Dissolvable microneedles in the skin: Determination the impact of barrier disruption and dry skin on dissolution.

Dissolvable microneedles (DMNs), fabricated from biocompatible materials that dissolve in both water and skin have gained popularity in dermatology. However, limited research exists on their application in compromised skin conditions. This study compares the hyaluronic acid-based DMNs penetration, formation of microchannels, dissolution, and diffusion kinetics in intact, barrier-disrupted (tape stripped), and dry (acetone-treated) porcine ear skin ex vivo. After DMNs application, comprehensive investigations including dermoscopy, stereomicroscope, skin hydration, transepidermal water loss (TEWL), optical coherence tomography (OCT), reflectance confocal laser scanning microscopy (RCLSM), confocal Raman micro-spectroscopy (CRM), two-photon tomography combined with fluorescence lifetime imaging (TPT-FLIM), histology, and scanning electron microscopy (SEM) were conducted. The 400 µm long DMNs successfully penetrated the skin to depths of ≈200 µm for dry skin and ≈200-290 µm for barrier-disrupted skin. Although DMNs fully inserted into all skin conditions, their dissolution rates were high in barrier-disrupted and low in dry skin, as observed through stereomicroscopy and TPT-FLIM. The dissolved polymer exhibited a more significant expansion in barrier-disrupted skin compared to intact skin, with the smallest increase observed in dry skin. Elevated TEWL and reduced skin hydration levels were evident in barrier-disrupted and dry skins compared to intact skin. OCT and RCLSM revealed noticeable skin indentation and pronounced microchannel areas, particularly in barrier-disrupted and dry skin. Additional confirmation of DMN effects on the skin and substance dissolution was obtained through histology, SEM, and CRM techniques. This study highlights the impact of skin condition on DMN effectiveness, emphasizing the importance of considering dissolvability and dissolution rates of needle materials, primarily composed of hyaluronic acid, for optimizing DMN-based drug delivery.

Characterization of a bioscaffold containing polysaccharide acemannan and native collagen for pulp tissue regeneration.

Dental pulp regeneration exploits tissue engineering concepts using stem cells/scaffolds/growth-factors. Extracted collagen is commonly used as a biomaterial-scaffold due to its biocompatibility/biodegradability and mimics the natural extracellular matrix. Adding biomolecules into a collagen-scaffold enhanced pulp regeneration. Acemannan, ß-(1-4)-acetylated-polymannose, is a polysaccharide extracted from aloe vera. Acemannan is a regenerative biomaterial. Therefore, acemannan could be a biomolecule in a collagen-scaffold. Here, acemannan and native collagen were obtained and characterized. The AceCol-scaffold's physical properties were investigated using FTIR, SEM, contact angle, swelling, pore size, porosity, compressive modulus, and degradation assays. The AceCol-scaffold's biocompatibility, growth factor secretion, osteogenic protein expression, and calcification were evaluated in vitro. The AceCol-scaffolds demonstrated higher hydrophilicity, swelling, porosity, and larger pore size than the collagen scaffolds (p < 0.05). Better cell-cell and cell-scaffold adhesion, and dentin extracellular matrix protein (BSP/OPN/DSPP) expression were observed in the AceCol-scaffold, however, DSPP expression was not detected in the collagen group. Significantly increased cellular proliferation, VEGF and BMP2 expression, and mineralization were detected in the AceCol-scaffold compared with the collagen-scaffold (p < 0.05). Computer simulation revealed that acemannan's 3D structure changes to bind with collagen. In conclusion, the AceCol-scaffold synergistically provides better physical and biological properties than collagen. The AceCol-scaffold is a promising material for tissue regeneration.

Delivery and diffusion of retinal in dermis and epidermis through the combination of prodrug nanoparticles and detachable dissolvable microneedles.

To minimize chemical degradation of retinal, we graft this aldehyde on chitosan chains to make them self-assemble into pro-retinal nanoparticles (PRNs), which we then load into detachable dissolvable microneedles (DDMNs) made of 1:1 (by weight) hyaluronic acid/maltose. The presence of PRNs in the hyaluronic acid-maltose needle matrix also helps improve the microneedles' mechanical strength. Ex vivo administration of PRN-loaded DDMNs on fresh porcine ear skin shows, as observed by stereomicroscopic and confocal fluorescence microscopic analyses of the cross-sectioned tissue pieces, complete deposition followed by dissolution of the needles and diffusion of the PRNs in epidermis and dermis. Rats administered with a single dose of PRN-loaded DDMNs show significantly increased epidermal thickness as compared to rats administered with control DDMNs (no PRN). Both the PRN-loaded DDMNs and the control DDMNs produce no skin irritation in rats.

Detachable dissolvable microneedles: intra-epidermal and intradermal diffusion, effect on skin surface, and application in hyperpigmentation treatment.

Delivering bioactive compounds into skin tissue has long been a challenge. Using ex vivo porcine and rat skins, here we demonstrate that a detachable dissolvable microneedle (DDMN) array, a special dissolvable microneedle that allows needle detachment from the base within 2 min post administration, can effectively embed a model compound into epidermis and dermis. Diffusion of the compound from the needle embedding sites to the nearby skin tissue is demonstrated at various post administration periods. The relationship between the time that a conventional dissolvable microneedle array is left on skin without needle detachment from the base and the degree of skin surface abrasion at each microneedle penetration spot is also demonstrated on skin of human volunteers. Co-loading glutathione with vitamin C (vitC) can stabilize vitC in the DDMN. DDMN loaded with vitC and glutathione can help erasing post-acne-hyperpigmentation spots.

Pathogenesis of Thai duck Tembusu virus in Cherry Valley ducks: The effect of age on susceptibility to infection.

Several duck Tembusu virus (DTMUV) clusters have been identified since its first emergence in 2010. However, the pathogenesis evaluation of DTMUV has been restricted to cluster 2.2 Chinese DTMUVs. In this study, the pathogenesis of a cluster 2.1 Thai DTMUV was investigated in three ages of Cherry Valley ducks (1-, 4- and 27-week-old). In each age, 35 ducks were inoculated with a cluster 2.1 Thai DTMUV and evaluated for clinical signs, virus distribution and shedding, pathology and serological response. Our results demonstrated that all duck ages were susceptible to Thai DTMUV; however, Thai DTMUV induced greater disease severity in younger ducks (1- and 4-week-old) when compared to older ducks (27-week-old) reflected by higher morbidity and mortality rates, and higher degree of pathological severity. Corresponding to these results, longer-term viremia, higher levels of viral loads in tissues and lower neutralizing antibody titers were also observed in younger ducks compared to those in older ducks. However, it should be noted that a significant drop in egg production was found in older ducks, which also indicates the susceptibility to Thai DTMUV in older ducks. Interestingly, prolonged shedding period with high viral loads was observed in older ducks even without showing clinical signs, suggesting the potential role of the older ducks as the carriers of Thai DTMUV. This finding highlights the importance of monitoring DTMUV and preventing the transmission of DTMUV in adult ducks. Overall, this study provides insights into the pathogenesis and infection dynamics of a cluster 2.1 Thai DTMUV in ducks.

Microneedle-Facilitated Intradermal Proretinal Nanoparticle Delivery.

Topical retinoid treatments stimulate biological activities in the skin. The main physical barrier, which limits the efficacy of transdermal drug delivery, is the stratum corneum. Proretinal nanoparticles (PRN) have already been proven to efficiently deliver retinal into the epidermis. In the present study, two transdermal drug delivery systems, microneedles (MN) and PRN, were combined to directly target the dermis. The microchannels induced by the MN, the PRN localization in the microchannels and the skin closure kinetics were investigated by non-invasive imaging techniques, such as dermoscopy, optical coherence tomography and multiphoton tomography. Additionally, the amount of retinal in the epidermis and dermis after application in three different forms (PRN-Loaded microneedles, PRN suspension or conventional retinal solution) was compared. All imaging techniques confirmed the formation of microchannels in the skin, which were partly still detectable after 24 h. Multiphoton tomography showed the release of PRN from the MN within the microchannels. The recovered retinal concentration in the dermis was significantly higher when applied via PRN-loaded microneedles. We hypothesized that this platform of PRN-loaded microneedles can provide a rapid and efficient administration of retinal in the dermis and could be of benefit in some skin conditions such as atrophic scar or photo-aged skin.

Solid Composite Material for Delivering Viable Cells into Skin Tissues via Detachable Dissolvable Microneedles.

Delivering cells to desired locations in the body is needed for disease treatments, tissue repairs, and various scientific investigations such as animal models for drug development. Here, we report the solid composite material that when embedded with viable cells, can temporarily keep cells alive. Using the material, we also show the fabrication of detachable dissolvable microneedles (DMNs) that can instantly deliver viable cells into skin tissue. B16-F10-murine-melanoma (B16-F10) and human-embryonic-kidney-293T (HEK293T) cells embedded in the solid matrix of the hyaluronic/polyvinylpyrolidone/maltose (HA/PVP/maltose) mixture show 50.6 ± 12.0 and 71.0 ± 5.96% survivals, respectively, when kept at 4 °C for 24 h. Detachable DMNs made of the HA/PVP/maltose mixture and loaded with B16-F10-cells were constructed, and the obtained DMN patches could detach the cell-loaded needles into the skin within 1 min of patch application. In vivo intradermal tumorgrafting mice with the DMNs containing 800 cells of B16-F10 developed tumors 10 times bigger in volume than tumors induced by hypodermic needle injection of suspension containing 100,000 cells. We anticipate this work to be a starting point for viable cell encapsulation in the solid matrix and viable cell delivery via DMNs.

Topical Proretinal Nanoparticles: Biological Activities, Epidermal Proliferation and Differentiation, Follicular Penetration, and Skin Tolerability.

Proretinal nanoparticles, the retinilidene-chitosan nanoparticles, have been developed to overcome the physicochemical instability of retinal and to lessen the dose-dependent cutaneous irritation, through sustaining the release of retinoid. Compared to conventional retinal at the same concentration, proretinal nanoparticles had no cytotoxicity and could induce a spontaneously immortalized human keratinocyte line to express more cellular retinoic acid binding protein-2. Compared to rats topically applied with conventional retinal which showed clear skin irritation and inflammation, daily topical application of proretinal nanoparticles to rats for 28 consecutive days produced neither irritation nor inflammation but significantly increased epidermal proliferation, epidermal thickness, cellular retinoic acid binding protein- 2 expression, and up-regulation of various differentiation markers including keratin 5, keratin 10, keratin 14, cellular retinoic acid binding protein-2, and proliferating cell nuclear antigen. Through the use of confocal laser scanning microscopy, we observed the in vivo follicular penetration of proretinal nanoparticles with the depth of penetration independent of postapplication time. Proretinal nanoparticles provide better biological activities of retinoids on epidermis and could eliminate the side effect of retinoid dermatitis.

Increasing the percutaneous absorption and follicular penetration of retinal by topical application of proretinal nanoparticles.

Topical retinoids are frequently applied for therapeutic and cosmeceutical reasons although their bioavailability is low due to their chemical and photochemical instability. Moreover, skin irritation is a common side effect. Therefore, proretinal nanoparticles (PRN) as a novel formulation of topical retinoids, which are based on chitosan grafted with retinal through reversible linkage, were developed and their skin penetration behavior was studied. As nanoparticles preferably penetrate into the hair follicles, the follicular penetration depths of PRN at different time points were investigated. Moreover, the release capacity of the nanoparticulate system was studied using fluorescein as a model drug. Additionally, the concentration of retinal in the stratum corneum and in the hair follicles was quantified after application in particulate and non-particulate form. The results showed that the nanocarriers reached the infundibular area of the hair follicles, irrespective of the incubation time. The nanoparticles were able to release their model drug within the hair follicle. The retinal concentration delivered to the stratum corneum and the hair follicles was significantly higher when retinal was applied in the particulate form. In conclusion, the presented proretinal nanoparticle system may help to overcome the main problems of topical retinoid therapy, which are skin irritation, chemical and photochemical instability and low bioavailability, thus improving the topical retinoid therapy.

Proretinal nanoparticles: stability, release, efficacy, and irritation.

Despite many potent biological activities, retinoids such as retinoic acid (RA) and retinal possess dose-related broad side effects. In this study, we show that this problem, which has been unsolvable for a long time, can be tackled through a controlled release strategy in which retinal is continuously delivered to the skin via sustained release from proretinal nanoparticles. The water dispersible proretinal nanoparticles are stable when kept in water at neutral pH and at room temperature for 8 months under light-proof conditions, and show sustained release of retinal into human synthetic sebum at a pH of 5. In the daily topical application tests performed for 4 weeks on rats' skin, the nanoparticles showed superior ability to increase epidermal thickness compared to RA and retinal, with no skin irritation observed for the proretinal particles, but severe skin irritation observed for RA and free retinal. When tested under occlusion conditions in human volunteers, insignificant skin irritation was observed for the proretinal nanoparticles. The 12-week, double-blind, split-face study on human volunteers indicates better antiaging efficacy of the particles as compared to the free RA.