An overview on Nanomaterials for Biomedical Application

By

Jagannath Sardar
(Roll No.: 09610309) PhD Scholar

Department of Mechanical Engineering

Indian Institute of Technology Guwahati October, 2009

Contents
1. 2. Introduction Classification of Nanomaterials
2.1 Carbon Based Materials 2.2 Metal Based Materials 2.3. Dendrimers 2.4. Quantum dots

Page
1 3
3 4 4 6

3.

Structures of Nanomaterials
3.1 Some general structures of well known nanomaterials

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7

4. 5.

Important nanomaterials for biomedical application Synthesis of Nanomaterials

5.1 Gold nanomaterials synthesis 5.2 Carbon nanomaterials synthesis 5.3 Synthesis of Quantum Dots

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10 12 15

6.

Nanomaterials in Biomedical Applications

6.1 Drug Delivery 6.2 Disease detection 6.3 Tissue Engineering 6.4 Diagnosis and treatment

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17 25 26 28

7. Conclusion 8. References

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1. Introduction:
The National Nanotechnology Initiative (NNI) subcommittee has given a definition of Nanotechnology as: nanotechnology is the understanding and control of matter at dimensions of roughly 1 to 100 nanometers, where unique phenomena enable novel applications. A nanometer is one billionth of a mater; a sheet of paper is about 100000 nanometers thick. Encompassing nanoscale science, engineering and technology, nanotechnology involves imaging, measuring, modeling and manipulating matter at this length scale [1, 2].

Figure 1.1. Nanoparticles in comparison with other biological entities Nanotechnology has been developed in many areas over the decades, one of the most important areas of this technology is nanomaterials level, which plays an important role in biomedical applications. Biomedical Nanotechnology is the unification of biotechnology and nanotechnology, including biomedical nanometrics and nano-materials. Diagnostics, drugs delivery, and prostheses & implants are three major areas where nanotechnology is entering biomedical areas. Biomedical Nanotechnology is attracting increasing interest as an emerging interdisciplinary research field straddling nanotechnology and biotechnology. As tools for combining nanotechnology with biomedical science, information techniques and algorithms appear to be gaining importance in biomedical nanometrics and nano-materials, with applications from the storage and reproduction 2

of genetic information to the control of developmental processes to sophisticated DNA-based computation and engineering [3]. Venkat S. Kalambur and John C. Bischof had explain in their presentation as a Magnetic nanoparticles can be potentially used in the targeted delivery of therapeutic agents in vivo, in the hyperthermic treatment of tumors, in magnetic resonance imaging (MRI) as contrast agents and in the biomagnetic separations of biomolecules. An understanding of the movement, heating and visualization of these nanoparticles in physiological systems in vitro and in vivo is required to tailor these nanoparticles for a few of these applications. Secondly, a characterization of the biodistribution and injury in vitro and in vivo are required when these nanoparticles are used as therapeutic agents in hyperthermia and drug delivery [4]. Many researchers have discussed about the affection of nano biomaterials in pharmaceutical and biological applications. For instance some of the recent developments for medicine applications are; tissue engineering, detection of protein and cancer therapy, drug delivery systems and medical imaging for cancer diagnosis. One of growing area is drug delivery system with benefit of targeting a specific cell for delivery with more therapy efficacy. For biomedical point of view, the cell parts of the organism are the small entity, about10um or sometimes even smaller which requires small dimensions of nano size application of materials which is enormous characteristics. Some important applications of the nano size materials are as - Fluorescent biological labels - Drug and gene delivery - Bio detection of pathogens - Detection of proteins - Probing of DNA structure - Tissue engineering - Tumour destruction via heating (hyperthermia) - Separation and purification of biological molecules and cells - MRI contrast enhancement - Phagokinetic studies etc. [5]. In this scope, we will discuss about nanomaterial for medical and biological applications along with salient overview with recent developments of the nanomaterials.

2. Classification of Nanomaterials:
Nanoparticles are constituted of several tens or hundreds of atoms or molecules and can have a variety of sizes and morphologies (amorphous, crystalline, spherical, needles, etc.). Some kinds of nanoparticles are already available commercially in the form of dry powders or liquid dispersions. The latter is obtained by combining nanoparticles with an aqueous or organic liquid to form a suspension or paste. It may be necessary to use chemical additives (surfactants, dispersants) to obtain a uniform and stable dispersion of particles. With further processing steps, nanostructured powders and dispersions can be used to fabricate coatings, components or devices that may or may not retain the nanostructure of the particulate raw materials. Industrial scale production of some types of nanoparticulate materials like carbon black, polymer dispersions or micronised drugs have been established for a long time [6]. Nanomaterials can be classified in different ways. Here we can classify as nanomaterials are four main types: 1. Carbon Based Materials 2. Metal Based Materials 3. Dendrimers 4. Quantum Dots

2.1 Carbon Based Materials:

These nanomaterials are composed mostly of carbon, most commonly taking the form of a hollow spheres, ellipsoids, or tubes. Spherical and ellipsoidal carbon nanomaterials are referred to as fullerenes, while cylindrical ones are called nanotubes. These particles have many potential applications, including improved films and coatings, stronger and lighter materials, and applications in electronics. Apart from that carbon nanofibres are also in this category. Linear nanostructures such as nanowires, nanotubes or nanorods can be generated from different material classes e.g. metals, semiconductors or carbon by means of several production techniques. As one of the most promising linear nanostructures, carbon nanotubes can be mentioned, which can occur in a variety of modifications (e.g. single- or multi-walled, filled or surface modified). Carbon nanotubes are expected to find a broad field of application in nanoelectronics (logics, data storage or wiring, as well as cold electron sources for flat panel displays and microwave amplifiers), and also as fillers for nanocomposites for materials with 4

special properties. At present, carbon nanotubes can be produced by Chemical vapor deposition (CVD) methods on a several tons per year scale and the gram quantities are already available commercially [7].

2.2 Metal Based Materials:

These nanomaterials include quantum dots, nanogold, nanosilver and metal oxides, such as titanium dioxide. A quantum dot is a closely packed semiconductor crystal comprised of hundreds or thousands of atoms, and whose size is on the order of a few nanometers to a few hundred nanometers. Changing the size of quantum dots changes their optical properties [6]. In this category, another commercially important class of nanoparticulate materials are metal oxide nanopowders, such as silica (SiO2), titania (TiO2), alumina (Al2O3) or iron oxide (Fe3O4, Fe3O3). But also other nanoparticulate substances like compound semiconductors (e.g. cadmium telluride, CdTe, or gallium arsenide, GaAs) metals (especially precious metals such as Ag, Au) and alloys are finding increasing product application.

2.3. Dendrimers:
These nanomaterials are nanosized polymers built from branched units. The surface of a dendrimer has numerous chain ends, which can be tailored to perform specific chemical functions. This property could also be useful for catalysis. Also, because three-dimensional dendrimers contain interior cavities into which other molecules could be placed, they may be useful for drug delivery.

Figure 2.1. Dendrimers Structures Dendrimers have unique characteristics including monodispersity and modifiable surface functionality, along with highly defined size and structure. This makes these polymers attractive candidates as carriers in drug delivery applications. Drug delivery can be achieved by coupling a drug to polymer through one of two approaches. Hydrophobic drugs can be complexed within the hydrophobic dendrimer interior to make them water-soluble or drugs can be covalently coupled onto the surface of the dendrimer. Using both methods we compared the efficacy of generation 5PAMAM (Poly amidoamine) dendrimers in the targeted drug delivery of methotrexate coupled to the polymer. The amine-terminated dendrimers bind to negatively charged membranes of cells in a non-specific manner and can cause toxicity in vitro and in vivo [8]. We will discuss in details later on the Dendrimers in drug delivery.

2.4. Quantum dots:

Quantum dots are spherical nano-sized crystals. They can be made of nearly every semiconductor metal (e.g., CdS, CdSe, CdTe, ZnS, PbS), but alloys and other metals (e.g. Au) can also be used. The prototypical quantum dot is cadmium selenide (CdSe). Quantum dots range between 2 and 10 nm in diameter (10 to 50 atoms). Generally, quantum dots consist of a semiconductor core, over coated by a shell (e.g., ZnS) to improve optical properties, and a cap enabling improved solubility in aqueous buffers (figure 2.1).

Figure 2.1. Schematic representation of a quantum dot. The cadmium selenide core is surrounded by a shell of zinc sulphide. Finally, a cap can be encapsulate the binary quantum dot by different material such as silica. The diameter of quantum dots ranges between 2-10 nm Apart from those major classifications there are several categories of nanomaterials are available, such as: Nanopowders, Nanocapsules, Nanoporous materials, Nanocomposites, Thin Films etc.

3. Structures of Nanomaterials:
Nanoparticles are designed and prepared with molecule and particle structures that can exibit functions desired for applications. The size and shape of the prepared particles are primarily important in the function adaptation [9]. The formation of condensed particles, the volume universally shows quite a pronounced tendency to approach a minimum, which is best achieved through the formation of one crystal lattice or another. Various pseudoclose packings can also be formed, especially when crystalline or quasicrystalline nanoparticles have sufficiently small sizes [10]. Knowledge on the structural features of nanoparticles is important for any technical and sophisticated application. These structural features may be determined by the analysing the

Fourier transformation of the diffraction pattern, which yields the radial distribution function, also called the pair distribution function [11].

3.1 Some general structures of well known nanomaterials:

Gold Nanoparticles

Silver Nanoparticles Silver Nanoparticles

Nanosilica

Dendrimer

Silicon-carbide nanowires

DNA strand

Quantum Dot B-Cd, Y-Se, RElectrons

Figure 3.1. Different Nanostructures [1, 12]

4. Important nanomaterials for biomedical applications:

For biomedical application there are a number of nanomaterials are being used. Some of the important nanomaterials are listed below and has reviewed on their characteristics and surface fictionalization in brief. One of the most important materials is metal nanoparticles, such as Gold (Au) and Silver (Ag) nonparticles are excellent nanomaterials providing a powerful platform in biomedical applications of biomolecular recognition and sensing, drug delivery, and imaging. Metal nanoparticles conjugates with activated enzyme through functionalized Thiol Ligands (figure 4.1). Similarly Partha Ghosh et al has shown an image on an enzymatic activity of gold nanoparticles in figure 6.

Figure 4.1 Metal NPs are used as a chemical scaffold to regulate the enzymatic activity Numerous examples can be found in the literature where bioconjugated Au nanoparticles are used as colorimetric biosensors detecting proteins, viruses, and bacteria at an extremely sensitive level. An additional advantage of nanoparticles is that the multiple ligands presented on the nanoparticle surface could drastically enhance affinities of specific monovalent interactions via the multivalent binding between NPs and the biological target [13]. Partha Ghosh et.al [14]. has reviewed Gold nanoparticles provide non-toxic carriers for drug and gene delivery applications.

Figure 4.2. Various applications of gold nanoparticles in therapy With these systems, the gold core imparts stability to the assembly, while the monolayer allows tuning of surface properties such as charge and hydrophobicity [14]. Another important material is Quantum dots (QDs). It has zero-dimension materials exhibiting quantum confinement in all three spatial dimensions. They are semiconductor nanocrystals whose bandgap depends on the size of the QDs. The energy gap increases with decreasing particle size, and therefore smaller QDs emit light at higher energy, ie, lower wavelength and blue-shift, whereas larger QDs absorb and fluoresce at longer wavelengths and red shift. QDs have broad excitation spectra yet narrow and tunable emissions, and have thus been widely used as optical labels in a wide range of biomedical applications including immunoassays for proteins, nucleic acids, bacteria and toxin analysis [15]

Magnetic nanoparticles of iron oxides have a long history of investigation and have shown
remarkable potentials in biomedical research. A unique characteristics of magnetic particles is their ability to move simply by the influence of an external magnetic field. Magnetic nanoparticles with the appropriate surface chemistry have thus attracted increasingly interests and have been widely used in the life sciences [16] including magnetic resonance imaging (MRI) contrast enhancement [17], drug delivery, hyperthermia, cell separation [18], and tissue repair. Both inherent properties of magnetic nanoparticles (magnetic, non-porous, controllable size and high stability) and modification of their surfaces play crucial roles in such applications. 10

Superparamagnetic iron oxide nanoparticles can furthermore improve the diagnostic value by enhancing the MRI contrast on surrounding healthy and pathological tissues, increasing the MRI resolution at the microscopic-level [19]. One of the most important materials is Carbon nanotubes (CNTs) which are well-ordered, hexagonal lattice networks of carbon atoms, which can be viewed as one or more layers of graphene sheets rolled up into a cylinder. The diameter of CNTs varies depending on the number of layers, and CNTs of high aspect ratios can be fabricated [1]. CNTs have been the subject of intense interest due to their superb electrical and thermal conductivities, exceptional mechanical strength, and excellent chemical and thermal stability [20]. Biomedical applications of CNTs require the functionalization of the materials with appropriate ligands rendering them bioactive and at the same time compatible with the biological environment (58-60). A number of methods have been developed to conjugate carbohydrates, peptides, and proteins on CNTs [21]. Apart from those materials there are many important materials are also used in biomedical sectors such as nanosilica, dendrimers, nanogel, nanoclay and so on so forth.

5. Synthesis of Nanomaterials:
In this occasion we will discus the synthesis process of some important nanomaterials.

5.1 Synthesis of Gold nanomaterials:

Several research groups have fabricated delivery systems based on GNPs bearing functional moieties, which are anchored with thiol-linkers, in their monolayers. A wide variety of monolayer protected clusters (MPCs) can be formed rapidly and in scalable fashion using the one-pot protocol developed by Schiffrin et al. in 1994 [22]. In this preparation, AuCl4- salts are reduced with NaBH4 in the presence of the desired thiol capping ligand or ligands (figure 5.1). The core size of the particles can be varied from 1.5 nm to ~6 nm by varying the thiolgold stoichiometry [14].

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Figure 5.1. Formation of MPCs using the Schiffrin reaction and MMPCs using the Murray's place-exchange reaction. Ashkan Tavakoli Naeini et al. [23] said that one of the most widely used methods is the reduction of tetrachloroaurate ions (AuCl4-) in aqueous medium using sodium citrate to generate particles with diameters typically ranging from 10 to 100 nm. Although this method has good control over producing a particular particle size, it is limited to the synthesis of larger particles. The Brust method and various modifications are useful for the generation of Au NPs having core sizes ranging from 1 to 4 nm. In the Brust method, the transfer of AuCl4 into toluene or chloroform is performed using tetraalkylammonium bromide followed by reduction with sodium borohydride in the presence of alkylthiols. On the other hand Linear-dendritic copolymers containing hyperbranched poly (citric acid) and linear poly(ethyleneglycol) blocks (PCA-PEGPCA) are used as reducing and capping agents to synthesize and support gold nanoparticles (Au NPs). Another method of synthesis of ultra fine gold nanoparticles created by the Liverpool University team. They created a gold nanoparticle one thousand times smaller in diameter than a human red blood cell, and attached it to a gold substrate by means of a 'molecular wire'. The wire was synthesised by Professor Don Bethell [24], who explains: "Its role was to act as a 'spacer' between the nanoparticle and the conducting substrate, and also as a chemical glue, since both ends were composed of sulphur-containing thiols which adhere readily to gold." They have given a schematic to synthesis gold nanoparticles in the figure 5.2.

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Figure 5.2. Nanoscale redox gate constructed with a single gold nanoparticle

5.2 Synthesis of Carbon nanomaterials:

Synthesis of carbon nanotubes can be done by different methods. The three most commonly used methods are the arc discharge, laser ablation, and chemical vapor deposition (CVD) techniques. While the arc and laser methods can produce large quantities of carbon nanotubes they lead to resilient contaminants, including pyrolytic and amorphous carbon [25], which are difficult to remove from the sample. Such impurities result in low recovery yield for the carbon nanotube product [26]. However, recent advances in scaling up these methods, as well as development new fabrication methods such as high pressure carbon monoxide (HiPCO), have created commercial supplies of carbon nanotubes with more than 90% purity with competitive prices. In contrast, the less scalable CVD process offers the best chance of obtaining controllable routes for the selective production of carbon nanotubes with defined properties [27]. CVD is catalytically driven, wherein a metal catalyst is used in conjunction with the thermal decomposition of hydrocarbon feedstock gases to produce carbon nanotubes. In most cases, the resultant growth of nanotubes occurs on a fixed substrate within the process. Figure 5.3 illustrates a typical CVD process for the generation of SWNTs. SWNTs are synthesized by the reaction of a hydrocarbon (e.g., CH4) vapor over a dispersed Fe catalyst. The synthesis apparatus consists of a quartz tube reactor inside a combined preheater and furnace set-up. The preheat section is operated 13

at~2000C. The catalysts are deposited and then hydrocarbon vapors are carried into the reaction zone of the furnace.

Figure 5.3. Schematic of a chemical vapor deposition (CVD) reactor that uses a two-zone furnace. Carbon nanotubes grow on the substrate placed inside the quartz tube [25]. Carbon nanofibres are a form of vapor-grown carbon fibre which is a discontinuous graphite filament produced in the gas phase from the pyrolysis of hydrocarbons [1]. Carbon nanofibres have transport and mechanical properties that approach the theoretical values of single crystal graphite, similar to the fullerness, but they can be made in high volumes at low cost ultimately lower than that of conventional carbon fibres. Yan Yan et al [28], explained that the carbon nanoparticles are prepared in solution by two methods: solvothermal synthesis and hot injection. In solvothermal synthesis amphiphilic triblock copolymer P123 (0.54.5 g) was dissolved in 40 mL of toluene with stirring until a homogeneous solution was formed. Mesophase pitches (MPs) (0.15 g) were added to the solution, followed by sonicate for the dispersion and stirring for 1 h. The mixture was centrifuged to separate the insoluble species and a dark brown homogenous solution was obtained for the reaction. Twenty-five milliliters of the above homogenous solution was added into a teflon-lined stainless steel autoclave with capacity of 30 mL. The autoclave was kept at 2000 C for 40 min and then cooled to room temperature naturally. The resultants were collected and centrifuged to get black powders. The products were rinsed several times with acetone and dried in ambient temperature. In Hot injection synthesis, MPs (0.25 g) were triturated and dissolved in 5 mL of oleic acid. The indiscerptible solid was then removed by centrifugation (ca. 50% of the MPs could be dissolved in oleic acid). This cold MP stock solution was quickly injected into 50 ml of oleic acid solvent 14

containing 0.763.04 mmol of H2SO4 at 1800C with N2 flowing. The reaction mixture was stirred vigorously at 1800C for 530 min. The products were then collected by centrifugation and washing as described above. Carbon nanofibres are manufactured by pyrolysis method which is illustrated in the figure 5.4.

Figure 5.4. Pyrolysis for carbon nanofibres [1] At the same time Ben Wong et al. [29] has given a synthesis procedure to produce gold nanoparticle composite. They have shown One gram of substrate, either silica or polymer was placed into a sintered container and loaded within a 10 cm3 autoclave. Approximately 100 mg of gold complex was added into the autoclave outside of the sintered container. There was no direct contact between the substrate and the gold precursor. The autoclave was filled with CO2 and the pressure was increased to 27.58MPa (4000 psi) at 313 K. CO2 was supplied to the autoclave by means of a refrigerator/compressor pump (NWA PM-101) connected to a chromatography grade CO2 cylinder. The temperature was controlled by means of an externally situated thermocouple connected via a feedback loop to a set of heating cartridges. The heating cartridges were located within an aluminium heating block surrounding the autoclave. After 24 h of processing, the CO2 was released from the autoclave over a period of approximately 20 s.

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5.3 Synthesis of Quantum Dots:

Quantum dots are specially engineered nanoscale crystals of semiconductor compounds. The name comes from the fact that their infinitesimal size enables a quantum electronics effect that causes the crystals to fluoresce brilliantly at specific, sharply defined colors. Bright, stable, tiny, and tunable across a broad spectrum of colors, quantum dots that are engineered to attach themselves to particular proteins have become a popular research tool in areas such as cancer research for detecting, labeling, and tracking specific biomarkers and cells. Making good quantum dots for biological research is complex. First a semiconductor compoundtypically a mixture of cadmium and seleniummust be induced to crystallize into discrete nanocrystals of just the right size. Cadmium is toxic, and the compound also can oxidize easily (ruining the effect), so the nanocrystals must be encapsulated in a protective shell such as zinc sulfide. To make them water soluble for biological applications, a short organic molecule called a ligand is attached to the zinc atoms. The organic ligand also serves as a tether to attach additional functional molecules that cause the dot to bind to specific proteins. The accepted commercial method uses a high-temperature reaction (about 300 degrees Celsius) that must be carefully controlled under an inert gas atmosphere for the crystallization and encapsulation stages. An intermediate ligand material that can tolerate the high temperature is used to promote the crystallization process, but it must be chemically swapped afterward for a different compound that makes the material water soluble. The ligand exchange stepas well as several variations on the processis known to significantly alter the luminescence and stability of the resulting quantum dots. Biomedical applications require high-quality water-soluble quantum dots. Quantum dots could be made directly in water but often have narrow available size ranges and wide size distribution. On the contrary, quantum dots produced from high temperature organic solvent synthetic strategies are monodisperse (leads to narrow FWHM) with very wide emission color ranging from ultraviolet to near infrared (3002500 nm) by simply changing the size, composition and/or structure. However, these quantum dots synthesized in organic solvents are insoluble in water. William W. Yu et al. [30] has given a method to make the high-quality hydrophobic quantum dots soluble in water and also active in bioconjugate reactions. The quantum dots synthesized in organic solvents have hydrophobic surface ligands such as trioctylphosphine oxide (TOPO), trioctylphosphine (TOP), tetradecylphosphonic acid (TDPA) or oleic acid. These hydrophobic ligands could be replaced by some water-soluble bifunctional 16

molecules in which one end connects to quantum dot surface atoms and the other end is hydrophilic and may also be reactive to biomolecules (Fig. 5.5). Examples of some water-soluble bifunctional molecules used are mercaptocarbonic acids [HS-(CH2)n-COOH, n = 115], 2aminoethanethiol, dithiothreitol, dihydrolipoic acid, oligomeric phosphines, peptides, and crosslinked dendrons.

Figure 5.5. Quantum dot (QD) water solubilization strategies. At the same time they found that quantum dots could enter the cells through endocytosis, and the cell death was highly related to the uptake quantity no matter what surface coating is. Nonetheless, the surface coating did affect the uptake quantity and in turn influenced the intracellular cytotoxicity [31].

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Figure 5.6. In vitro cytotoxicity of water-soluble quantum dots with different PEG coatings (the numbers of 750 and 6000 are the molecular weights of PEG polymers) to SK-BR-3 human breast cells. Full growth medium of confluent SK-BR-3 cells were incubated with water-soluble quantum dots in the dark over 24 h.

6. Nanomaterials in Biomedical Applications:

Application of nanomaterials in biomedical sectors can be divided into various areas, such as Drug delivery, disease detection, Tissue Engineering, biosensor, MRI contrast enhancement, antimicrobial treatment, protein detection, Manipulation of cells and biomolecules, Cancer and tumor therapy, etc. We will discuss some of the areas where nanomaterials are indispensable.

6.1 Drug Delivery:

Drug delivery systems (DDSs) provide positive attributes to a free drug by improving solubility, in vivo stability, and biodistribution. They can also alter unfavorable pharmacokinetics of some free drugs. Moreover, huge loading of pharmaceuticals on DDSs can render drug reservoirs for controlled and sustained release to maintain the drug level within

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therapeutic window. For example, a gold nanoparticle with 2 nm core diameter could be, in principle, conjugated with ~100 molecules to available ligands (n=~108) in the monolayer [14]. Zubarev et al. have recently succeeded in coupling of ~70 molecules of paclitaxel, a chemotherapeutic drug, to a GNP with 2 nm core diameter [32]. Drug delivery and related pharmaceutical development in the context of nanomedicine should be viewed as science and technology of nanometer scale complex systems (101000 nm), consisting of at least two components, one of which is a pharmaceutically active ingredient, although nanoparticle formulations of the drug itself are also possible. The whole system leads to a special function related to treating, preventing or diagnosing diseases sometimes called smart-drugs or theragnostics. The primary goals for research of nano-bio-technologies in drug delivery include:

More specific drug targeting and delivery, Reduction in toxicity while maintaining therapeutic effects, Greater safety and biocompatibility, and Faster development of new safe medicines [33].

The main issues in the search for appropriate carriers as drug delivery systems pertain to the following topics that are basic prerequisites for design of new materials. They comprise knowledge on (i) drug incorporation and release, (ii) formulation stability and shelf life (iii) biocompatibility, (iv) biodistribution and targeting and (v) functionality. In addition, when used solely as carrier the possible adverse effects of residual material after the drug delivery should be considered as well. One of the major challenges in drug delivery is to get the drug at the place it is needed in the body thereby avoiding potential side effects to non diseased organs. This is especially challenging in cancer treatment where the tumor may be localized as distinct metastases in various organs. The non restricted (cyto) toxicity of chemotherapeutics thus limits the full use of their therapeutic potential. Local drug delivery or drug targeting results in increased local drug concentrations and provides strategies for more specific therapy. Nanoparticles have specific particles as tools to enable these strategies. These include benefits such as their small size which allows penetration of cell membranes, binding and stabilization of proteins, and lysosomal escape after endocytosis.

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Ajay kumar gupta et al [34]. has shown that paramagnetic nanoparticles are also important for drug delivery. Recent work by Kannan and Katti et al [35]. investigated gum arabic labeled radioactive AuNPs that localize in liver. This study combines the therapeutic property of radioactive gold 198Au (max = 0.96 MeV, t1/2 = 2.7 days) and target specific biomolecule to form a powerful radiopharmaceutical for targeted drug delivery. This gum Arabic labeled radioactive gold can be used to treat liver cancers with higher radiation dose inherent to radioactive nanoparticles that contain thousands of radioactive atoms. Patri AK et al. has shown that drug delivery can be proceeding by dendrimers also [36]. Barrett E. Rabinow [37] has shown a drug delivery system by nanoparticles in the figure 6.1 a and 6.1 b below:

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Figure 6.1 a. Trafficking of drug nanoparticles by macrophages, monocytes and neutrophils. Tao Xu et al. [38] explained solid nanoparticles, polymeric nanoparticles, and polymeric selfassemblies, have attracted increasing attention for use as potential drug delivery systems. The 21

advantages of using nanoparticle systems for drug delivery result from their two basic properties. First, nanoparticulates, due to their small size, can penetrate small capillaries and be taken up by cells, which allows for efficient drug accumulation at target sites in the body. Second, the use of biodegradable materials for nanoparticulate preparation allows for sustained drug release within the target over a period of days or even weeks after administration.

Figure 6.1 b. schematic illustration of drug delivery via active and passive targeting, solid and dotted line respectively. Gold nanoparticles are capable of delivering large biomolecules, without restricting themselves as carriers of only small molecular drugs. Tunable size and functionality make them a useful scaffold for efficient recognition and delivery of biomolecules. They have shown the success in 22

delivery of peptides, proteins, or nucleic acids like DNA or RNA. Gold nanoparticles can likewise be nanocarriers of peptides and proteins of interest. Rotello et al. have reported that cationic tetraalkyl ammonium functionalized GNPs recognize the surface of an anionic protein through complementary electrostatic interaction and inhibit its activity (Fig. 6.2) [14]. The activity was recovered due to release of free protein by treating the proteinparticle complex with GSH, showing GNPs as potential protein transporters.

Figure 6.2. Schematic representation of glutathione (GSH)-mediated disruption of nanoparticle -galactosidase protein interactions. Gold Nanoparticles provide attractive candidates for gene delivery. They can be made quite small to provide a high surface-to-volume ratio, maximizing the payload/carrier ratio. Perhaps equally important, the monolayer coverage of MPCs and MMPCs systems allow tuning of the charge and hydrophobicity to maximize transfection efficiency while minimizing toxicity. We have shown in our earlier studies that gold nanoparticles functionalized with cationic quaternary ammonium groups (a) bind plasmid DNA through electrostatic interactions. Many researcher reveals that gold nanoparticles have good drug delivery capability. Nevertheless, dendrimers are also very efficient for drug delivery point of view. Sonke Svenson 23

[39] has shown how drug delivery takes place by with the help of dendrimer nanoparticles. He also reviewed the bio compatability of the dendrimers. The therapeutic areas for dendrimerdrug formulations include anticancer, anti-inflammatory, and antimicrobial treatments. In many cases, dendrimers have been functionalized with poly(ethylene glycol) (PEG) chains in order to enhance their container properties and to improve the biocompatibility of these carrier molecules.

Figure 6.3. Mechanism of dimeric prodrug activation by a single enzymatic cleavage [40] The biocompatibility of dendrimers follows patterns known from other small particles such as micelles and liposomes. Cationic surfaces show cytotoxicity; however, derivatization with fatty acid or PEG chains, reducing the overall charge density and minimizing contact between cell surfaces and dendrimers, reduces these toxic effects [38]. In the figure 6.4 represents the dendritic structure and the effect of PEG. 24

Figure6.4. (A) Structure of a poly(etherhydroxylamine) (PEHAM) dendrimer G1 conjugated with 25% of PEG550 to its surface. (B) Solubility enhancement of camptothecin formulated with PEHAM dendrimer G1-PEG550 in water (light) and wateralcohol (dark) compared to drug as received. (C) Dissolution curves of free drug and lyophilized drug formulations with PEHAM dendrimers G1-PEG550 (2) and G3-OH (3) [41].

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6.2 Disease detection:

A number of researchers discovered that a nanoparticle has disease detection capability in human body. For disease detection different nanomaterials are being used. Such as cancer cell detectionat early stages, tumor detection as well as treatment, bacterial protein detection and destruction, etc. Optical and electronic properties of AuNPs can be utilized to enhance the contrast in molecular imaging for the detection of cancer at early stages. For example, AuNPs labeled with monoclonal antibodies against EGFR (epidermal growth factor receptor) that are over-expressed in skin cancer were shown to localize on the abnormal cervical SiHa cell lines by imaging via ndoscopecompatible microscopies, such as optical coherence tomography and reflectance confocal microscopy [42]. Kenneth E. Scarberry et al. [43] says in their one research paper that Magnetic cobalt spinel ferrite nanoparticles coated with biocompatible polygalacturonic acid were functionalized with ligands specific for targeting expressed EphA2 receptors on ovarian cancer cells. By using such magnetic nanoparticle-peptide conjugates, targeting and extraction of malignant cells were achieved with a magnetic field. Targeting ovarian cancer cells with receptor specific peptidemodified magnetic nanoparticles resulted in cell capture from a flow stream in vitro and from the peritoneal cavity of mice in vivo. Successful removal of metastatic cancer cells from the abdominal cavity and circulation using magnetic nanoparticle conjugates indicate the feasibility of a dialysis-like treatment and may improve long term survival rates of ovarian cancer patients. This approach can be applied for fighting other cancers, such as leukemia, once the receptors on malignant cells are identified and the efficacy of targeting ligands is established. Apart from that Magnetic iron oxide nanoparticles encased in a biocompatible material can make detecting cancer cells easier, even if the cancer cells are small and clearer so there is less mistakes in the detecting process. These particles stick to the tumor cells turning them into little magnets which are then attracted to the tip of a biopsy needle [44].

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6.3 Tissue Engineering:

As emerging areas, tissue and implant engineering are evolving to address the shortage of human tissue and organs. The core of the tissue engineering and regenerative medicine are the fabrication of scaffolds, in which a given cell population is seeded, proliferated, and differentiated with the introduction of functional cell types from many different sources [45]. Nanostructured materials and their modified forms offer some attractive possibilities in the fields of tissue and implant engineering [34]. Bone-like nanostructure scaffolds have been developed using the technology of composites to imitate natural bone in bone tissue engineering [46]. Magnetic nanoparticles have been used in cell isolation and purification for tissue engineering and other applications. Nanoparticles can be conjugated with intense, stable fluorescence for tracing cells through several generations, which is ideal for the study of migration, motility, morphology and other cell functions.

Figure 6.5. (A) Schematic drawing of long bone microstructure (compact bone). (BD) Schematic drawing of honeycomb scaffold for guided and biomimetic long bone tissue regeneration. (E) nano-hydroxyapatite (NanoHA)- and degradable polymer based honeycomb scaffold. (F) A honeycomb scaffold was implanted into radius defect rabbits to regenerate long bone. 27

Biomimetic processing is based on the idea that biological systems store and process information at the molecular level. Extending this concept to the processing of nanocomposites for biomedical devices and tissue engineering, such as scaffold for bone regeneration, has been brought out in the last few years [46]. Several research groups have reported the synthesis of novel bone nanocomposites of HA and collagen, gelatin, or chondroitin sulfate, through a selfassembly mechanism. Indeed, hierarchical self-assembly of nano-fibrils is ubiquitous in nature, as in bone, muscle, and intestine, etc, and can lead to the creation of specific shapes and conformations in macromolecular structures. The mechanical property of CNT polystyrene composite is increased through covalent crosslinkage between CNTs and polymer matrix (fig. 6.6) [37].

Figure 6.6. High-resolution-TEM showing the ultra-thin (2 nm) coating on CNT (A) increased the tensile strength of the CNT-polystyrene composite.

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6.4 Diagnosis and treatment:

The biological application of nanoparticles is a rapidly developing area of nanotechnology that raises new possibilities in the diagnosis and treatment of human cancers. In cancer diagnostics, fluorescent nanoparticles can be used for multiplex simultaneous profiling of tumor biomarkers and for detection of multiple genes and matrix RNA with fluorescent in-situ hybridisation. Maksym V Yezhelyev et al. [47] says that some breast cancers express protein biomarkers (eg: oestrogen receptor, progesterone receptor, and ERBB2) on which therapeutic decisions are made. Semiconductor fluorescent nanocrystals, such as quantum dots, have been conjugated to antibodies, allowing for simultaneous labelling and accurate quantification of these target proteins in one breast tumour section. The use of nanoparticlesnot only quantum dots of different sizes and emission spectra, but also gold-containing nanoparticles (i.e., Raman probes)will allow the simultaneous detection and quantification of several proteins on small tumour samples, which will ultimately allow the tailoring of specific anticancer treatment to an individual patients specific tumor protein profile. The ability to detect molecular targets simultaneously on individual tumor samples could allow correlation between gene products and proteins in real time. In addition, the effects of an individual treatment on expression of the target protein can be monitored before and after treatment, and provide a rapid method to measure the efficacy of a targeted therapy [48]. As earlier we have discussed, Quantum dots are fluorescent nanoparticles with sizes of 210 nm that contain a core of hundreds to thousands of atoms of group II and VI elements (e.g., cadmium, technetium, zinc, and selenide) or group III (e.g., tantalum) and V elements (e.g., indium).22,23 Quantum dots containing a cadmium selenide core and a zinc sulphide shell, surrounded by a coating of a coordinating ligand and an amphiphilic polymer, are most commonly used for biological application. This structure enables quantum dots to emit powerful fluorescence that differs in nature from organic dyes. Quantum dots can be tuned to emit waves at between 450 nm and 850 nm (i.e., from ultraviolet to near infrared) by changing the size or chemical composition of the nanoparticle [47]. This so-called quantum confinement effect produces many quantum-dot colours, which can be visualized simultaneously with one light source. Quantum dots emit narrow symmetrical emission peaks with minimum overlap between spectra, allowing unique resolution of their spectra and measurement of fluorescent intensity from several multicolour fluorophores by real-time quantitative spectroscopy. These key advantages make it possible to label multiple molecular targets simultaneously by use of 29

quantum dots both in vitro and in vivo. However, use of quantum dots in imaging and therapeutics in vivo is limited by the toxic effects of the heavy-metal core (fig. 6.7).

Figure 6.7. Methods for conjugating quantum dots to biomolecules EDAC=ethyl-3-dimethylamino-propyl-carbodiimide. SMCC=succinimidyl-4-N-maleimidomethyl-cyclohexane carboxylate. COOH=carboxyl group. NH2= amine group. SH=sulohydryl group. (A) Traditional covalent crosslinking chemistry with EDAC as catalyst. (B) Conjugation of antibody fragments to quantum dots via reduced sulphhydrylamine coupling. The level of fluorescent emission from these conjugated nanoparticles correlates with expression of the protein (fig.6.8). The bright fluorescence of quantum dots enables identification of targets in low levels in cancer cells, resulting in increased sensitivity [49].

30

Figure 6.8: Use of quantum dots to detect protein expression in tumour expressing oestrogen receptor and progesterone receptor (top) or ERBB2 (bottom) (A) Paraffin-embedded human breast tumours stained with human antibodies against oestrogen receptor (ER), ERBB2, and progesterone receptor (PR) conjugated with quantum dots (565 nm, 655 nm, and 605 nm, respectively). (B) Fluorescent intensity from quantum dots shows level of labelled biomarker expression in each tumour. au=arbitrary units.

Alf Lamprecht et al [50]. has shown how the nanoparticles acts to delivers the drug and at the same time how does it acts to treatment of inflammatory Bowel disease. All nanoparticles were prepared with poly[DL-lactide-coglycolide], a biocompatible and biodegradable polymer that is now well established for use in humans. Nanoparticles were characterized in terms of size, polydispersity, surface potential, encapsulation efficiency, and drug release. The biodegradable polymer poly[DL-lactide-co-glycolide] 50/50 (PLGA) (mol. wt. 5,000 or 20,000) was purchased from Wako (Osaka, Japan). Rolipram was received as a gift from Schering AG (Berlin, Germany). Trinitrobenzenesulfonic acid (TNBS) and o-dianisidine hydrochloride were obtained from Sigma Chemical (Deisenhofen, Germany) and hexadecyltrimethylammonium bromide was obtained from Fluka (Deisenhofen, Germany). All

31

other chemical reagents were purchased from Sigma Chemical (Steinheim, Germany), Merck AG (Darmstadt, Germany), or Nacalai Tesque Inc. (Kyoto, Japan) and were of analytical grade. To evaluate the therapeutic value of rolipram-containing nanoparticles, the effect of the carrier system was studied on preexisting colitis. On day 3, all animals received an intrarectal application of TNBS except the healthy control group. Before this time point, animals showed no clinical problems. After inducing the experimental colitis the clinical score increased rapidly and consistently for the next 3 days for all groups. The inflamed tissue showed an extremely increased mucus production in the area of distal colon compared with the histology of healthy gut sections from the control group (fig. 6.9). Significant damages of the intestinal tissue, e.g., ulceration, have been observed [51].

Figure 6.9. Determination of colon/body ratio (a) and myeloperoxidase activity (b) after final drug administration (day 11) and washout phase (day 15). Healthy control group ( ), colitis control group (t), rolipram solution receiving group (s), PLGA nanoparticles (mol. wt. 20,000)receiving group (p), and PLGA nanoparticles (mol. wt. 5,000)-receiving group (o). From the above discussion we have seen that the disease detection and treatment according to the degree of inflammation of the disease, different nanomaterials can be used. Fortunately it is revealed that the efficiency for the purposes of treatment of the disease nanomaterials are indispensable for the advance generation of human being.

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7. Conclusion:
Nanotechnology is a very vast area. In this review it is not possible to elaborate all the aspects of a big technology. Some of the important points have been discussed in this space. As we know that nanotechnology is the understanding and control of matter at dimensions of roughly 1 to 100 nanometers, where unique phenomena enable novel applications. A nanometer is one billionth of a mater (10-9m). These very small materials have typical characteristics. With the help of those characteristics nanomaterials are being used in biomedical sectors. Huge applications of nanomaterials have been takes place in that sector due to its different merits to different acts. Some of the applications in biomedical have been enlisted which are overviewed in this occasion. Here we have discussed about

Types of nanomaterials Carbon Based Materials Metal Based Materials Dendrimers Nanocomposites

Different Nanostructures Synthesis of the nanomaterials Importance of nanomaterials Activities of the nanomaterials Application of the nanomaterials in different biomedical sectors Such as: Drug Delivery Disease detection Tissue Engineering Diagnosis and treatment For those applications different types of nanomaterials are being used, such as, Gold/Silver nanoparticles, carbon nanotubes, quantum dots, dendrimers, metallic magnetic nanoparticles and different metal oxide etc.

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