Biological Buffers Research Articles

Buffering capacity is determinant for restoring early α-synuclein aggregation.

For disordered proteins, including α-synuclein (Syn), the aggregation of which is implicated in Parkinson's disease, it is known that at mild acidic and at the pI solution conditions the use of either strong or weak electrolytes minimized Syn aggregation. The mechanism is driven by electrostatic forces but remains, however, poorly understood. To address this issue, we used two biological buffers as weak electrolytes, at a low concentration (10mM) and monitored the aggregation of Syn solutions from pH7 to pH2, by means of light scattering techniques. When the citrate buffer was used, in which there is buffering capacity in the pH range studied, the maximum of Syn aggregation was very close to the isoelectric point (pI=4.7). When using tris-HCl, in which there is almost no buffering capacity in the pH range studied, it was for the first time observed a slow transition of the pI (of ca. 1h) from 4.7 to 4-3, for a 33.5μM protein concentration, as an example. We also observed in the protein solutions (in tris-HCl) the very early formation of large Syn aggregates. When there is buffering capacity, such as pH7, these early large Syn aggregates dissociate, followed by association/aggregation. When there is no buffering capacity, such as pH3, the referred early large Syn aggregates only dissociate. Overall, early large Syn aggregates dissociation can cause entropy in the protein solutions and Syn aggregation is only restored by the altered electrostatic forces due to the existing buffering capacity. Finally, by using an innovative strategy based in the ANS dye fluorescence intensity variation, we determined of the occurrence of the liquid-liquid phase separation process at pH7 Syn solutions.

Insight into Sonoluminescence Augmented by ZnO-Functionalized Nanoparticles.

Recent advances in optical imaging techniques rely on the use of nanosized contrast agents for in vitro and in vivo applications. We report on an imaging method based on the inertial cavitation of ultrasound-irradiated water solutions that lead to sonoluminescence (SL), here, newly proposed in combination with semiconductor nanoparticles, in particular, aminopropyl-functionalized zinc oxide nanocrystals. The obtained measurements confirm the ability of such nanocrystals to increase the sonoluminescence emission, together with the ability to modify the SL spectrum when compared to the pure water behavior. In particular, it is shown that the UV component of SL is absorbed by the semiconductor behavior that is also confirmed in different biologically relevant media. Finally, optical images of nanocrystal-assisted SL are acquired for the first time, in particular, in biological buffers, revealing that at low ultrasound intensities, SL is measurable only when the nanocrystals are present in solution. All of these results witness the role of amine-functionalized zinc oxide nanocrystals for sonoluminescence emission, which makes them very good candidates as efficient nanocontrast agents for SL imaging for biological and biomedical applications.

Exploring the versatility of pentafulvene-maleimide cycloaddition as a ligation strategy: buffer and pH effects.

Ligation chemistries are often required to perform under stringent conditions that preserve the integrity and function of (bio)conjugates, including specific biological buffers. To explore the versatility of the pentafulvene-maleimide ligation for (bio)conjugations, we studied the stability of the coupling partners and their Diels-Alder cycloaddition (DAC) in buffers used commonly in biological assays, protein chemistry and bioconjugates. The stability of 6,6-dialkylpentafulvene and maleimide derivatives to a panel of buffers with pH values between 3.7 and 10.1 was monitored via1H NMR spectroscopy. While the pentafulvene displayed excellent stability, hydrolysis of the maleimide was observed in several cases, although the extent of hydrolysis did not correlate with pH. For almost all buffers the pentafulvene-maleimide DAC proceeded efficiently and at a significantly faster rate than maleimide hydrolysis under the conditions studied. The buffer composition nor pH appeared to have a significant effect on the kinetics of the DAC with second-order rate constants (k2) ranging from 0.14 to 0.33 M-1 s-1 (23 ± 1 °C). This study highlights the versatility of the pentafulvene-maleimide ligation to proceed under a wide range of conditions relevant for (bio)conjugations and that maleimide hydrolysis in aqueous system may be promoted or inhibited by certain buffers.

Computational study of the acetic acid extraction process from an aqueous solution with the aid of biological buffer

The liquid-liquid extraction of acetic acid from an aqueous solution with 1-heptanol as an extraction solvent in the extraction column and mixer-decanter at 30°C and atmospheric pressure was simulated using Aspen Plus. A Non-Random Two-Liquid (NRTL) based model was developed by minimizing maximum-likelihood objective function. In the simulation of the extraction column and mixer-decanter, the effect of the number of stages and the flow rate of the solvent on the percent recovery can be seen. In addition, a comparison of the percent recovery value between acetic acid extraction systems using EPPS (4-(2-hydroxyehtyl)-1-piperazine propanesulfonic acid) and HEPES (4-(2-hydroxyethyl)-1 piperazineethanesulfonic acid) buffers was also carried out with systems without buffers. In this study, an economic analysis was also carried out for the acetic acid extraction system using an extraction column and a mixer-decanter. Based on the simulation results, the acetic acid extraction system with the addition of HEPES buffer using extraction column with the number of stages = 8 and solvent to feed mass ratio = 1.6 was the most optimal and efficient extraction process obtained in this study. This system has the capital cost of 180,729,262.67 USD with the percent recovery up to 99.83% and the mass fraction of acetic acid in the raffinate phase is 0.0002 which is also extremely low.

Hofmeister effects of anions on self-assembled thermogels

Thermogels are temperature-responsive soft biomaterials with numerous biomedical applications. They possess high water content and can spontaneously gelate by forming non-covalent physical crosslinks between their constituent amphiphilic polymers when warmed. However, despite the ubiquity of salts in biological fluids and buffer media, the influence of salts on thermogelling polymers and the overall physical properties of the resulting hydrogels are poorly understood. Herein, we elucidate the effects of common inorganic salts on the gelation and micellization properties of a thermogelling polymer containing poly(ethylene glycol), poly(propylene glycol), and poly(caprolactone) components. The identity of the salts' anions and their concentrations was found to exhibit significant effects on the thermogel properties, in some cases being able to decrease the sol-to-gel phase transition by up to 10 °C. We demonstrate that these notable influences are likely brought about by the changes in solvation of both the polymer's hydrophobic and hydrophilic segments, as well as by direct interactions of poorly hydrated anions with the hydrophobic polymer segments. Our findings show that the effects of salts on amphiphilic thermogelling polymers are non-negligible and hence need to be taken into account for engineering and optimization of thermogel properties for different biomedical applications.

Bicarbonate modulates delafloxacin activity against MDR Staphylococcus aureus and Pseudomonas aeruginosa.

To investigate the utility of recently approved delafloxacin and other fluoroquinolones against leading MDR bacterial pathogens under physiologically relevant conditions. MIC and MBC assays were conducted for MDR strains of Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae in the standard antibiotic susceptibility testing medium CAMHB, amended Roswell-Park Memorial Institute tissue culture medium (RPMI+) or 20% fresh human whole blood. In vivo correlation of in vitro findings was performed in a murine P. aeruginosa pneumonia model. Mechanistic bases for the findings were explored by altering media conditions and with established fluoroquinolone accumulation assays. Fluoroquinolone MICs were increased in RPMI+ compared with CAMHB for all four MDR pathogens. Specifically, delafloxacin MICs were increased 32-fold versus MDR S. aureus and 8-fold versus MDR P. aeruginosa. MBC assays in 20% human whole blood and a murine MDR P. aeruginosa pneumonia model both confirmed that delafloxacin activity was reduced under physiological conditions. Bicarbonate (HCO3-), a key component of host physiology found in RPMI+ but absent from CAMHB, dictated delafloxacin susceptibility in CAMHB and RPMI+ by impairing its intracellular accumulation. Standard in vitro antibiotic susceptibility testing conditions overpredicted the effectiveness of delafloxacin against MDR pathogens by failing to capture the role of the biological buffer HCO3- to impair delafloxacin accumulation. This work showcases limitations of our current antibiotic susceptibility testing paradigm and highlights the importance of understanding host microenvironmental conditions that impact true clinical efficacy.

A Novel Polyvinylpyrrolidone-Stabilized Illite Microparticle with Enhanced Antioxidant and Antibacterial Effect.

Illite is a clay mineral that shows antioxidant and antibacterial activities because of the abundance of important clay elements in its structure. However, illite has low bioactivity due to its low solubility and electron-donating ability in aqueous solutions. Therefore, we aimed to develop polyvinylpyrrolidone (PVP)-stabilized illite microparticles (P-lite MPs) via polymer adsorption on illite surfaces. An increasing amount of PVP was used to coat a fixed amount of illite to prepare P-lite MPs of different hydrodynamic diameters in the range of 4–9 μm. These sizes were maintained for 2 weeks during storage in a biological buffer without any noticeable changes. The stabilization of illite microparticles using a hydrophilic PVP polymer improved their aqueous dispersity and free radical-scavenging activity. Since the large surface area of microparticles provides several sites for interactions, the smallest P-lite MP exhibited the highest antioxidant and antibacterial activities. More importantly, the MPs showed effective free radical-scavenging activity in vitro without any cytotoxicity. Therefore, P-lite MPs with improved bioavailability may represent a suitable bioactive material for various industrial and biomedical applications.

Stable in Biocompatible Buffers Silver Nanoisland Films for SERS.

We investigated the stability of silver nanoisland films, which were formed on glass surface by the method of out-diffusion, in biocompatible buffers and the applicability of the films in surface enhanced Raman scattering (SERS). We have shown that silver nanoisland films are stable in one of the most widespread in biological studies buffer—phosphate buffer saline (PBS), and in 1:100 water-diluted PBS, in the PBS-based buffer, in which NaCl is replaced by the same amount of NaClO4, and in acidic phosphate buffer. At the same time, the replacement of NaCl in PBS by N(CH3)4Cl leads to the degradation of the nanoislands. It was shown that after exposure to PBS the nanoisland films provided a good SERS signal from a monolayer of 1,2-di(4-pyridyl)ethylene (BPE), which makes silver nanoisland films promising for biosensor applications. Additionally, in our experiments, we registered for the first time that silver nanoparticles formed in the bulk of the samples dissolved after exposing to PBS, while nanoislands on the glass surface stayed unchanged. We associate this phenomenon with the interaction of ions contained in PBS solution with silver, which results in the shift of corresponding chemical equilibrium.

The Bioinorganic Chemistry of Mammalian Metallothioneins.

The functions, purposes, and roles of metallothioneins have been the subject of speculations since the discovery of the protein over 60 years ago. This article guides through the history of investigations and resolves multiple contentions by providing new interpretations of the structure-stability-function relationship. It challenges the dogma that the biologically relevant structure of the mammalian proteins is only the one determined by X-ray diffraction and NMR spectroscopy. The terms metallothionein and thionein are ambiguous and insufficient to understand biological function. The proteins need to be seen in their biological context, which limits and defines the chemistry possible. They exist in multiple forms with different degrees of metalation and types of metal ions. The homoleptic thiolate coordination of mammalian metallothioneins is important for their molecular mechanism. It endows the proteins with redox activity and a specific pH dependence of their metal affinities. The proteins, therefore, also exist in different redox states of the sulfur donor ligands. Their coordination dynamics allows a vast conformational landscape for interactions with other proteins and ligands. Many fundamental signal transduction pathways regulate the expression of the dozen of human metallothionein genes. Recent advances in understanding the control of cellular zinc and copper homeostasis are the foundation for suggesting that mammalian metallothioneins provide a highly dynamic, regulated, and uniquely biological metal buffer to control the availability, fluctuations, and signaling transients of the most competitive Zn(II) and Cu(I) ions in cellular space and time.

Liquid Metal Particles and Polymers: A Soft–Soft System with Exciting Properties

ConspectusGallium-based liquid metal alloys are a special type of material that is in the liquid state at (or near) room temperature. They are particularly attractive due to their unique combination of a fluidic and metallic body, together with a chemically reactive and functionable surface. As a fluid, liquid metals provide the best union of stretchability, deformability, and electrical conductivity among all soft materials. Such an advantage in combination with their low toxicity and relatively good biocompatibility have imparted liquid metals with unique features that can be harnessed for versatile applications in fields such as electronics, energy, chemistry, and biomedical research. More importantly, the fluidic nature of liquid metals allows them to be readily processed using shear for making particles with variable sizes (from nm to mm), which is not possible with solid materials. These particles have a liquid metal core–solid metal oxide shell (conductor–semiconductor) structure, allowing them to merge, transform shape, change phase, respond to stimuli, and self-heal.Despite these unique features, limited surface stability and functionality, unpredictable reactivity, and uncontrollable hydrophilicity of liquid metal particles niche their wider applications in biomedical fields. To bestow liquid metal particles with desirable surface properties while taking the benefits offered by soft features, another important soft material—polymers—can be synthesized and engineered on an on-demand basis to coat or embed liquid metal particles. This leads to the formation of liquid metal–polymer soft composites with versatile surface properties. More specifically, polymer segments with corresponding functions for surface anchoring, tuning solubility, enhancing biocompatibility, providing stimuli-responsive properties, and further bioconjugation can be linked together, thereby forming macromolecules to graft liquid metal particles for yielding soft–soft systems with exciting properties.Herein, we provide a concise review of our contributions to the production, investigation, characterization, and application of liquid metal particle–polymer composites. First, we summarize various top-down techniques developed for producing micro- to nanosized liquid metal particles. We highlight two platforms we developed for tackling long-existing problems encountered by sonication—the most widely adopted method for producing liquid metal particles. Second, we discuss the design of polymers for surface modification of particles. Various grafting strategies for polymers synthesized using different approaches are elaborated. We also discuss factors that affect the colloidal and chemical stability of the composite in biological buffers. Methods for further surface functionalization of the composite are presented, followed by providing examples of biomedical and sensing applications for the system. Next, we introduce the fabrication, unique properties, and applications of elastomeric hybrid composites incorporating liquid metal particle fillers. Finally, we offer a perspective on the opportunities and challenges for the future development of this exciting soft–soft system for realizing synergistic outcomes.

Polyculture in fish farming as a method of preventing fish diseases

Methods of prevention of fish diseases are aimed at destroying parasites in the host body and in the fish habitat — in the reservoir (chemoprophylaxis, increasing fish resistance), at preventing parasites from entering the reservoir (limiting the import of fish, import of healthy fish, quarantine measures, etc.), as well as preventing their spread and entry into the host body. It is shown that polyculture is not only the most effective method of using the natural forage base of the reservoir and, as a result, increasing fish productivity, but also a means of preventing mass infectious and other fish diseases. To prevent mass infectious diseases of fish and death from them both in safe and unfavorable ponds of carp fish farms, it is advisable to grow other fish species together with carp that do not suffer from diseases peculiar to carp. As a result, so-called sparse species plantings of fish are created in the ponds, while the total biomass remains high. At the same time, the natural food supply of ponds is most fully used and a kind of biological buffer is created that prevents the emergence and spread of contagious diseases. Polyculture can play a particularly important role in pond farms that already have certain problems with rubella, filometroidosis, botryocephalosis, carp pox, etc. Long-term studies of breeding herds in pond farms have shown that most of them have single foci of diseases, which, if there are optimal conditions for the pathogen, are ready to immediately respond with an outbreak of the disease or are generally quarantined for a particular fish disease. When determining the objects of polyculture, it is necessary to reduce the proportion of fish species that have a significant number of common diseases.

Silica@zirconia Core@shell Nanoparticles for Nucleic Acid Building Block Sorption.

The development of delivery systems for the immobilization of nucleic acid cargo molecules is of prime importance due to the need for safe administration of DNA or RNA type of antigens and adjuvants in vaccines. Nanoparticles (NP) in the size range of 20–200 nm have attractive properties as vaccine carriers because they achieve passive targeting of immune cells and can enhance the immune response of a weakly immunogenic antigen via their size. We prepared high capacity 50 nm diameter silica@zirconia NPs with monoclinic/cubic zirconia shell by a green, cheap and up-scalable sol–gel method. We studied the behavior of the particles upon water dialysis and found that the ageing of the zirconia shell is a major determinant of the colloidal stability after transfer into the water due to physisorption of the zirconia starting material on the surface. We determined the optimum conditions for adsorption of DNA building blocks, deoxynucleoside monophosphates (dNMP), the colloidal stability of the resulting NPs and its time dependence. The ligand adsorption was favored by acidic pH, while colloidal stability required neutral-alkaline pH; thus, the optimal pH for the preparation of nucleic acid-modified particles is between 7.0–7.5. The developed silica@zirconia NPs bind as high as 207 mg dNMPs on 1 g of nanocarrier at neutral-physiological pH while maintaining good colloidal stability. We studied the influence of biological buffers and found that while phosphate buffers decrease the loading dramatically, other commonly used buffers, such as HEPES, are compatible with the nanoplatform. We propose the prepared silica@zirconia NPs as promising carriers for nucleic acid-type drug cargos.

A Novel Chitosan Nanosponge as a Vehicle for Transepidermal Drug Delivery.

Transepidermal drug delivery achieves high drug concentrations at the action site and ensures continuous drug delivery and better patient compliance with fewer adverse effects. However, drug delivery through topical application is still limited in terms of drug penetration. Chitosan is a promising enhancer to overcome this constraint, as it can enhance drug diffusion by opening the tight junctions of the stratum corneum. Therefore, here, we developed a novel chitosan nanosponge (CNS) with an optimal ratio and molecular weight of chitosan to improve drug penetration through skin. To prepare the CNS, two types of chitosan (3 and 10 kDa) were each conjugated with poloxamer 407 using para-nitrophenyl chloroformate, and the products were mixed with poloxamer 407 at ratios of 5:5, 8:2, and 10:0. The resulting mixtures were molded to produce flexible soft nanosponges by simple nanoprecipitation. The CNSs were highly stable in biological buffer for four weeks and showed no toxicity in human dermal fibroblasts. The CNSs increased drug permeability through human cadaver skin in a Franz-type diffusion cell, with substantially higher permeability with 3 kDa chitosan at a ratio of 8:2. This suggests the applicability of the novel CNS as a promising carrier for efficient transepidermal drug delivery.

Isolation, characterization and in-vitro antigenicity studies of outer membrane proteins (OMPs) of Salmonella gallinarum coated gold nanoparticles (AuNPs)

The development of biomimetic nanoparticles by combining the natural cellular material with synthetic nanoparticles has inspired innovative therapeutic strategies as compared to traditional line of therapeutics. Therefore, the research was conducted to prepare the outer membrane proteins (OMPs) from Salmonella gallinarum and coated on to the surface of synthetic gold nanoparticles (AuNPs). The size of the final product was characterized by Zeta sizer and Zeta potential. The coating of outer membrane proteins onto the gold nanoparticles (AuNPs) was confirmed by transmission electron microscopy (TEM). The sterility, stability and safety of the prepared antigen was confirmed by standard culture media, in biological buffer solution and in-vitro pyrogenic testing, respectively. In-vitro antigenicity of outer membrane protein alone and coated with gold nanoparticles were observed through macrophage phagocytosis assay. On the basis of results, it was anticipated that bacterial outer membrane protein has immunogenic antigenic properties and coating of bacterial outer membrane protein on the surfaces of synthetic gold nanoparticles not only preserved the complex biological characteristics of bacteria but also provided greater immune responses as compared to outer membrane protein alone. So, it was concluded that the coating of bacterial outer membrane proteins on the surface of synthetic gold nanoparticles have synergistic effects to induce the immune responses and promising potential to develop the effective antibacterial vaccine against salmonellosis in poultry birds.

Robust and Versatile Coatings Engineered via Simultaneous Covalent and Noncovalent Interactions

AbstractInterfacial modular assembly has emerged as an adaptable strategy for engineering the surface properties of substrates in biomedicine, photonics, and catalysis. Herein, we report a versatile and robust coating (pBDT‐TA), self‐assembled from tannic acid (TA) and a self‐polymerizing aromatic dithiol (i.e., benzene‐1,4‐dithiol, BDT), that can be engineered on diverse substrates with a precisely tuned thickness (5–40 nm) by varying the concentration of BDT used. The pBDT‐TA coating is stabilized by covalent (disulfide) bonds and supramolecular (π‐π) interactions, endowing the coating with high stability in various harsh aqueous environments across ionic strength, pH, temperature (e.g., 100 mM NaCl, HCl (pH 1) or NaOH (pH 13), and water at 100 °C), as well as surfactant solution (e.g., 100 mM Triton X‐100) and biological buffer (e.g., Dulbecco's phosphate‐buffered saline), as validated by experiments and simulations. Moreover, the reported pBDT‐TA coating enables secondary reactions on the coating for engineering hybrid adlayers (e.g., ZIF‐8 shells) via phenolic‐mediated adhesion, and the facile integration of aromatic fluorescent dyes (e.g., rhodamine B) via π interactions without requiring elaborate synthetic processes.

The oxidative degradation of Calmagite using added and in situ generated hydrogen peroxide catalysed by manganese(II) ions: Efficacy evaluation, kinetics study and degradation pathways

Manganese (II) ions (Mn(II)) catalyse the oxidative degradation of Calmagite (CAL, 2-hydroxy-1-(2-hydroxy-5methylphenylazo)-4-naphthalenesulfonic acid) at room temperature using added and in situ generated hydrogen peroxide (H2O2), using 1,2-dihydroxybenzene-3,5-disulfonate, disodium salt and monohydrate (Tiron) as the co-catalyst for the in situ generation of H2O2. The percentage of CAL degradation with the in situ generated H2O2 was 91.1 % after 30 min which is lower than that in the added H2O2/Mn(II) system (96.0 %). A one-eighth-lives method was applied to investigate the kinetic parameters in the added H2O2 system, with and without Mn(II), involving phosphate, carbonate, and two biological buffers at different pHs. Percarbonate (HCO4−) was found to be the main reactive species for CAL degradation in the added H2O2 system buffered by carbonate in the absence of Mn(II). Manganese (IV) = O (Mn(IV) = O) and manganese(V) = O (Mn(V) = O) are the main reactive species in the added H2O2/Mn(II) system buffered by carbonate and non-carbonate buffers respectively. pH 8.5 was the optimum pH for CAL degradation when buffered by carbonate, while pH 10.0 is the best pH for the systems not using carbonate buffer. Using a high performance liquid chromatography/electrospray ionisation mass spectrometer (HPLC/ESI-MS), the degradation intermediates of CAL were identified as 1-amino-2-naphthol-4-sulfonate ion, 1-amino-2-naphthol-4-sulfinic ion, 1-amino-2-naphthol, and 1-nitroso-2-naphthol.

Magnetic and GRPR-targeted reduced graphene oxide/doxorubicin nanocomposite for dual-targeted chemo-photothermal cancer therapy

Herein, we design a rGO-based magnetic nanocomposite by decorating rGO with citrate-coated magnetic nanoparticles (CMNP). The magnetic rGO (mrGO) was modified by phospholipid-polyethylene glycol to prepare PEGylated mrGO, for conjugating with gastrin-releasing peptide receptor (GRPR)-binding peptide (mrGOG). The anticancer drug doxorubicin (DOX) was bound to mrGO (mrGOG) by π-π stacking for drug delivery triggered by the low pH value in the endosome. The mrGOG showed enhanced photothermal effect under NIR irradiation, endorsing its role for dual targeted DOX delivery. With efficient DOX release in the endosomal environment and heat generation from light absorption in the NIR range, mrGOG/DOX could be used for combination chemo-photothermal therapy after intracellular uptake by cancer cells. We characterized the physico-chemical as well as biological properties of the synthesized nanocomposites. The mrGOG is stable in biological buffer solution, showing high biocompatibility and minimum hemolytic properties. Using U87 glioblastoma cells, we confirmed the magnetic drug targeting effect in vitro for selective cancer cell killing. The peptide ligand-mediated targeted delivery increases the efficiency of intracellular uptake of both nanocomposite and DOX up to ~3 times due to the over-expressed GRPR on U87 surface, leading to higher cytotoxicity. The increased cytotoxicity using mrGOG over mrGO was shown from a decreased IC50 value (0.70 to 0.48 μg/mL) and an increased cell apoptosis rate (19.8% to 47.1%). The IC50 and apoptosis rate changed further to 0.19 μg/mL and 76.8% in combination with NIR laser irradiation, with the photothermal effect supported from upregulation of heat shock protein HSP70 expression. Using U87 tumor xenograft model created in nude mice, we demonstrated that magnetic guidance after intravenous delivery of mrGOG/DOX could significantly reduce tumor size and prolong animal survival over free DOX and non-magnetic guided groups. Augmented with NIR laser treatment for 5 min, the anti-cancer efficacy significantly improves with elevated cell apoptosis and reduced cell proliferation. Together with safety profiles from hematological as well as major organ histological analysis of treated animals, the mrGOG nanocomposite is an effective nanomaterial for combination chemo-photothermal cancer therapy.

The Structure-Properties-Cytotoxicity Interplay: A Crucial Pathway to Determining Graphene Oxide Biocompatibility.

Interest in graphene oxide nature and potential applications (especially nanocarriers) has resulted in numerous studies, but the results do not lead to clear conclusions. In this paper, graphene oxide is obtained by multiple synthesis methods and generally characterized. The mechanism of GO interaction with the organism is hard to summarize due to its high chemical activity and variability during the synthesis process and in biological buffers’ environments. When assessing the biocompatibility of GO, it is necessary to take into account many factors derived from nanoparticles (structure, morphology, chemical composition) and the organism (species, defense mechanisms, adaptation). This research aims to determine and compare the in vivo toxicity potential of GO samples from various manufacturers. Each GO sample is analyzed in two concentrations and applied with food. The physiological reactions of an easy model Acheta domesticus (cell viability, apoptosis, oxidative defense, DNA damage) during ten-day lasting exposure were observed. This study emphasizes the variability of the GO nature and complements the biocompatibility aspect, especially in the context of various GO-based experimental models. Changes in the cell biomarkers are discussed in light of detailed physicochemical analysis.

Controlling Nucleopeptide Hydrogel Self-Assembly and Formation for Cell-Culture Scaffold Applications.

Hydrogels made from self-assembling peptides have significant advantages in tissue engineering, namely a biocompatible nature and large molecular repertoire. Short peptides in particular allow for straightforward synthesis, self-assembly, and reproducibility. Applications are currently limited, however, due to potential toxicity of the chemical modifications that drive self-assembly and harsh gelation conditions. Peptides conjugated to nucleobases present one opportunity for a naturally derived species to minimize cytotoxicity. We have developed a hydrogel-formation environment for nucleopeptide gelation modulated entirely by biological buffers and salts. Self-assembly in this system is dependent on buffer and ion identity mediated by pKa and formulation in the former and by valency and ionicity in the latter. Solutions at physiological pH and osmolarity, and in turn compatible with cell culture, initiate hydrogel formation and analytical and computational methods are used to explore pH and salt effects at the molecular and structural level. The mechanism of nucleopeptide self-assembly enables tuning of mechanical properties through the addition of divalent cations and one order of magnitude increase in hydrogel storage modulus. The stability of these constructs therefore provides an opportunity for long-term cell culture, and we demonstrate survival and proliferation of fibroblasts on hydrogel surfaces. This novel, biological buffer-mediated gelation methodology expands opportunities for tissue engineering applications of short peptides and their derivatives.

Stable and scalable SERS tags conjugated with neutravidin for the detection of fibroblast activation protein (FAP) in primary fibroblasts

SERS tags are a class of nanoparticles with great potential in advanced imaging experiments. The preparation of SERS tags however is complex, as they suffer from the high variability of the SERS signals observed even at the slightest sign of aggregation. Here, we developed a method for the preparation of SERS tags based on the use of gold nanostars conjugated with neutravidin. The SERS tags here obtained are extremely stable in all biological buffers commonly employed and can be prepared at a relatively large scale in very mild conditions. The obtained SERS tags have been used to monitor the expression of fibroblast activation protein alpha (FAP) on the membrane of primary fibroblasts obtained from patients affected by Crohn’s disease. The SERS tags allowed the unambiguous identification of FAP on the surface of cells thus suggesting the feasibility of semi-quantitative analysis of the target protein. Moreover, the use of the neutravidin–biotin system allows to apply the SERS tags for any other marker detection, for example, different cancer cell types, simply by changing the biotinylated antibody chosen in the analysis.