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Nanotechnology on Duty in Medical Applications Current Pharmaceutical Biotechnology, 2005, 6, 17-33 17 Nanotechnology on Duty in Medical Applications T. Kubik1,*, K. Bogunia-Kubik2 and M. Sugisaka3 1Institute of Cybernetics Engineering, Wroclaw University of Technology, Janiszewskiego 11/17, 50-372, Wroclaw, Poland; 2L. Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, R. Weigla 12, 53-114 Wroclaw, Poland and 3Faculty of Engineering, Oita University, Dannoharu 700, 870-1192 Oita, Japan Abstract: At the beginning of 21st century, fifty years after discovery of deoxyribonucleic acid (DNA) double helix structure, scientific world is faced with a great progress in many disciplines of biological research, especially in the field of molecular biology and operating on nucleid acid molecules. Many molecular biology techniques have been implemented successfully in biology, biotechnology, medical science, diagnostics, and many more. The introduction of polymerase chain reaction (PCR) resulted in improving old and designing new laboratory devices for PCR amplification and analysis of amplified DNA fragments. In parallel to these efforts, the nature of DNA molecules and their construction have attracted many researchers. In addition, some studies concerning mimicking living systems, as well as developing and constructing artificial nanodevices, such as biomolecular sensors and artificial cells, have been conducted. This review is focused on the potential of nanotechnology in health care and medicine, including the development of nanoparticles for diagnostic and screening purposes, the manufacture of unique drug delivery systems, antisense and gene therapy applications and the enablement of tissue engineering, including the future of nanorobot construction. Key Words: Nanotechnology, nanoparticles, diagnostics, medicine, drug delivery. INTRODUCTION: 50 YEARS OF DNA DISCOVERY aspects of DNA diagnostics have been noticed. These ranging from PCR-based techniques, DNA microarrays and Year 2003 marks the 50th anniversary of the discovery of microfluidics to fluorescent in situ hybridisation (FISH), the double helix structure of the deoxyribonucleic acid single nucleotide polymorphism (SNP) or short tandem (DNA) molecule. The double helix structure of DNA was repeat (STR) detection, microelectrochemical systems first described by biologist James Watson (who in 1951, then (MEMS) and MEMS-based microfluidics (lab-on-a-chip) 23 years old, travelled from the USA to work with Francis technology for DNA analysis, and many others (reviewed by Crick) and Francis Crick, an English physicist at the Demidov [4]). In parallel, ribonucleic acid (RNA)-based University of Cambridge. This discovery was the result of approaches have been developed allowing an in vitro gene work by James Watson and Francis Crick at the Cavendish expression analysis due to the use of reverse transcriptase Laboratory at the University of Cambridge, and also Maurice catalysing the synthesis of copy cDNA. Wilkins and Rosalind Franklin at King’s College, London. They found that DNA is composed of two anti-parallel More recently, introduction of DNA analogues detection strands, which wind about a common axis to form a double systems, have attracted considerable attention. These helix, and two DNA strands are held together by weak bonds systems are based on sequence-specific base paring between between the basis of each strand, forming base pairs (bp): A- the DNA strands according to the Watson-Crick model. T pairs and G-C pairs. The DNA structure model and (A-T; They include, for example, the use of peptide nucleic acid G-C) complementarity rules were published on 25th of April (PNA) technology [5], or a new class of bicyclic high 1953 in the British journal Nature [1]. This revolutionary affinity DNA analogs called locked nucleic acid (LNA) [6]. discovery of DNA led to Watson and Crick being awarded the Nobel Prize in Medicine in 1962. It is also worthy to mention the applications of triple- stranded nucleic acid structures [7] and cell-free DNA and RNA in plasma as a new tools for molecular diagnostics [8]. DNA Based Diagnostic Procedures Starting from the DNA double helix discovery 50 years All these approaches are the milestones in the development of RNA/DNA diagnostics. In the 21st century ago, through the discovery and isolation of DNA polymerase they will play an important role in many areas of diagnostics in late sixties (1958-1959), description of polymerase chain reaction (PCR) over 25 years later [2, 3], discovery of and science, including the health care, medicine or pharmaceutical industry. This is especially true, since their restriction enzymes, hybridisation, and sequencing progress is supported by nanotechnology solutions, which techniques leading to the human genom sequencing, a great have been already successfully introduced. In addition the progress in the development of DNA technology and various knowledge and understanding of the mechanisms of biological processes, such as gene expression and protein *Address correspondence to this author at the Institute of Cybernetics production (DNA synthesis, transcription into messenger Engineering, Wroclaw University of Technology, Janiszewskiego 11/17, 50- mRNA and translation into protein) help not only in the 372 Wroclaw, Poland; Tel: +48 71 3202745, +48 71 3212677; Fax: +48 71 3203408; E-mail: [email protected] development of medical diagnostic procedures but may be 1389-2010/05 $50.00+.00 © 2005 Bentham Science Publishers Ltd. 18 Current Pharmaceutical Biotechnology, 2005, Vol. 6, No. 1 Kubik et al. also applied in medical treatment, by introducing gene and [12]. Nanotechnology (in other words molecular antisense therapy. nanotechnology, molecular manufacturing) also comprises In this review we will present the recent achievements of the building of nanostructures and manufacturing nanometre- nanoscience giving some examples of their potential scale objects. Therefore, the manipulation of nanoscale applications in the fields of molecular diagnostics, particles, different kinds of microscopy, the manipulation of development of drug delivery systems, and nucleic acid molecules, constructing nanometre resolution diagnostic and based therapies. analytical devices, and nanomachines and nanorobots, can all be described by this term - nanotechnology. ANALYSIS AND OPERATIONS WITH THE USE APPLICATION IN DIAGNOSTICS NANOTECHNOLOGY; MACHINES, MATERIALS BASED ON DNA MOLECULES Biosensors The term nanotechnology is derived from the Greek Biosensors are chemical sensors, in which recognition word ‘nano’, meaning ‘dwarf’, and applies to the principles processes rely on biochemical mechanisms utilisation. They of engineering and manufacturing at a molecular level. The consist of a biological element (responsible for sampling), most common definition of nanotechnology is that of and a physical element (often called transducer, transmitting manipulation, observation and measurement at a scale of less sampling results for further processing), Fig. (1) [13-16]. than 100 nanometres. Nanotechnology is intrinsically multidisciplinary, reliant on the basic science, analytical The biological element of a biosensor contains a bio- techniques and methodologies of a number of disciplines sensitive layer, which can either contain bioreceptors or be including: chemistry, physics, electrical engineering, made of bioreceptors covalently attached to the transducer. material science and molecular biology. Nanotechnology is Bioreceptors are responsible for binding the analyte of often represented by two fundamentally different interest to the sensor for the measurement. They might be approaches: ‘top-down’ and ‘bottom-up’. ‘Top-down’ refers biological molecular species (such as antibodies, enzymes, to making nanoscale structures by machining, templating and proteins or nucleic acids) or living biological systems (e.g., lithographic techniques, whereas ‘bottom-up’, or molecular cells, tissues, or whole organisms). The type of biological element defines biological specificity conferring mechanism nanotechnology, applies to building organic and inorganic materials into defined structures, atom-by-atom or molecule- used. by-molecule, often by self-assembly or self-organisation. The physical element translate information from the Biologists/chemists are involved in the synthesis of biological element into a chemical or physical output signal inorganic, organic and hybrid nanomaterials for the use in with a define sensitivity. Thus in the biosensor the chemical nanodevices, the development of novel nanoanalytical information, ranging from the concentration of a specific techniques, and the manipulation of biological molecules sample component to a total composition analysis, is such as DNA and the evolution of molecular machines. transformed into an analytically useful signal. Thus nanotechnology includes a wide range of different Biosensors may be classified according to the type of scientific fields, from lithography (with line widths less than biological element, or to the mode of signal transduction or, one micron) to nanomachines and nanorobots. The term alternatively, a combination of the two. Concerning the nanotechnology can be understood as altering individual biological specificity conferring mechanism used one can atoms and molecules at a precise location, either chemically distinguish 5 major categories of biosensors (see Fig. (2)): or physically. Nanotechnology also seeks to develop devices 1) antibody/antigen based, that can scan and to manipulate
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