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Über dieses Buch

This book characterizes how to design and synthesize nanomaterials of an organic and mineral nature. The book also discusses the visualization of developed nanomaterials and their bio-applications, as well as describes the biomedical effects and environmental impact of nanomaterials.

This is an ideal book for students studying biomedicine or the life sciences, as well as researchers and professionals in medicine, environmental protection, biotechnology, agriculture, and the food industry. More specifically, this book addresses the important nanomaterials and nanobiotechnologies that are used in those fields in biomedicine and life sciences.

Inhaltsverzeichnis

Frontmatter

Introduction

Frontmatter

Principal Trends in Nanobiotechnology

Abstract
The creation of multifunctional “smart” nanomaterials is one of the major trends in the modern pharmaceutical market, whose greatest priorities are (1) carriers for addressed drug delivery that can also diminish the negative side effects of drug action in the treated organism; (2) drug forms that are capable of overcoming biological barriers in the body, in particular multiple drug resistance, which blocks the action of traditional drugs; (3) nanoplatforms for the delivery of a big proportion of drugs that are poorly soluble in water; (4) carriers for delivery of DNA and miRNA for use in biotechnology and gene therapy; (5) biocompatible labels for visualization of drug-delivery routes, as well as for monitoring of their action and clearance in the body; and (6) effective diagnostic approaches, especially for early diagnostics and detection of specific biomolecules and targeted cells in the body. In this introductory chapter, I briefly describe the above-listed trends in nanobiotechnology. Because of limited space, new materials for transplantation and for detection and isolation of specific cells, especially stem and immune cells, which are also based on application of novel multifunctional nanomaterials, are out of this book’s scope.
Rostyslav Stoika

Design and Synthesis of Nanomaterials

Frontmatter

Molecular Design, Synthesis, and Properties of Surface-Active Comb-Like PEG-Containing Polymers and Derived Supramolecular Structures for Drug Delivery

Abstract
This chapter is devoted to the tailored synthesis and study of the properties of novel surface-active polymeric drug carriers containing side chains of PEG and other, including polyelectrolyte, chains for waterborne delivery systems. The strategy of synthesis of PEG-containing polymeric carriers via reactions of radical polymerization and further polymer-analogous transformations involving epoxide (so called “grafting to”) and peroxide ("grafting from") fragments of epoxide-containing polyperoxides of various composition and fine structure were developed and studied. The dependence of PEG grafting degree on the length of the blocks of GMA links in the backbone was shown. Water-soluble surfactants combining grafted side PEG and anionic polyelectrolyte chains were synthesized using polymerization initiated by the comb-like PEG-containing macro-initiator with side peroxide groups. An another promising way of “grafting through” synthesis of the comb-like polymeric drug carriers with side PEG chains via controlled polymerization of PEG methacrylate macromers in the presence of functional chain transfer agents was developed. The molecular weight characteristics, functionality, and surface activity of the developed polymers were studied using SEC and GPC techniques, FT-IR, NMR spectroscopy, and elementary analysis. The binding of water-soluble and water-insoluble anticancer drugs with PEGylated carriers via combination of different mechanisms was studied by using luminescent, RAMAN, UV-spectroscopy, and surface tension measurement techniques. Stable waterborne drug delivery systems based on the polymeric micelles loaded with water-soluble and water-insoluble drugs were developed and studied using SAXS, TEM, SEM, and DLS methods. The developed PEG-containing comb-like polymeric carriers and derived drug delivery systems were shown to be nontoxic in vitro (cell cultures) and in vivo (laboratory mice and rats). Their use enhances drug delivery to tumor cells, reduces the effective drug therapeutic dose, and offers a possibility to circumvent acquired resistance of tumor cells to drug action.
Nataliya Mitina, Anna Riabtseva, Olena Paiuk, Nataliya Finiuk, Miroslav Slouf, Ewa Pavlova, Lesya Kobylinska, Roman Lesyk, Orest Hevus, Vasyl Garamus, Rostyslav Stoika, Alexander Zaichenko

A Novel Water-Soluble C60 Fullerene-Based Nano-Platform Enhances Efficiency of Anticancer Chemotherapy

Abstract
Noncovalent water-soluble nanocomplexes of C60 fullerene (C60) with chemotherapeutic drugs (Doxorubicin (Dox), Cisplatin (Cis), and herbal alkaloid Berberine (Ber)) were created. Their anticancer action toward various tumor cells was studied in vitro, addressing specifically their biological synergy, compared with the action of these drugs in the non-immobilized form. Different theoretical and experimental (SEM and AFM microscopy, UV-Vis, DLS, NMR and SANS spectroscopy, ITC calorimetry) methods were applied for getting insight into the nature of the nanocomplexes with drug molecules, as well as into the physical forces enabling stabilization of these complexes. Physicochemical mechanisms were proposed for drug interaction with C60.
An enhancement of the toxic action of the created water-soluble C60-drug nanocomplexes toward cancer cells, compared to the action of free drug, was found. Specifically, the C60-Dox nanocomplexes demonstrated ~3.5 higher cytotoxic potential in the leukemic cell lines (CCRF-CEM, Jurkat, THP1, and Molt-16) in comparison with free Dox in the nanometer range of concentrations. Besides, C60 doubled the intracellular level of the up-taken Dox, which also evidenced its function as a nanocarrier. The toxic effect of C60-Cis nanocomplex toward Lewis lung carcinoma (LLC) cells was shown to be higher with IC50 values 3.3 and 4.5 times at 48 h and 72 h, respectively, as compared to the IC50 of free drug. 12.5 μМ Cis had no effect on LLC cells’ viability. The C60-Cis nanocomplex in Cis-equivalent concentration substantially decreased the viability of tumor cells, impaired their shape and adhesion, inhibited migration, and induced their accumulation in the pro-apoptotic sub-G1 phase of cell cycle. An induction of apoptosis by the C60-Cis nanocomplex was confirmed by the activation of caspase 3/7 and externalization of phosphatidylserine on the outer membrane of LLC cells after their double staining with the Annexin V-FITC/PI. The complexation with C60 promoted intracellular uptake of the Ber. An increase in C60 concentration in the C60-Ber nanocomplexes was accompanied by the elevation of their antiproliferative potential toward CCRF-CEM cells in the order: free Ber ˂ 1:2 ˂ 1:1 ˂ 2:1.
These findings suggest a universal potential of water-soluble pristine C60 as a unique nano-platform for the delivery of the chemotherapeutic drugs in cytotoxic effect of these drugs.
Yuriy Prylutskyy, Olga Matyshevska, Svitlana Prylutska, Anna Grebinyk, Maxim Evstigneev, Sergii Grebinyk, Larysa Skivka, Vsevolod Cherepanov, Anton Senenko, Rostyslav Stoika, Uwe Ritter, Peter Scharff, Thomas Dandekar, Marcus Frohme

Magnetic Iron Oxide Particles for Theranostics

Abstract
For many years, magnetic particles attracted a lot of interest in many fields, mainly including biomedicine. Recently, development of innovative strategies to tune the unique properties of magnetic nanoparticles for specific applications as theranostic agents has become one of the most challenging goals. This paper provides an overview of the synthesis, modification, and functionalization of magnetic nano- and microparticles. Special attention was paid to iron oxide composites used in cell separations and theranostics, such as anticancer drug delivery, diagnosis, and/or therapy of autoimmune and brain disorders. Last, but not the least, phenolic compound-modified magnetic particles were used as antioxidants or silver-containing carriers as antibacterial agents in in vitro and in vivo studies. Magnetic microparticles can be also incorporated in biosensors, exemplified by thionine-conjugated magnetic poly(carboxymethyl methacrylate-co-ethylene dimethacrylate) particles that showed higher enzymatic activity than nonmagnetic particles. Magnetic nano- and microparticles were characterized by a range of physicochemical methods, including transmission electron microscopy, dynamic light scattering, vibrating sample magnetometry, and Fourier-transform infrared spectroscopy, in terms of determination of particle morphology (size and its distribution), specific surface area, magnetic, and chemical properties. The chemical composition and crystallinity were confirmed by X-ray photoelectron spectroscopy. Moreover, the interactions between the magnetic particles and cells and/or other biological species were described.
Beata Zasońska, Daniel Horák

Biological Applications of Nanomaterials

Frontmatter

Controlled Delivery and Reduced Side Effects of Anticancer Drugs Complexed with Polymeric Nanocarrier

Abstract
A big challenge in the development of pharmaceutical drugs is to eliminate or at least reduce the side effects of highly active drugs, especially the antitumor agents demonstrating general toxicity in the body that significantly restricts their use. An efficient way to overcome this problem is to use a multifunctional nanocarrier of the drug that will allow the toxic antitumor agent to act at the site of its delivery to targeted cells in specific organs or tissues. The use of nanoparticles permits to optimize efficiency, minimize side effects, and improve cancer chemotherapy. Polymeric nanoparticles possess a huge potential in cancer chemotherapy, being one of the most widely tested nano-platforms and offering more effective and less toxic options to patients. Nanoconjugates can influence apoptotic mechanisms and enhance the cytotoxic action of drugs conjugated to the nanoparticles. The side effects of such action in the organism strongly depend upon the balance between the reactive oxygen species and the activity of the antioxidant system in the targeted cells. Drug delivery systems provide prolonged circulation of drug in blood, are able to accumulate in the pathological loci, and are capable of transferring active drug molecules effectively into specific cells and their organelles. Thus, the conjugation of the antitumor drug with a polymeric nanocarrier and application of drug in the form of a stable drug delivery system can reduce its general toxicity in the organism, compared with drug action in free form.
Lesya Kobylinska, Nataliya Mitina, Alexander Zaichenko, Rostyslav Stoika

Nano- and Microparticles and Their Role in Inflammation and Immune Response: Focus on Neutrophil Extracellular Traps

Abstract
Nano- and microparticles have become a normal part of our life, starting from medical drugs and cosmetics and ending in the television screens. Many of the nanoparticles are quite dangerous since the materials they use are not easily compatible with human tissue and provoke inflammation and immune responses. However, our body has developed the response on how to cope with those nanoparticles which are causing damaging effects to cells and tissues. This response is mediated by the neutrophilic granulocytes producing neutrophil extracellular traps—a weapon used to isolate and sequester particular matter in the safe deposits in the body. This chapter focuses on the mechanisms of how nanoparticles interact with neutrophils. It will provide an example of such interaction for main groups of nanoparticles—naturally occurring in the body, those formed during pathological conditions, and artificial pollutants. Examples of beneficial use of nanoparticle-induced inflammation are described for novel nanoadjuvants.
Galyna Bila, Andrii Rabets, Rostyslav Bilyy

Basic Principles of Nanotoxicology

Abstract
The need for using multifunctional nanomaterials and nanobiotechnologies is determined by a broad development of modern industries like agriculture, pharmaceutics, and medicine. However, the non-addressed action of anticancer drugs causes severe adverse effects in the body, which is a big problem in antitumor chemotherapy. This problem might be circumvented via immobilization of traditional antitumor drugs on the nanoplatforms of various structure that might increase the effectiveness of action of antitumor drugs, as well as reduce their overall toxicity in the organism. Another problem that appears upon using traditional antitumor chemotherapy is rapid development of multiple drug resistance of malignant neoplasms, mainly caused by functioning of special transport system in plasma membrane of cells. Developed nanocarriers could make the anticancer drugs “invisible” for this system.
At first glance, the problem of poor water solubility of many natural (e.g., taxol) and synthetic (various heterocyclic compounds) antitumor substances appears simple, but in fact, it is quite difficult to solve. These substances are well soluble only in organic solvents (e.g., dimethyl sulfoxide) that are highly toxic to the body. The use of specific nanoplatforms for immobilization of biologically active substances, poorly soluble in water, has a number of advantages, including biocompatibility, the ability to choose the desired size, architecture of the molecule, and its chemical functionalization. Chemical and physicochemical properties of these nanoplatforms are designed to make them convenient for conjugation of various medicines, including antitumor drugs.
Up until recent decades, the environmental impact of nanomaterials widely used for the biomedical goals got out of the scope of investigators. However, presently, it is clearly understandable that due to an increased effectiveness of the biological action, the biomedical nanomaterials might be even more dangerous for the living organisms, including humans, than the traditional medicines.
Rostyslav Stoika

Bioimaging, Biocompatibility, and Functioning of Polymeric Nanocarriers for Gene Delivery

Abstract
Materials for targeted drug delivery and gene transfer, in particular for DNA and siRNA delivery into cells, are currently the two most relevant in the international pharmacological market. The leading positions in the field of drug delivery belong to the biocompatible and biodegradable multifunctional nanoscale materials capable of forming water-soluble forms of drugs, providing their addressed delivery, and crossing the biological barriers in the organism. Labeling these materials with a biocompatible dye is also important for monitoring drug action or gene expression, biodistribution, and excretion from the treated organism.
In this chapter, principal approaches used in the synthesis of materials for gene delivery are shown, and their biocompatibility and functions are characterized for cells of different origin, namely, bacteria, yeast, plants, and mammals. Usually, the DNA that should be delivered to the target cells is “labeled” by gene coding for the green fluorescent protein (GFP); the gene delivery platform might be additionally labeled with a specific fluorescent dye. This helps monitoring the uptake, biodistribution, and location of the action of the transported nucleic acid in the cell (tissue, organ, organism), as well as the ways of possible clearance of the gene delivery platform from the body.
The chemical structure of materials for delivery of nucleic acids used in gene therapy should contain a positively charged group, usually an amine. Some advanced platforms also contain covalently conjugated vector elements (e.g., antibody or other ligand for specific receptors on plasma membrane of the targeted cells) for the addressed action. The hybrid platforms for simultaneous delivery of anticancer drugs and specific siRNA were created, and such multifunctionality is an important trend in the development of novel nanoscale materials used for disease treatment, gene therapy, and other biomedical purposes.
Nataliya Finiuk, Nataliya Mitina, Alexander Zaichenko, Rostyslav Stoika

Environmental Impacts of Nanomaterials

Frontmatter

Uptake, Biodistribution, and Mechanisms of Toxicity of Metal-Containing Nanoparticles in Aquatic Invertebrates and Vertebrates

Abstract
The increasing production and use of nanoparticles (NPs) have raised concerns with regard to their environmental accumulation and toxicity in nontarget organisms. Aquatic ecosystems receive high inputs of nanopollutants from riverine and terrestrial sources, and thus aquatic organisms are potentially vulnerable to the off-target toxic effects of nanoparticles. The reactivity and the toxicological profile of the metal-based NPs strongly depend on their physicochemical properties, including the particle size, shape, surface area, surface charge and coating, aggregation, dissolution, and protein corona effects. This chapter focuses on nano-ZnO as model nanopollutants with high production volume, broad use, and relatively well-studied toxicity, and summarizes the current state of knowledge about the toxic effects of nanoparticles and their mechanisms in aquatic invertebrates and lower vertebrates. We discussed the mechanisms of uptake and accumulation of Zn and ZnO in aquatic organisms and considered the ZnO-specific toxic mechanisms (related to the release of ionic Zn) as well as the toxic mechanisms shared between nano-ZnO and other nanoparticles such as redox activity, immunotoxicity and cytotoxicity reflected in the oxidative stress, lysosomal and mitochondrial damage, inflammation, and programmed cell death. We also identified the existing gaps in our knowledge with regard to the toxicity and environmental risk assessment of the nanopollutants including nano-ZnO and discussed possible research approaches to close these gaps.
Halina Falfushynska, Inna Sokolova, Rostyslav Stoika

Metallothioneins’ Responses on Impact of Metal-Based Nanomaterials for Biomedical Use

Abstract
Development and application of nanomaterials, specifically metal-containing nanoparticles (Me-NPs), are rapidly increasing in various technology and industry sectors, especially in medicine. Most frequently, these Me-NPs contain zinc (Zn), titanium (Ti), silver (Ag), copper (Cu), cadmium (Cd), as well as some other metals or their cations, noted in the Introduction. Therefore, the evaluation of the bioavailability of these particles seems to be crucial for understanding of the mechanisms of their activity and general toxicity. Broadening of their utilization also leads to the environmental pollution. Aquatic organisms are not only suitable experimental models for elucidation of the impact of the Me-NPs on biological systems, but also frequent evident targets of their toxic action. Biological effects of the Me-NPs can be realized by nanoparticles per se or by their constituents released after the biodestruction. Metallothioneins (MTs) are low molecular weight proteins uniquely composed of ~30% of cysteine that are widely distributed in all animal species. They bind cations of transition/post-transition metals, mainly Zn, Cd, and Cu, and participate in their distribution within the cell. Besides, they act as highly inducible stress proteins. Therefore, the evaluation of the expression and metal-binding function of MTs might be important for the analysis of the biodestruction of the Me-NPs. MTs as highly reactive thiols can also be involved in the scavending of the reactive oxygen species (ROS) induced by Me-NPs. On the contrary, there were attempts to define MTs response on the application of Me-NPs.
In this chapter, the role of MTs responses to the Me-NPs in the aquatic and other organisms was evaluated taking into account the previous experience of the authors and data reviewed in literature. More detailed attention was paid to the analysis of such responses in invertebrates, fish, and amphibia. The effects of the Me-NPs at the combined exposures have been also characterized. Results have shown that the ability of MTs to release metals from the nanoparticles is particular for different nanoparticles (n-ZnO, n-TiO2, Zn- or Co-containing nanonized vinyl-pyrrolidone-derived polymeric substances) and dependent on the animal species: bivalve mollusk, cyprinidae fish, and frog. Thus, the role of the MTs system of the aquatic and other organisms should be considered to monitor the environmental consequences of water pollution with the Me-NPs.
Oksana Stoliar, Rostyslav Stoika

Environmental Nanoparticles: Focus on Multipollutant Strategy for Environmental Quality and Health Risk Estimations

Abstract
The effect of environmental pollutants and factors on human organisms is complex. A combination of various chemical, physical, and biological environmental agents and factors can lead to combined health effects that may significantly differ from the effects of particular single agent or factor. Individual chemical compounds are naturally present at low concentrations as the environmental pollutants; however, they can interact with each other that can result in additive and synergistic combined effects of the pollutant mixture. Among different modes of interaction of environmental pollutants and factors, synergism of pollutant health effects is of special attention because it can result in underestimation of health hazard feasibility. The multipollutant strategies, prediction of feasible synergistic toxic effects of different environmental pollutants, and determination of a quantitative index of their synergism, as well as an assessment of additivity, antagonism, inhibition, potentiation and masking can significantly improve or worsen health hazard prognosis. Here, a feasibility of additive and synergistic effects of combinations of heavy metals, persistent organic pollutants, such as polychlorinated biphenyls, and insecticide neonicotinoids and air pollution particulate matter on nervous system health outcomes was analyzed. Recognizing multipollutant strategies may lead to the development of multi-pollutant models, and risk assessment approaches, which may address more efficiently the overall air quality objectives. Taking into consideration the outcomes of conducted analysis, new information technologies overlapping neurotoxic multipollutant distribution and expected harmful health effects can be developed.
Tatiana Borisova

Backmatter

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