UNIT 5

NANOBIOTECHNOLOGY ITS APPLICATIONS

Nanotechnology

Nano is obtained from latin word nanus which means dwarf Nanotechnology, or nanotech, is the study and design of

machines on the molecular and atomic level. To be considered nanotechnology, these structures must be anywhere from 1 to 100 nanometers in size. Nanotechnology deals with phenomena whose physics or chemistry differs from that of bulk materials of the same composition. It uses a basic unit of measure called a "nanometer Bionanotechnology and nanobiotechnology are terms that refer to the intersection of nanotechnology and biology Nanobiotechnology, on the other hand, refers to the ways that nanotechnology is used to create devices to study biological systems Bionanotechnology is essentially miniaturized biotechnology

Father of nanotechnology:
Richard Rick Errett

Smalley Nobelist in chemistry for co-discovering fullerenes The US Senate passed a resolution to honor Smalley, crediting him as the Father of Nanotechnology Eric Drexler, the father of nanotechnology

Nanoparticles
Nanoparticles are particles that have one

dimension that is 100 nanometers or less in size. Major change in the properties occurs. Two significant effects are Increase in the ratio of surface area to volume Quantum effects are predominant Nanoparticles present several different morphologies (flakes, spheres, dendritic shapes, etc.).

N a n o te ch n o lo g y B a se d D ia g n o stics In v itro Im a g in g

Introduction
The application of nanobiotechnology in

medical diagnostics can be grouped into two areas, 1.Invitro

Biosensors Integrated devices

2.In-vivo
Implantable devices Medical imaging applications.

Traditional in vitro diagnosis:

In-vitro diagnosis for medical applications has

traditionally been a laborious task Blood and other body fluids or tissue samples are sent to a laboratory for an analysis, which could take hours, days or weeks, depending on the technique used, and be highly labor intensive. The many disadvantages include sample deterioration, cost, lengthy waiting times (even for urgent cases), inaccurate results for small sample quantities, difficulties in integrating parameters obtained by a wide variety of methods and poor standardization of sample collection.

Nano imaging:
Miniaturization, parallelization and integration

of different functions on a single device, based on techniques derived from the electronics industry, have led to the development of a new generation of devices that are smaller, faster and cheaper, do not require special skills, and provide accurate readings. These analytical devices require much smaller samples and will deliver more complete (and more accurate) biological data from a single measurement.

Contd.,
The requirement for smaller samples also means less invasive

and less traumatic methods of extraction. Nanotechnology enables further refinement of diagnostic techniques, leading to high throughput screening (to test one sample for numerous diseases, or screen large numbers of samples for one disease) and ultimately pointof-care diagnostics. Today, through the application of nanotechnology, both imaging tools and marker/contrast agents are being dramatically refined towards the end goals of detecting disease as early as possible, eventually at the level of a single cell, and monitoring the effectiveness of therapy. The convergence of nanotechnology and medical imaging opens the doors to a revolution in molecular imaging (also called nano-imaging) in the foreseeable future, leading to the detection of a single molecule or a single cell in a complex biological environment.

In-vitro Diagnostics
An in-vitro diagnostic tool can be a single biosensor, or

an integrated device containing many biosensors. A biosensor is a sensor that contains a biological element, such as an enzyme, capable of recognising and signalling (through some biochemical change) the presence, activity or concentration of a specific biological molecule in solution. A transducer is used to convert the biochemical signal into a quantifiable signal. Key attributes of biosensors are their specificity and sensitivity. Nanoanalytical tools like scanning probe microscopy or imaging mass spectrometry offer new opportunities for in-vitro diagnostics, like molecular pathology or reading out highly integrated ultra-sensitive biochips.

Techniques derived from the electronics industry

have enabled the miniaturisation of biosensors, allowing for smaller samples and highly integrated sensor arrays, which take different measurements in parallel from a single sample. Higher specificity reduces the invasiveness of the diagnostic tools and simultaneously increases their effectiveness significantly in terms of providing biological information such as phenotypes, genotypes or proteomes. Several complex preparation and analytical steps can be incorporated into lab-on-a-chip devices, which can mix, process and separate fluids, realising sample analysis and identification.

Integrated devices can measure tens to thousands of

signals from one sample, thus providing the general practitioner or the surgeon with much more complementary data from his patients sample. Some nanobiodevices for diagnostics have been developed to measure parts of the genome or proteome using DNA fragments or antibodies as sensing elements and are thus called gene or protein chips. Cellson-chips use cells as their sensing elements, employed in many cases for pathogen or toxicology screening. Integrated devices can be used in the early diagnosis of disease and for monitoring the progress of therapy. Recent developments aim at developing in-vitro diagnostic tools to be used in a standard clinical environment or e.g. as point-of-care devices.

Basis For A Strategic Research Agenda In-vitro Diagnostics

The ultimate goal is the fast, reliable, specific and cost-effective detection of

a few molecules (or even a single molecule) in a complex, non amplified and unlabelled biological sample. The improvement of invitro diagnostics towards this goal requires:

Nanoanalytical instruments of the highest spatial resolution, sensitivity and

range of information and integrated, combined instruments

Better sensitivity of screening methods, enabling the sample size to be

decreased or for the early detection of low concentrations of disease markers

Higher specificity, for quantitative detection of markers in complex samples

Stronger reliability, simplicity of use and robustness

Faster analysis

Nanoparticles and their medical applications

Applications
A list of some of the applications of nanomaterials to

biology or medicine is given below: - 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

Tissue engineering
Natural bone surface is quite often contains features that are about 100

nm across. If the surface of an artificial bone implant were left smooth, the body would try to reject it. Because of that smooth surface is likely to cause production of a fibrous tissue covering the surface of the implant. This layer reduces the bone-implant contact, which may result in loosening of the implant and further inflammation. It was demonstrated that by creating nano-sized features on the surface of the hip or knee prosthesis one could reduce the chances of rejection as well as to stimulate the production of osteoblasts. The osteoblasts are the cells responsible for the growth of the bone matrix and are found on the advancing surface of the developing bone. The effect was demonstrated with polymeric, ceramic and, more recently, metal materials. More than 90% of the human bone cells from suspension adhered to the nanostructured metal surface . In the end this findings would allow to design a more durable and longer lasting hip or knee replacements and to reduce the chances of the implant getting loose.

Contd.,
Titanium is a well-known bone repairing material

widely used in orthopaedics and dentistry. It has a high fracture resistance, ductility and weight to strength ratio. Unfortunately, it suffers from the lack of bioactivity, as it does not support sell adhesion and growth well. Apatite coatings are known to be bioactive and to bond to the bone.

Contd.,
Several techniques were used in the past to

produce an apatite coating on titanium. Those coatings suffer from thickness nonuniformity, poor adhesion and low mechanical strength. In addition, a stable porous structure is required to support the nutrients transport through the cell growth. It was shown that using a biomimetic approach a slow growth of nanostructured apatite film from the simulated body fluid resulted in the formation of a strongly adherent, uniform nanoporous layer The layer was found to be built of 60 nm

Cancer therapy
Photodynamic cancer therapy is based on the destruction of

the cancer cells by laser generated atomic oxygen, which is cytotoxic. A greater quantity of a special dye that is used to generate the atomic oxygen is taken in by the cancer cells when compared with a healthy tissue. Hence, only the cancer cells are destroyed then exposed to a laser radiation. Unfortunately, the remaining dye molecules migrate to the skin and the eyes and make the patient very sensitive to the daylight exposure. This effect can last for up to six weeks. To avoid this side effect, the hydrophobic version of the dye molecule was enclosed inside a porous nanoparticle The dye stayed trapped inside the nanoparticle and did not spread to the other parts of the body. At the same time, its oxygen generating ability has not been affected and the pore size of about 1 nm freely allowed for the oxygen to diffuse out.

Contd.,
Nanoparticles composed of polymers, lipids,

and inorganic materials loaded with drugs have been developed for targeted-therapy delivery systems. Additional efforts have focused on developing novel materials for the early detection of cancer, based on optically active agents such as quantum dots (QDs, semiconductor nanocrystals), gold, or magnetic (for example, iron oxide) nanoparticles. Advantages in using QDs and gold nanoparticles over traditional fluorophores include their enhanced optical properties, stability in biological fluids, and potential for

Multicolor optical coding for biological assays

The ever increasing research in proteomics and

genomic generates escalating number of sequence data and requires development of high throughput screening technologies. Realistically, various array technologies that are currently used in parallel analysis are likely to reach saturation when a number of array elements exceed several millions. A three-dimensional approach, based on optical "bar coding" of polymer particles in solution, is limited only by the number of unique tags one can reliably produce and detect.

Contd.,
Single quantum dots of compound

semiconductors were successfully used as a replacement of organic dyes in various biotagging applications. This idea has been taken one step further by combining differently sized and hence having different fluorescent colours quantum dots, and combining them in polymeric microbeads . A precise control of quantum dot ratios has been achieved. The selection of nanoparticles used in those experiments had 6 different colours as well as 10 intensities. It is enough to encode over 1 million combinations. The uniformity and reproducibility of beads was high letting for the bead identification accuracies

Manipulation of cells and bimolecules

Functionalized magnetic nanoparticles have found

many applications including cell separation and probing Most of the magnetic particles studied so far are spherical, which somewhat limits the possibilities to make these nanoparticles multifunctional Alternative cylindrically shaped nanoparticles can be created by employing metal electrodeposition into nanoporous alumina template Depending on the properties of the template, nanocylinder radius can be selected in the range of 5 to 500 nm while their length can be as big as 60 m. By sequentially depositing various thicknesses of different metals, the structure and the magnetic properties of individual cylinders can be tuned widely.

Contd .,

As surface chemistry for functionalisation of metal

surfaces is well developed, different ligands can be selectively attached to different segments. For example, porphyrins with thiol or carboxyl linkers were simultaneously attached to the gold or nickel segments respectively. Thus, it is possible to produce magnetic nanowires with spatially segregated fluorescent parts. In addition, because of the large aspect ratios, the residual magnetisation of these nanowires can be high. Hence, weaker magnetic field can be used to drive them. It has been shown that a self-assembly of magnetic nanowires in suspension can be controlled by weak external magnetic fields. This would potentially allow controlling cell assembly in different shapes and forms.

Protein detection
Proteins

are the important part of the cell's language, machinery and structure, and understanding their functionalities is extremely important for further progress in human well being. Gold nanoparticles are widely used in immunohistochemistry to identify protein-protein interaction. However, the multiple simultaneous detection capabilities of this technique are fairly limited. Surface-enhanced Raman scattering spectroscopy is a well-established technique for detection and identification of single dye molecules. By combining both methods in a single nanoparticle probe one can drastically improve the multiplexing capabilities of protein probes.

 The

nanoparticles are coated with hydrophilic oligonucleotides containing a Raman dye at one end and terminally capped with a small molecule recognition element (e.g. biotin). Moreover, this molecule is catalytically active and will be coated with silver in the solution of Ag(I) and hydroquinone. After the probe is attached to a small molecule or an antigen it is designed to detect, the substrate is exposed to silver and hydroquinone solution. A silver-plating is happening close to the Raman dye, which allows for dye signature detection with a standard Raman microscope. Apart from being able to recognise small molecules this probe can be modified to contain antibodies on the surface to recognise proteins. When tested in the protein array format against both

Contd.,

Commercial exploration
Some of the companies that are involved in the

development and commercialisation of nanomaterials in biological and medical applications Several companies exploit quantum size effects in semiconductor , nanocrystals for tagging biomolecules, or use bio-conjugated gold nanoparticles for labelling various cellular parts. A number of companies are applying nano-ceramic materials to tissue engineering and orthopaedics.

Nanosystem and its medical applications

Nanosystems
Nanosystem, a set of nanoscale components,

characterized by precise molecular order, working together to serve a set of purposes Complex nanosystems can be of macroscopic size.

Nanosystems for Simultaneous Imaging and Drug Delivery to T Cells

Most noninvasive tracking approaches for T cells in

the body aim for isolation of those cells; ex vivo labeling with molecular beacons such as bioluminescent markers, PET, or MR contrast agents; and then injection of labeled cells into the whole body and analysis of their trafficking dynamics and localization. This information is important for diagnostic assessment of the many disease states that involve aberrant T-cell migration to tissue sites. Dextran-coated superparamagnetic iron oxide and other iron oxidebased nanoparticulates have emerged as leading candidates for this approach because of their high magnetic susceptibility coupled with the versatility in their coating with ligands that facilitate binding to target cells.

Contd.,
This is a productive step forward in noninvasive

cellular imaging; however, for tracking pathogenic lymphocytes combined with the potential for therapeutic intervention, other targeted nanosystems considerable potential. These new capabilities may allow restoration of immune competence involving pathogenic lymphocytes and offer the potential for tracking the location and as well as localizing the drug depot to target T cells. Challenges in using these systems for routine monitoring and therapy include properly optimizing the surface to evade nonspecific endocytosis, understanding their clearance, and further evaluating their systemic toxicity and pharmacokinetics.

Multifunctional nanosystems for cancer theragnostics

Cancer remains one of the most challenging issues in modern

medicine. In developed countries, its mortality has remained relatively constant for the past 35 years Nanotechnology-based cancer treatments have received considerable attention in recent decades as they may provide unique approaches for early prediction, prevention, and diagnosis, as well as personalized therapy An exciting concept in nanomedicine is the fusion of therapy with diagnostics, known as theragnostics Designed to increase the efficiency and safety of treatment, the nanosystems used in theragnostics have multiple functions:

diagnosis, delivery of targeted therapy, and monitoring the therapeutic response

Contd.,
Because

of their combinatorial character, theragnostic nanoparticles require a suitable carrier able to encapsulate many active agents as well as accommodate appropriate surface functionalization to facilitate cellular recognition. Aside from being useful tools for studying the drug-delivery process and efficiency of chemotherapy at the cellular level, these nanosystems offer the possibility of identifying patients who do not respond to specific therapy, thus helping to move personalized medicine forward.

cosmetics
Water or hydro alcoholic nanosystems can be applied in the following cosmetic-hygienic preparations: Cosmetic and hygienic lotions; Foam-washing means: shampoos, liquid soaps, gels for bathe, etc.; Cosmetic creams; Cosmetic high filled systems: tooth-pastes, masks, scrubs, decorative cosmetics.

Nanomotors
The use of biomolecular motors offers an

interesting alternative to silicon based systems. A number of enzymes, such as kinesin, myosin and adenosine triphosphate (ATP) synthase act as nanosized linear or rotary biological motors. The integration of biomolecular motors with nanoscale engineered systems enables the development of hybrid organic-inorganic devices capable of using ATP as an energy source. This approach may enable the creation of a

Functionalized nano particle systemes:

The interactions of functionalized nanoparticles

with biomaterial structures can control the chemical reactivity of the biomolecules. Functionalized nanoparticles can operate as a biopromoter /bioinhibitor or a reporter, with some advantages over their molecularsized counterparts due to their unique photophysical and electronic properties. The solubility of nanoparticles in water can be greatly improved by functionalization of their surfaces with highly hydrophilic biomolecules.

Contd.,
Nanoparticles functionalized with groups that

provide site for the affinity binding of biomolecules have been used for the specific attachment of proteins and nucleotides. Nanoparticle-antibody conjugates were used for affinity binding of their respective antigens. Nanoparticles are already employed in several areas of drug delivery and cosmetics.

DRUG DELIVERY SYSTEM

Introduction:
Any drug delivery system may be defined as a system

comprising of:

Drug formulation Medical device or dosage form/technology to carry the

drug inside the body Mechanism for the release

The method by which a drug is delivered can have a

significant effect on its efficacy. Some drugs have an optimum concentration range within which maximum benefit is derived, and concentrations above or below this range can be toxic or produce no therapeutic benefit at all. On the other hand, the very slow progress in the efficacy of the treatment of severe diseases, has suggested a growing need for a multidisciplinary approach to the delivery of therapeutics to targets in tissues.

Drug delivery system concerns with

Cost Efficacy in patients Optimum drug delivery Bioavailability

Conventional drug delivery systems:

Conventional drug delivery involves the

formulation of the drug into a suitable form, such as a compressed tablet for oral administration or a solution for intravenous administration. These dosage forms have been found to have serious limitations in terms of higher dosage required, lower effectiveness, toxicity and adverse side effects. In traditional drug delivery systems such as oral ingestion or intravascular injection, the medication is distributed throughout the body through the systemic blood circulation. For most therapeutic agents, only a small portion of the medication reaches the organ to be affected.

Steps:
First, drug absorptionfrom the site of administration

permits entry of the therapeutic agent (either directly or indirectly) into plasma (input). Absorption is defined as the passage of a drug from its site of administration into the plasma. Second, the drug may then reversibly leave the blood stream and distribute into the interstitial and intracellular fluids (distribution). Third, the drug may be metabolized by the liver, kidney, or other tissues. Finally, the drug and its metabolites are eliminated from the body (output) in urine, bile, or feces.

Routes of drug administration

The route of administration is determined

primarily by the properties of the drug (such as water or lipid solubility, ionization, etc.) and by the therapeutic objectives (for example, the desirability of a rapid onset of action or the need for long-term administration or restriction to a local site). The route of administration (ROA) that is chosen may have a profound effect upon the speed and efficiency with which the drug acts.

The main routes of drug administration are:

Enteral:

oral, sublingual, recta Intravascular (intravenous), intramuscular, Subcutaneous inhalation, intranasal, intrathecal (in CSF), topical, transderma

Parenteral:

Other:

Enteral routes:
Drug placed directly in the GI tract: Oral- swallowing Sublingual- placed under the tongue Rectum- absorption through the rectum

Oral
Giving a drug by mouth is the most common

route of administration but it is also the most variable, and requires the most complicated pathway to the tissues. Little absorption occurs until the drug enters the small intestine. Drug absorption from the intestine: The drug absorbed by passive transport mechanism in intestine at a rate determined by the ionization and lipid solubility of the drug molecules. Strong bases of pKa10 or higher are poorly absorbed, as are strong acids of pKaless than 3, because they are fully ionized.

Advantages:
Convenient- can be self-administered, pain

free, easy to take Absorption- takes place along the whole length of the GI tract Cheap- compared to most other parenteral routes

Targeted drug delivery,

Targeted drug delivery, sometimes called smart drug delivery, It is a method of delivering medication to a patient in a

manner that increases the concentration of the medication in some parts of the body relative to others. The goal of a targeted drug delivery system is to prolong, localize, target and have a protected drug interaction with the diseased tissue. The conventional drug delivery system is the absorption of the drug across a biological membrane, whereas the targeted release system is when the drug is released in a dosage form. The advantages to the targeted release system is

The reduction in the frequency of the dosages taken by the patient, Having a more uniform effect of the drug, Reduction of drug side effects, and Reduced fluctuation in circulating drug levels.

The disadvantage of the system is high cost which makes

productivity more difficult and the reduced ability to adjust the dosages.

Contd.,
Targeted drug delivery systems have been

developed to optimize regenerative techniques. The system is based on a method that delivers a certain amount of a therapeutic agent for a prolonged period of time to a targeted diseased area within the body. This helps maintain the required plasma and tissue drug levels in the body. Therefore, avoiding any damage to the healthy tissue via the drug. The drug delivery system is highly integrated and requires various disciplines, such as chemists, biologist and engineers, to join forces to optimize this system

Types
There are two kinds of targeted drug delivery, Active targeted drug delivery, such as

some antibody medications; and Passive targeted drug delivery, such as the enhanced permeability and retention effect

Delivery vehicles
There are different types of drug delivery vehicles,

such as,

polymeric micelles, liposomes, lipoprotein-based drug carriers, nano-particle drug carriers, dendrimers etc. non-toxic, biocompatible, non-immunogenic and biodegradable

An ideal drug delivery vehicle must be

Liposomes
The most common vehicle currently used for

targeted drug delivery is the liposome. Liposomes are non-toxic, non-hemolytic and nonimmunogenic even upon repeated injections; They are biocompatible and biodegradable and can be designed to avoid clearance mechanisms (reticuloendothelial system (RES), renal clearance, chemical or enzymatic inactivation, etc.) Lipid based, ligand coated nanocarriers can store their payload in the hydrophobic shell or the hydrophilic interior depending on the nature of the drug/contrast agent being carried

Advantages
The advantages of the liposomes are : Liposomes can be metabolisedin vivo since they are made of phosphollipids. They are non toxic and non antigenic because they are made of phospholipids. Liposomes allow the sustained delivery of drugs with low therapeutic index by altering their pharmacokinetic pattern. They facilitate site specific drug uptake and delivery. Liposomes can protect encapsulated drugs from degradation. They can increase the convenience of therapy by reducing frequency of drug administration. They can alter tissue distribution of drugs in a therapeutically favourable way. Lioposomes show compatibility with both lipophilic and hydrophilic drugs. Liposomes surfaces can be modified to provide

Disadvantages
The disadvantages of liposomes are : Their liquid crystal structure hinders a true membrane transfer and causes physical instability. Their loading capacity is quite weak. Allergic reactions are possible. Poor encapsulation of drugs could be a limitation.

Disease categories for Liposomal Drug Delivery

Tuberculosis Leprosy (bacterial) Malaria (protozoal) Hepatitis Diabetes Arthritis Metal Detoxification

Micelles and dendrimers

Another type of drug delivery vehicle used is

polymeric micelles. They are prepared from certain amphiphilic copolymers consisting of both hydrophilic and hydrophobic monomer units. They can be used to carry drugs that have poor solubility. Techniques have been developed that utilize reactive polymers along with a hydrophobic additive to produce a larger micelle which create a range of sizes. Dendrimers are also polymer-based delivery vehicles. They have a core that branches out in regular intervals to form a small, spherical and very

Biodegradable particles
Biodegradable particles have the ability to

target diseased tissue as well as deliver their payload as a controlled release therapy. Biodegradable particles bearing ligands to Pselectin, endothelial selectin (E-selectin) and ICAM-1 have been found to adhere to inflamed endothelium. Therefore the use of biodegradable particles can be also be used for cardiac tissue

Artificial DNA nanostructures

The success of DNA nanotechnology in constructing

artificially designed nanostructures out of nucleic acids such as DNA, combined with the demonstration of systems for DNA computing, has led to speculation that artificial nucleic acid nanodevices can be used to target drug delivery based upon directly sensing its environment. These methods make use of DNA solely as a structural material and a chemical, and do not make use of its biological role as the carrier of genetic information. Nucleic acid logic circuits have been demonstrated that could potentially be used as the core a system which releases a drug only in response to a stimulus such as a specific mRNA. Additionally, a DNA "box" with a controllable lid has been synthesized using the DNA origami method. This structure could encapsulate a drug in its close state, and open to release it only in response to a desired stimulus.

Monoclonal antibodies
Monoclonal antibodies are artificially produced

proteins which exhibit specificity for one single antigen. The inherent specificity of the monoclonal antibodies for antigens provides the rationale for their use in drug targeting for therapeutic applications. The purpose is to destroy diseased tissues while leaving healthy tissues unharmed, thereby also reducing side effects of drugs.

Contd.,
Two aspects of monoclonal antibodies need to

be understood for successful development of therapeutic products. One is the possible susceptibility of their binding properties to even minor modifications in the environment. The second involves the kinetics of antibody binding. It may be necessary, for example, that the antigenic determinant recognised by a given antibody is in a particular conformation before binding can take place.

Contd.,
The major application for which monoclonals

are being studied for therapeutic use is cancer. Much of the work relates to the conjugation of different chemotherapeutic agents with antibodies and studying their effects in various tumor models. Clinical trials are being carried out for various products catering to different diseases like melanoma,, sepsis, graft-vsheart disease ovarian and colorectal cancer as also breast and lung cancer.

Nanoparticulate systems
Nanoparticles are colloidal particulate systems in the

sub-micron size range acting as carriers of drug molecules. These carriers are solid spheres and their surface is amorphous and lipophilic with a negative charge. Depending on the manufacturing procedure the size varies between 10 nm to 1000 nm. The porosity varies between 3 nm and 6 nm and the wall thickness in the case of nano capsules could be between 15 nm and 60 nm. The drug is dissolved, entrapped and/or adsorbed to the macromolecular material. However, it is possible to alter the distribution of nanoparticles in the body either by coating them with serum components or by attaching antibodies to their surfaces. Alternatively, magnetic nanoparticles could be used to enhance site specificity.

Nanomedical capsule in cancer treatment

Advantages:
Nanoparticulate systems provide a number of benefits which are : They control both the site and rate of drug delivery Adverse effects and toxic reactions are minimised They enhance the therapeutic efficacy of the drug The particles are non toxic and bio-degradable Reproducibility is quite easily achieved They are quite stable and less costly than other

Limitations:
Nanoparticulate delivery systems suffer from

their targetability is limited to the liver, the spleen and to a small extent, the bone marrow.

Medical applications
Medical applications using nanoparticles are : Treatment of infections of the Reticulo Endothelial system. Enzyme replacement therapy in the liver. Treatment of Cancer. Vaccination.

Applications of targeted drug delivery:

Targeted drug delivery can be used to treat

many diseases, such as the cardiovascular diseases and diabetes. However, the most important application of targeted drug delivery is to treat cancerous tumors.

Nanoparticles synthesis
Nanoparticles synthesis is done by two major

routes, which include classical synthesis and green synthesis. The green synthesis techniques generally utilize relatively non-toxic chemicals non-toxic solvents, biological extracts and systems. Biological methods are considered safe and ecologically sound for the nanomaterial fabrication as an alternative to conventional physical and chemical methods.

Biological synthesis of Nanoparticles

Currently there are several methods for the production

of Nanoparticles like Chemical and pysical methods. But there are evidences for the harmfulness of these nanoparticles to the environment. Now there is a need to grow more and more ecofriendly nanoparticles that are best suited to environment. Recently it has been shown that microbes and plants can synthesize nanoparticles in them. Moreover shape, size and nature can be controlled by these biological approaches through just modifying pH, temperature and nutrient media composition. Here we are utilizing general metabolic properties of cells and used to develop nanoparticles. This is an ecofriendly approach and with lots of benefits to human kind.

Nanoparticles
Gold, silver, and copper have been used mostly

for the synthesis of stable dispersions of nanoparticles, which are useful in areas of photography, catalysis, biological labeling, photonics, optoelectronics and surfaceenhanced Raman scattering (SERS) detection.

Biological route
Biological routes to the synthesis of these

particles have been proposed by exploiting microorganisms and by vascular plants. The functions of these materials depend on their composition and structure. Plants have been reported to be used for synthesis of metal nanoparticles of gold and silver and of a gold-silver-copper alloy. Of this colloidal silver is of particular interest because of its distinctive properties such as good conductivity, chemical stability, and catalytic and antibacterial activity

Biomolecules
Researchers report that biomolecules like

protein, phenols and flavonoids not only play a role in reducing the ions to the nanosize, but also play an important role in the capping of the nanoparticles. The reduction of Ag+ ions by combinations of biomolecules found in these extracts such as vitamins, enzymes/proteins, organic acids such as citrates, amino acids, and polysaccharides is environmentally benign, yet chemically complex.

Growing nanomaterials in plants

the term "green nanotechnology In a broad sense, this term includes a wide range of possible applications,

from nanotechnology-enabled, environmentally friendly manufacturing processes that reduce waste products (ultimately leading to atomically precise molecular manufacturing with zero waste); the use of nanomaterials as catalysts for greater efficiency in current manufacturing processes by minimizing or eliminating the use of toxic materials (green chemistry principles); the use of nanomaterials and nanodevices to reduce pollution (e.g. water and air filters); and the use of nanomaterials for more efficient alternative energy production (e.g. solar and fuel cells). Unfortunately, there is a flip side to these benefits. As scientists experiment with the development of new chemical or physical methods to produce nanomaterials, the concern for a negative impact on the environment is also heightened: some of the chemical procedures involved in the synthesis of nanomaterials use toxic solvents, could potentially generate hazardous byproducts, and often involve high energy consumption (not to mention the unsolved issue of the potential toxicity of certain nanomaterials). This is leading to a growing awareness of the need to develop clean, nontoxic and environmentally friendly procedures for synthesis and assembly of nanoparticles. Scientists are now exploring the use of biological organisms to literally grow nanomaterials.

In nature, numerous inorganic materials are synthesized

by living organisms. These bioinorganic materials can be extremely complex both in structure and function, and also exhibit exquisite hierarchical ordering from the nanometer to macroscopic length scales which has not even remotely been achieved in laboratory-based syntheses. Inorganic materials in the form of hard tissues are an integral part of most multicellular biological systems. Hard tissues (e.g. bone or nacre) are generally biocomposites containing structural biomacromolecules and some 60 different kinds of minerals that perform a variety of vital structural, mechanical, and physiological function

Bacteria:
Imitating nature and performing biosynthesis of nanoparticles

has been explored by scientists for several years now. Initially this was explored for the possibility of using live bacteria and yeast for the remediation of metalcontaminated waters. It has been shown that many bacteria and plants can actively uptake and bioreduce metal ions from soils and solutions during the detoxification process, thereby forming insoluble complexes with the metal ion in the form of nanoparticles. A well-known example of bioreduction and nanoparticle production is the magnetostatic bacteria that can synthesize magnetic nanoparticles ("Nanostructured magnetism in living systems"). Another early example of bacterial biosynthesis methods is the growth of metallic nanoparticles in bacterial cells ("Biologically Produced Silver-Carbon Composite Materials for Optically Functional Thin-Film Coatings").

Contd.,
Some well-known examples of microbial systems

synthesizing inorganic materials include magnetotactic bacteria for magnetite nanoparticles S-layer bacteria for gypsum and calcium carbonate layers silver mine- inhabiting Pseudomonas sp. that reduces silver ions to form silver nanoparticles Nanocrystals of gold, silver and their alloys have been synthesized within the cells of lactic acid bacteria Fungus and actinomycete species were reported to synthesize silver or gold nanoparticles of different shapes and sizes , but use of green plants for similar nanoparticle biosynthesis methodologies is an exciting possibility that is largely

Plants
The first report of live plants synthesizing

nanoparticles appeared in 2002 when it was shown that gold nanoparticles, ranging in size from 2 to 20 nm, could form inside alfalfa seedlings ("Formation and Growth of Au Nanoparticles inside Live Alfalfa Plants"). Subsequently it was shown that alfalfa also could form silver nanoparticles when exposed to a silver rich solid medium ("Alfalfa Sprouts: A Natural Source for the Synthesis of Silver Nanoparticles").

Contd.,
There are also cases of synthesizing metallic

nanoparticles from dead biomass. A recent example is the fabrication of silver nanoparticles ranging from 55 to 80 nm in size, and triangular or spherical shaped gold nanoparticles, from the sundried biomass of Cinnamomum camphora leaf (commonly know as camphor tree) with aqueous silver or gold precursors at ambient temperature ("Biosynthesis of silver and gold nanoparticles by novel sundried Cinnamomum camphora leaf").

 The first report of plants synthesizing gold or silver

nanoparticles appeared when alfalfa seedlings were shown to uptake gold or silver from metals-enriched nutrient media . These studies demonstrated that Au(III) or Ag(I) ions were reduced in the solid media to Au(0) or Ag(0) by alfalfa plants, and then the metal atoms were absorbed into the plant, where growth of nanoparticles took place. Another dimension was added to the green chemistry approach for pure metal synthesis with the use of plant broths . Shankar et al. used lemongrass and geranium plant extracts to induce the formation of gold nanoparticles or structures when reacted with aqueous chloroauric acid . Here we demonstrate uptake of high amounts of gold(III) ions by a leguminous shrub, Sesbania drummondii, with subsequent reduction of Au(III) ions to Au(0) inside plant cells or tissues. What makes our report different from other studies is the intracellular formation and growth of spherical gold nanoparticles (620 nm) and their in situ catalytic function. The nanoparticle-bearing biomatrix of Sesbania has the ability to reduce a hazardous and toxic pollutant, aqueous

 The optoelectronic and physicochemical properties of

nanoscale materials are a strong function of particle size. Nanoparticle shape also contributes significantly to modulating their electronic properties . Therefore, producing biologically-inspired nanostructures of a desirable size and morphology is critical to their commercial applicability. The shape and size of biogenic nanoparticles depends on the biological species involved, for example, geranium leaf broth reacted with aqueous chloroaurate ions induced a variety of gold nanoparticle shapes that included rods, flat sheets and triangles, while its endophytic fungus (Colletotrichum sp.) produced essentially spherical nanoparticles under the same conditions . Yet another plant, lemongrass (leaf broth) induced the formation of a high percentage of single-crystalline gold nanotriangles under similar conditions . Even the parts of a plant species affect the nature of nanoparticle synthesis. The root broth prepared from geranium exhibited a propensity towards forming flat nanostructures in comparison with the spherical nanoparticles that formed with geranium stem broth .

Contd.,
Scientists are already working on finding out what exactly the

biochemical processes are that lead to nanoparticle formation in plants; if and how nanoparticle shape modulation and size control can be managed; what range of chemical composition of nanoparticles can be biosynthesized (so far it is mostly confined to metals, but the synthesis of oxides, nitrides and carbides etc. is also of great interest); and whether plant extracts can be used in nanobiosynthesis. The big question of course is whether these methods could be used for large scale fabrication of nanomaterials. At this point there just is not enough knowledge to answer the question if biosynthesis techniques will ever be able to compete with existing physical and chemical synthesis processes. But once scientists understand the processes involved in biosynthesis, and with some genetic engineering, who knows, maybe one day we will see nanomaterial production in microbe farms and truly green nanotechnology vegetable

Mechanism
The mechanism for the reduction of Ag ions to silver

could be due to the presence of water-soluble antioxidative substances like ascorbate. This acid is present at high levels in all parts of plants. Ascorbic acid is a reducing agent and can reduce, and thereby neutralize, reactive oxygen species leading to the formation of ascorbate radical and an electron. This free electron reduces the Ag+ ion to Ag0. It has been reported that ionic silver strongly interacts with thiol group of vital enzymes and inactivates them. Experimental evidence suggests that DNA loses its replication ability once the bacteria have been treated with silver ions. The antibacterial effect of nanoparticles can be attributed to their stability in the medium as a colloid, which modulates the phosphotyrosine profile of the bacterial proteins and

Synthesis from herb

Indian researchers at Patna University have

biosynthesised silver nanoparticles from Desmodium triflorum. Desmodium triflorum is a wild much branched slender diffused herb with trifoliate leaves occurring as small under herb found in grasslands, fields, and agricultural lands forming a green turf on the ground. The dry plant was powdered, added with distilled water, heated and the extract was added to AgNO3 solutions.

Contd.,
The bioreduction of Ag+ ions took place . The solution containing the signatory color of AgNPs

(dark brown) was dryed in oven to get powders of silver nanoparticles. Thus stable and spherically shaped nanoparticles of average size ~10nm were synthesized using desmodium plant. The green synthesis of AgNPs fulfills all the three main steps, which must be evaluated based on green chemistry perspectives, including selection of solvent medium, selection of environmentally benign reducing agent and selection of nontoxic substances for the AgNPs stability. The study further showed that Ag nanoparticles presented good antibacterial performance against common pathogens. The nanoparticles when combined with the antibiotics show synergic effect in

Biosynthesis of Crystalline Silver and Gold Nanoparticles by Extremophilic Yeasts

It has been demonstrated that extremophilic yeasts, isolated from

acid mine drainage in Portugal, are able to grow in presence of Ag and Au ions and can be exploited to produce nanocrystalline silver and gold particles. By using microorganisms for NP synthesis, advantage of a natural detoxification bioprocess can be withdraw which can be considered as an eco-friendly contrary to the classic AuNP synthesis methods that require a myriad of expensive and toxic reducing, stabilizing, and functionalizing chemicals. The resulting Ag and Au nanoparticles displayed controllable structural and optical properties as demonstrated by UV-vis spectroscopic studies, TEM observations, and XRD data. The results indicate that the Ag and Au nanoparticles formation could take place intracellularly and/or extracellularly. The data suggests that the cell wall plays an important role for the biosynthesis of both metal nanoparticles whereas the compounds released by the yeasts were only able to reduce the silver ions. A gNPs displayed an average diameter of 20nm whereas the size range of the AuNPs was ca. 20 to 100nm. The nanoparticles were well dispersed, indicating that they are capped by stabilizing agents, most probably by proteins.

Green synthesis of lead sulfide nanoparticles by the lead resistant marine yeast, Rhodosporidium diobovatum
The intracellular synthesis of stable lead sulfide

nanoparticles by a marine yeast, Rhodosporidium diobovatum the PbS nanoparticles were characterized by UVvisible absorption spectroscopy, X-ray diffraction (XRD) and energy dispersive atomic spectroscopy (EDAX). UV-visible absorption scan revealed a peak at 320 nm, a characteristic of the nanosize range. XRD confirmed the presence of PbS nanoparticles of cubic structure. Crystallite size as determined from transmission electron microscopy was found to be in the range