity towards a human keratinocyte cell line. Furthermore, in vivo tests using a silkworm animal model indicated the biocompatibility of the phytosynthesized AgNPs, as they had no adverse effects on insect larvae viability. These findings emphasize the potential of targeted AgNPs synthesized from viticultural waste as environmentally friendly antimicrobial agents with minimal impact on higher organisms.
- MeSH
- antiinfekční látky farmakologie chemie MeSH
- biofilmy účinky léků MeSH
- bourec MeSH
- buněčné linie MeSH
- gramnegativní bakterie účinky léků MeSH
- keratinocyty účinky léků MeSH
- kovové nanočástice * chemie MeSH
- kvasinky účinky léků MeSH
- larva účinky léků MeSH
- lidé MeSH
- mikrobiální testy citlivosti * MeSH
- rostlinné extrakty farmakologie chemie MeSH
- stříbro * farmakologie chemie metabolismus MeSH
- technologie zelené chemie MeSH
- velikost částic MeSH
- viabilita buněk účinky léků MeSH
- Vitis * chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Stilbenes in food and medicinal plants have been described as potent antiphlogistic and antioxidant compounds, and therefore, they present an interesting potential for the development of dietary supplements. Among them, macasiamenene F (MF) has recently been shown to be an effective anti-inflammatory and cytoprotective agent that dampens peripheral and CNS inflammation in vitro. Nevertheless, this promising molecule, like other stilbenes and a large percentage of drugs under development, faces poor water solubility, which results in trickier in vivo administration and low bioavailability. With the aim of improving MF solubility and developing a form optimized for in vivo administration, eight types of conventional liposomal nanocarriers and one type of PEGylated liposomes were formulated and characterized. In order to select the appropriate form of MF encapsulation, the safety of MF liposomal formulations was evaluated on THP-1 and THP-1-XBlue-MD2-CD14 monocytes, BV-2 microglia, and primary cortical neurons in culture. Furthermore, the cellular uptake of liposomes and the effect of encapsulation on MF anti-inflammatory effectiveness were evaluated on THP-1-XBlue-MD2-CD14 monocytes and BV-2 microglia. MF (5 mol %) encapsulated in PEGylated liposomes with an average size of 160 nm and polydispersity index of 0.122 was stable, safe, and the most promising form of MF encapsulation keeping its cytoprotective and anti-inflammatory properties.
- Publikační typ
- časopisecké články MeSH
In this work oleuropein and lentisk oil have been co-loaded in different phospholipid vesicles (i.e., liposomes, transfersomes, hyalurosomes and hyalutransfersomes), to obtain a formulation capable of both inhibiting the production of different markers connected with inflammation and oxidative stress and promoting the skin repair. Liposomes were prepared using a mixture of phospholipids, oleuropein and lentisk oil. Tween 80, sodium hyaluronate or their combination have been added to the mixture to obtain transfersomes, hyalurosomes and hyalutransfersomes. Size, polydispersity index, surface charge and stability on storage was evaluated. The biocompatibility, anti-inflammatory activity and wound healing effect were tested using normal human dermal fibroblasts. Vesicles were small (mean diameter ∼ 130 nm) and homogeneously dispersed (polydispersity index ∼ 0.14), highly negatively charged (zeta potential 02053-64 mV) and capable of loading 20 mg/mL of oleuropein and 75 mg/mL of lentisk oil. The freeze-drying of dispersions with a cryoprotectant permitted to improve their stability on storage. The co-loading of oleuropein and lentisk oil in vesicles inhibited the overproduction of inflammatory markers, especially MMP-1 and IL-6, counteracted the oxidative stress induced in cells using hydrogen peroxide, and promoted the healing of a wounded area performed in vitro in a cell monolayer of fibroblasts. The proposed co-loading of oleuropein and lentisk oil in natural-based phospholipid vesicles may hold promising therapeutic value especially for the treatment of a wide variety of skin disorders.
- MeSH
- cytokiny metabolismus MeSH
- fosfolipidy * metabolismus MeSH
- hojení ran MeSH
- kůže metabolismus MeSH
- lidé MeSH
- liposomy * metabolismus MeSH
- oxidační stres MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Most modern wound dressings assist the wound-healing process. In contrast, conventional wound dressings have limited antibacterial activity and promote sporadic fibroblast growth. Therefore, wound dressings with prolonged substance release must be improved. This research aimed to develop hydrogel films. These were synthesized from alginate and pectin, incorporated with mangosteen extract (ME), and encapsulated in niosomes (ME-loaded niosomes). Subsequently, we examined the in vitro release and physical characteristics of ME-loaded niosomes. These characteristics included particle pH, size, charge, polydispersity index (PDI), and drug loading properties. These properties included drug loading content (DLC), entrapment efficiency (EE), and yield (Y). Additionally, we examined the swelling ratio and biological characteristics of the hydrogel film. These characteristics included antibacterial activity, cytotoxicity (L929), cell attachment to the tested materials, cell migration, hemocompatibility, and in vivo irritation. Significant results were obtained using a 2:1 niosome preparation containing Span60 and cholesterol. Ratio influenced size, charge, PDI, DLC, EE, and Y. The results were 225.5 ± 5.83 nm, negatively charged, 0.38, 16.2 ± 0.87%, 64.8 ± 3.49%, and 87.3 ± 3.09%, respectively. Additionally, the release of encapsulated ME was pH sensitive because 85% of the ME can be released at a pH of 5.5 within seven days and decrease to 70% at a pH of 7.4. The maximum swelling ratios of patches with 0.5% and 1% Ca2+ crosslinking were 867 wt% and 1,025 wt%, respectively, after 30 min. These results suggested that a medium dose (15 mg) of niosomal ME incorporated in a hydrogel film provided better bacterial inhibition, cell migration, and cell adhesion in an in vitro model. Additionally, no toxicity was observed in the fibroblasts and red blood cells. Therefore, given the above-mentioned advantages, this product can be a promising candidate for wound dressing applications.
- Publikační typ
- časopisecké články MeSH
Poly(lactic-co-glycolic acid) (PLGA) nanoparticle-based drug delivery systems are known to offer a plethora of potential therapeutic benefits. However, challenges related to large-scale manufacturing, such as the difficulty of reproducing complex formulations and high manufacturing costs, hinder their clinical and commercial development. In this context, a reliable manufacturing technique suitable for the scale-up production of nanoformulations without altering efficacy and safety profiles is highly needed. In this paper, we develop an inline sonication process and adapt it to the industrial scale production of immunomodulating PLGA nanovaccines developed using a batch sonication method at the laboratory scale. The investigated formulations contain three distinct synthetic peptides derived from the carcinogenic antigen New York Esophageal Squamous Cell Carcinoma-1 (NY-ESO-1) together with an invariant natural killer T-cell (iNKT) activator, threitolceramide-6 (IMM60). Process parameters were optimized to obtain polymeric nanovaccine formulations with a mean diameter of 150 ± 50 nm and a polydispersity index <0.2. Formulation characteristics, including encapsulation efficiencies, release profiles and in vitro functional and toxicological profiles, are assessed and statistically compared for each formulation. Overall, scale-up formulations obtained by inline sonication method could replicate the colloidal and functional properties of the nanovaccines developed using batch sonication at the laboratory scale. Both types of formulations induced specific T-cell and iNKT cell responses in vitro without any toxicity, highlighting the suitability of the inline sonication method for the continuous scale-up of nanomedicine formulations in terms of efficacy and safety.
- Publikační typ
- časopisecké články MeSH
Green methods have become vital for sustainable development of the scientific and commercial sphere; however, they can bring new challenges, including the need for detailed characterization and elucidation of efficacy of their products. In this study, green method of silver nanoparticles (AgNPs) production was employed using an extract from grapevine canes. The aim of the study was to contribute to the knowledge about biosynthesized AgNPs by focusing on elucidation of their antifungal efficiency based on their size and/or hypothesized synergy with bioactive substances from Vitis vinifera cane extract. The antifungal activity of AgNPs capped and stabilized with bioactive compounds was tested against the opportunistic pathogenic yeast Candida albicans. Two dispersions of nanoparticles with different morphology (characterized by SEM-in-STEM, DLS, UV-Vis, XRD, and AAS) were prepared by modification of reaction conditions suitable for economical production and their long-term stability monitored for six months was confirmed. The aims of the study included the comparison of the antifungal effect against suspension cells and biofilm of small monodisperse AgNPs with narrow size distribution and large polydisperse AgNPs. The hypothesis of synergistic interaction of biologically active molecules from V. vinifera extracts and AgNPs against both cell forms were tested. The interactions of all AgNPs dispersions with the cell surface and changes in cell morphology were imaged using SEM. All variants of AgNPs dispersions were found to be active against suspension and biofilm cells of C. albicans; nevertheless, surprisingly, larger polydisperse AgNPs were found to be more effective. Synergistic action of nanoparticles with biologically active extract compounds was proven for biofilm cells (MBIC80 20 mg/L of polydisperse AgNPs in extract), while isolated nanoparticles suspended in water were more active against suspension cells (MIC 20 mg/L of polydisperse AgNPs dispersed in water). Our results bring new insight into the economical production of AgNPs with defined characteristics, which were proven to target a specific mode of growth of significant pathogen C. albicans.
- MeSH
- antibakteriální látky farmakologie MeSH
- antifungální látky metabolismus farmakologie MeSH
- biofilmy MeSH
- Candida albicans metabolismus MeSH
- kovové nanočástice * MeSH
- mikrobiální testy citlivosti MeSH
- rostlinné extrakty metabolismus farmakologie MeSH
- stříbro * metabolismus farmakologie MeSH
- voda metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Hyaluronan (HA) is a major component of peri- and extra-cellular matrices and plays important roles in many biological processes such as cell adhesion, proliferation and migration. The abundance, size distribution and presentation of HA dictate its biological effects and are also useful indicators of pathologies and disease progression. Methods to assess the molecular mass of free-floating HA and other glycosaminoglycans (GAGs) are well established. In many biological and technological settings, however, GAGs are displayed on surfaces, and methods to obtain the size of surface-attached GAGs are lacking. Here, we present a method to size HA that is end-attached to surfaces. The method is based on the quartz crystal microbalance with dissipation monitoring (QCM-D) and exploits that the softness and thickness of films of grafted HA increase with HA size. These two quantities are sensitively reflected by the ratio of the dissipation shift (ΔD) and the negative frequency shift (- Δf) measured by QCM-D upon the formation of HA films. Using a series of size-defined HA preparations, ranging in size from ~ 2 kDa tetrasaccharides to ~ 1 MDa polysaccharides, we establish a monotonic yet non-linear standard curve of the ΔD/ - Δf ratio as a function of HA size, which reflects the distinct conformations adopted by grafted HA chains depending on their size and surface coverage. We demonstrate that the standard curve can be used to determine the mean size of HA, as well as other GAGs, such as chondroitin sulfate and heparan sulfate, of preparations of previously unknown size in the range from 1 to 500 kDa, with a resolution of better than 10%. For polydisperse samples, our analysis shows that the process of surface-grafting preferentially selects smaller GAG chains, and thus reduces the average size of GAGs that are immobilised on surfaces comparative to the original solution sample. Our results establish a quantitative method to size HA and other GAGs grafted on surfaces, and also highlight the importance of sizing GAGs directly on surfaces. The method should be useful for the development and quality control of GAG-based surface coatings in a wide range of research areas, from molecular interaction analysis to biomaterials coatings.
Despite the efficacy and potential therapeutic benefits that poly(lactic-co-glycolic acid) (PLGA) nanomedicine formulations can offer, challenges related to large-scale processing hamper their clinical and commercial development. Major hurdles for the launch of a polymeric nanocarrier product on the market are batch-to-batch variations and lack of product consistency in scale-up manufacturing. Therefore, a scalable and robust manufacturing technique that allows for the transfer of nanomedicine production from the benchtop to an industrial scale is highly desirable. Downstream processes for purification, concentration, and storage of the nanomedicine formulations are equally indispensable. Here, we develop an inline sonication process for the production of polymeric PLGA nanomedicines at the industrial scale. The process and formulation parameters are optimized to obtain PLGA nanoparticles with a mean diameter of 150 ± 50 nm and a small polydispersity index (PDI < 0.2). Downstream processes based on tangential flow filtration (TFF) technology and lyophilization for the washing, concentration, and storage of formulations are also established and discussed. Using the developed manufacturing and downstream processing technologies, production of two PLGA nanoformulations encasing ritonavir and celecoxib was achieved at 84 g/h rate. As a measure of actual drug content, encapsulation efficiencies of 49.5 ± 3.2% and 80.3 ± 0.9% were achieved for ritonavir and celecoxib, respectively. When operated in-series, inline sonication and TFF can be adapted for fully continuous, industrial-scale processing of PLGA-based nanomedicines.
- Publikační typ
- časopisecké články MeSH
The cell wall chitosan was extracted from fungi belonging to different taxonomic classes, namely, Benjaminiella poitrasii (Zygomycetes, dimorphic), Hanseniaspora guilliermondii, Issatchenkia orientalis, Pichia membranifaciens, and Saccharomyces cerevisiae (Ascomycetes, yeasts), and Agaricus bisporus and Pleurotus sajor-caju (Basidiomycetes). The maximum yield of chitosan was 60.89 ± 2.30 mg/g of dry mycelial biomass of B. poitrasii. The degree of deacetylation (DDA) of chitosan extracted from different fungi, as observed with 1H NMR, was in the range of 70-93%. B. poitrasii chitosan exhibited the highest DDA (92.78%). The characteristic absorption bands were observed at 3450, 1650, 1420, 1320, and 1035 cm-1 by FTIR. Compared to chitosan from marine sources (molecular weight, MW, 585 kDa), fungal chitosans showed lower MW (6.21-46.33 kDa). Further, to improve the efficacy of B. poitrasii chitosan (Bp), nanoparticles (Np) were synthesized using the ionic gelation method and characterized by dynamic light scattering (DLS). For yeast and hyphal chitosan nanoparticles (BpYCNp and BpHCNp), the average particle size was <200 nm with polydispersity index of 0.341 ± 0.03 and 0.388 ± 0.002, respectively, and the zeta potential values were 21.64 ± 0.34 and 24.48 ± 1.58 mV, respectively. The B. poitrasii chitosans and their nanoparticles were further evaluated for antifungal activity against human pathogenic Candida albicans ATCC 10231, Candida glabrata NCYC 388, Candida tropicalis ATCC 750, Cryptococcus neoformans ATCC 34664, and Aspergillus niger ATCC 10578. BpHCNps showed lower MIC90 values (0.025-0.4 mg/mL) than the chitosan polymer against the tested human pathogens. The study suggested that nanoformulation of fungal chitosan, which has low molecular weight and high % DDA, is desirable for antifungal applications against human pathogens. Moreover, chitosans as well as their nanoparticles were found to be hemocompatible and are therefore safe for healthcare applications.
- MeSH
- antifungální látky farmakologie MeSH
- chitosan * farmakologie MeSH
- houby MeSH
- lidé MeSH
- Mucorales * chemie MeSH
- nanočástice * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The molecular weight (Mw) of dextran derivatives, such as regioselectively oxidized dicarboxydextran (DXA), is greatly influencing their faith in an organism, which could be possibly used to improve anticancer drug delivery. Here we present a modified method of sulfonation-induced chain scission allowing direct and accurate control over the Mw of DXA without increasing its polydispersity. Prepared DXA derivatives (Mw = 10-185 kDa) have been conjugated to cisplatin and the Mw of the carrier found to have a significant impact on cisplatin release rates, in vitro cytotoxicity, and migrastatic potential. Conjugates with the high-Mw DXA showed particularly increased anticancer efficacy. The best conjugate was four times more effective against malignant prostatic cell lines than free cisplatin and significantly inhibited the ovarian cancer cell migration. This was traced to the characteristics of spontaneously formed cisplatin-crosslinked DXA nanogels influenced by Mw of DXA and amount of loaded cisplatin.
- MeSH
- adenokarcinom farmakoterapie metabolismus MeSH
- buňky A549 MeSH
- cisplatina chemie farmakologie MeSH
- dextrany chemie MeSH
- lékové transportní systémy metody MeSH
- lidé MeSH
- molekulová hmotnost MeSH
- nádory prostaty farmakoterapie metabolismus MeSH
- nádory vaječníků farmakoterapie metabolismus MeSH
- nádory farmakoterapie metabolismus MeSH
- nanogely chemie MeSH
- nosiče léků chemie MeSH
- oxidace-redukce MeSH
- pohyb buněk účinky léků MeSH
- protinádorové látky chemie farmakologie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
