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Vashist, J Nanomater Mol Nanotechnol 2013, 2:2http://dx.doi.org/10.4172/2324-8777.1000109

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Journal of Nanomaterials & Molecular Nanotechnology

Nanomaterials-Based Health Care and Bioanalytical Applications: Trend and ProspectsSandeep Kumar Vashist1*

AbstractThere has been a rapidly growing trend towards the use of nanomaterials (NM) in healthcare and bioanalytical sciences during the last decade. It has led to a wide range of prospective applications of NM in biosensors, diagnostics, therapeutics, drug delivery, medicine, biomedical imaging, signal enhancement, naked-eye assays, water purification and environmental monitoring. Several potential techniques have been developed for the cost-effective production, characterization, surface modification, functionalization, formation of nanocomposites and toxicity analysis of NM. The formulation of international regulatory guidelines to assess the NM toxicity is still a challenge for the scientific community and policy makers. However, it is expected that several NM-based products will be commercially-available in the coming years, after demonstrating compliance with the healthcare and bioanalytical requirements, and the nanotechnology regulatory guidelines that will be made in the next few years. We describe here the trend and prospects of highly prospective NM-based healthcare and bioanalytical applications.

KeywordsNanomaterials; Healthcare; Bioanalytical sciences; Trends; Prospects

IntroductionThe phenomenal advances in the field of nanomaterials (NM)

during the last two decades have resulted in numerous applications in healthcare and bioanalytical sciences [1-5]. They have been extensively used in biosensors [1,3-6], diagnostics [7-10], drug delivery [2,11,12], therapeutics [13-15], assays [16], medicine [17], biomedical imaging, environmental monitoring [18], and food packaging/safety [19]. These technological developments have led to transformative changes in the scientific landscape and emergence of new scientific disciplines, such as plasmonics, nanotoxicology [20], and environmental health and safety.

The National Nanotechnology Initiative was established by President Clinton in 2000, with the main objective of building, characterizing and understanding the nanoscale devices. The estimated economic impact of >1 trillion US dollars for the next two

*Corresponding author: Sandeep Kumar Vashist, PhD, The Head of Immunodiagnostics HSG-IMIT- Institut für Mikro- und Informationstechnik, Georges-Koehler-Allee 103, 79110 Freiburg, Germany, E-mail: sandeep.kumar.vashist@hsg-imit.de

Received: February 05, 2013 Accepted: March 06, 2013 Published: March 12, 2013

decades pulled the international scientific community and policy makers into the field of nanotechnology [21], as demonstrated by exponentially increased number of publications (Figure 1). The multi-billion dollar microelectronics industry has clearly demonstrated the potential of productive nanotechnology.

The most widely used NM are carbon nanotubes (CNT), graphene, quantum dots (QD), nanoparticles (NP) and nanocomposites, which have been extensively employed for various healthcare and bioanalytical applications. Due to the huge multi-billion dollar glucose monitoring market [22,23], the first intended application of any NM has always been the diabetic glucose sensing. Several techniques have been developed for the production and characterization of NM, while the environmental health and safety of NM are also being critically evaluated. Similarly, a wide range of prospective strategies have also been devised for modifying the surface of NM, inducing specific functional groups and development of nanocomposites. The potential developments in NM-based healthcare and bioanalytical applications are discussed here along with the challenges involved.

Nanomaterials and their Applications In Healthcare and Bioanalytical Sciences

NM have been widely employed in in vitro diagnostics [1,3-10], imaging, drug delivery [2,11-13], and therapeutics [5,13-15]. The NM-based diagnostic formats have been demonstrated to have ultra-high sensitivity, lower limit of detection, rapid response, high cost-effectiveness, signal enhancement, long-term stability, high-throughput, minimal sample requirement and capability for multiplex detection [24]. The use of NM has enabled the diagnosis of diseases at a very early stage [9,13], and the detection of ultra-trace concentrations of target analytes, using minimum sample volume. QD [25] and NP are much better imaging agents for targeting the specific disease sites in the body by conjugating them to biomarker-specific vectors. The plasmonic NP-based drug delivery has greater

Figure 1: Number of peer-reviewed articles published on nanomaterials during the past two decades, as determined from ISI Web of Knowledge on Feb 04, 2013 using the search terms of “carbon nanotubes”, “graphene”, “nanoparticles”, “quantum dots”, “chitosan” and “nanocomposites”.

Publication Period

1991-2000

2001-2005

2006-2010

2011-Present

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0

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100000

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160000 NanocompositesChitosanQuantum dotsGrapheneCarbon nanotubesNanoparticles

Citation: Vashist SK (2013) Nanomaterials-Based Health Care and Bioanalytical Applications: Trend and Prospects. J Nanomater Mol Nanotechnol 2:2.

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potential for targeted therapeutics to treat cancer (and other diseases), as the use of NP improves the bioavailability and pharmacokinetics of therapeutics. It will lead to targeted therapy at specific disease sites, and will avoid the exposure of healthy tissues.

The most widely-used NM (Figure 2), along with their healthcare and bioanalytical applications is described below.

Carbon Nanotubes (CNT)

CNT [26] are hollow cylindrical tubes having one, two or several concentric graphite layers capped by fullerenic hemispheres, which are referred to as single-, double- and multi-walled CNT, respectively. They have been widely used in biosensors [1,3,5], diagnostics, electrochemical sensors [1], drug delivery systems [2], tissue engineering, cell tracking and labelling, and other bioanalytical applications [27]. The technology developments have enabled the cost-effective bulk production of CNTs, by critically improving the synthesis and functionalization of CNTs. Several strategies have been devised for functionalizing CNTs with different chemical groups; conjugating them to biomolecules; and preparing the CNT-based electrodes. They have high mechanical strength, high thermal conductivity, high chemical stability, remarkable electrocatalytic activity, minimal surface fouling, low overvoltage and high aspect ratio (surface-to-volume), which makes them ideal for electrochemical sensing. The CNT-based electrochemical sensors [1,3], having rapid response and lower limit of detection, have been used to detect glucose, neurotransmitters/neurochemicals, proteins, cells, DNA, microorganisms, pharmaceutical substances and other biomolecules. The direct electron transfer between the enzyme and the CNT-based electrode has further enabled mediator less electrochemical sensing, which detects analytes with high precision, without any interference from electroactive physiological substances and drug metabolites. Similarly, CNT have tremendous potential for developing next-generation of highly efficient delivery systems for drugs (anti-cancer, anti-inflammatory, and other drugs) and biomolecules (DNA, RNA and proteins), due to their large surface area; unique structural,

electrical and optical properties; well-defined physico-chemical properties; and no toxicity (in case of functionalized CNT). Several conjugation strategies have already been developed to bind drugs/biomolecules to CNT.

However, several challenges still need to be tackled in order to make the CNT-based products commercially-viable [5]. The cost-effective synthesis of ultrapure CNTs without any metallic impurities is the foremost challenge. The development of biocompatible CNTs by surface modification, surface functionalization or bioconjugation is another critical requirement for several applications. The formulation of Nanotechnology Regulation, comprising the International guidelines to determine the toxicity of nanomaterials, is still in progress [28]. However, it is expected that the compliance of CNT-based products with these guidelines will provide the desired momentum for commercialization in near future.

Graphene

Graphene is the most widely used nanomaterial since its discovery in 2004 [4,5,29]. It is a two-dimensional planar sheet of sp2-bonded carbon atoms that are densely packed in a honeycomb crystal lattice. It has high surface-to-volume ratio, unique optical properties, extremely high mechanical strength, excellent electrical conductivity, remarkably high carrier mobility, high carrier density, high thermal conductivity, room temperature Hall Effect, ambipolar field-effect characteristics, and high signal-to-noise ratio. The large surface area of graphene leads to high surface loading of biomolecules, while it’s excellent conductivity and small band gap enables the conduction of electrons between the biomolecules and the electrode surface. It is a transparent material with very low production cost, low environmental impact, and about two-fold higher effective surface area than CNT. Moreover, the homogeneity enables the uniform and efficient functionalization of graphene. Chemical vapor deposition is used to produce large areas of single layer graphene, while chemical or thermal reduction of graphene oxide is the most commonly used mass production method for the cost-effective production of graphene. A wide range of chemical modification and biomolecular binding strategies [30], have been developed for inducing specific functional groups on graphene, and binding it to biomolecules. Moreover, numerous nanocomposites have also been developed by conjugating graphene to nanomaterials, and/or polymers.

Graphene has been widely used in healthcare and bioanalytical sciences [4,5,31,32], for the detection of a wide range of analytes, such as glucose, glutamate, hydrogen peroxide, benzene, ethyl benzene, xylenes, cyclohexane, nicotinamide adenine dinucleotide, hemoglobin, cholesterol, protein biomarkers, saccharides and cancer cells. It has immense potential for the development of electrochemical biosensors, based on the direct electron transfer between the enzyme and the electrode surface [6]. The graphene-functionalized electrodes have superior analytical performance, negligible interference from biological substances and drug metabolites, and excellent anti-fouling ability [33]. Graphene-based non-enzymatic electrodes have been used for the detection of ascorbic acid, uric acid, dopamine and hydrogen peroxide. Graphene-based nano-electronic devices have also been employed for DNA sensors, gas sensors, pH sensor, detection of environmental contaminants, detection of pharmaceutical compounds, detection of bacteria (Escherichia coli), and development of field-effect transistors (FET). Graphene oxide is ideal for food-packaging, to keep the foods fresher for extended periods of time due

Figure 2: Structures of most widely used nanomaterials in healthcare and bioanalytical sciences i.e. (A) Carbon nanotubes (left–single-walled; right–multi-walled), (B) Graphene (left–single-layered; right–multi-layered), (C) Nanoparticles, (D) Quantum dots, and (E) Chitosan. The figures (A-E) are reproduced with permission from OMICS Publishing Group [3,4], Royal Society of Chemistry [72], AZoNano [73], American Chemistry Society [74], and Elsevier B.V. [75], respectively.

Citation: Vashist SK (2013) Nanomaterials-Based Health Care and Bioanalytical Applications: Trend and Prospects. J Nanomater Mol Nanotechnol 2:2.

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to its potential anti-bacterial properties. Graphene-based membranes are impermeable to all gases and liquids, but allow water vapours to pass through. So, they will be highly beneficial for the alcoholic beverage industry and the biofuel production for the cost-effective distillation of ethanol at room temperature. Similarly, graphene filters have superior analytical performance for desalination, with respect to the conventional techniques. Moreover, graphene nanoribbons and graphene quantum dots have also been developed and have tremendous potential for ultrasensitive detection, due to their high sensitivity to the field effect and the chemical disruption at edges. However, the formulation of International regulatory guidelines for determining the safety of nanomaterials, together with strict compliance with the healthcare and bioanalytical requirements, are critical for the development of commercially-viable graphene-based products.

Nanoparticles (NP)

NP has been extensively used for healthcare and bioanalytical applications [34,35], due to their unique optical and other properties, and their ability to change color in response to the binding of molecules to their surface. The properties of NP change according to variation in their size or shape. Gold NP (GNP) is the most widely used NP due to their non-toxicity, biocompatibility and inert core [36-38]. They are employed for the early stage detection and photothermal therapy of cancer and other diseases, because of their plasmon absorption and scattering properties. They have been used for the highly sensitive detection of analytes [39,40]. As they accumulate preferentially at the tumor sites, they are potential nanocarriers of drugs, DNA and genes for the therapy of cancer and other diseases. They are also widely used in biomedical imaging, as they enhance the Raman and Rayleigh signals, thereby providing more analytical information. Therefore, they act as potential diagnostic drug delivery and therapeutic agents. Magnetic NP are the second most widely used NP that have been used for the detection of proteins, enzymes, DNA, mRNA, drugs, metabolites, pathogens, and tumor cells [24,41-43]. They have also been used for the photothermal destruction of tumor cells [44]. Various types of magnetic sensors based on different signal transduction mechanisms have been developed [41]. The diagnostic magnetic resonance technology has also been employed extensively for magnetic biosensing [42], highly sensitive analyte detection, multiplex analysis and development of point-of-care diagnostics.

Other nanomaterials

QD are inorganic nanocrystals that have unique optical properties, such as broad excitation range, narrow size-tunable emission, high photochemical stability and no photobleaching. They have been widely used as fluorescent probes [25,45], for the optical detection of ions, organic compounds, pharmaceutical analytes and biomolecules, in addition to the in vivo detection of target sites in cancer and in therapeutics [46]. They have enormous potential for multiplex analysis, mainly due to their small size and size-dependent emission [25,47].

Chitosan is another NM that has been widely used in biosensors, diagnostics, lab-on-a-chip devices, and other healthcare/bioanalytical applications [48], due to its biocompatible, biodegradable and non-toxic nature [49]. It is obtained by the deacetylation of chitin, which is the second most abundant natural polymer found in the shells of crustaceans, insects’ cuticles, and fungal cell walls. Being transparent, it is appropriate for optical sensors. Moreover, it is suitable for

electrochemical sensors due to its porous nature and highly ion permeability. It forms stable films under neutral and basic pH conditions. The amine groups of chitosan aid in the covalent binding of biomolecules, and the formation of nanocomposites with polymers or NPs. However, chitosan needs to be chemically modified to make it soluble in water and other common solvents.

Dendrimers are nanometer-scalethree-dimensional macromolecules, with a very high density of surface functional groups. They are hyperbranched, monodispersed and star-shaped structures that have a distinct core, interior dendron and exterior surface, with terminal functional groups. They have been used extensively in various biosensors/diagnostics [50,51], based on electrochemistry, fluorescence, surface enhanced Raman scattering, impedimetry and surface plasmon resonance. The use of dendrimers provides numerous analytical advantages, in terms of increased analytical sensitivity, high stability, high reproducibility and reduced non-specific binding. They have also been used in drug delivery, gene transfection and catalysis [51].

Lipid vesicles, thin lipid films and liposomes are biological NM, having composition very similar to the cell membrane [52,53]. They are highly biocompatible and can embed the biocomponents (such as receptors and proteins), under non-denaturing conditions. Moreover, they are highly sensitive to pH and temperature, and can effectively encapsulate hydrophilic or hydrophobic drugs. They have been used for the development of biosensors, diagnostics and controlled drug-delivery systems [52-54]. They have also been used for signal amplification in optical, electrochemical and acoustic biosensors. The hybrid nanoparticles, composed of lipids and polydiacetylene (PDA), have also been prepared and used for developing smart colorimetric biosensors, which are based on the chromatic transition, resulting from the change in conformation of PDA after specific biomolecular interactions [55].

A wide range of other NM [56-58] and nanocomposites [59-61], with unique properties have also been prepared, and used in various applications. The last two decades have witnessed significant developments in NM and their extensive applications in healthcare and bioanalytical sciences, which have generated tremendous technology push, as evident from the continuously increasing number of publications (Figure 1).

Challenges

Despite considerable technology push, there are still several challenges that need to be tackled in order to make the NM-based products commercially-viable. The reproducible and cost-effective production of NM is critical for all NM-based applications and products. There is also a need for stringent characterization, to obtain analytical information pertaining to the storage, functionalization, modification and usage of NM under optimum conditions. Moreover, the nature of metallic impurities in NM (such as in CNT) should be investigated, as they can substantially affect the properties and toxicity of NM. As each NM is unique, the material safety has to be evaluated individually for each NM [62]. The critical physiological parameters, i.e. absorption, distribution, metabolism, excretion and toxicity, must be determined if the NMis intended for in vivo applications. The risk of cytotoxicity increases if the NM contains heavy metals, and has prolonged tissue retention. The NM toxicity [63-67], should be analysed as per the international regulatory guidelines, that are still under formulation. However, the NP handling guidelines provided

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by the National Institute for Occupational Safety and Health [68], can be employed for developing new manufacturing processes with minimized workplace exposure risks. The drafting of claims for the Nanotechnology Regulation [28], is still in progress, with the main focus on the development of nanotechnology and evaluation of NM safety. The developed NM-based products and applications should be correlated with the established technologies, so that their advantages and limitations are apparent to all [5]. Moreover, the potential end-user trials of NM-based products should be done in the actual healthcare and bioanalytical settings, using the “real-world” samples, which will clearly demonstrate their utility and analytical merits.

There are potential concerns about the safety of NM, especially for therapeutic [69] and in vivo biomedical applications. The conflicting reports by researchers due to the lack of regulatory guidelines for NM toxicity analysis have exaggerated these concerns, and led to misconceptions about NM. However, these can be effectively addressed with information outreach, where the safety of NMs for various applications can be clearly addressed. Besides, the scientific community is intensively determining the toxicological profiles and potential adverse effects of NMs [63-66]; understanding their biological interaction mechanisms; developing robust and widely acceptable analytical tools and tests for characterizing NMs in various environments[67]; and determining the safety of NM throughout its life cycle, i.e. research and development, production, use, disposal, and/or recycle. A tiered testing system for assessing the NM toxicity has also been suggested [70]. It is based on the physico-chemical characterization of NM, prior to and during its subsequent testing in cell-free, cellular and in vivo assays. Recently, the use of an integrated multi-faceted approach has been suggested for the determination of NM safety [71]. This is based on the full life cycle assessment of NM and empirically derived risk assessment. It has also stressed the need for research, public education and media coverage, apart from the integrated legislation, in order to prevent the undesirable effect due to NM exposure.

Most of the healthcare and bioanalytical applications have employed non-porous NM, as they are free from the diffusion problems that are usually encountered with porous NM. Moreover, it is essential for NM-based biosensors to have an inert surface that do not contribute to the specific detection of the analyte [72,73]. Additionally, it is necessary to use an appropriate immobilization strategy for the efficient immobilization of biomolecules to the surface [74-76], as it can lead to much better assays with superior analytical performance [77-81]. The sample preparation [82] and other bioanalytical parameters also need to be optimized for the specific NM-based application.

Finally, the need of employing NM should be critically assessed for a particular application. They should be used in applications where they provide considerable analytical improvements, while the use of NM should be discouraged when they do not provide any major analytical improvement, but only adds to the cost and complexity. As an example, many NM have always been used initially for diabetic blood glucose monitoring due to its enormous market potential [83]. However, it is has always been clear to the experts and investors that the existing commercially-available blood glucose meters are already much better, and can detect the pathophysiological glucose without any technical concern [22,23,84]. Therefore, the use of NM in glucose monitoring only adds to the cost, without providing any considerable analytical merit.

The recent patent trend in NM clearly shows the growing scientific and industrial interest in the use of NM, and the continuously emerging highly prospective applications of NM in healthcare and bioanalytical sciences [85-88]. The commercial potential for NM has been estimated to be >300 billion Euros [89]. There is no doubt that the use of NM in certain applications has critically improved the technology by providing distinct analytical features, which has led to the emergence of new technologies, such as naked-eye Enzyme-linked Immunosorbent Assay (ELISA) [90] and alphaLISA [91]. It is expected that the next decade will witness the successful commercialization of many NM-based products, in addition to the continuous technology developments.

ConclusionsThe significant advances in NM have generated tremendous

technology push. The on-going research efforts are focussed on evaluating the safety of NM and formulating the international regulatory guidelines for the same, which is critical for further technology advancement. The analytical advantages of employing NM for various healthcare and bioanalytical applications are already apparent to the scientific community, as demonstrated by numerous publications. NM have been widely employed in numerous applications, as they provide enhanced signal, higher analytical sensitivity, lower limit of detection and better analytical characteristics. However, the commercial success of various NM-based products will be determined by the key technology differentiators, cost-effectiveness, reliability and market pull. The development of commercially-viable NM-based products requires critical investigation of their production uniformity, functional reproducibility, analytical parameters, toxicity, biocompatibility, and compliance with healthcare and bioanalytical requirements. Presently, the researchers are looking into the development of cost-effective procedures for producing reproducible, stable and biocompatible NM. Similarly, various surface modification and functionalization strategies are being employed to develop biocompatible and non-toxic NM. Based on the continuous technology developments and dedicated research efforts, it is expected that many NM-based products in healthcare and bioanalytical sciences will be commercialized successfully in the coming years. References

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91. AlphaLISA.

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