Maryna Ornatska

Interaction of nanoparticles with lipid membrane.

A nanoscale range of surface feature curvatures where lipid membranes lose integrity and form pores has been found experimentally. The pores were experimentally observed in the l-alpha-dimyristoyl phosphatidylcholine membrane around 1.2-22 nm polar nanoparticles deposited on mica surface. Lipid bilayer envelops or closely follows surface features with the curvatures outside of that region. This finding provides essential information for the understanding of nanoparticle-lipid membrane interaction, cytotoxicity, preparation of biomolecular templates and supported lipid membranes on rough and patterned surfaces.

Paper bioassay based on ceria nanoparticles as colorimetric probes.

We report the first use of redox nanoparticles of cerium oxide as colorimetric probes in bioanalysis. The method is based on changes in the physicochemical properties of ceria nanoparticles, used here as chromogenic indicators, in response to the analyte. We show that these particles can be fully integrated in a paper-based bioassay. To construct the sensor, ceria nanoparticles and glucose oxidase were coimmobilized onto filter paper using a silanization procedure. In the presence of glucose, the enzymatically generated hydrogen peroxide induces a visual color change of the ceria nanoparticles immobilized onto the bioactive sensing paper, from white-yellowish to dark orange, in a concentration-dependent manner. A detection limit of 0.5 mM glucose with a linear range up to 100 mM and a reproducibility of 4.3% for n = 11 ceria paper strips were obtained. The assay is fully reversible and can be reused for at least 10 consecutive measurement cycles, without significant loss of activity. Another unique feature is that it does not require external reagents, as all the sensing components are fixed onto the paper platform. The bioassay can be stored for at least 79 days at room temperature while maintaining the same analytical performance. An example of analytical application was demonstrated for the detection of glucose in human serum. The results demonstrate the potential of this type of nanoparticles as novel components in the development of robust colorimetric bioassays.

Biocomputing Security System: Concatenated Enzyme-Based Logic Gates Operating as a Biomolecular Keypad Lock

A biomolecular security system mimicking a keypad lock device was developed using enzyme-based concatenated AND logic gates resulting in the implication logic network.

Colorimetric Paper Bioassay for the Detection of Phenolic Compounds

A new type of paper based bioassay for the colorimetric detection of phenolic compounds including phenol, bisphenol A, catechol and cresols is reported. The sensor is based on a layer-by-layer (LbL) assembly approach formed by alternatively depositing layers of chitosan and alginate polyelectrolytes onto filter paper and physically entrapping the tyrosinase enzyme in between these layers. The sensor response is quantified as a color change resulting from the specific binding of the enzymatically generated quinone to the multilayers of immobilized chitosan on the paper. The color change can be quantified with the naked eye but a digitalized picture can also be used to provide more sensitive comparison to a calibrated color scheme. The sensor was optimized with respect to the number of layers, pH, enzyme, chitosan and alginate amounts. The colorimetric response was concentration dependent, with a detection limit of 0.86 (±0.1) μg/L for each of the phenolic compounds tested. The response time required for the sensor to reach steady-state color varied between 6 and 17 min depending on the phenolic substrate. The sensor showed excellent storage stability at room temperature for several months (92% residual activity after 260 days storage) and demonstrated good functionality in real environmental samples. A procedure to mass-produce the bioactive sensors by inkjet printing the LbL layers of polyelectrolyte and enzyme on paper is demonstrated.

Biochemically controlled bioelectrocatalytic interface.

A switchable bioelectrocatalytic system for glucose oxidation controlled by external biochemical signals exemplifies interfacing between bioelectronic and biochemical ensembles.

Boolean logic gates that use enzymes as input signals.

Biochemical systems that demonstrate the Boolean logic operations AND, OR, XOR, and InhibA were developed by using soluble compounds, which represent the chemical “devices”, and the enzymes glucose oxidase (GOx), glucose dehydrogenase (GDH), alcohol dehydrogenase (AlcDH), and microperoxidase‐11 (MP‐11), which operated as the input signals that activated the logic gates. The enzymes were used as soluble materials and as immobilized biocatalysts. The studied systems are proposed to be a step towards the construction of “smart” signal‐responsive materials with built‐in Boolean logic.

Interaction of lipid membrane with nanostructured surfaces.

Tiny details of the phospholipid (DMPC) membrane morphology in close vicinity to nanostructured silica surfaces have been discovered in the atomic force microscopy experiments. The structural features of the silica surface were varied in the experiments by the deposition of silica nanoparticles of different diameter on plane and smooth silica substrates. It was found that, due to the barrier function of the lipid membrane, only particles larger than 22 nm in diameter with a smooth surface were completely enveloped by the lipid membrane. However, nanoparticles with bumpy surfaces (curvature diameter of bumps as that of particles <22 nm) were only partially enveloped by the lipid bilayer. For the range of nanostructure dimensions between 1.2 and 22 nm, the lipid membrane underwent structural rearrangements by forming pores (holes). The nanoparticles were accommodated into the pores but not enveloped by the lipid bilayer. The study also found that the lipid membrane conformed to the substrate with surface structures of dimensions less than 1.2 nm without losing the membrane integrity. The experimental results are in accord with the analytical free energy model, which describes the membrane coverage, and numerical simulations which evaluate adhesion of the membrane and dynamics as a function of surface topology. The results obtained in this study are useful for the selection of dimensions and shapes for drug-delivery cargo and for the substrate for supported lipid bilayers. They also help in qualitative understanding the role of length scales involved in the mechanisms of endocytosis and cytotoxicity of nanoparticles. These findings provide a new approach for patterning supported lipid membranes with well-defined features in the 1.2-22 nm range.

Langmuir-Blodgett monolayers from lower generation amphiphilic monodendrons

Abstract Amphiphilic monodendrons of lower generations, AD12-N, containing a benzyl-15-crown-5 polar focal point, photochromic spacer and different number of dodecyl tails as peripheral groups ( n =1–8) have been investigated for their ability to form uniform monolayers at solid surfaces. The surface pressure–area behavior, photomechanical behavior and the morphology of the monomolecular films were investigated. We observed that all compounds studied are capable of forming stable Langmuir and Langmuir Blodgett monolayers, with virtually flat packing of molecules. Higher generation dendrimers form very uniform monolayers, without the usual domain microstructure. For AD12-4 monolayer on solid support, we observed stripped microstructure with several layers (3–6) bundled together. The periodicity of this structure of 8 nm was close to layered spacing, obtained from X-ray data for bulk material. For this compound, we proposed the model of double-layered packing of the molecules, with partial overlapping of the central segments and suggested that deposition on a solid substrate resulted in changing orientation of molecular fragments. Fast reversible photochromic response was observed for all monolayers with a conversion level of 50%.

Biomedical Applications of Metal Oxide Nanoparticles

Metal oxide nanoparticles have a unique structure, interesting and unusual redox and catalytic properties, high surface area, good mechanical stability and are biocompatible. For these reasons, metal oxide nanoparticles have attracted considerable interest in the field of biomedical therapeutics, bio-imaging and biosensing. This chapter discusses properties and biomedical applications of selected nanometer size metal oxides. These materials have become important components in medical implants, cancer diagnosis and therapy and in neurochemical monitoring. For example, titania is the material of choice in medical implants; it provides an excellent biocompatible surface for cell attachment and proliferation. Ceria-based nanoparticles, on the other hand, have recently received a great deal of attention because of their redox, auto-catalytic and antioxidant properties. Several other metal oxides have been used as gas sensing nanoprobes for cell labeling and separation, as contrast agents for magnetic resonance imaging (MRI) and as carriers for targeted drug delivery. New and emerging applications of nanoceria as neuroprotective agents possessing antioxidant/free radical scavenging properties are emerging in the biomedical field, and ceria-based nanoparticles may be used as therapeutic agents in the treatment of medical diseases related to reactive oxygen species, such as spinal cord repair, stroke and degenerative retinal disorders. Issues related to biocompatibility and toxicity of these nanoparticles for in vivo biomedical applications remain to be fully explored.

Biomolecular Oxidative Damage Activated by Enzymatic Logic Systems: Biologically Inspired Approach

Logical, responsible, practical. Enzymatic logic gates that process chemical input signals were used to trigger the release of redox‐active iron ions, which produce reactive oxygen species in a catalytic cascade, and thus result in oxidative damage in biomolecules. Functional coupling between enzymatic logic gates and oxidative damage systems resulted in “smart” biochemical ensembles that are activated upon receiving a certain pattern of biochemical signals.