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Publications (61)
Despite extensive study of extracellular vesicles (EVs), specifically exosomes (EXs) as biomarkers, important modulators of physiological or pathological processes, or therapeutic agents, relatively little is known about nonconventional sources of EXs, such as invertebrate or plant EXs, and their uses. Likewise, there is no clear information on the...
Supramolecular structures, such as micelles, liposomes, polymerosomes or dendrimerosomes, are widely studied and used as drug delivery systems. The behavior of amphiphilic building blocks strongly depends on their spatial distribution and shape of polar and nonpolar component. This report is focused on the development of new versatile synthetic pro...
Zebrafish (Danio rerio) serves as a popular animal model for in vivo acute toxicity evaluation with the Fish embryo test (FET). Over the last few years there has been an effort to develop various systems for a high-throughput zebrafish embryo cultivation and FET. In this paper, we present a novel design of a millifluidic system fabricated by 3D pri...
E-Shell 300 3D-printed material demonstrated a considerable negative impact on cell proliferation and severe developmental toxicity due to release of surfactant residues. Post-treatment with ethanol improved the biocompatibility of the material.
Glycodendrimers are a novel group of dendrimers (DDMs) characterized by surface modifications with various types of glycosides. It has been shown previously that such modifications significantly decrease the cytotoxicity of DDMs. Here, we present an investigation of glucose-modified carbosilane DDMs (first-third-generation, DDM1-3Glu) interactions...
Non-viral gene delivery vectors studied in the gene therapy applications are often designed with the cationic nitrogen containing groups necessary for binding and cell release of nucleic acids. Disadvantage is a relatively high toxicity which restricts the in vivo use of such nanoparticles. Here we show, that the 3rd generation carbosilane dendrime...
Glycodendrimers (Glyco-DDMs) represent a rapidly growing class of nanoparticles with promising properties for biomedical applications but concerns regarding the impact on human health and environment are still justified. Here we report, for the first time, the comparative study of in vivo developmental toxicity of carbosilane Glyco-DDMs and their c...
The influence of three generations of five different phosphonium carbosilane dendrimers and one ammonium carbosilane dendrimer as a reference (PMe3, PBu3, P(Et2)2(CH2)3OH, PPh3, P(MeOPh)3 and NMe3, peripheral functional groups) on dimyristoylphosphatidylcholine (DMPC) or a lipid mixture dimyristoylphosphatidylcholine/dimyristoylphosphatidylglycerol...
Recent developments in molecular evolution technologies have led to novel types of high-affinity recombinant protein binders (PB) able to substitute antibodies in many diagnostic and therapeutic applications. Despite almost a decade of research, they have so far only been sporadically used for biosensor construction. Here, we present a proof-of-pri...
Herein, we report a novel concept of low-cost flexible platform for fluorescence-based biosensor. The surface of polyethylene naphthalate (PEN) foil was exposed to KrF excimer laser through a photolitographic contact mask. Laser initiated surface modification resulted in micro-patterned areas with surface functional groups available for localized c...
We report the synthesis and cytotoxicity evaluation of a completely new class of cationic carbosilane dendrimers functionalized with several different phosphonium peripheral groups and an ammonium functionalised one as a reference. The carbosilane dendrimers with NMe3, PMe3, P(Et2)2(CH2)3OH, PBu3, P(C6H4-OMe)3 and P(Ph)3 peripheral substituents wer...
Regardless of the route of administration, some or all of a therapeutic agent will appear in the blood stream, where it can act on blood cells and other components of the plasma. Recently we have shown that poly(ethylene imines) (PEIs) which interact with plasma proteins are taken up into erythrocyte membranes. These observations led us to investig...
For the design of a biohybrid structure as a ligand-tailored drug delivery system (DDS), it is highly sophisticated to fabricate a DDS based on smoothly controllable conjugation steps. This article reports on the synthesis and the characterization of biohybrid conjugates based on noncovalent conjugation between a multivalent biotinylated and PEGyla...
Interactions between maltose functionalized hyperbranched poly(ethylene imine)s (95% maltose decoration denoted as Mal-PEI A; 33% maltose decoration denoted as Mal-PEI B) and red blood cells (RBCs) and between red blood cell membranes were investigated. We monitored the degree of hemolysis, the change in cell shape, the influence of polymers on the...
The influence of maltose-modified poly(propylene imine) (PPI) dendrimers on dimyristoylphosphatidylcholine (DMPC) or dimyristoylphosphatidylcholine/dimyristoylphosphatidylglycerol (DMPC/DMPG) (3%) liposomes was studied. Fourth generation (G4) PPI dendrimers with primary amino surface groups were partially (open shell glycodendrimers - OS) or comple...
Polymers with functionalized surfaces have attracted a lot of attention in the last few years. Due to the progress in the techniques of polymer micro-patterning, miniaturized bioanalytical assays and biocompatible devices can be developed. In the presented work, we performed surface modification of polyethylene naphthalate (PEN) foil by an excimer...
The nanopatterning method applied through micrometer slit for polyethylene naphthalate (PEN) substrate was proposed in this paper. Surface roughness, formation of nanoscale ripple-like structures and the dependence of their dimensions on the value of laser fluence was determined by atomic force and laser confocal microscopy, and compared with value...
In the last few years many anti-viral drugs have been developed to fight against H1V-1 infection and have been proved to control infection. Highly active antiretroviral therapy (HAART) has made an immense impact on fighting the HIV/AIDS pandemic, however, it still possesses major drawbacks such as the need of keeping greater than 95% adherence to b...
Tooth enamel, the hardest tissue in the body, is formed by the evolutionarily highly conserved biomineralization process that is controlled by extracellular matrix proteins. The intrinsically disordered matrix protein ameloblastin (AMBN) is the most abundant nonamelogenin protein of the developing enamel and a key element for correct enamel formati...
In this work, a unique high-performance liquid chromatographic method was developed and applied for monitoring the synthesis of polyethyleneglycol surface modified poly(amidoamine) cystamine core dendrimers (PEG-PAMAMs) and PEG-PAMAM-alkynes with a single alkyne moiety attached to the core of a dendron through a unique sulfhydryl group. The separat...
The dendrimer chemistry reported offers a route to synthetic targets molecules with spherical shape, well-defined surface chemistries and dimensions that match the size of virus particles. The largest target, a generation 13 dendrimer comprising triazines linked by diamines, is stable across ranges of concentration, pH, temperature, solvent polarit...
Scanning electron microscopy and atomic force microscopy were used to study the silk spinning apparatus and silks of Harpactea rubicunda spiders. Three types of silk secretions that are produced by three kinds of silk spinning glands (ampullate, piriform, and pseudaciniform) and released through three types of spigots, were confirmed for both adult...
Scanning electron microscopy and atomic force microscopy were used to study the silk spinning apparatus and silks of Harpactea rubicunda spiders. Three types of silk secretions that are produced by three kinds of silk spinning glands (ampullate, piriform, and pseudaciniform) and released through three types of spigots, were confirmed for both adult...
Dendrimers are artificial polymeric macromolecules which are widely considered to be a promising tool for future gene therapy applications. They have been used as efficient delivery vehicles for antisense oligonucleotides targeting the interior of cells. We demonstrate that dendriplexes formed from anti-HIV oligodeoxynucleotides ANTI-TAR, GEM91, an...
Recombinant ligands derived from small protein scaffolds show promise as robust research and diagnostic reagents and next generation protein therapeutics. Here, we derived high-affinity binders of human interferon gamma (hIFNγ) from the three helix bundle scaffold of the albumin-binding domain (ABD) of protein G from Streptococcus G148. Computation...
The success of gene therapy depends on the development of suitable carriers, and because of their architecture dendrimers are promising tools for gene delivery. This research concerns the use of second generation carbosilane dendrimers as carriers for anti-HIV oligodeoxynucleotides (ODNs). The aim was to characterize complexes formed by positively...
Granular non-fibrillar aggregates (GNAs) are identified as possible toxic species in Alzheimer's disease. GNAs form on the surface of negatively charged biological membranes and as a consequence of an acidic environment, off the polymerization pathway at neutral pH. Aβ (1-40) GNAs disturb the bilayer structure of model membranes and seem to be more...
The principle of synchronous detection (SD) has been applied to biosensor measurement. SD principle achieves significant increases in the signal to noise ratio, limit of detection and overall measurement robustness. Application of SD in biosensor measurement improves the analysis of the response and avoids the influence of interference/noise produc...
We compared a novel PSII-biosensor assay with a standard algal growth inhibition test for detection of photosynthetic herbicides--diuron, atrazine and isoproturon in liquid samples. To evaluate the convenience and sensitivity, values of the parameters EC50 and LOD and the duration of assays were compared. The biosensor assay was made with an electr...
Cocoons of Theridiosoma gemmosum consist of two main parts, the egg sac case and the stalk. The inner space of the egg sac case is filled with nonsticky flocculent silk. Measuring 600-800 nm in diameter, the flocculent threads are never made up of bundles of longitudinally oriented nanofibrils. The egg case wall consists of a lower layer of highly...
This paper presents a synthesis of a novel nanoparticle label with selective biorecognition properties based on a biotinylated silver-dendrimer nanocomposite (AgDNC). Two types of labels, a biotin-AgDNC (bio-AgDNC) and a biotinylated AgDNC with a poly(ethylene)glycol spacer (bio-PEG-AgDNC), were synthesized from a generation 7 (G7) hydroxyl-termina...
ABSTRACT: Cocoons of Theridiosoma gemmosum consist of two main parts, the egg sac case and the stalk. The inner space of the egg sac case is filled with nonsticky flocculent silk. Measuring 600-800 nm in diameter, the flocculent threads are never made up of bundles of longitudinally oriented nanofibrils. The egg case wall consists of a lower layer...
Nanostructured materials have attracted considerable attention in many fields of science, for their extremely peculiar properties due to the down-scaling. Amongst thelarge number of nanomaterials applications in biosensing and bioanalyticaltechniques, this chapter is focused on the modification of sensitive surfaces withnanostructured materials (na...
Metal nanowires were synthesized according 10 an elcctroless deposition method in polycarbonate template, obtaining nano electrodes ensembles with specific features and well defined diameter size. Nano electrode ensembles were coupled with disposable screen printed substrate in order to increase the sensitivity of the commonly used carbon graphite...
This paper reports on the synthesis and characterization of a novel electrochemical label for sensitive electrochemical stripping metalloimmunoassays based on silver dendrimer-encapsulated nanoparticles (NPs). Silver dendrimer nanocomposites (Ag-DNCs) were synthesized from a generation 5-7 (G5-7) hydroxyl-terminated ethylenediamine-core-type (2-car...
Two methods for monolayer immobilization of photosystem II (PSII) isolated from thermophilic cyanobacteria Synechococcus elongatus and prospects for its use as a biosensor for detection of herbicides are reported: (i) immobilization based on recombinant (His)6‐tagged PSII coupled with nickel‐nitrilotriacetic acid (Ni‐NTA) chelator monolayer on Au e...
Gold nanowires were synthesized within polycarbonate membranes according to an electroless deposition method, obtaining nanoelectrode ensembles (NEEs) with special electrochemical features. NEEs were coupled with home-produced carbon graphite screen printed electrodes and the electrochemical properties of the original nanoelectrode ensemble on scre...
Photosystem II (PSII) modified gold electrodes have been prepared providing mediatorless electron transport on the basis of electrodeposited conductive layer poly-mercapto-p-benzoquinone (polySBQ). Such electrodes are suitable in construction of biosensors for PSII inhibiting herbicides. PolySBQ layer was synthesized on (i) screen-printed gold elec...
A micro-cell consisting of a planar array of Au working electrodes, covered by a glass reservoir (600nl capacity), connected with perpendicular capillaries and provided with an Ag/AgCl wire reference electrode (RE) has been manufactured. Addressed on-chip immobilization and sensing of biomolecules was demonstrated by the immobilization of histidine...
We have examined the persistence and movement of a urea-type herbicide, isoproturon [IPU; 3-(4-isopropylphenyl)-1,1'-dimethylurea], in soil using a novel herbicide-detection device, the prototype of a portable electrochemical biosensor based on Photosystem II particles immobilized on printed electrodes, and evaluated its results against two other m...
Rome (I) *Pilloton@mclink.it Electrochemical on-chip deposition of proteins for further biodetection of several analytes by multiplexed chronoamperometry in a continuous flow µ-chamber is presented. Recently, an original deposition procedure was developed obtaining oriented and reversible immobilisation of an engineered photosystem II with an histi...
Mass transport of the bulk of the analyte to the electrode and through the bioactive layer can be significantly improved by use of the nanoelectrode array and defined arrangement of protein film. This phenomenon has been studied by (i) atomic-force microscopy, (ii) electrochemical measurements of PSII activity, and (iii) digital simulations for an...
The expansion of miniaturization of analytical devices, due mostly to the reduction of analysis times, has imposed the evolution of sensitive detection methods. Fluorescence detection has been widely employed not only because its inherent high sensitivity is a good match for the small analyte volumes of fluorescent species but also because not fluo...
Rome (I) *Pilloton@mclink.it Electrochemical on-chip deposition of proteins for further biodetection of several analytes by multiplexed chronoamperometry in a continuous flow µ-chamber is presented. Recently, an original deposition procedure was developed obtaining oriented and reversible immobilisation of an engineered photosystem II with an histi...
A cross-branch approach inspired the birth of COSMIC (Coupling Smart Molecules into Chips) at ENEA. The activities have been focused on several aspects of the biosensing science, ranging from new functions or activities due to engineered molecules or genetically modified microorganisms, their oriented and reversible immobilisation on electrodes, op...
The interfacing of a on-chip microreactor with the external world is investigated from the microfluidic point of view. A solution for coupling of input and output capillaries perpendicular on the micro-reactor plane is presented. The micro-reactor consists of a planar array of 7 x 7 metallic electrodes covered by a borofloat® glass reservoir provid...
Photosystem II (PSII) modified gold electrodes providing mediatorless electron transport on the basis of an electrodeposited conductive layer of polySBQ have been prepared.
Such electrodes are suitable for construction of biosensors for PSII inhibiting herbicides.
Sulfobenzoquinone was prepared by organic synthesis. PolySBQ layer was synthesized on...
Electrochemical synthesis of nickel-nitrilotriacetic acid (Ni-NTA) chelators, for subsequent immobilization of (His)(6)-tagged proteins (Photosystem II (PSII) as model molecule), on Au or Au-graphite electrodes is compared to chemical synthesis. Results show: (i) higher Ni-NTA surface density, (ii) shorter treatment time (1-12 min vs. 16 h normally...
Monolayers of genetically modified proteins with an hexahistidine tag, (His)6, were obtained by using a Ni–NTA chelator synthesized on gold-sputtered surfaces (via sulphide bonds), or on gold and graphite (via sililating agents) working electrodes of screen-printed devices.Two kinds of proteins were produced and purified for this study:(a)a recombi...
A biosensor for the detection of triazine- and phenylurea-type herbicides was constructed using isolated Photosystem II (PS II) complexes as a biosensing element. PSII isolated from the thermophilic cyanobacterium Synechococcus elongatus was immobilized on the surface of a screen-printed sensor composed of a graphite working electrode and Ag/AgCl r...
A biosensor for the detection of triazine- and phenylurea-type herbicides was constructed using isolated Photosystem II (PS II) complexes as a biosensing element. PSII isolated from the thermophilic cyanobacterium Synechococcus elongatus was immobilized on the surface of a screen-printed sensor composed of a graphite working electrode and Ag/AgCl r...
Projects
Projects (3)
This project aims to introduce novel types of glycodendrimers based on carbosilane dendrimer core decorated with (oligo)sacharides and to investigate their interaction with model lipid membranes and biomolecules (anti-sense oligonucleotides, amyloid and prion peptides) by complex biophysical methods and computer modeling.
Project is focused on development of novel diagnostic device for high-throughtput in vivo studies of biological effects of molecules on fish model Danio rerio.
In this project, we are creating a research centre UniQSurf, which will exploit the years of the previous research activities focused in the field of bioactive surfaces, functional hybrid materials and surfaces for separation processes and catalysis and computer modelling of functional hybrid materials and surfaces.