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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. From this, new ideas on controlling the pharmacokinetics, pharmacodynamics, non-specific toxicity, immunogenicity, biorecognition, and efficacy of drugs were generated. These new strategies, often called drug delivery systems (DDS), are based on interdisciplinary approaches that combine polymer science, pharmaceutics, bioconjugate chemistry, and molecular biology. To minimize drug degradation and loss, to prevent harmful side-effects and to increase drug bioavailability and the fraction of the drug accumulated in the required zone, various drug delivery and drug targeting systems are currently under development. Among drug carriers one can name soluble polymers, microparticles made of insoluble or biodegradable natural and synthetic polymers, microcapsules, cells, cell ghosts, lipoproteins, liposomes, and micelles. The carriers can be made slowly degradable, stimuli-reactive (e.g., pH- or temperature-sensitive), and even targeted (e.g., by conjugating them with specific antibodies against certain characteristic components of the area of interest). Targeting is the ability to direct the drug-loaded system to the site of interest. Two major mechanisms can be distinguished for addressing the desired sites for drug release: (i) passive and (ii) active targeting. An example of passive targeting is the preferential accumulation of chemotherapeutic agents in solid tumors as a result of the enhanced vascular permeability of tumor tissues compared with healthy tissue. A strategy that could allow active targeting involves the surface functionalization of drug carriers with ligands that are selectively recognized by receptors on the surface of the cells of interest. Since ligandreceptor interactions can be highly selective, this could allow a more precise targeting of the site of interest. Controlled drug release and subsequent biodegradation are important for developing successful formulations. Potential release mechanisms involve: (i) desorption of surface-bound
/adsorbed drugs; (ii) diffusion through the carrier matrix; (iii) diffusion (in the case of nanocapsules) through the carrier wall; (iv) carrier matrix erosion; and (v) a combined erosion /diffusion process. The mode of delivery can be the difference between a drugs success and failure, as the choice of a drug is often influenced by the way the medicine is administered. Sustained (or continuous) release of a drug involves polymers that release the drug at a controlled rate due to diffusion out of the polymer or by degradation of the polymer over time. Palatial release is often the preferred method of drug delivery, as it closely mimics the way by which the body naturally produces hormones such as insulin. It is achieved by using drugcarrying polymers that respond to specific stimuli (e.g., exposure to light, changes in pH or temperature).
For over 20 years, researchers have appreciated the potential benefits of nanotechnology in providing vast improvements in drug delivery and drug targeting. Improving delivery techniques that minimize toxicity and improve efficacy offers great potential benefits to patients, and opens up new markets for pharmaceutical and drug delivery companies. Other approaches to drug delivery are focused on crossing particular physical barriers, such as the blood brain barrier, in order to better target the drug and improve its effectiveness; or on finding alternative and acceptable routes for the delivery of protein drugs other than via the gastro-intestinal tract, where degradation can occur.
Abstract:
The objective is to construct an apparatus to deliver a constant supply of protein containing drugs, including insulin, to the patients body. This device will be designed as small, disposable, and inexpensive form. Current devices for protein drug delivery are too costly and only deliver drugs at variable rates defined by computer systems. By delivering insulin at constant rate, the blood sugar level should not exhibit the fluctuations associated with the current methods. The theory behind this product is revolutionary, and similar devices do not currently exist on the market. Eventually this device would be used by individuals requiring protein drug treatments on a daily basis. The clients research specialty deals with pediatric diabetes, to which this device
could be applied. The ultimate goal of this project is to advance protein drug deliver technology and produce a more user-friendly, cost efficient apparatus.
Figure 1. Pharmaceutical carriers Micelles formed by self-assembly of amphiphilic block copolymers (5-50 nm) in aqueous solutions are of great interest for drug delivery applications. The drugs can be physically entrapped in the core of block copolymer micelles and transported at concentrations that can exceed their intrinsic water- solubility. Moreover, the hydrophilic blocks can form hydrogen bonds with the aqueous surroundings and form a tight shell around the micellar core. As a result, the contents of the hydrophobic core are effectively protected against hydrolysis and enzymatic degradation. In addition, the corona may prevent recognition by the reticuloendothelial system and therefore preliminary elimination of the micelles from the bloodstream. A final feature that makes amphiphilic block copolymers attractive for drug delivery applications is the fact that
their chemical composition, total molecular weight and block length ratios can be easily changed, which allows control of the size and morphology of the micelles. Functionalization of block copolymers with crosslinkable groups can increase the stability of the corresponding micelles and improve their temporal control. Substitution of block copolymer micelles with specific ligands is a very promising strategy to a broader range of sites of activity with a much higher selectivity.
Figure 2. Block copolymer micelles. Liposomes are a form of vesicles that consist either of many, few or just one phospholipids bilayers. The polar character of the liposomal core enables polar drug molecules to be encapsulated. Amphiphilic and lipophilic molecules are solubilized within the phospholipids bilayer according to their affinity towards the phospholipids. Participation of nonionic surfactants instead of phospholipids in the bilayer formation results in niosomes. Channel proteins can be incorporated without loss of their activity within the hydrophobic domain of vesicle membranes, acting as a size-selective filter, only allowing passive diffusion of small solutes such as ions, nutrients and antibiotics. Thus, drugs that are encapsulated in a nanocagefunctionalized with channel proteins are effectively protected from premature degradation by proteolytic enzymes. The drug molecule, however, is able to diffuse through the channel, driven by the concentration difference between the interior and the exterior of the nanocage.
Figure 4. A polymer-stabilized nanoreactor with the encapsulated enzyme. Dendrimers are nanometer-sized, highly branched and monodisperse macromolecules with symmetrical architecture. They consist of a central core, branching units and terminal functional groups. The core together with the internal units, determine the environment of the nanocavities
and consequently their solubilizing properties, whereas the external groups the solubility and chemical behaviour of these polymers. Targeting effectiveness is affected by attaching targeting ligands at the external surface of dendrimers, while their stability and protection from the Mononuclear Phagocyte System (MPS) is being achieved by functionalization of the dendrites with polyethylene glycol chains (PEG). Liquid Crystals combine the properties of both liquid and solid states. They can be made to form different geometries, with alternative polar and non-polar layers (i.e., a lamellar phase) where aqueous drug solutions can be included. Nanoparticles (including nanospheres and nanocapsules of size 10-200 nm) are in the solid state and are either amorphous or crystalline. They are able to adsorb and/or encapsulate a drug, thus protecting it against chemical and enzymatic degradation. Nanocapsules are vesicular systems in which the drug is confined to a cavity surrounded by a unique polymer membrane, while nanospheres are matrix systems in which the drug is physically and uniformly dispersed. Nanoparticles as drug carriers can be formed from both biodegradable polymers and nonbiodegradable polymers. In recent years, biodegradable polymeric nanoparticles have attracted considerable attention as potential drug delivery devices in view of their applications in the controlled release of drugs, in targeting particular organs / tissues, as carriers of DNA in gene therapy, and in their ability to deliver proteins, peptides and genes through the peroral route. Hydrogels are three-dimensional, hydrophilic, polymeric networks capable of imbibing large amounts of water or biological fluids. The networks are composed of homopolymers or copolymers, and are insoluble due to the presence of chemical crosslinks (tie-points, junctions), or physical crosslinks, such as entanglements or crystallites. Hydrogels exhibit a thermodynamic compatibility with water, which allows them to swell in aqueous media. They are used to regulate drug release in reservoir-based, controlled release systems or as carriers in swellable and swelling-controlled release devices. On the forefront of controlled drug delivery, hydrogels as enviro-intelligent and stimuli-sensitive gel systems modulate release in response to pH, temperature, ionic strength, electric field, or specific analyte concentration differences. In these systems, release can be designed to occur within specific areas of the body (e.g., within a certain pH of the digestive tract) or also via specific sites (adhesive or cell-receptor specific gels via
tethered chains from the hydrogel surface). Hydrogels as drug delivery systems can be very promising materials if combined with the technique of molecular imprinting.
Figure 5. Pegylated and pH sensitive micro- or nanogels. The molecular imprinting technology has an enormous potential for creating satisfactory drug dosage forms. Molecular imprinting involves forming a pre-polymerization complex between the template molecule and functional monomers or functional oligomers (or polymers) with specific chemical structures designed to interact with the template either by covalent, noncovalent chemistry (self-assembly) or both. Once the pre-polymerization complex is formed, the polymerization reaction occurs in the presence of a cross-linking monomer and an appropriate
solvent, which controls the overall polymer morphology and macro porous structure. Once the template is removed, the product is a heteropolymer matrix with specific recognition elements for the template molecule. Examples of MIP-based drug delivery systems involve: (i) rate-programmed drug delivery, where drug diffusion from the system has to follow a specific rate profile, (ii) activationmodulated drug delivery, where the release is activated by some physical, chemical or biochemical processes and (iii) feedback-regulated drug delivery, where the rate of drug release is regulated by the concentration of a triggering agent, such as a biochemical substance, the concentration of which is dependent on the drug concentration in the body. Despite the already developed interesting applications of MIPs, the incorporation of the molecular imprinting approach for the development of DDS is just at its incipient stage. Nevertheless, it can be foreseen that, in the next few years, significant progress will occur in this field, taking advantage of the improvements of this technology in other areas. Among the evolution lines that should contribute more to enhance the applicability of imprinting for drug delivery, the application of predictive tools for a rational design of imprinted systems and the development of molecular imprinting in water may be highlighted
Figure 6. The volume phase transition of the hydrogel -induced by an external stimuli (e.g., a change in pH, temperature or electrical field) modifies the relative distance of the functional groups inside the imprinted cavities. This alters their affinity for the template.
Figure 7. (A) Induced Swelling - As analyte (A) binds, the enzymatic reaction (E denotes covalently attached enzyme) produces a local pH decrease. For the cationic hydrogel, which is weakly basic, the result is ionization, swelling, and release of drug, peptide, or protein (filled circle). When A decreases in the bulk concentration, the gel shrinks. (B) Loss of Effective Crosslinks - Analyte competes for binding positions with the protein (P). As free analyte binds to the protein, effective cross-links are reversibly lost and release occurs. Conjugation of biological (peptides/proteins) and synthetic polymers is an efficient means to improve control over nanoscale structure formation of synthetic polymeric materials that can be used as drug delivery systems. Conjugation of suitable biocompatible polymers to bioactive peptides or proteins can reduce toxicity, prevent immunogenic or antigenic side reactions, enhance blood circulation times and improve solubility. Modification of synthetic polymers or polymer therapeutics with suitable oligopeptide sequences, on the other hand, can prevent random distribution of drugs throughout a patients body and allow active targeting. Functionalization of synthetic polymers or polymer surfaces with peptide sequences derived from extracellular matrix proteins is an efficient way to mediate cell adhesion. The ability of cationic peptide sequences to complex and condense DNA and oligonucleotides offers prospects for the development of non-viral vectors for gene-delivery based on synthetic polymeric hybrid materials.
Figure 8. Bioconjugates. The field of in-situ forming implants has grown exponentially in recent years. Liquid formulations generating a (semi-)solid depot after subcutaneous injection, also designated as implants, are an attractive delivery system for parenteral application because, they are less invasive and painful compared to implants. Localized or systemic drug delivery can be achieved for prolonged periods of time, typically ranging from one to several months. Generally, parenteral depot systems could minimize side effects by achieving constant, infusion-like plasma-level time profiles, especially important for proteins with narrow therapeutic indices. From a manufacturing point of view, in-situ forming depot systems offer the advantage of being relatively simple to manufacture from polymers. Injectable in-situ forming implants are classified into four categories, according to their mechanism of depot formation: (i) thermoplastic pastes, (ii) in-situ cross-linked polymer systems, (iii) in-situ polymer precipitation, and (iv) thermally induced gelling systems. The ultimate goal in controlled release is the development of a microfabricated device with the ability to store and release multiple chemical substances on demand. Recent advances in microelectro-mechanical systems (MEMS) have provided a unique opportunity to fabricate miniature biomedical devices for a variety of applications ranging from implantable drug delivery systems to lab-on-a-chip devices. The controlled release microchip has the following advantages: (i) multiple chemicals in any form (e.g., solid, liquid or gel) can be stored inside and
released from the microchip, (ii) the release of chemicals is initiated by the disintegration of the barrier membrane via the application of an electric potential, (iii) a variety of highly potent drugs can potentially be delivered accurately and in a safe manner, (iv) complex release patterns (e.g., simultaneous constant and pulsatile release) can be achieved, (v) the microchip can be made small enough to make local chemical delivery possible thus achieving high concentrations of drug at the site where it is needed while keeping the systemic concentration of the drug at a low level and (vi) water penetration into the reservoirs is avoided by the barrier membrane and thus the stability of protein-based drugs with limited shelf-life is enhanced.
Administration Routes
The choice of a delivery route is driven by patient acceptability, the properties of the drug (such as its solubility), access to a disease location, or effectiveness in dealing with the specific disease. The most important drug delivery route is the peroral route. An increasing number of drugs are protein- and peptide-based. They offer the greatest potential for more effective therapeutics, but they do not easily cross mucosal surfaces and biological membranes; they are easily denatured or degraded, prone to rapid clearance in the liver and other body tissues and require precise dosing. At present, protein drugs are usually administered by injection, but this route is less pleasant and also poses problems of oscillating blood drug concentrations. So, despite the barriers to successful drug delivery that exist in the gastrointestinal tract (i.e., acidinduced hydrolysis in the stomach, enzymatic degradation throughout the gastrointestinal tract by several proteolytic enzymes, bacterial fermentation in the colon), the peroral route is still the most intensively investigated as it offers advantages of convenience and cheapness of administration, and potential manufacturing cost savings. Pulmonary delivery is also important and is effected in a variety of ways - via aerosols, metered dose inhaler systems (MDIs), powders (dry powder inhalers, DPIs) and solutions (nebulizers), all of which may contain nanostructures such as liposomes, micelles, nanoparticles and dendrimers. Aerosol products for pulmonary delivery comprise more than 30% of the global drug delivery market. Research into lung delivery is driven by the potential for successful protein and peptide drug delivery, and by the promise of an effective delivery mechanism for gene therapy (for example, in the treatment of cystic fibrosis), as well as the need to replace chlorofluorocarbon propellants in MDIs. Pulmonary drug delivery offers both local targeting for
the treatment of respiratory diseases and increasingly appears to be a viable option for the delivery of drugs systemically. However, the pulmonary delivery of proteins suffers by proteases in the lung, which reduce the overall bioavailability, and by the barrier between capillary blood and alveolar air (air-blood barrier). Transdermal drug delivery avoids problems such as gastrointestinal irritation, metabolism, variations in delivery rates and interference due to the presence of food. It is also suitable for unconscious patients. The technique is generally non-invasive and aesthetically acceptable, and can be used to provide local delivery over several days. Limitations include slow penetration rates, lack of dosage flexibility and / or precision, and a restriction to relatively low dosage drugs. Parenteral routes (intravenous, intramuscular, subcutaneous) are very important. The only nanosystems presently in the market (liposomes) are administered intravenously. Nanoscale drug carriers have a great potential for improving the delivery of drugs through nasal and sublingual routes, both of which avoid first-pass metabolism; and for difficult-access ocular, brain and intraarticular cavities. For example, it has been possible to deliver peptides and vaccines systemically, using the nasal route, thanks to the association of the active drug macromolecules with nanoparticles. In addition, there is the possibility of improving the occular bioavailability of drugs if administered in a colloidal drug carrier. Trans-tissue and local delivery systems require to be tightly fixed to resected tissues during surgery. The aim is to produce an elevated pharmacological effect, while minimizing systemic, administration-associated toxicity. Trans-tissue systems include: drug-loaded gelatinous gels, which are formed in-situ and adhere to resected tissues, releasing drugs, proteins or gene-encoding adenoviruses; antibody-fixed gelatinous gels (cytokine barrier) that form a barrier, which, on a target tissue could prevent the permeation of cytokines into that tissue; cellbased delivery, which involves a gene-transduced oral mucosal epithelial cell (OMEC)implanted sheet; device-directed delivery - a rechargeable drug infusion device that can be attached to the resected site Gene delivery is a challenging task in the treatment of genetic disorders. In the case of gene delivery, the plasmid DNA has to be introduced into the target cells, which should get
transcribed and the genetic information should ultimately be translated into the corresponding protein. To achieve this goal, a number of hurdles are to be overcome by the gene delivery system. Transfection is affected by: (a) targeting the delivery system to the target cell, (b) transport through the cell membrane, (c) uptake and degradation in the endolysosomes and (d) intracellular trafficking of plasmid DNA to the nucleus.
Functions (active drug targeting, on-command delivery, intelligent drug release devices/ bioresponsive triggered systems, self-regulated delivery systems, systems interacting with the body, smart delivery); Virus-like systems for intracellular delivery; Nanoparticles to improve devices such as implantable devices/nanochips for nanoparticle release, or multi reservoir drug delivery-chips; Nanoparticles for tissue engineering; e.g. for the delivery of cytokines to control cellular growth and differentiation, and stimulate regeneration; or for coating implants with nanoparticles in biodegradable polymer layers for sustained release; Advanced polymeric carriers for the delivery of therapeutic peptide/proteins (biopharmaceutics), And also in the development of: Combined therapy and medical imaging, for example, nanoparticles for diagnosis and manipulation during surgery (e.g. thermotherapy with magnetic particles); Universal formulation schemes that can be used as intravenous, intramuscular or per oral drugs Cell and gene targeting systems. User-friendly lab-on-a-chip devices for point-of-care and disease prevention and control at home. Devices for detecting changes in magnetic or physical properties after specific binding of ligands on paramagnetic nanoparticles that can correlate with the amount of legend. Better disease markers in terms of sensitivity and specificity
total length of time the insulin continues to lower blood-glucose levels effectively, and in Figure I, the duration would span from B to D
These times can also be affected by the time of day and actions of the patient. During meal times, blood-glucose levels jump, and additional units of insulin may be needed to achieve the desired peak time performance and stability. It is always important for the patient toonitor their blood-glucose levels by testing their blood to determine if a higher dose of insulin is needed. In someone without diabetes, the pancreas releases a steady flow rate of insulin and increases insulin during meal times and other times when insulin is required. The average person will need 5 to 10 units of insulin per day to meet these daily needs. Many people have very high bloodglucose levels in the morning due to hormones and too little insulin received at night because
they are asleep. A design that delivers a constant flow of insulin overnight would solve this problem. One of the best ways to inject insulin is by using an insulin pump. An insulin pump works by injecting a basal rate of insulin over a 24 hour period; this is a constant rate of insulin that is determined for the specific individual. Basal rates are necessary to keep glucose at a safe level between meals and overnight A bolus dosage is a higher amount of insulin and is used to maintain safe glucose levels after an individual eats a meal or a snack. A bolus is given by pressing a button on the insulin pump, or having the pump programmed to deliver the bolus ata certain time
Skin Layers
The largest organ and primary shield of the body is skin tissue. As a result there are any different layers to help filter outside elements (i.e. chemicals, bacteria, temperature) as they are absorbed or repelled by each of the various layers. The top layer of the skin is called epidermis. It does not have any blood vessels but instead receives oxygen and other nutrients from the tissue layers below. The basement membrane lies at the bottom of the epidermis and helps connect it with the next layer of skin, the dermis. Blood vessels, nerves, sweat glands, and hair follicles can be found here. Just below the dermis lies a layer of subcutaneous fat, which rests on the muscles and bones and is attached by connective tissues. The subcutaneous fat contains larger blood vessels and nerves and is mostly made up of clumps of fat-filled cells called adipose cells These times can also be affected by the time of day and actions of the patient. During meal times, blood-glucose levels jump, and additional units of insulin may be needed to achieve the desired peak time performance and stability. It is always important for the patient to monitor their blood-glucose levels by testing their blood to determine if a higher dose of insulin is needed. In someone without diabetes, the pancreas releases a steady flow rate of insulin and increases insulin during meal times and other times when insulin is required. The average person will need 5 to 10 units of insulin per day to meet these daily needs. Many people have very high blood-glucose levels in the morning due to hormones and too little insulin received at night because they are asleep. A design that delivers a constant flow of insulin overnight would solve this problem. One of the best ways to inject insulin is by using an insulin pump. An insulin pump works by injecting a basal rate of insulin over a 24 hour period; this is a constant rate of insulin that is
determined for the specific individual. Basal rates are necessary to keep glucose at a safe level between meals and overnight A bolus dosage is a higher amount of insulin and is used to maintain safe glucose levels after an individual eats a meal or a snack. A bolus is given by pressing a button on the insulin pump, or having the pump programmed to deliver the bolus at a certain time
Figure II:
The three-dimensional structure of the skin, shown in this skin diagram(left) the thick hairless skin of the palm of the hand, (right) the thin hairy skin of the forearm
The thickness of each layer depends on the person and location on the body. The epidermis is only 35 to 50 m thick in most areas of the body. The palms and soles of a person are usually much thicker, while the tissue around the eyes is thinner and, therefore, more sensitive to the outside elements. The depth of the subcutaneous fat differs from one person to the next
Protein drug injections into the skin should be made into the subcutaneous tissue at a 90angle. Thin individuals or children should use short needles to inject at a 45 angle to avoid
intramuscular injection . Intramuscular injection is more painful and has a faster absorption rate which should be avoided unless it is an emergency There are four main sites for an injection: the triceps region, the upper thighs, the buttocks, and the abdomen. These sites generally contain a high percentage of fatty tissue and thus are ideal for injection. The buttocks region absorbs the injection slowest, while the abdomen absorbs the fastest and at the most consistent speed Absorption can be amplified by increased blood circulation resulting from exercise or the temperature of the environment
Figure III: Injection Sites: Abdomen, Thighs, Back of upper arms, Andbuttocks
Injection site rotation is important in order to prevent abnormal cell growth and fat deposits that can develop when you inject in the same spot all the time These growths are not harmful, but they can limit the absorption of the injection. Site rotation can help prevent hypertrophy and lip atrophy. Hypertrophy is an overgrowth of cells under the skin and lip atrophy is the disappearance of fatty cells that leaves depressions in the skin surface
move when electric rrent is applied to them. NanoMuscles behave similar to human muscles; they contract when a current is applied to them and will stay contracted unless a tensile force is applied returning the actuator to the relaxed state [18]. They are very small and do not require a gear box like electric motors. NanoMuscles can be more precisely controlled than electric motors by controlling the magnitude and duration of the current applied to the SMA wire. NanoMuscles can act in a linear or otational motion and can produce force independent of speed, unlike electric motors. They are also virtually silent in operation
Figure IV: Left: A linear NanoMuscle shown with a paper clip for size comparison. Right: A linear NanoMuscle shown with the digital interface [18].
Metering valves are common components of various analytical chemistry systems including high pressure liquid chromatography applications. Californias Western Analytical Products produces biocompatible micro-metering valves capable of delivering 3 L/min. The P- 445,446, and 447 series are rated to 800 psi and connect via 1/16 tubing.
EXECUTIVE SUMMARY
1. Any drug delivery system may be defined as a system comprising of: a) drug formulation b) medical device or dosage form/technology to carry the drug inside the body c) mechanism for the release
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.
New drug delivery systems have been developed or are being developed t overcome the limitation of the conventional drug delivery systems to meet the need of the healthcare profession. These systems can be characterized as controlled drug release systems and targeted drug delivery systems.
2.
be based on two basic parameters : Route of entry (A) and Dosage form (B).
Such a definition implies that there are a vast number of members in this group. Many of them may not even be feasible, while many others may not be relevant. So, the set of most relevant new drug delivery systems is deduced as follows :
CLASSIFICATION OF DRUG DELIVERY SYSTEMS a) Transdermal Delivery Systems b) Carrier Based Delivery Systems
y y y y Liposomes Monoclonal Antibodies Nanoparticles Microspheres
Localize drug action by placing a rate controlled system near or at the desired tissue or organ.
Target drug action by using carriers or chemical derivatisation to deliver drug to a particular site.
3. Several approaches to the design oi controlled release systems have been developed. Majorapproaches include the following :
Several factors affect the successful design of controlled release products. These require careful ideation in the developmental process.
y y
In order to fulfill the two functions of controlled release technologies, viz., site specific targeted drug delivery and/or rate controlled drug delivery, the devices/products must meet two basic requirements :
(i) The device must hold the entire dosage of drug required for complete therapy; and (if) The device must be able to control the rate of drug release from the system.
Polymers play an important role in meeting both these requirements. They can be fabricated to act as reservoirs of the total amount of drug. Also their properties can be suitably modified to control the rate of release to the desired level.
For the regulatory approval of controlled release products, three critical issues in polymer formulation are involved. a) Non toxicity b) Bio-degradability; and c) Controlled release of drug. 4. Although new delivery systems have a number of therapeutic benefits, but they do have certain limitations such as : a) If systems fail, overdosing occurs due to doses dumping. b) The large physical size of the dosage unit poses problems in usage. c) Often sub optimum bio-availability is observed. d) Variability in drug levels is also observed.
5. A few controlled release systems have already been commercialized abroad. Their echnologies too are well established. These systems are : Transdermal Patches Implants Nasal Systems
6. Transdermal Drug Delivery Systems are laminated patches which adhere to the skin and permit absorption of drugs from the skin surface through its layers into the general blood circulation, at controlled rates, resulting in sustained blood levels. The benefits from transdermal drug delivery are : a) it avoids the risk and inconvenience of intravenous therapy. b) allows treatment to be discontinued if so desired. c) the dosing regimen is simplified and the system is convenient to use, leading to excellent patient compliance. d) lower dosages are sufficient, minimising side effects. e) avoids the variable metabolism and absorption associated with oral drug administration.
7.
Since, drug release from patches (Transdermal Drug Delivery System) is constant over very
long periods, the homoastasis of the body is affected and the possibility of some undesirable effects on the abrupt removal of transdermal patches cannot be ruled out. It causes side-effects like dryness of the mouth or drowsiness. Anticholinergics like scopolamine can also interfere with orientation, cognition and memory and may cause delirium, particularly in geriatric patients. Therefore, it should be used with caution. A washout period could be allowed in such cases before the next patch is applied.
8. Major Companies manufacturing transdermal patches are Ciba Geigy, Alza Corporation, earle. Schwartz Pharma, Boehringer Ingelheim andToaeiyo.The main products which have been ommercialised include Transderm- Scop, Nitro-Dur, Nitrodisc and Deposit for delivering copolamine,Transderm - Nitro for deliverying nitroglycerin as also Catarpress TTS for
delivering clonidine. 9 Implants are sterile polymeric devices of varied shapes containing one or
more medicaments for introduction into body tissues for release in a controlled manner. The advantages of these devices are :
a) Implants can provide uninterrupted treatment for prolonged time periods e.g. several years or even a lifetime. b) The treatment is often reversible since the device can be removed if any undesirable effects occur. c) Implants provide the dual advantages of location and rate control.
a) Surgical intervention required either for implantation/sub sequent removal. b) Possibility of infection at implant sites. c) The size of the device may pose acceptability problems. d) In the case of hydrogel implants, because of the high water content, low molecular weight drugs are able to diffuse out quickly, making sustained delivery difficult. e) The cross linking agents used may increase the frictional irritation of the implants.
11.
cardiovascular diseases and brain diseases. Major companies involved in the manufacture of these systems include Alza Corporation, Meditronic Inc. Becton Dickinson and Infusaid Inc. Variety of systems have so far been commercialised. Some of these are the Progestasert, Norplant, Today, Vaginal ring, and Dual release ring for contraception, the lacrisert for artificial tear therapy, the Alzet osmotic pump for various drugs and the Ocusert for glaucoma management 12. Conventional nasal formulations in the form of spray and drops have used delivery
systems like the rhinyle catheters, single dose pipettes, metered dose spray pumps (nonpressurised) and metered dose aerosol valve devices Out of these, the spray pumps and the aerosol valve devices lend themselves to controlled delivery of nasal formulations. The spray pump operates on the mechanical energy provided by the depression of the actuator, while the
aerosol valve device operates on the energy provided by the propellant in the system. Both systems are simple to use and provide multiple dosing facility.
The relevant criteria to assess which device is more suitable in a given case are : y y dose accuracy dose reproducibility for the same device, comparatively for different devices and from batch to batch for each device. y y y y tailing off or emptying characteristics. rapid priming of the dose especially for pump systems. absorption or adsorption of ingredients from the formulation into/ onto the device. leaching out of materials from the device into the formulation.
13. The advantages of Nasal Drug Delivery Systems are : y y y y y y y Ease of administration. Rapid absorption and onset of action. Bypass of presystemic clearance. It is a non invasive route. Chronic self administration is possible. Simple formulations are sufficient. The nasal mucosa offers a large surface area for absorption and a favourable metabolic environment. y y y y Bio-availability appears to be satisfactory. Reduced hospital out patient care. Accurate consistent dosing. Greater efficiency with smaller doses leading to lower manufacturing costs.
14. The disadvantages associated with Nasal Drug Delivery Systems are :
y y y Untoward immunogenic reactions may occur. Inadequate availability of toxicity data for penetration enhancers. Nasal pathology may adversely affect product effectiveness.
15.
A few nasal products have been commercialised. These include calitonin for metabolic
bone disease, desmopressin acetate for primary mocturnal enuresis. A few others are also developed for which approval is awaited e.g. Vitamin B12 for pernicious anemia, "Nasacort" for steroid delivery.
16. The oral route of administration for controlled release systems, has received a lot of attention. Most oral systems are either tablets or capsules, and the nature of the release mechanisms loyed involve either diffusion or dissolution.
The system usually consist of two parts comprising of an immediately released dose and a sustaining portion that contains several times the therapeutic dose for maintaining drug levels at desired levels.
17. Microencapsulation is the process of applying relatively thin reproducible coatings to small particles of solids or droplets of liquids and dispersions to produce microcapsules. The various considerations in selecting a micro encapsulation process are :
The route of administration of microcapsules primarily determines the procedure used for microencapsulation. For the oral route, a single coating could often suffice while the equirements for implantable microcapsules could be more stringent involving issues like the bio-degradability of the polymer implant and the antigenic and thrombogenic properties of the capsules.
The physico-chemical properties of the drug would be another important consideration in choosing a microencapsulation process. It may not be possible to encapsulate solids by the same process as liquids. Also the acidity or alkalinity of the drug may preclude the use of certain wall materials.
The physical and chemical properties of the wall materials need to be considered arefully before, during and after encapsulation. As an example, waxes or long chain fatty acids are soluble in many organic solvents. This property makes them good candidates for deposition about an aqueous drug core by spray drying. However, this very same property precludes their use in encapsulating organic liquids.
Microencapsulation provides a means for converting liquids to solids, of altering colloidal and surface properties, of providing environmental protection and of controlling the secharacteristics (or availability) of coated materials.
Uniqueness of microencapsulation is the smallness of coated particles and their subsequent use and adaptation to a wide variety of dosage forms and product applications
Because of the smallness of the particles, drug moieties can be widely distributed throughout the gastro-intestinal tract, thus potentially improving drug absorption.
y y y y y
The release characteristic of coated products are not always reproducible. The coating may often be discontinuous. No single technique can be adapted to all core materials. In the case of very sensitive Pharmaceuticals the shelf life would be inadequate. Economic limitation too may be a stumbling block in its application.
21. The technology for osmotic pumps exploits the tendency of a fluid to equalise the concentration of substances on both sides of a semipermeable membrane. The osmotic pump system is a variable release system in that it can be designed to deliver different drugs at different rates. Osmotic devices can be employed for two purposes :
y y
To be used as research tools for animal pharmacology and chemical studies. To be used as dosage forms for drug delivery. As drug delivery systems, for pharmaceutical applications they could be used as oral
22. The unique proprietary technology of osmotic drug delivery offers several benefits.
y y
The delivery rates provided are higher than diffusion controlled systems. The biological environmental influences like pH, gastro-intestinal motility, etc., on the system are minimised.
y y y y y y
The system functions without being significantly affected by drug properties. The delivery rates are predictable and programmable. These systems are satisfactory in vitro and in vivo release profiles. The side effects of drugs administered are minimised. Patient compliance is improved since the frequency of dosing is reduced. The device is tamper resistant
There appear to be no apparent disadvantages of these systems. These pumps are used widely in animal pharmacology studies to administer drugs locally to distant targets to study their effects. The study mainly involves optimisation of dosing frequency and rates as also the bio-availability of drugs. The versatility of these systems is evident from the fact that numerous
drugs including peptides, neurotransmitters and nerve growth factors have been studied using these pumps.
24.
to release drugs phenyl propanolamine and Vitamin C, respectively, both developed by Alza orporation.
25.
Ion Exchange Resin System involves preparation of drug charged resin and its drying to
form beads. In the environment of the Gastro-intestinal tract the drug molecule is exchanged for an appropriately charged ion and so the drug is released at a controlled rate.
y y y
Reduction in the total drug dosage used. Fluctuations in blood levels are reduced. Frequency of dosing can be reduced making the treatment simpler for the patient
Besides these, additional benefits from ion exchange delivery systems are
The release of drugs is governed by certain factors like pH and ion strength and these conditions are fixed in the GI tract, making the formulation of such systems much more similar.
A variety of ion exchange resins are available providing a wider choice to control resin characteristics like porosity and particle size.
27. The disadvantages associated with Ion Exchange Delivery System are :
y The possible risk of drug accumulation, should the balance between rate of release of drug and its excretion is not maintained. y It is not possible to interrupt the treatment, once the drug is administered. " Prolonged administration may disturb the ionic strength of the y gastro-intestinal fluids.
The major ion exchange resin system commercialised todate is the Pennkinetic System, which has been used to deliver a variety of antitussives.
28. Pellets in Capsule System is a relatively simple technology and has been developed to deliver controlled amounts of the drug. The main systems commercialised so far include SODAS (Naproxen and Verpanil), ASTRIX (aspirin), DORRYX (doxycycline) SRYC (Erythromycin) andELANTANLASO (Isosorbide mononitrate).
29.
enclosing an aqueous compartment. Liposomes resemble cell membranes in structure and composition.
In order that drugs be efficacious, they need to be administered in certain concentrations. However, dilution in blood, metabolism and uptake by healthy tissues can all occur, resulting in poor delivery at the diseased site in the body, on the one hand and toxicity effects on the other.
Liposomes provide the ideal means to tackle all these issues. Drugs incorporated in liposomes and administered to the body can be delivered at the desired site, in the needed concentrations without being toxic.
y y y
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 bioadhesion which can be used to enhance the residence time at a particular site.
Liposomes are basically stable structures since they represent the favourable ermodyiiamic
state of phospholipids in water. However there are many circumstances in which liposome instability can occur.
Liposomal stability during storage influences their stability and performance in vivo. Stability problems with liposomes arise on two accounts, viz..
The stability problems during processing arise mainly on account of the phospholipids used. These get degraded mainly by peroxidation and hydrolysis reactions. Stability relating to the drug need to be handled on a case to case basis. Since, the drug is generally less stable than the lipids, conditions which ensure drug integrity will also ensure lipid stability.
y y y y y y y y y y
b) Immune System
c) Use as Adjuvants
y y
Major companies developing liposomal products include the Liposome Company, Liposome Technology Inc., Squibb, Vestar Inc and Bristol Myers.
Several products undergoing clinical trials are doxorubicin, amphoterin B, gentamicin and daunorobicin.
y y
y y y y
Products for dry eye syndrome, glaucoma and allergies Albuterol for bronchodilation Products for preventing post surgical adhesions Immunomodulators including mirranryl tripeptide - phosphatidyl ethanolamine for cancer and viral infections.
34.
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.
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.
35. The major application for which monoclonals are being studied for therapeutic use is cancer. Muchofthe 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 arious products catering to different diseases like melanoma,, sepsis, graft-vsheart disease ovarian and colorectal cancer as also breast and lung cancer.
36. 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 ran 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. The preferred mode of administration is parenteral. However, it is possible to alter the distribution of anoparticles in the body either by coating them with serum components or by attaching ntibodies to their surfaces. Alternatively, magnetic nanoparticles could be used to enhance site specificity.
y y y y y y
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 colloidal systems.
Nanoparticulate delivery systems suffer from their targetability is limited to the liver, the spleen and to a small extent, the bone marrow. 38. Medical applications using nanoparticles are : y y y y Treatment of infections of the Reticulo Endothelial system. Enzyme replacement therapy in the liver. Treatment of Cancer. Vaccination.
39.
Microspheres are small solid particulate carriers containing dispersed drug particles either
in solution or crystalline form. The importance of Microspheres has been growing because of their use as carriers for drugs or other therapeutic agents. Microspheres are made from natural and synthetic polymers. Different materials have been used for microsphere systems like albumin, gelatin, starch, ethylcellulose and synthetic polymers such as polylactic acid polycyano acrylates and polyhydroxybutyrate.
Routes of administration are by injection, i.e. intravenous, intramascular and intraarticular or by the nasal route. They reach the target site by some suitable passive mechanism or by direct administration into the relevant body compartment. The size of the microspheres can range from tens of nanometers to one hundred microns or more smaller particles below 500 nm are often termed nanoparticles or nanospheres.
The physical characteristics of these systems differ and depend on their applications. The feasibility of a microsphere system of drug delivery is determined largely by :
y y
the physico-chemical characteristics and dose of the drug used; and the required release pattern of the drug.
These aspects need to be considered carefully early in the development of an application. 43. Research and Development World
Major thrust areas for research & development in the world, the various drug delivery systems are :
a) Liposomes Drug liposome formulations have been investigated for a variety of applications, including the treatment of cancer and infectious diseases, vaccines for influenza and AIDS, peptide delivery,
ophthalmic, cardiovascular, and nonsteroidal anti-inflammatory drugs. Doxorubicin, mphotericin B and Gentamicin are the most widely studied drugs. Ampicillin, eisomycin, Streptomycin and Penicillin have also been studied and increased therapeutic index obtained
In order to broaden the range of potential applications and to increase the duration of liposomal formulations in the bloodstream, a technology is being developed to coat liposomes with carbohydrate moieties and target them.
and targeting liposomes to migratory cell populations is also being explored. y Studies to assess the potential of neosomes and virosomes as drug carriers are under way.
b) Transdermal Systems
Research efforts are on to increase the permeability of drugs by this route, using several techniques. Electrically induced transport or iontophoresis is being tried in dental and physical therapy applications. One project involves the use of square, triangular or sinusoidal waveforms of a periodic DC current. Electro osmosis is another technique which uses less current than iontophoresis. This is being tried for drug with almost no ionic strength and with low viscosity. For better permeation of high molecular weight drugs, an ultrasound technique is being investigated.
Multichannel devices e.g. the omniflow (Abbott Laboratories), a four channel pump is being developed to take care of independent and simultaneous delivery of several infusions. Efforts to provide versatility in programmable devices, e.g. Open loop systems in which drug delivery will be based on precise biological or pharmaco-kinetic parameters. An external source
could be the doctor himself or a computer programme and an element is provided which would calculate the dose required. Closed loop systems are being developed for automatic drug dosing upon feedback from biosensors. The biosensors have not yet been developed adequately. An area for closed loop systems applications being studied is control of sodium nitro prusside infusion rates to lower blood pressure after heart surgery. The IVAC 560 is one such device which is not yet available. Research on chrono-biological applications to determine whether circadian rhythms affect the metabolism of drugs, and whether diurnal variations exist in pain perception and narcotic algesic requirements, are also being carried out. Developmental work by different manufacturers to provide several programmable options like optional alarms, drug dose tapering ability, different drugs delivery etc. are going on. Other areas of developmental work are remote controlled reprogramming of implants and treatment of implants with antimicrobial agents to provide prevention of post surgical infections.
d) Monoclonal Antibodies The main thrust lies in the development of bifunctional monoclonals which have one binding site for the antigen and the other for the drug.
The basic research efforts are directed to attempt the delivery of peptides and proteins via this route. f) Microspheres/Nanoparticles
The developmental work for microspheres relates mainly to trying out different drug formulations especially for cancer therapy.
44. In India major thrust areas of research and development for each of these systems are :
a) Liposomes
Investigation of suitability of liposomes as vehicles for homing of drug/enzymes to specific sites in the biophase. Evaluation of liposomes as models for study of membrane structure and function. Targeted drug delivery through liposomes for the treatment of tuberculosis. Studies to enhance effectiveness of liposomised sodium stibogluconate against L. donovani infection by giving the drug more than once. Use of plant glycosides for drug delivery systems. Drug development and selective targeting of anti leishmanial drug. Development of recombinant toxins for site specific targeted delivery for diagnosis of infectious diseases. Evaluation of virosomes as a potential tool for targeting of drugs. Receptor mediated Endocytosis of Macromolecular conjugates in Selective Drug Delivery.
b) Nanoparticles
Study of nanoparticles as a drug delivery system for primaquine and metronidazole.
Current Device
An insulin pump is a small reservoir of insulin, usually stored in a small syringe, a batteryperated pump, and a computer programmable device that allows the user to program how much insulin the pump should deliver. This entire device is usually no larger than a cell phone or pager (shown below) and can be worn on the inside of the pants or on the belt. The pump delivers the insulin via an infusion set. The pump delivers a basal rate of insulin between meals and overnight, this is called background insulin. However, when the patient eats a meal more insulin is needed in the blood and the patient must tell the pump to deliver more insulin based on blood sugar levels (taken by the patient) [21]. The infusion set consists of a small tube that runs from the pump to a metal needle or a soft needle called the cannula. The needle, or cannula, is inserted and left in the skin usually on the abdomen, but sometimes on the thighs or the buttocks. There are many different infusion sets that differ mainly in size, shape, angle of needle insertion, and length of the needle used [12].
Figure VIII: MiniMed 508 Insulin Pump Figure VII is a picture of an insulin pump and the tube that would run to the infusion set. The small syringe holds the insulin and a small, battery-operated motor drives the syringe, which forces the insulin into the tube. The insulin pump does have some drawbacks. They are quite costly and range from $5,000 and $7,500. Frequent use of the pump could cause weight gain [12]. In addition, it may be bothersome being attached to the pump all day and night. Furthermore, an overnight stay at the hospital may be necessary in order to be trained on using the pump [13].
Competition:
Currently Professor Beebe of the UW-Madison BME department is performing research using hydrogels to address the same subject matter as this project. Hydrogels are expanding solids that can expand by a wide array of stimuli [20]. Some stimuli include amino acids, temperature, and different pH levels. A working idea of this expanding solid is of a small valve. When a certain pH travels through the valve, the solid swells cutting off flow. However, when a 13 different pH is introduced, the solid then contracts. Currently the gel is toxic due to the monomers that stick around after polymerization and are released when the gel is stimulated. The gels matrix spreads apart allowing for the monomers to fall out and contaminate the fluid or surrounding areas. However, hydrogels that are soaked in drugs can be used as wound dressings. When applied to the wound, the drug seeps out and onto the wound. The expansion rate of the gels is non-linear leading to difficult control issues. An advantage of hydrogels is the fact that they may be formed in any shape. Figure IX shows a simplified model of the function a hydrogel. [20].
opposed to having injections four times per day. The freedom to eat on his schedule is also offered by the insulin pumps. His blood sugar is managed better than ever before and it adjusts to fit his needs. The pumps are getting quieter and they offer flexibility for his busy schedule. However, Jayson does have some complaints about the current insulin pumps. The training for his pump lasted for approximately 16 hours and proved to be more of a nuisance than help. For the first two weeks with the pump, Jayson was required to take prick blood sugar tests every two hours. He was also instructed to fast for a day to view his blood sugar rates. The main problem with the current insulin pump is that it is always connected. Jayson cannot remove the pump for more than four hours; otherwise the pump does not deliver his needed basal rate of insulin. When sleeping, Jayson is always conscious of the pump, especially when he turns over. The size of the pump is also a limiting factor, as it becomes bothersome when playing sports. The pump is expensive and has a variety of hidden costs associated with it. Insulin and infusions sets are additional weekly costs besides the initial cost of the pump itself. Without health insurance, an insulin pump would not be a possibility for Jayson. A cheaper
Figure IX: A simplified drawing of a microvalve using a hydgrogel as the flow controller. As a fluid flows through the opening, it reacts with the gel and induces it to close.
d) Implants
Cefazoline sustained release bio-degradable microbeads (in vitro and in vivo animal studies). Sustained release tablet (SKI) formulation of rifampicin. (In vivo studies) Development of sustained release oral dosage from of pyrazinamide. (In vivo studies)
Optimisation of an indigeneous spheronization technology to develop controlled release products. Prolonged Release Liquid Formulation using ion exchange resins. f) Microencapsulation/microspheres Possibilities of controlled and targeted release of drugs are under examination. NCL can undertake work for specific industrial sponsors in microencapsulation technology for drugs and Pharmaceuticals. In vitro release studies on microcapsules for controlled release of diphtheria toxoid. g) Polymers in drug delivery Novel bases for rectal administration as suppositories. The work relates to improving the properties of bases. Study of controlled release of drug from radiation cured polymeric systems. A radiation cured transdermal system (852 EHA 15 MNA) has been studied on radiation polymerized beads. Future Research The next step for the research is to perform a more extensive materials search. Valve meters, NanoMuscles, and other construction materials will be the focus for the remainder of the semester. Beyond the materials search, the group will perform some preliminary engineering calculations to determine the needed volume, flow rates, and acceptable pressures involved with the design. Design Constraints The device will eventually be used by the patient directly. Therefore, the device must be sterilized for use and sharp edges must be covered when not in use. It will be used for 8 to 24 hour intervals and should be able to hold at least 0.48 mL of liquid medication. The finalized device should deliver 60 l/hour ( 12 l/hour) at a constant rate. It should be a disposable design that operates for a single use. The device will be attached to the abdomen of the patient, so it must operate between 55 and 90 F under normal atmospheric pressure and humidity. The finalized device should not be bulky, but rather similar in size and shape to a wrist watch. Ideally, it will have a 3 cm diameter, .5 cm thickness, and a .5 cm needle length. The materials for construction must be light enough to remain attached to the patient and must maintain accuracy after at least one year after production. At this time, one working model is desired and the research group would like to keep the cost of this device around $500. The clients major concern is that a functional device is developed; however, a scaled up initial model is requested at this time. A full product design specification is available in the Appendix.
Appendix: References [1] Ahern J, Mazur ML. Apr 2001. Site Rotation. Diabetes Forecast. 54,4:66-68. [2] Anonymous. Jan 2004. Insulin Administration. Diabetes Care. 27:S106-S109. [3] The Basics of Insulin. American Diabetes Association. 16 Sept 2004. <http://www.diabetes.org/type-1-diabetes/basics.jsp>. [4] Campbell, R. Keith. Update on Insulin Injection Devices. U.S. Pharmacist. 16 Sept 2004. <http://www.uspharmacist.com/oldformat.asp?url=newlook/files/Feat/may00insul in.cfm>. [5] Diabetes Mellitus. Microsoft Encarta Online Encyclopedia 2004. 10 Sept 2004. <http://encarta.msn.com>. [6] Flow Rate Calculations. 15 Sept. 2004. <http://www.ecsel.psu.edu/~tac167/flowcalcs.htmL>. [7] Fluid Systems. 15 Sept 2004. <http://claymor.engineer.gvsu.edu/jackh/books/model/chapters/fluids.pdf>. [8] Fomo Products, Inc. 24 Sept 2004. <http://www.fomo.com/msds/a16150.pdf >. [9] Hydrostatic Pressure in a Liquid. 15 Sept 2004. <http://www.ac.wwu.edu/~vawtr/PhysicsNet/Topics/Pressure/Hydrostatic.htmL>. [10] Insulin Routines. American Diabetes Association. 16 Sept 2004 <http://www.diabetes.org/type-1-diabetes/injections.jsp>. [11] Insulin Storage and Syringe Safety Information. American Diabetes Association. 16 Sept 2004. <http://www.diabetes.org/type-1-diabetes/safety.jsp>. [12] Insulin Pump 101. 19 Sept 2004. <http://www.banting.com/tcenter/pump101.htmL>. [13] Insulin Pumps. American Diabetes Association. 10 Sept 2004. http://www.diabetes.org/type-1-diabetes/insulin-pumps.jsp [14] Lumber T. Jul 2004. Tips for Site Rotation. Diabetes Forecast. 57,7:68-70. 36 [15] Kempinger, Jayson. Personal Interview: Austin Ramme. 25 Sept 2004. [16] Micro-Forged Metering Valves. Hoke, Inc. 15 Sept 2004. <http://www.hoke.com/pdf/1600_series.pdf>.
[17] Micro Needle Valves. Scientific Instrument Services. 15 Sept 2004. <http://www.sisweb.com/catalog/?page=D64>. [18] NanoMuscles. 21 Sept 2004. <http://www.nanomuscle.com/products/tech.htmL>. [19] Protein-Based Drugs: A Look Through The Crystal Ball. American Diabetes Association. 20 Sept 2004. <http://www.diabetesincontrol.com/modules.php?name=News&file=article&sid= 599>. [20] Puccinelli, John. Personal Interview: Christopher Westphal. 30 Sept 2004. [21] Pump Basics. 18 Sept 2004. <http://www.childrenwithdiabetes.com/pumps/basics.htm>. [22] P-445,P-446 Micro-Metering Valve. Western Analytical Products. 15 Sept 2004. <http://www.westernanalytical.com/upchurch/micro-metering.htm>. [23] Tight Diabetes Control. American Diabetes Association. 16 Sept 2004. <http://www.diabetes.org/type-1-diabetes/tight-control.jsp>. [24] The three skin layers: epidermis, dermis, subcutaneous fat. 18 Sept 2004. <www.pg.com/science/skincare/Skin_tws_10.htm>.
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