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Abstract
Conventional drug delivery using pills or injection is often not suitable for new protein, DNA, and other therapies.¹⁻³ An attractive alternative involves transdermal delivery from a patch, which avoids (i) degradation in the gastrointestinal tract and first-pass effects of the liver associated with oral delivery and (ii) the pain and inconvenience of intravenous injection.⁴⁻⁷ Delivery across skin also offers the possibility to continuously control the delivery rate, in contrast to conventional methods that deliver a large, discrete bolus. Despite these advantages, transdermal drug delivery is severely limited by the poor permeability of human skin; most drugs do not cross skin at therapeutic rates. Chemical,⁸ electrical,⁹ ultrasonic,¹⁰ and other methods have been developed to increase rates of transdermal transport, but have made only limited clinical impact to date.
... They consist of micron-sized projections that pierce the stratum corneum allowing the drugs to bypass the main barrier to diffusion. Studies have shown that the microneedle arrays cause no or little pain and are well tolerated by users, making it preferential to injection by syringe (Simonsen et al. 1999;Nir et al. 2003;Prausnitz et al. 2005). The microneedles can also offer a number of other benefits over other drug delivery methods. ...
The technology of fabricating microneedle arrays to deliver high molecular weight drugs across skin in a minimally invasive manner is receiving increasing attention. Microneedle arrays with different geometries have been manufactured using materials such as glass, polymer, metal, etc. However, a framework that can identify the optimum designs of these arrays seems to be lacking. This is important since by optimizing the microneedles dimensions (e.g., surface area of the patch, microneedle radius, etc.) the permeability of drugs in skin can be increased. To address this issue, this study presents an optimization framework for transdermal delivery of high molecular weight drug from microneedle. The optimization process is based on determining an optimization function (g) for various microneedles patterns (e.g., square, diamond, triangular, etc.). We argue that higher the value of g is the higher the drug permeability in skin is. The outputs of the developed framework have allowed us to identify the optimum design of both solid and hollow microneedles. In particular, the results have been used to predict skin permeability of high molecular weight using microneedle system. Also, optimum designs based on different classifications of skin thickness (e.g., race, age, etc.) for transdermal delivery of drugs are suggested.
... The cost constraints of the medical device industry suggest that microneedles should cost well under US$1.00 and ideally less than US$0.10 per microneedle array when mass produced [20]. Given this constraint, a challenge of the microneedle fabrication process was not only to meet device performance benchmarks, but also to keep the fabrication process as simple as possible. ...
Administration of protein and DNA biotherapeutics is limited by the need for hypodermic injection. Use of micron-scale needles to deliver drugs in a minimally invasive manner provides an attractive alternative, but application of this approach is limited by the need for suitable microneedle designs and fabrication methods. To address this need, this paper presents a conical polymer microneedle design that is fabricated using a novel integrated lens technique and analyzed for its ability to insert into the skin without mechanical failure. Microneedle master structures were fabricated using microlenses etched into a glass substrate that focused light through SU-8 negative epoxy resist to produce sharply tapered structures. Microneedle replicates were fabricated out of biodegradable polymers by micromolding. Because microneedle mechanical properties are critical to their insertion into the skin, we theoretically modeled two failure modes (axial mode and transverse mode), and analytical models were compared with measured data showing general agreement. Guided by this analysis, polymer microneedles were designed and demonstrated to insert to different depths into porcine skin in vitro. "Long" polymer microneedles were also demonstrated in human subjects to insert deeply without failure
Delivery of drugs through skin is obstructed by the excellent barrier properties of the outermost skin layer, the stratum corneum (SC). A strategy employing microneedles have recently emerged as a minimally invasive device for disrupting the SC structure and creating holes for molecules to pass through. Hollow-typed microneedles permit drug delivery which can be modulated over time via active delivery controlled by hand or pump. In this study, the potential of hollow microneedle for overcoming the outermost skin barrier and facilitating drug delivery into skin was investigated. Fluorescein isothiocyanate (FITC)-dextrans (4.3 kDa), FD-4, was used as a model large molecular compound. The effects of injection volume and formulation on drug release behavior from skin were determined. FD-4 was favorably loaded into the lower epidermis as well as the superficial dermis of the skin by a hollow microneedle. The release profiles of FD-4 were analyzed by Higuchi model based on Fick’s law of diffusion. The higher the volume of FD-4 solution injected, the faster the FD-4 release rate from skin. Liposome formulation exhibited no difference on drug release profiles compared with the solution. The results provide information for designing an effective hollow microneedles system.
Skin makes an excellent site for drug and vaccine delivery due to easy accessibility, immuno-surveillance functions, avoidance of macromolecular degradation in the gastrointestinal tract and possibility of self-administration. However, macromolecular drug delivery across the skin is primarily accomplished using hypodermic needles, which have several disadvantages including accidental needle-sticks, pain and needle phobia. These limitations have led to extensive research and development of alternative methods for drug and vaccine delivery across the skin. This review focuses on the recent trends and developments in this field of micro-scale devices for transdermal macromolecular delivery. These include liquid jet injectors, powder injectors, microneedles and thermal microablation. The historical perspective, mechanisms of action, important design parameters, applications and challenges are discussed for each method.
The purpose of this research was to examine the pharmacokinetics (PK) of drug uptake for microneedle-based intradermal (ID) delivery of several classes of protein drugs compared to standard subcutaneous (SC) administration.
Systemic absorption kinetics of various proteins were analyzed following microneedle-based ID delivery and standard injection methods in the swine model. Comparative PK data were determined using standard non-compartmental techniques based on blood serum levels.
Delivery of proteins using microneedles resulted in faster systemic availability, measured via t(max,) and increased maximal drug concentration, C(max,) over SC delivery for all proteins tested. Some agents also exhibited increased bioavailability for the ID route. Imaging studies using reporter dyes showed rapid lymphatic-mediated uptake.
Microneedle delivery is applicable to a wide variety of protein drugs and is capable of effective parenteral administration of therapeutic drug dosages. This delivery route alters absorption kinetics via targeting a tissue bed better perfused with lymphatic and blood vessels than the SC space. Microneedle delivery may afford various advantages, including a robust method to increase the absorption rate and bioavailability of proteins that have been challenging to deliver at therapeutic levels or with physiologically relevant profiles.
Hypodermic needles are in widespread use, but patients are unhappy with the pain, anxiety, and difficulty of using them. To increase patient acceptance, smaller needle diameters and lower insertion forces have been shown to reduce the frequency of painful injections. Guided by these observations, fine needles and microneedles have been developed to minimize pain and have found the greatest utility for delivery of vaccines and biopharmaceuticals such as insulin. However, pain reduction must be balanced against limitations of injection depth, volume, and formulations introduced by reduced needle dimensions. In some cases, needle-free delivery methods provide useful alternatives.
The threat of pandemic influenza and other public health needs motivate the development of better vaccine delivery systems. To address this need, microneedles have been developed as micron-scale needles fabricated using low-cost manufacturing methods that administer vaccine into the skin using a simple device that may be suitable for self-administration. Delivery using solid or hollow microneedles can be accomplished by (1) piercing the skin and then applying a vaccine formulation or patch onto the permeabilized skin, (2) coating or encapsulating vaccine onto or within microneedles for rapid, or delayed, dissolution and release in the skin, and (3) injection into the skin using a modified syringe or pump. Extensive clinical experience with smallpox, TB, and other vaccines has shown that vaccine delivery into the skin using conventional intradermal injection is generally safe and effective and often elicits the same immune responses at lower doses compared to intramuscular injection. Animal experiments using microneedles have shown similar benefits. Microneedles have been used to deliver whole, inactivated virus; trivalent split antigen vaccines; and DNA plasmids encoding the influenza hemagglutinin to rodents, and strong antibody responses were elicited. In addition, ChimeriVax-JE against yellow fever was administered to nonhuman primates by microneedles and generated protective levels of neutralizing antibodies that were more than seven times greater than those obtained with subcutaneous delivery; DNA plasmids encoding hepatitis B surface antigen were administered to mice and antibody and T cell responses at least as strong as hypodermic injections were generated; recombinant protective antigen of Bacillus anthracis was administered to rabbits and provided complete protection from lethal aerosol anthrax spore challenge at a lower dose than intramuscular injection; and DNA plasmids encoding four vaccinia virus genes administered to mice in combination with electroporation generated neutralizing antibodies that apparently included both Th1 and Th2 responses. Dose sparing with microneedles was specifically studied in mice with the model vaccine ovalbumin. At low dose (1 microg), specific antibody titers from microneedles were one order of magnitude greater than subcutaneous injection and two orders of magnitude greater than intramuscular injection. At higher doses, antibody responses increased for all delivery methods. At the highest levels (20-80 microg), the route of administration had no significant effect on the immune response. Concerning safety, no infections or other serious adverse events have been observed in well over 1,000 microneedle insertions in human and animal subjects. Bleeding generally does not occur for short microneedles (<1 mm). Highly localized, mild, and transient erythema is often observed. Microneedle pain has been reported as nonexistent to mild, and always much less than a hypodermic needle control. Overall, these studies suggest that microneedles may provide a safe and effective method of delivering vaccines with the possible added attributes of requiring lower vaccine doses, permitting low-cost manufacturing, and enabling simple distribution and administration.
Despite the advantages of drug delivery through skin, transdermal drug delivery is only used with a small subset of drugs because most compounds cannot cross the skin at therapeutically useful rates. Recently, a new concept known as microneedle was introduced and could be used to pierce effectively to deliver drugs using micron-sized needles in a minimally invasive and painless manner. In this study, the polymer microneedle-roller was fabricated so that it can be applied into the permeation of L-ascorbic acid. Moreover, a recent publication suggested the possibility of ascorbic acid 2-phosphate as a hair restorer; hence, this study was carried out to check the effect of L-ascorbic acid itself on the hair growing rate in rats according to the presence of various application frequencies of the polymer microneedle-roller. When the polymer microneedle-roller was applied nine times with four directions into rat's shaved skin, the permeation of L-ascorbic acid increased by 10.54-fold compared to that of the absence of the polymer microneedle-roller. The histological examination revealed that the skin pretreated with various application frequencies of the polymer microneedle-roller had more transport pathways. The faster hair growing phenomenon was observed in the presence of polymer microneedle-roller compared to the absence of the polymer microneedle-roller.
Non-invasive transdermal delivery using microneedle arrays was recently introduced to deliver a variety of large and hydrophilic compounds into the skin, including proteins and DNA. In this study, a microneedle array was applied to the delivery of a hydrophobic drug, ketoprofen, to determine if transdermal delivery in rats can be improved without the need for permeation enhancers. The ability of a microneedle to increase the skin permeability of ketoprofen was tested using the following procedure. A microneedle array was inserted into the lower back skin of a rat using a clip for 10 min. Subsequently, 24 mg/kg of a ketoprofen gel was loaded on the same site where the microneedle had been applied. Simultaneously, the microneedle was coated with 24 mg/kg of a ketoprofen gel, and inserted into the skin using a clip for 10 min. As a negative control experiment, only 24 mg/kg of the ketoprofen gel was applied to the shaved lower back of a rat. Blood samples were taken at the indicated times. The plasma concentration (C(p)) was obtained as a function of time (t), and the pharmacokinetic parameters were calculated using the BE program. The group loaded with the microneedle coated with ketoprofen gel showed a 1.86-fold and 2.86-fold increase in the AUC and C((max)) compared with the ketoprofen gel alone group. These results suggest that a microneedle can be an ideal tool for transdermal delivery products.
Interstitial fluid (ISF) is a specimen of increasing interest for glucose measurements because it can be obtained in a minimally invasive manner. Our previous study showed that sufficient ISF can be obtained using microneedles to measure glucose with a conventional electrochemical glucose monitor. The aim of this study was to assess the trueness of this glucose monitor using split-sample comparison with whole blood. We used ISF as specimen and our gas chromatography/mass spectrometry (GC/MS) method as reference.
We obtained 50 ISF samples and 40 whole blood samples from hairless Sprague- Dawley rats and analyzed for glucose by both methods.
For whole blood, a non-significant bias of 5.7% (+/-2 SD: -54.9% to 66.3%) was determined. ISF glucose measurements showed a significant constant bias of 29.5% (+/-2 SD: -85.0% to 144%), which seems to be caused in part by the lack of red blood cells in ISF. The correlation coefficients were 0.782 and 0.679 for whole blood and ISF, respectively.
The assessed electrochemical glucose monitor shows a close agreement with our GC/MS reference method for whole blood, for which this monitor was optimized. When glucose measurements are performed with ISF as matrix, the observed bias needs to be taken into consideration. Further studies are necessary to elucidate the reasons for the wide dispersion of data for ISF.
Previous work has shown that infusion flow rates can be increased by an order of magnitude by partially retracting microneedles after insertion into the skin. This study sought to determine the mechanism by which retraction increases fluid infusion by piercing human cadaver skin with single microneedles, fixing the skin after retracting microneedles to different distances, and examining skin microstructure by histology. We found that microneedle insertion to 1080 microm from the skin surface resulted primarily in skin indentation and only 100-300 microm penetration into the skin. This caused significant compaction of the skin, which probably pressed out most water and thereby dramatically lowered the flow conductivity of skin beneath the needle tip. Retraction of the microneedle allowed the skin to recoil back toward its original position, which relieved the skin compaction and increased local flow conductivity. Altogether, these results suggest that microneedle insertion to penetrate into the skin followed by microneedle retraction to relieve skin compaction is an effective approach to infuse fluid into the skin in a minimally invasive manner.
As an alternative to hypodermic injection or implantation of controlled-release systems, this study designed and evaluated biodegradable polymer microneedles that encapsulate drug for controlled release in skin and are suitable for self-administration by patients.
Arrays of microneedles were fabricated out of poly-lactide-co-glycolide using a mold-based technique to encapsulate model drugs--calcein and bovine serum albumin (BSA)--either as a single encapsulation within the needle matrix or as a double encapsulation, by first encapsulating the drug within carboxymethylcellulose or poly-L: -lactide microparticles and then encapsulating drug-loaded microparticles within needles.
By measuring failure force over a range of conditions, poly-lactide-co-glycolide microneedles were shown to exhibit sufficient mechanical strength to insert into human skin. Microneedles were also shown to encapsulate drug at mass fractions up to 10% and to release encapsulated compounds within human cadaver skin. In vitro release of calcein and BSA from three different encapsulation formulations was measured over time and was shown to be controlled by the encapsulation method to achieve release kinetics ranging from hours to months. Release was modeled using the Higuchi equation with good agreement (r2 > or = 0.90). After microneedle fabrication at elevated temperature, up to 90% of encapsulated BSA remained in its native state, as determined by measuring effects on primary, secondary, and tertiary protein structure.
Biodegradable polymer microneedles can encapsulate drug to provide controlled-release delivery in skin for hours to months.
Coated microneedles have been shown to deliver proteins and DNA into the skin in a minimally invasive manner. However, detailed studies examining coating methods and their breadth of applicability are lacking. This study's goal was to develop a simple, versatile and controlled microneedle coating process to make uniform coatings on microneedles and establish the breadth of molecules and particles that can be coated onto microneedles. First, microneedles were fabricated from stainless steel sheets as single microneedles or arrays of microneedles. Next, a novel micron-scale dip-coating process and a GRAS coating formulation were designed to reliably produce uniform coatings on both individual and arrays of microneedles. This process was used to coat compounds including calcein, vitamin B, bovine serum albumin and plasmid DNA. Modified vaccinia virus and microparticles of 1 to 20 micro m diameter were also coated. Coatings could be localized just to the needle shafts and formulated to dissolve within 20 s in porcine cadaver skin. Histological examination validated that microneedle coatings were delivered into the skin and did not wipe off during insertion. In conclusion, this study presents a simple, versatile, and controllable method to coat microneedles with proteins, DNA, viruses and microparticles for rapid delivery into the skin.
With the limitations of oral drug delivery and the pain and needle phobias associated with traditional injections, drug delivery research has focused on the transdermal delivery route. A formidable barrier to transdermal drug delivery is the stratum corneum, the superficial layer of the skin. In the last 10 years, microneedles were proposed as a mechanical tool to pierce through the stratum corneum, in order to create drug delivery channels without stimulating underlying pain nerves. Since then, the field of microneedles has rapidly evolved to spawn a plethora of potential transdermal applications. In this review, the authors provide an overview of the progress in microneedle research and design, and the advancements that have been made in employing this technology for transdermal applications.
To develop a rational basis for designing coating solution formulations for uniform and thick coatings on microneedles and to identify coating strategies to form composite coatings, deliver liquid formulations, and control the mass deposited on microneedles.
Microneedles were fabricated using laser-cutting and then dip-coated using different aqueous, organic solvent-based or molten liquid formulations. The mass of riboflavin (vitamin B(2)) coated onto microneedles was determined as a function of coating and microneedle parameters. Coated microneedles were also inserted into porcine cadaver skin to assess delivery efficacy.
Sharp-tipped microneedles, including pocketed microneedles, were fabricated. Excipients that reduced coating solution surface tension improved coating uniformity, while excipients that increased solution viscosity improved coating thickness. Evaluation of more than 20 different coating formulations using FDA approved excipients showed that hydrophilic and hydrophobic molecules could be uniformly coated onto microneedles. Model proteins were also uniformly coated on microneedles using the formulations identified in the study. Pocketed microneedles were selectively filled with solid or liquid formulations to deliver difficult-to-coat substances, and composite drug layers were formed for different release profiles. The mass of riboflavin coated onto microneedles increased with its concentration in the coating solution and the number of coating dips and microneedles in the array. Coatings rapidly dissolved in the skin without wiping off on the skin surface.
Microneedles and coating formulations can be designed to have a range of different properties to address different drug delivery scenarios.
To test the hypothesis that coated microneedles can deliver drugs into the eye via intrascleral and intracorneal routes in a minimally invasive manner.
Solid metal microneedles measuring 500 to 750 microm in length were coated with model drugs, protein, and DNA; inserted into nonpreserved human cadaveric sclera; and imaged. Microneedles coated with sodium fluorescein were then inserted into rabbit cornea in vivo. After needle removal, fluorescein concentration in the anterior segment of the rabbit eye was measured for 24 hours. Similar experiments were performed using pilocarpine-coated microneedles, and the rabbit pupil size was monitored afterward.
In vitro insertion tests showed that microneedles were mechanically strong enough to penetrate into human cadaveric sclera and that the drug coating rapidly dissolved off the needles within the scleral tissue within 30 seconds after insertion. In vivo delivery from fluorescein-coated microneedles showed that fluorescein concentrations in the anterior chamber were 60 times greater than those achieved by topical application without microneedles. Similarly, microneedle delivery of pilocarpine caused rapid and extensive rabbit pupil constriction. There were no measurable inflammatory responses caused by microneedle insertion.
This study demonstrated for the first time that coated microneedles can deliver drugs into the eye via intrascleral and intracorneal routes. This minimally invasive approach may avoid the complications associated with intraocular injection and systemic administration.
Despite the advantages of drug delivery through the skin, such as easy accessibility, convenience, prolonged therapy, avoidance of the liver first-pass metabolism and a large surface area, transdermal drug delivery is only used with a small subset of drugs because most compounds cannot cross the skin at therapeutically useful rates. Recently, a new concept was introduced known as microneedles and these could be pierced to effectively deliver drugs using micron-sized needles in a minimally invasive and painless manner. In this study, biocompatible polycarbonate (PC) microneedle arrays with various depths (200 and 500 microm) and densities (45, 99 and 154 ea/cm2) were fabricated using a micro-mechanical process. The skin permeability of a hydrophilic molecule, calcein (622.5D), was examined according to the delivery systems of microneedle, drug loading, depth of the PC microneedle, and density of the PC microneedle. The skin permeability of calcein was the highest when the calcein gel was applied to the skin with the 500 microm-depth PC microneedle, simultaneously. In addition, the skin permeability of calcein was the highest when 0.1g of calcein gel was coupled to the 500 microm-depth PC microneedle (154 ea/cm2) as well as longer microneedles and larger density of microneedles. Taken together, this study suggests that a biocompatible PC microneedle might be a suitable tool for transdermal drug delivery system of hydrophilic molecules with the possible applications to macromolecules such as proteins and peptides.
The purpose of this investigation was to evaluate the in vitro microneedle (MN) enhanced percutaneous absorption of naltrexone hydrochloride salt (NTX x HCl) compared to naltrexone base (NTX) in hairless guinea pig skin (GP) and human abdominal skin. In a second set of experiments, permeability of the major active metabolite 6-beta-naltrexol base (NTXOL) in the primarily unionized (unprotonated) form at pH 8.5 was compared to the ionized form (pH 4.5).
In vitro fluxes of NTX, NTX.HCl and ionized and unionized NTXOL were measured through microneedle treated or intact full thickness human and GP skin using a flow through diffusion apparatus. Solubility and diffusion samples were analyzed by HPLC.
Both GP and human skin show significant increases in flux when treated with 100 MN insertions as compared to intact full thickness skin when treated with NTX.HCl or ionized NTXOL (pH 4.5; p < 0.05). MN increased GP skin permeability for the hydrophilic HCL salt of NTX by tenfold and decreased lag time by tenfold too. Similar results were found using human skin, such that skin permeability to NTX.HCl was elevated to 7.0 x 10(-5) cm/h. Permeability of the primarily unionized (unprotonated) form of NTXOL at pH 8.5 was increased by MN only threefold and lag time was only modestly reduced. However, MN treatment with the primarily ionized (protonated) form of NTXOL at pH 4.5 increased skin permeability fivefold and decreased lag time fourfold.
Enhancement was observed in vitro in both GP and human skin treated with MN compared to intact skin with the salt form of NTX and the ionized form of NTXOL. We conclude that transdermal flux can be optimized by using MN in combination with charged (protonated) drugs that have increased solubility in an aqueous patch reservoir and increased permeability through aqueous pathways created by MN in the skin.
This paper presents a novel fabrication process for a tapered hollow metallic microneedle array using backside exposure of SU-8, and analytic solutions of critical buckling of a tapered hollow microneedle. An SU-8 mesa was formed on a Pyrex glass substrate and another SU-8 layer, which was spun on top of the SU-8 mesa, was exposed through the backside of the glass substrate. An array of SU-8 tapered pillar structures, with angles in the range of 3.1 • –5 • , was formed on top of the SU-8 mesa. Conformal electrodeposition of metal was carried out followed by a mechanical polishing using a planarizing polymeric layer. All organic layers were then removed to create a metallic hollow microneedle array with a fluidic reservoir on the backside. Both 200 µm and 400 µm tall, 10 by 10 arrays of metallic microneedles with inner diameters of the tip in the range of 33.6–101 µm and wall thickness of 10–20 µm were fabricated. Analytic solutions of the critical buckling of arbitrary-angled truncated cone-shaped columns are also presented. It was found that a single 400 µm tall hollow cylindrical microneedle made of electroplated nickel with a wall thickness of 20 µm, a tapered angle of 3.08 • and a tip inner diameter of 33.6 µm has a critical buckling force of 1.8 N. This analytic solution can be used for square or rectangular cross-sectioned column structures with proper modifications.
A two-wafer polysilicon micromolding process has been developed for the fabrication of hollow tubes useful for microfluidic applications. These small tubes can be fabricated with a pointed end, resulting in a micro hypodermic injection needle. Microneedles are desired because they reduce both insertion pain and tissue damage in the patient. Such microneedles may be used for low flow rate, continuous drug delivery, such as the continuous delivery of insulin to a diabetic patient. The needles would be integrated into a short term drug delivery device capable of delivering therapeutics intradermally for about 24 hours. In addition, microneedles can be used for sample collection for biological analysis, delivery of cell or cellular extract based vaccines, and sample handling providing interconnection between the microscopic and macroscopic world.
The strength of microneedles was examined analytically, experimentally and by finite element analysis. Metal coatings provide significant increases in the achievable bending moments before failure in the needles. For example, a 10 μ m platinum coating increased the median bending moment of a 160 μ m wide, 110 μ m high microneedle with a 20 μ m wall from 0.25 to 0.43 mNm. In addition, fluid flow in microneedles was studied experimentally. Microneedles 192 μ m wide, 110 μ m high and 7 mm long have flow rates of 0.7 ml/sec under a 138 kPa inlet pressure. This flow capacity exceeds previous microneedle capacities by an order of magnitude.
Microfabrication technology, more commonly applied to the manufacture of integrated circuits, can be used to build devices useful for mechanical delivery of drugs and genes. Microprobes fabricated using silicon micromachining have been used to deliver DNA into cells as an alternative to bombardment and microinjection. This idea can be extended to intravascular stents with integrated microprobes capable of piercing compressed plaque and delivering anti-restenosis therapies into coronary arteries. Preliminary experiments using filleted rabbit arteries have demonstrated transection of the internal elastic lamina. New nonplanar microfabrication technologies are necessary for creating practical devices with cylindrical symmetry; a promising possibility is to use microfabricated structures of anodic metal oxides.
We evaluated the Macroflux microprojection array patch technology as a novel system for intracutaneous delivery of protein antigens.
Macroflux microprojection array systems (330-microm micro-projection length, 190 microprojections/cm2, 1- and 2-cm2 area) were coated with a model protein antigen, ovalbumin (OVA), to produce a dry-film coating. After system application, microprojection penetration depth, OVA delivery, and comparative immune responses were evaluated in a hairless guinea pig model.
Macroflux microprojections penetrated into hairless guinea pig skin at an average depth of 100 microm with no projections deeper than 300 microm. Doses of I to 80 microg of OVA were delivered via 1- or 2-cm2 systems by varying the coating solution concentration and wearing time. Delivery rates were as high as 20 microg in 5 s. In a prime and boost dose immune response study, OVA-coated Macroflux was most comparable to equivalent doses injected intradermally. Higher antibody titers were observed when OVA was administered with the microprojection array or intradermally at low doses (1 and 5 microg). Macroflux administration at 1- and 5-microg doses gave immune responses up to 50-fold greater than that observed after the same subcutaneous or intramuscular dose. Dry coating an adjuvant, glucosaminyl muramyl dipeptide, with OVA on the Macroflux resulted in augmented antibody responses.
Macroflux skin patch technology provides rapid and reproducible intracutaneous administration of dry-coated antigen. The depth of skin penetration targets skin immune cells; the quantity of antigen delivered can be controlled by formulation, patch wearing time, and system size. This novel needle-free patch technology may ultimately have broad applications for a wide variety of therapeutic vaccines to improve efficacy and convenience of use.
Arrays of micrometer-scale needles could be used to deliver drugs, proteins, and particles across skin in a minimally invasive manner. We therefore developed microfabrication techniques for silicon, metal, and biodegradable polymer microneedle arrays having solid and hollow bores with tapered and beveled tips and feature sizes from 1 to 1,000 microm. When solid microneedles were used, skin permeability was increased in vitro by orders of magnitude for macromolecules and particles up to 50 nm in radius. Intracellular delivery of molecules into viable cells was also achieved with high efficiency. Hollow microneedles permitted flow of microliter quantities into skin in vivo, including microinjection of insulin to reduce blood glucose levels in diabetic rats.
The past twenty five years have seen an explosion in the creation and discovery of new medicinal agents. Related innovations in drug delivery systems have not only enabled the successful implementation of many of these novel pharmaceuticals, but have also permitted the development of new medical treatments with existing drugs. The creation of transdermal delivery systems has been one of the most important of these innovations, offering a number of advantages over the oral route. In this article, we discuss the already significant impact this field has made on the administration of various pharmaceuticals; explore limitations of the current technology; and discuss methods under exploration for overcoming these limitations and the challenges ahead.
The purpose of this study was to design and fabricate arrays of solid microneedles and insert them into the skin of diabetic hairless rats for transdermal delivery of insulin to lower blood glucose level.
Arrays containing 105 microneedles were laser-cut from stainless steel metal sheets and inserted into the skin of anesthetized hairless rats with streptozotocin-induced diabetes. During and after microneedle treatment, an insulin solution (100 or 500 U/ml) was placed in contact with the skin for 4 h. Microneedles were removed 10 s, 10 min, or 4 h after initiating transdermal insulin delivery. Blood glucose levels were measured electrochemically every 30 min. Plasma insulin concentration was determined by radioimmunoassay at the end of most experiments.
Arrays of microneedles were fabricated and demonstrated to insert fully into hairless rat skin in vivo. Microneedles increased skin permeability to insulin, which rapidly and steadily reduced blood glucose levels to an extent similar to 0.05-0.5 U insulin injected subcutaneously. Plasma insulin concentrations were directly measured to be 0.5-7.4 ng/ml. Higher donor solution insulin concentration, shorter insertion time, and fewer repeated insertions resulted in larger drops in blood glucose level and larger plasma insulin concentrations.
Solid metal microneedles are capable of increasing transdermal insulin delivery and lowering blood glucose levels by as much as 80% in diabetic hairless rats in vivo.
Microneedles are promising microfabricated devices for minimally invasive drug delivery applications. Needles can be integrated into a variety of devices. However, any portable drug delivery device with integrated microneedles will need an equally compact means to deliver therapeutics. This work presents microneedles integrated with an on-chip MEMS positive displacement micropump for continuous drug delivery applications. The generation and collapse of thermally generated bubbles with flow rectified by directional check valves are used to achieve net pumping through the device. Visualization methods have observed net flow rates of water out of a microneedle at approximately 2.0 nl/s with a pressure of 3.9 kPa. In addition, continuous pumping was achieved for more than 6 hours with the heaters actuating for over 18 hours (15,000 cycles) without failing.
A new anthrax vaccine under clinical investigation is based on recombinant Bacillus anthracis protective antigen (rPA). Here, we investigated microneedle-based cutaneous and nasal mucosal delivery of rPA in mice and
rabbits. In mice, intradermal (id) delivery achieved up to 90% seroconversion after a single dose, compared with 20% after
intramuscular (im) injection. Intranasal (inl) delivery of a liquid formulation required 3 doses to achieve responses that
were comparable with those achieved via the id or im routes. In rabbits, id delivery provided complete protection against
aerosol challenge with anthrax spores; in addition, novel powder formulations administered inl provided complete protection,
whereas a liquid formulation provided only partial protection. These results demonstrate, for the first time, that cutaneous
or nasal mucosal administration of rPA provides complete protection against inhalational anthrax in rabbits. The novel vaccine/device
combinations described here have the potential to improve the efficacy of rPA and other biodefense vaccines.
This paper presents a novel process for the fabrication of out-of-plane hollow microneedles in silicon. The fabrication method consists of a sequence of deep-reactive ion etching (DRIE), anisotropic wet etching and conformal thin film deposition, and allows needle shapes with different, lithography-defined tip curvature. In this study, the length of the needles varied between 150 and 350 micrometers. The widest dimension of the needle at its base was 250 μm. Preliminary application tests of the needle arrays show that they are robust and permit skin penetration without breakage. Transdermal water loss measurements before and after microneedle skin penetration are reported. Drug delivery is increased approximately by a factor of 750 in microneedle patch applications with respect to diffusion alone. The feasibility of using the microneedle array as a blood sampler on a capillary electrophoresis chip is demonstrated.
We describe the microfabrication, packaging and testing of a
micromachined dry biopotential electrode, (i.e., where electrolytic gel
is not required). It consists of an array of micro-dimensioned, very
sharp spikes, (i.e., needles) designed for penetration of human skin
which circumvent high impedance problems associated with layers of the
outer skin. The spikes are etched in silicon by deep reactive ion
etching and are subsequently covered with a silver-silverchloride
(Ag-AgCl) double layer. The electrode-skin-electrode impedance of dry
spiked electrodes having a size of 4×4 mm<sup>2</sup> is reduced
compared to standard electrodes using electrolytic gel and having a
comparable size. Recorded low amplitude biopotentials resulting from the
activity of the brain, (i.e., EEG signals) are of high quality, even for
spiked electrodes as small as 2×2 mm<sup>2</sup>. The spiked
electrode offers a promising alternative to standard electrodes in
biomedical applications and is of interest in research of new biomedical
methods
The goal of this study was to design, fabricate, and test arrays of hollow microneedles for minimally invasive and continuous delivery of insulin in vivo. As a simple, robust fabrication method suitable for inexpensive mass production, we developed a modified-LIGA process to micromachine molds out of polyethylene terephthalate using an ultraviolet laser, coated those molds with nickel by electrodeposition onto a sputter-deposited seed layer, and released the resulting metal microneedle arrays by selectively etching the polymer mold. Mechanical testing showed that these microneedles were sufficiently strong to pierce living skin without breaking. Arrays containing 16 microneedles measuring 500 μm in length with a 75 μm tip diameter were then inserted into the skin of anesthetized, diabetic, hairless rats. Insulin delivery through microneedles caused blood glucose levels to drop steadily to 47% of pretreatment values over a 4-h insulin delivery period and were then approximately constant over a 4-h postdelivery monitoring period. Direct measurement of plasma insulin levels showed a peak value of 0.43 ng/ml. Together, these data suggest that microneedles can be fabricated and used for in vivo insulin delivery.
To overcome the skin's barrier properties that block transdermal delivery of most drugs, arrays of microscopic needles have been microfabricated primarily out of silicon or metal. This study addresses microneedles made of biocompatible and biodegradable polymers, which are expected to improve safety and manufacturability. To make biodegradable polymer microneedles with sharp tips, micro-electromechanical masking and etching were adapted to produce beveled- and chisel-tip microneedles and a new fabrication method was developed to produce tapered-cone microneedles using an in situ lens-based lithographic approach. To replicate microfabricated master structures, PDMS micromolds were generated and a novel vacuum-based method was developed to fill the molds with polylactic acid, polyglycolic acid, and their co-polymers. Mechanical testing of the resulting needles measured the force at which needles broke during axial loading and found that this failure force increased with Young's modulus of the material and needle base diameter and decreased with needle length. Failure forces were generally much larger than the forces needed to insert microneedles into skin, indicating that biodegradable polymers can have satisfactory mechanical properties for microneedles. Finally, arrays of polymer microneedles were shown to increase permeability of human cadaver skin to a low-molecular weight tracer, calcein, and a macromolecular protein, bovine serum albumin, by up to three orders of magnitude. Altogether, these results indicate that biodegradable polymer microneedles can be fabricated with an appropriate geometry and sufficient strength to insert into skin, and thereby dramatically increase transdermal transport of molecules.
Toward achieving painless injections and other microfluidic applications, we microfabricated conically tapered needles of micron dimensions. The relationship between pressure drop and flow rate through microneedles was experimentally quantified as a function of fluid viscosity and microneedle length, diameter, and cone half-angle. At Reynolds numbers ≲ 100, dimensionless pressure drop (2ΔP/ρv2) sharply decreased with increasing Reynolds number, indicating the importance of viscous forces. At larger Reynolds numbers, the flow was almost inviscid, as indicated by a weak dependency of dimensionless pressure drop on Reynolds number. Numerical simulations showed good agreement with experimental data and predicted that flow through conically tapered microneedles is primarily controlled by the diameter and taper angle at the microneedle tip. A characteristic feature of flow through conically tapered microneedles is a favorable axial pressure gradient that accelerates fluid through the microneedle, thus inhibiting growth of the viscous boundary layer on the microneedle wall. © 2005 American Institute of Chemical Engineers AIChE J, 2005
Vaccination is one of the major achievements of modern medicine. As a result of vaccination, diseases such as polio and measles have been controlled and small pox has been eradicated. However, despite these successes there are still many microbial diseases that cause tremendous suffering because there is no vaccine or the vaccines available are inadequate. In addition, even if vaccines were available for all infectious diseases there is no guarantee that people would use them routinely. One of the major impediments to ensuring vaccine efficacy and compliance is that of delivery. Presently most vaccines are given by intramuscular administration. Unfortunately this is often traumatic, especially in infants. Thus, if it was possible to replace intramuscular immunization by mucosal (oral/intranasal) or transdermal delivery it may be possible to both enhance mucosal immunity as well as improve overall compliance rates. The transdermal route has been used by the pharmaceutical industry for the delivery of various low molecular weight drugs. Some of the approaches used for smaller compounds may also have potential for delivery of either protein or polynucleotide vaccines. However, there is a greater challenge to delivering large molecular weight molecules through the skin due to size, charge and other physicochemical properties. This review will describe the recent advances that have been made in dermal and topical delivery as related to vaccines.
Enough information has been gained from clinical trials to allow the conclusion that human gene transfer is feasible, can evoke biologic responses that are relevant to human disease, and can provide important insights into human biology. Adverse events have been uncommon and have been related to the gene delivery strategies, not to the genetic material being transferred. Human gene transfer still faces significant hurdles before it becomes an established therapeutic strategy. However, its accomplishments to date are impressive, and the logic of the potential usefulness of this clinical paradigm continues to be compelling.
Although modern biotechnology has produced extremely sophisticated and potent drugs, many of these compounds cannot be effectively delivered using current drug delivery techniques (e.g., pills and injections). Transdermal delivery is an attractive alternative, but it is limited by the extremely low permeability of skin. Because the primary barrier to transport is located in the upper 10-15 micron of skin and nerves are found only in deeper tissue, we used a reactive ion etching microfabrication technique to make arrays of microneedles long enough to cross the permeability barrier but not so long that they stimulate nerves, thereby potentially causing no pain. These microneedle arrays could be easily inserted into skin without breaking and were shown to increase permeability of human skin in vitro to a model drug, calcein, by up to 4 orders of magnitude. Limited tests on human subjects indicated that microneedles were reported as painless. This paper describes the first published study on the use of microfabricated microneedles to enhance drug delivery across skin.
Micromachined needle arrays have been designed, fabricated, and characterized. The design includes arrays of 25 needles with fluid coupling channels and dual structural supports. Numerical modeling of fluid flow characteristics was performed, demonstrating that the needle coupling channels redistribute flow when the input or output ports are fully restricted. Micromachining technologies have been used to batch fabricate hollow metallic fluid coupled needle arrays. The significance of this work includes the development of the hollow metallic micromachined needle arrays for biomedical applications, as well as a discussion of structural, fluidic, and biological design considerations. The micromachined needle array has many advantages, including (a) reduced trauma at penetration site (small size), (b) greater freedom of patient movement (minimal penetration), (c) a practically pain-free drug delivery device (distribution of force), (d) precise control of penetration depth (needle extension length), and (e) they can be stacked and packaged into a 3-D device for fluid transfer.
Microscopic needles previously shown capable of transdermal delivery of drugs and proteins are demonstrated to be painless when pressed into the skin of human subjects.
A unique minimally invasive system for painless blood testing is now being commercialized for measurement of blood glucose concentration by diabetics. The novel component of this system, a consumable microsampling and assay device, consists of a tough, flexible silicon microneedle comparable in cross-section to a human hair integrated with a silicon microcuvette. This microneedle is capable of reliably taking a very small sample of whole blood completely painlessly, unlike sticks with the much larger metal lancet that must be used in all other current systems. The device permits a one-step process that avoids the need to transfer blood from a skin puncture to a test strip, thus minimizing blood required and possible mess. The small hand-held instrument containing the consumable is touched to the skin of the arm or any other part of the body, not necessarily the tip of the finger, and held there for one second. During this time, the microneedle is advanced and then withdrawn under microprocessor control, puncturing the skin and drawing less than 200 nanoliters of blood into the microcuvette, where the assay is performed automatically. The instrument calculates the blood glucose concentration, displays the result, and holds it in memory for recall. The consumable is produced by silicon microelectromechanical systems technology and can be produced in high volume at low unit cost. This technology shows promise of being extended to other analytes and to continuous monitoring.
A new microfabrication technology, microelectromechanical systems (MEMS), is envisioned for improved insulin delivery in the context of a device currently being developed for the Defense Advanced Research Projects Agency (DARPA). The drug delivery system utilizes MEMS technology to move and control fluids at the microscale, making possible the reconstitution and delivery of extremely small amounts of drug with extreme precision. In this article, the required microscale components that are currently being developed for the system are described. MEMS are made using fabrication methods similar to that utilized in microelectronics. Consequently, MEMS technology can be used to fabricate devices that are extremely small. The fundamental difference is that MEMS devices can either move themselves or control the movement of other materials, such as fluids. Furthermore, this manufacturing method is intrinsically low-cost and therefore is ideal for drug delivery systems. The current development of a new drug delivery system for controlled drug reconstitution and delivery system for DARPA is described as are the MEMS-based components for the required fluidic control. The adaptation of the system for insulin delivery is addressed and is envisioned to be a fully self-contained parenteral drug delivery system about the size of a 4-mm thick credit card.
Skin is an attractive target for delivery of genetic therapies and vaccines. However, new approaches are needed to access this tissue more effectively. Here, we describe a new delivery technology based on arrays of structurally precise, micron-scale silicon projections, which we term microenhancer arrays (MEAs). In a human clinical study, these devices effectively breached the skin barrier, allowing direct access to the epidermis with minimal associated discomfort and skin irritation. In a mouse model, MEA-based delivery enabled topical gene transfer resulting in reporter gene activity up to 2,800-fold above topical controls. MEA-based delivery enabled topical immunization with naked plasmid DNA, inducing stronger and less variable immune responses than via needle-based injections, and reduced the number of immunizations required for full seroconversion. Together, the results provide the first in vivo use of microfabricated devices to breach the skin barrier and deliver vaccines topically, suggesting significant clinical and practical advantages over existing technologies.
With the technological advances in biomedical sciences and the better understanding of how the immune system works, new immunisation strategies and vaccine delivery options, such sprays, patches, and edible formulations have been developed. This has opened up the possibility of administering vaccines without the use of needles and syringes. Already topical immunisation is a reality and it has the potential to make vaccine delivery more equitable, safer, and efficient. Furthermore, it would increase the rate of vaccine compliance and greatly facilitate the successful implementation of worldwide mass vaccination campaigns against infectious diseases. This review gives a brief account of the latest developments of application of candidate vaccine antigens onto bare skin and describes some of our recent observations using peptide and glycoconjugate vaccines as immunogens.
The enhancement of transdermal transport by ultrasound is reviewed. After a brief discussion of the physics of ultrasound and its medical applications, the effects of high- and low-frequency ultrasound on the transport of substances across the skin are examined. The impact of low-frequency sonophoresis appears to be much more important, with significant increases in transport into and from the skin following its application. Although the mechanism of action remains incompletely defined, cavitation and thermal processes are strongly implicated.
The skin represents an accessible somatic tissue for therapeutic gene transfer. The superficial lipophilic layer of the skin, the stratum corneum, however, constitutes a major obstacle to the cutaneous delivery of charged macromolecules such as DNA.
To determine whether silicon-based microneedles, microfabricated via a novel isotropic etching/BOSCH reaction process, could generate microchannels in the skin of sufficient dimensions to facilitate access of lipid : polycation : pDNA (LPD) nonviral gene therapy vectors.
Scanning electron microscopy was used to visualize the microconduits created in heat-separated human epidermal sheets after application of the microneedles. Following confirmation of particle size and particle surface charge by photon correlation spectrocopy and microelectrophoresis, respectively, the diffusion of fluorescent polystyrene nanospheres and LPD complexes through heat-separated human epidermal sheets was determined in vitro using a Franz-type diffusion cell. In-vitro cell culture with quantification by flow cytometry was used to determine gene expression in human keratinocytes (HaCaT cells).
The diffusion of 100 nm diameter fluorescent polystyrene nanospheres, used as a readily quantifiable predictive model for LPD complexes, through epidermal sheets was significantly enhanced following membrane treatment with microneedles. The delivery of LPD complexes either into or through the membrane microchannels was also demonstrated. In both cases considerable interaction between the particles and the epidermal sheet was observed. In-vitro cell culture was used to confirm that LPD complexes mediated efficient reporter gene expression in human keratinocytes in culture when formulated at the appropriate surface charge.
These studies demonstrate the utility of silicon microneedles in cutaneous gene delivery. Further studies are required to elucidate fully the influence of the physicochemical characteristics of gene therapy vectors, e.g. particle diameter and surface charge, on their diffusion through microchannels and to quantify gene expression in vivo.
In order to achieve enhanced topical drug delivery, it is necessary to make physical or biomolecular structural alterations to the stratum corneum by suitable techniques or by the use of specific chemical agents or drug carriers. The role of the chemical penetration enhancer is to reversibly alter the barrier properties of the stratum corneum by disruption of the membrane structures or by maximizing drug solubility within the skin. Alternatively, permeant delivery to the dermal vasculature using one of several physical methods to reduce diffusional resistance within the skin may be used to promote drug penetration. In the present article, we summarize the major facets of the diverse spectrum of penetration enhancement techniques that include modification of the stratum corneum, lipid-based delivery systems, drug/vehicle interactions, bypassing the stratum corneum, and electrical techniques of enhancement.
As a hybrid between a hypodermic needle and transdermal patch, we have used microfabrication technology to make arrays of micron-scale needles that transport drugs and other compounds across the skin without causing pain. However, not all microneedle geometries are able to insert into skin at reasonable forces and without breaking. In this study, we experimentally measured and theoretically modeled two critical mechanical events associated with microneedles: the force required to insert microneedles into living skin and the force needles can withstand before fracturing. Over the range of microneedle geometries investigated, insertion force was found to vary linearly with the interfacial area of the needle tip. Measured insertion forces ranged from approximately 0.1-3N, which is sufficiently low to permit insertion by hand. The force required to fracture microneedles was found to increase with increasing wall thickness, wall angle, and possibly tip radius, in agreement with finite element simulations and a thin shell analytical model. For almost all geometries considered, the margin of safety, or the ratio of fracture force to insertion force, was much greater than one and was found to increase with increasing wall thickness and decreasing tip radius. Together, these results provide the ability to predict insertion and fracture forces, which facilitates rational design of microneedles with robust mechanical properties.
The effect of vibratory actuation on microneedle insertion force was investigated. Hollow micro hypodermic injection needles were fabricated by a two-wafer polysilicon micromolding process. A vibratory actuator operating in the kHz range was coupled with the hypodermic microneedles. The force to insert microneedles into excized animal tissue was measured with a load cell. Results showed a greater than 70% reduction in microneedle insertion force by using vibratory actuation. The application of vibratory actuation provides a promising method to precisely control the microneedle insertion forces to overcome microneedle structural material limitations, minimize insertion pain, and enhance the efficiency of drug delivery.
One long-standing approach for improving transdermal drug delivery uses penetration enhancers (also called sorption promoters or accelerants) which penetrate into skin to reversibly decrease the barrier resistance. Numerous compounds have been evaluated for penetration enhancing activity, including sulphoxides (such as dimethylsulphoxide, DMSO), Azones (e.g. laurocapram), pyrrolidones (for example 2-pyrrolidone, 2P), alcohols and alkanols (ethanol, or decanol), glycols (for example propylene glycol, PG, a common excipient in topically applied dosage forms), surfactants (also common in dosage forms) and terpenes. Many potential sites and modes of action have been identified for skin penetration enhancers; the intercellular lipid matrix in which the accelerants may disrupt the packing motif, the intracellular keratin domains or through increasing drug partitioning into the tissue by acting as a solvent for the permeant within the membrane. Further potential mechanisms of action, for example with the enhancers acting on desmosomal connections between corneocytes or altering metabolic activity within the skin, or exerting an influence on the thermodynamic activity/solubility of the drug in its vehicle are also feasible, and are also considered in this review.
A combination of surface- and bulk-micromachining techniques is
used to demonstrate the feasibility of fabricating microhypodermic
needles. These microneedles, which may be built with on-board fluid
pumps, have potential applications in the chemical and biomedical fields
for localized chemical analysis, programmable drug-delivery systems, and
very small, precise sampling of fluids. The microneedles are fabricated
in 1, 3, and 6 mm lengths with fully enclosed channels formed of silicon
nitride. The channels are 9 μm in height and have one of two widths,
30 or 50 μm. Access to the channels is provided at their shank and
distal ends through 40-μm square apertures in the overlying silicon
nitride layer. The microneedles are found to be intact and undamaged
following repetitive insertion into and removal from animal-muscle
tissue (porterhouse steak)
A bulk-micromachined multichannel silicon probe capable of selectively delivering chemicals at the cellular level as well as electrically recording from and stimulating neurons in vivo has been developed. The process buries multiple flow channels in the probe substrate, resulting in a hollow-core device, Microchannel formation requires only one mask in addition to those normally used for probe fabrication and is compatible with on-chip signal-processing circuitry. Flow in these microchannels has been studied theoretically and experimentally. For an effective channel diameter of 10 μm, a channel length of 4 mm, and water as the injected fluid, the flow velocity at 11 torr is about 1.3 mm/s, delivering 100 pl in 1 s. Intermixing of chemicals, with the tissue fluid due to natural diffusion through the outlet orifice becomes significant for dwell times in excess of about 30 min, and a shutter is proposed for chronic use. The probe has been used for acute monitoring of the neural responses to various chemical stimuli in guinea pig superior and inferior colliculus.
Microneedles and other structures have been developed for introducing therapeutic agents into tissues and cells. Microstructures for transdermal delivery hold the promise of pain-free drug injection. Electrodes integrated with microneedles can sense and monitor the effects of injected materials on tissues. Microprobes have been shown to be effective in transfecting cells through the delivery of DNA in experiments with both plants and animals. Microfabricated delivery devices have great potential for local delivery of drugs and genes where systemic administration presents serious safety concerns. In this paper, we review recent progress in microdevices for delivering therapeutic agents, including microneedles, DNA transfection schemes, and intravascular drug and gene delivery systems.
- Madou M.J.