Improving the reach of vaccines to low-resource regions, with a needle-free vaccine delivery device and long-term thermostabilization

Dry-coated microprojections can deliver vaccine to abundant antigen-presenting cells in the skin and induce efficient immune responses and the dry-coated vaccines are expected to be thermostable at elevated temperatures. In this paper, we show that we have dramatically improved our previously reported gas-jet drying coating method and greatly increased the delivery efficiency of coating from patch to skin to from 6.5% to 32.5%, by both varying the coating parameters and removing the patch edge. Combined with our previous dose sparing report of influenza vaccine delivery in a mouse model, the results show that we now achieve equivalent protective immune responses as intramuscular injection (with the needle and syringe), but with only 1/30th of the actual dose. We also show that influenza vaccine coated microprojection patches are stable for at least 6 months at 23 degrees C. inducing comparable immunogenicity with freshly coated patches. The dry-coated microprojection patches thus have key and unique attributes in ultimately meeting the medical need in certain low-resource regions with low vaccine affordability and difficulty in maintaining "cold-chain" for vaccine storage and transport. (C) 2011 Elsevier B.V. All rights reserved

Protocol to evaluate a pilot program to upskill clinicians in providing genetic testing for familial melanoma

Introduction Genetic testing for hereditary cancers can improve long-term health outcomes through identifying high-risk individuals and facilitating targeted prevention and screening/surveillance. The rising demand for genetic testing exceeds the clinical genetic workforce capacity. Therefore, non-genetic specialists need to be empowered to offer genetic testing. However, it is unknown whether patient outcomes differ depending on whether genetic testing is offered by a genetics specialist or a trained non-genetics clinician. This paper describes a protocol for upskilling non-genetics clinicians to provide genetic testing, randomise high-risk individuals to receive testing from a trained clinician or a genetic counsellor, and then determine whether patient outcomes differed depending on provider-type. Methods An experiential training program to upskill dermatologically-trained clinicians to offer genetic testing for familial melanoma is being piloted on 10–15 clinicians, prior to wider implementation. Training involves a workshop, comprised of a didactic learning presentation, case studies, simulated sessions, and provision of supporting documentation. Clinicians later observe a genetic counsellor led consultation before being observed leading a consultation. Both sessions are followed by debriefing with a genetic counsellor. Thereafter, clinicians independently offer genetic testing in the clinical trial. Individuals with a strong personal and/or family history of melanoma are recruited to a parallel-group trial and allocated to receive pre- and post- genetic testing consultation from a genetic counsellor, or a dermatologically-trained clinician. A mixed method approach measures psychosocial and behavioural outcomes. Longitudinal online surveys are administered at five timepoints from baseline to one year post-test disclosure. Semi-structured interviews with both patients and clinicians are qualitatively analysed. Significance This is the first program to upskill dermatologically-trained clinicians to provide genetic testing for familial melanoma. This protocol describes the first clinical trial to compare patient-reported outcomes of genetic testing based on provider type (genetic counsellors vs trained non-genetic clinicians)

Protocol to evaluate a pilot program to upskill clinicians in providing genetic testing for familial melanoma.

IntroductionGenetic testing for hereditary cancers can improve long-term health outcomes through identifying high-risk individuals and facilitating targeted prevention and screening/surveillance. The rising demand for genetic testing exceeds the clinical genetic workforce capacity. Therefore, non-genetic specialists need to be empowered to offer genetic testing. However, it is unknown whether patient outcomes differ depending on whether genetic testing is offered by a genetics specialist or a trained non-genetics clinician. This paper describes a protocol for upskilling non-genetics clinicians to provide genetic testing, randomise high-risk individuals to receive testing from a trained clinician or a genetic counsellor, and then determine whether patient outcomes differed depending on provider-type.MethodsAn experiential training program to upskill dermatologically-trained clinicians to offer genetic testing for familial melanoma is being piloted on 10-15 clinicians, prior to wider implementation. Training involves a workshop, comprised of a didactic learning presentation, case studies, simulated sessions, and provision of supporting documentation. Clinicians later observe a genetic counsellor led consultation before being observed leading a consultation. Both sessions are followed by debriefing with a genetic counsellor. Thereafter, clinicians independently offer genetic testing in the clinical trial. Individuals with a strong personal and/or family history of melanoma are recruited to a parallel-group trial and allocated to receive pre- and post- genetic testing consultation from a genetic counsellor, or a dermatologically-trained clinician. A mixed method approach measures psychosocial and behavioural outcomes. Longitudinal online surveys are administered at five timepoints from baseline to one year post-test disclosure. Semi-structured interviews with both patients and clinicians are qualitatively analysed.SignificanceThis is the first program to upskill dermatologically-trained clinicians to provide genetic testing for familial melanoma. This protocol describes the first clinical trial to compare patient-reported outcomes of genetic testing based on provider type (genetic counsellors vs trained non-genetic clinicians)

Elongate microparticles for enhanced drug delivery to ex vivo and in vivo pig skin

The delivery of therapeutics and cosmaceuticals into and/or through the skin is hindered by epidermal barriers. To overcome the skin's barriers we have developed a novel cutaneous delivery method using high aspect ratio elongate microparticles (EMPs). Using ex vivo and in vivo pig skin we assess the penetration and delivery characteristics of the elongate microparticles. With reflectance confocal microscopy we observed that the elongate microparticles successfully penetrated the epidermis and upper dermis. Delivery was then assessed using two different length populations of EMPs, comparing their delivery profile to topical alone using sodium fluorescein and confocal microscopy. We observed a relatively uniform and continuous delivery profile in the EMP treated area within the upper layers of the skin - up to seven times greater than topical alone. Finally, we delivered two therapeutically relevant compounds (Vitamins A and B3), showing enhanced delivery using the EMPs. To our knowledge this is the first report using high aspect ratio elongate microparticles in this manner for enhanced topical delivery to the skin

Indicatori economico-ambientali per lo sviluppo sostenibile della pesca in laguna Veneta.

The buccal mucosa (inner cheek) is an attractive site for delivery of immunotherapeutics, due to its ease of access and rich antigen presenting cell (APC) distribution. However, to date, most delivery methods to the buccal mucosa have only been topical - with the challenges of: 1) an environment where significant biomolecule degradation may occur; 2) inability to reach the APCs that are located deep in the epithelium and lamina propria; and 3) salivary flow and mucous secretion that may result in removal of the therapeutic agent before absorption has taken place. To overcome these challenges and achieve consistent, repeatable targeted delivery of immunotherapeutics to within the buccal mucosa (not merely on to the surface), we utilised microprojection arrays (Nanopatches - 110 μm length projections, 3364 projections, 16 mm surface area) with a purpose built clip applicator. The mechanical application of Nanopatches bearing a dry-coated vaccine (commercial influenza vaccine, as a test case immunotherapeutic) released the vaccine to a depth of 47.8 ± 14.8 μm (mean ± SD, n = 4), in the mouse buccal mucosa (measured using fluorescent delivered dyes and CryoSEM). This location is in the direct vicinity of APCs, facilitating antigenic uptake. Resultant systemic immune responses were similar to systemic immunization methods, and superior to comparative orally immunised mice. This confirms the Nanopatch administered vaccine was delivered into the buccal mucosa and not ingested. This study demonstrates a minimally-invasive delivery device with rapid (2 min of application time), accurate and consistent release of immunotherapeutics in to the buccal mucosa - that conceptually can be extended in to human use for broad and practical utility

Microbiopsy engineered for minimally invasive and suture-free sub-millimetre skin sampling

We describe the development of a sub-millimetre skin punch biopsy device for painless and suture-free skin sampling for molecular diagnosis and research. Conventional skin punch biopsies range from 2-4 mm in diameter. Local anaesthesia is required and sutures are usually used to close the wound. Our microbiopsy is 0.50 mm wide and 0.20 mm thick. The microbiopsy device is fabricated from three stacked medical grade stainless steel plates tapered to a point and contains a chamber within the centre plate to collect the skin sample. We observed that the application of this device resulted in a 0.21 ± 0.04 mm wide puncture site in volunteer skin using reflectance confocal microscopy. Histological sections from microbiopsied skin revealed 0.22 ± 0.12 mm wide and 0.26 ± 0.09 mm deep puncture sites. Longitudinal observation in microbiopsied volunteers showed that the wound closed within 1 day and was not visible after 7 days. Reflectance confocal microscope images from these same sites showed the formation of a tiny crust that resolved by 3 weeks and was completely undetectable by the naked eye. The design parameters of the device were optimised for molecular analysis using sampled DNA mass as the primary end point in volunteer studies. Finally, total RNA was characterized. The optimised device extracted 5.9 ± 3.4 ng DNA and 9.0 ± 10.1 ng RNA. We foresee that minimally invasive molecular sampling will play an increasingly significant role in diagnostic dermatology and skin research

Influence of channel width and velocity of microbiopsy on DNA, extraction, RNA extraction and pain scores in volunteers

<p>Channel width DNA extracted: total DNA extracted (ng) from different channel widths (mm) in 20 volunteers (v1-v20).</p> <p>Channel width pain scores: the level of pain scored (10 point Likert scale) by 20 volunteers (v1-v20) when applied with different channel widths (mm).</p> <p>Velocity DNA extracted: total DNA (ng) extracted using microbiopsy at different velocities (m/s) in 20 volunteers (v1-v20).</p> <p>Velocity pain scores: the level of pain (10 point Likert scale) scored by 20 volunteers (v1-v20) in response to different velocities (m/s).</p> <p>Roughness amp. DNA extracted: the total DNA (ng) extracted using microbiopsy with different roughness amplitude in 20 volunteers (v1-v20).</p> <p>RNA extracted: the total RNA extracted (ng) from excised AK lesions using 0.15 mm channel width microbiopsies (n=5).</p

Imaging of skin penetration by Nanopatch microprojections.

<p>(A) The size of a single Nanopatch relative to a forefinger. (B+C) SEM images of microprojection morphology after dry-etch fabrication. (D) Representative cryo-SEM images of the mouse ear skin surface following application of a single Nanopatch. D(i) shows a far field view of the corner of the patched area, with micro-channel openings characteristic of microprojection penetration, adjacent to unbroken skin. D(ii) shows perforated area in higher magnification, with a single micro-channel next to a hair follicle inset. Scale bar inset = 10 µm. (E) Representative micrographs of ear tissue sections following delivery of Nanopatch coated with a fluorescent dye. BF: brightfield image, F: fluorescence image, BF+F: both brightfield and fluorescent images overlaid. SC = <i>stratum corneum</i>; VE = viable epidermis; D = dermis.</p

CD8⁺ T cell immunogenicity of Nanopatch-delivered viral vector vaccines in prime boost schedules.

<p>Mice (n = 5/6) were primed with 5×10⁷ PFU MVA.PbCSP (no TH or SC) (A) or 5×10⁹ VP ChAd63.ME-TRAP +10% ^w/_v TH+SC (B), either by coated Nanopatch or ID injection. Two weeks post-MVA.PbCSP priming, a boost immunisation of MVA.PbCSP was given and 8 weeks after ChAd63.METRAP a boost immunisation of MVA.ME-TRAP (no TH or SC) was given (dose 5×10⁷ PFU, given either ID or by Nanopatch). One week post-MVA (A) or 3 weeks post-ChAd63 (B) or 2 weeks post-boost (C+D), blood was taken for analysis of Pb9-specific IFN-γ secreting cells. SFC = spot forming cells. PBMC = peripheral blood mononuclear cells.</p

Viral viability throughout formulation and during Nanopatch dry-coating.

<p>(A+B) ChAd63.ME-TRAP (1×10⁹ VP) and MVA.GFP (1×10⁷ PFU) were mixed with combinations of MC and PS20, with or or without the disaccharides TH+SC (10% ^w/_v each sugar). Formulations were added to DF-1 cell monolayers (A; MVA, n = 4) or HEK-293A cells (B; ChAd63, n = 5) to evaluate viral titre, which was compared to unformulated virus. (C+D) Formulations containing ChAd63.ME-TRAP (C; 1×10⁹ VP) and MVA.GFP (D; 1×10⁷ PFU) were coated onto Nanopatch and immediately eluted into D-MEM. Eluates (n = 4/5) were added to cell monolayers in infectivity assays as before. Eluted viral titres were compared against unformulated, liquid virus. Negative control wells contained D-MEM only. NS = not significant. nd = no data. IFU = infectious units, PFU = plaque forming units.</p