T Cell Transcriptional Profiling and Immunophenotyping Uncover LAG3 as a Potential Significant Target of Immune Modulation in Multiple Myeloma.

Autologous stem cell transplant (ASCT) is the standard of care for patients with multiple myeloma (MM). The clinical significance of peripheral blood T lymphocyte (PBTL) immunologic changes associated with ASCT is poorly understood. Here we evaluated T cell transcriptional messenger RNA profiles and immunophenotypes to correlate immunologic senescence, exhaustion, and anergy with clinical endpoints in a cohort of patients with MM undergoing ASCT. ASCT induced global transcriptional T cell changes and altered molecular levels of markers of T cell subtypes, T cell activation, and exhaustion. These included reduced CD4/CD8 ratio, skewing toward the Th1 subset, reduced expression of costimulatory receptors CD27 and CD28, heightened T cell activation, and increased expression of immune modulatory molecules LAG3 and PD1. Multicolor flow cytometry experiments confirmed altered circulating CD4 and CD8 subsets and skewing toward differentiated effector cells. Moreover, ASCT promoted an exhausted immunophenotype in CD3+CD4+ subsets and a senescent immunophenotype in CD3+CD8+ subsets. Subset-specific altered expression was also seen for surface molecules with immunomodulatory function. ASCT affected soluble levels of molecules with immunomodulatory function by increasing plasma HVEM and TIM3. High molecular LAG3 level was associated with inferior event-free survival post-ASCT (hazard ratio = 5.44; confidence interval, 1.92 to 15.46; P = .001; adjusted P [controlling for false discovery rate] = .038). Using a comprehensive evaluation of PBTLs on a molecular and phenotypic level, we have identified that ASCT induces global T cell alterations with CD4 and CD8 subset-specific changes. Moreover, LAG3 emerged as an early biomarker of adverse events post-ASCT. These findings will support the development of treatment strategies targeting immune defects in MM to augment or restore T cell responses.

Wnt/ß-catenin signaling modulates corneal epithelium stratification via inhibition of Bmp4 during mouse development.

The development of organs with an epithelial parenchyma relies on reciprocal mesenchymal-epithelial communication. Mouse corneal epithelium stratification is the consequence of a coordinated developmental process based on mesenchymal-epithelial interactions. The molecular mechanism underlying these interactions remains unclear. The Wnt/ß-catenin signaling pathway is involved in fundamental aspects of development through the regulation of various growth factors. Here, we show that conditional ablation of either ß-catenin (Ctnnb1(cKO)) or co-receptors Lrp5/6 (Lrp5/6(cKO)) in corneal stromal cells results in precocious stratification of the corneal epithelium. By contrast, ectopic expression of a murine Ctnnb1 gain-of-function mutant (Ctnnb1(cGOF)) retards corneal epithelium stratification. We also discovered that Bmp4 is upregulated in the absence of ß-catenin in keratocytes, which further triggers ERK1/2 (Mapk3/1) and Smad1/5 phosphorylation and enhances transcription factor p63 (Trp63) expression in mouse corneal basal epithelial cells and in a human corneal epithelial cell line (HTCE). Interestingly, mouse neonates given a subconjunctival BMP4 injection displayed a phenotype resembling that of Ctnnb1(cKO). Conditional ablation of Bmp4 eradicates the phenotype produced in Ctnnb1(cKO) mice. Furthermore, ChIP and promoter-luciferase assays show that ß-catenin binds to and suppresses Bmp4 promoter activity. These data support the concept that cross-talk between the Wnt/ß-catenin/Bmp4 axis (in the stromal mesenchyme) and Bmp4/p63 signaling (in the epithelium) plays a pivotal role in epithelial stratification during corneal morphogenesis.

An acryl resin-based swellable microneedles for controlled release intradermal delivery of granisetron.

Swellable microneedles (SMNs) are made of hydrogels and can deliver drug with controlled delivery rate by the cross-link density of the hydrogel. In this study, an acryl resin-based SMNs was developed for poorly water-soluble drugs. The making process of the SMNs is very simple and only need 60 min. The SMNs has high mechanical strength and is not easily broken. In-vitro release of SMNs-loaded model drug, granisetron base (GRB), was investigated. The results showed that seven days controlled release of GRB was obtained when SMNs contained pore-foaming agents (1.5% dicalcium phosphate (CaHPO4) and 1.5% polyvinylpyrrolidone (PVP)). The maximum amount delivered into skin was 86.158 ± 7.82% of the initial GRB (2.1 mg) loaded on SMNs preparation. Pharmacokinetics study in rats indicated a dose-dependent profile of plasma GRB concentrations and that the controlled release of 2.1 mg dose was observed for 144 hours. In conclusion, these SMNs provided a potential minimally invasive route for controlled-release systemic delivery of poorly water-soluble drugs.

Dissolving microneedle-based intradermal delivery of interferon-α-2b.

The dermal and transdermal delivery of protein pharmaceuticals faces enormous challenges, and at the same time, has very significant potential for the non-invasive treatment of both localized and systemic diseases. To demonstrate the pharmaceutical usefulness of dissolving microneedles (MNs) containing interferon-α-2b (IFN), IFN MNs were prepared using a new method. IFN were encapsulated in MNs with dose from 4.94 ± 0.64 to 23.79 ± 2.48 µg, and in vitro release test showed the efficiency reached 49.2%. After percutaneous administration of IFN MNs to rats, serum IFN levels were measured for 12 h. The peak serum IFN level, maximum drug concentration (Cmax), and the time to reach maximum concentration (Tmax), were 11.58 ± ng/ml and 40 min, respectively, for high-dose MNs group. The area under the curve (AUC) of MNs group was 28.85 ng·h/ml, while intramuscular injection (IM) group with equal dose was 31.17 ng·h/ml. Immunogenicity analysis showed the anti-IFN antibody got back to normal level at ninth week, and there was no difference between male and female rats. IFN MNs showed good stability for 2 months and no damage to the administered rats' skin. The results demonstrated the IFN MNs have a great potential to provide an alternative to IM.

Controllable coating of microneedles for transdermal drug delivery.

Coated microneedles have been paid much attention recently, and several coating strategies have been developed to address the problems during coating process. However, there are still some unresolved issues, such as, precise control requirements, microneedle substrate contamination and high processing temperature. The purpose of this study was to develop a simple and controllable method to make uniform coatings on microneedles at room temperature. This novel method avoids the contamination of microneedle substrate by providing both the adsorption force of thickener and micro-scale coating film produced by a newly design device. Thickeners were screened to enhance the mass of coatings. The parameters that influence the coatings were tested systematically, which made coating process controllable. Finally, three model drugs were coated onto microneedles to prove the method is applicable more broadly. In addition, insertion experiments were carried out to test the drug delivery feasibility of the coated microneedles. In conclusion, this study presents a simple and controllable method to coat microneedles with small molecular chemical drugs or large proteins for rapid skin drug delivery.

Industrializable approach for preparing hydrogel microneedles and their application in melanoma treatment.

Microneedles (MNs) technology has been studied in transdermal drug delivery for more than 20 years with hundreds of clinical trials conducted. However, there are currently no commercially available MNs in medicine due to challenges in materials safety, cost-effective fabrication, and large-scale manufacturing. Herein, an approach for rapid and green fabrication of hydrogel microneedles (HMNs) based on infrared irradiation process was proposed for the first time. The optimized formulation consisted of polyvinyl alcohol (PVA) and polyvinyl pyrrolidone (PVP), which acted as cross-linked materials and pore-forming agents, respectively. The manufacturing method involved placing MNs patches under infrared irradiation at 70 °C for 2 min and annealing to obtain HMNs with excellent swelling behavior, mechanical strength, and biocompatibility. When model drugs azelaic acid (AZA) and matrine (MAT) were loaded into HMNs systems, the chemical stability of MAT was significantly improved. Ex vivo transdermal delivery experiments indicated that HMNs could achieve synchronous release of AZA and MAT, and the 24-hour percutaneous permeability rates of both drugs were 73.09 ± 0.48 % and 71.56 ± 1.23 %, respectively. In-vivo pharmacokinetic studies, HMNs administration presented dose-dependent stable blood drug concentrations for both drugs. Additionally, prominent anti-tumor efficacy and biosecurity were observed in the drug-loaded HMNs group in the pharmacodynamic evaluation. In summary, the efficient, convenient, and low-cost fabrication method based on infrared irradiation offers the possibility of mass production of drug-loaded HMNs, showing potential for industrial manufacturing development.

Nanosuspension-Loaded Dissolving Microneedle Patches for Enhanced Transdermal Delivery of a Highly Lipophilic Cannabidiol.

Purpose: Transdermal Drug Delivery System (TDDS) offers a promising alternative for delivering poorly soluble drugs, challenged by the stratum corneum's barrier effect, which restricts the pool of drug candidates suitable for TDDS. This study aims to establish a delivery platform specifically for highly lipophilic drugs requiring high doses (log P > 5, dose > 10 mg/kg/d), to improve their intradermal delivery and enhance solubility. Methods: Cannabidiol (CBD, log P = 5.91) served as the model drug. A CBD nanosuspension (CBD-NS) was prepared using a bottom-up method. The particle size, polydispersity index (PDI), zeta potential, and concentration of the CBD-NS were characterized. Subsequently, CBD-NS was incorporated into dissolving microneedles (DMNs) through a one-step manufacturing process. The intradermal dissolution abilities, physicochemical properties, mechanical strength, insertion depth, and release behavior of the DMNs were evaluated. Sprague-Dawley (SD) rats were utilized to assess the efficacy of the DMN patch in treating knee synovitis and to analyze its skin permeation kinetics and pharmacokinetic performance. Results: The CBD-NS, stabilized with Tween 80, exhibited a particle size of 166.83 ± 3.33 nm, a PDI of 0.21 ± 0.07, and a concentration of 46.11 ± 0.52 mg/mL. The DMN loaded with CBD-NS demonstrated favorable intradermal dissolution and mechanical properties. It effectively increased the delivery of CBD into the skin, extended the action's duration in vivo, and enhanced bioavailability. CBD-NS DMN exhibited superior therapeutic efficacy and safety in a rat model of knee synovitis, significantly inhibiting TNF-α and IL-1ß compared with the methotrexate subcutaneous injection method. Conclusion: NS technology effectively enhances the solubility of the poorly soluble drug CBD, while DMN facilitates penetration, extends the duration of action in vivo, and improves bioavailability. Furthermore, CBD has shown promising therapeutic outcomes in treating knee synovitis. This innovative drug delivery system is expected to offer a more efficient solution for the administration of highly lipophilic drugs akin to CBD, thereby facilitating high-dose administration.

Effective transcutaneous immunization against hepatitis B virus by a combined approach of hydrogel patch formulation and microneedle arrays.

Transcutaneous immunization (TCI) has many advantages compared with needle-based administrations. But the conventional TCI shows poor permeation of antigens across the skin barrier. In this study, Functional MicroArray (FMA) system was used to poke the skin and increase the permeability, and the hydrogel patch formulation was used as the carrier for transdermal delivery of hepatitis B surface antigen (HBsAg) and cholera toxin B (CTB) as an adjuvant. In vitro permeation of antigen was studied using porcine ear skin and rat abdominal skin. The results showed that FMA system could significantly increase the permeation of HBsAg across skin compared with conventional TCI. HBsAg loaded hydrogel formulation exhibited better antigenic thermostability than the liquid formulation. In vivo immunization studies were performed in mice, and the serum IgG titer, IgG2a/IgG1 ratio were measured. The results showed that TCI with FMA induced more potent immune responses than the groups without FMA pretreatment. CTB adjuvanted TCI group could induce higher IgG titers compared with the group without CTB. Furthermore, TCI group can maintain a longer duration of stable IgG titers compared with the intramuscular injection (IM) group. In conclusion, the FMA/hydrogel system was proved to be a potential vaccination strategy against hepatitis B virus.

Clinical Development and Evaluation of a Multi-Component Dissolving Microneedle Patch for Skin Pigmentation Disorders.

Excessive melanin deposition in the skin leads to various skin pigmentation diseases, such as chloasma and age spots. The deposition is induced by several factors, including tyrosinase activities and ultraviolet-induced oxidative stress. Herein, we propose a multi-component, multi-pathway drug combination, with glabridin, 3-O-ethyl-L-ascorbic acid, and tranexamic acid employed as, respectively, a tyrosinase inhibitor, an antioxidant, and a melanin transmission inhibitor. Considering the poor skin permeability associated with topical application, dissolving microneedles (MNs) prepared with hyaluronic acid/poly(vinyl alcohol)/poly(vinylpyrrolidone) were developed to load the drug combination. The drug-loaded microneedles (DMNs) presented outstanding skin insertion, dissolution, and drug delivery properties. In vitro experiments confirmed that DMNs loaded with active ingredients had significant antioxidant and inhibitory effects on tyrosinase activity. Furthermore, the production of melanin both in melanoma cells (B16-F10) and in zebrafish was directly reduced after using DMNs. Clinical studies demonstrated the DMNs' safety and showed that they have the ability to effectively reduce chloasma and age spots. This study indicated that a complex DMN based on a multifunctional combination is a valuable depigmentation product worthy of clinical application.

Soluble Polymer Microneedles Loaded with Interferon Alpha 1b for Treatment of Hyperplastic Scar.

To achieve the painless administration of interferon alpha 1b (rhIFNα-1b), a double-layered soluble polymer microneedle (MN) patch loaded with rhIFNα-1b was used to deliver rhIFNα-1b transdermally. The solution containing rhIFNα-1b was concentrated in the MN tips under negative pressure. The MNs punctured the skin and delivered rhIFNα-1b to the epidermis and dermis. The MN tips implanted in the skin dissolved within 30 min and gradually released rhIFNα-1b. The rhIFNα-1b had a significant inhibitory effect on the abnormal proliferation of fibroblasts and excessive deposition of collagen fibers in the scar tissue. The color and thickness of the scar tissue treated using the MN patches loaded with rhIFNα-1b were effectively reduced. The relative expressions of type I collagen (Collagen I), type III collagen (Collagen III), transforming growth factor beta 1 (TGF-ß1), and α-smooth muscle actin (α-SMA) were significantly downregulated in scar tissues. In summary, the MN patch loaded with rhIFNα-1b provided an effective method for the transdermal delivery of rhIFNα-1b.

Aspirin microcrystals deposited on high-density microneedle tips for the preparation of soluble polymer microneedles.

To reduce mucosal damage in the gastrointestinal tract caused by aspirin, aspirin microcrystals were loaded in soluble polymeric microneedle (MN) tips. Aspirin was prepared into aspirin microcrystals by jet milling. Aspirin microcrystals with particle sizes of 0.5-5 µm were loaded on MN tips with a height of 250 µm or 300 µm. The aspirin microcrystals suspended in a polymer solution were concentrated in the MN tips under negative pressure. The aspirin microcrystals had high stability in the MNs since they were not dissolved in solution during the fabrication process. The MN patch packaged in an aluminum-plastic bag containing silica gel desiccant can be stored at 4 °C. The MN tips implanted in the skin of Institute of Cancer Research (ICR) mice dissolved within 30 min. Isolated porcine ear skin was punctured by MNs with heights of 300 µm and 250 µm to depths of 130 µm and 90 µm, respectively. The fluorescent red (FR) release from MNs reached 98.59% within 24 h. The MNs delivered aspirin microcrystals to the epidermis and dermis, providing a smooth plasma concentration in rats. The MNs loaded with aspirin microcrystals did not evoke primary irritation on the dorsal skin of Japanese white rabbits. In summary, MNs loaded with aspirin microcrystals provide a new approach to improve the stability of aspirin in MN patches.

Dissolvable polymeric microneedles loaded with aspirin for antiplatelet aggregation.

To reduce mucosal damage in the gastrointestinal tract caused by aspirin, we developed a dissolvable polymeric microneedle (MN) patch loaded with aspirin. Biodegradable polymers provide mechanical strength to the MNs. The MN tips punctured the cuticle of the skin and dissolved when in contact with the subcutaneous tissue. The aspirin in the MN patch is delivered continuously through an array of micropores created by the punctures, providing a stable plasma concentration of aspirin. The factors affecting the stability of aspirin during MNs fabrication were comprehensively analyzed, and the hydrolysis rate of aspirin in the MNs was less than 2%. Compared to oral administration, MN administration not only had a smoother plasma concentration curve but also resulted in a lower effective dose of antiplatelet aggregation. Aspirin-loaded MNs were mildly irritating to the skin, causing only slight erythema on the skin and recovery within 24 h. In summary, aspirin-loaded MNs provide a new method to reduce gastrointestinal adverse effects in patients requiring aspirin regularly.

Thermal stability of exenatide encapsulated in stratified dissolving microneedles during storage.

As low-temperature storage and transportation of peptides require high costs, improving the dosage form of peptides can reduce costs. We developed a thermostable and fast-releasing stratified dissolving microneedle (SDMN) system for delivering exenatide (EXT) to patients with type 2 diabetes. Among the tested polymers, dextran and polyvinyl alcohol (PVA) were the best at stabilizing EXT under high-temperature storage for 9 weeks. The two polymers possess a relatively high glass transition temperature (Tg) and weak hydrogen bonding between PVA and EXT. Additionally, zinc sulfate (ZnSO4) had a stabilizing effect on EXT among the selected stabilizers, suggesting that EXT formed a dimer after coordination with zinc ions (Zn2+). In addition, the denaturation temperature (Tm) of EXT was increased by adding ZnSO4, thus stabilizing EXT. Accordingly, SDMNs consisting of a tip layer (dextran encapsulating the Zn2+-EXT complex) and a base layer (PVA) were fabricated. Within 2 min of implantation, the EXT loaded on the patch was quickly released into the skin. Transdermal pharmacokinetics studies showed that manufactured SDMNs generated comparable efficacy to subcutaneous injection. Significantly, the remaining EXT amount was not significantly different under storage at 40 °C and -20 °C for 3 months, supporting that the SDMN system had excellent delivery efficiency and stability, thus reducing the dependence on the cold chain.

Simultaneous basal-bolus delivery of fast-acting insulin and its significance in diabetes management.

Insulin delivery relies on subcutaneous or intravascular injection, leading to reduced patient compliance. Transdermal delivery of insulin has been successfully demonstrated but dose accuracy and skin irritation are problematic in addition to the complex basal-bolus delivery profile required by insulin therapy. Here we present a novel intraepidermal delivery technology (delivered site at epidermis layer, <150 µm) by combining skin pretreatment with short microneedles (<150 µm in length) and iontophoresis transdermal patch (enhanced transport via electrical field) that can provide a continuous basal dose and on-demand bolus dosing for mealtime insulin needs. To our knowledge, this is the first demonstration of therapeutic equivalence between fast-acting human regular insulin and long-acting insulin with possibilities for on-demand dose adjustment. This new intraepidermal delivery technology is likely to change the therapy regimen of patients suffering from insulin-dependent diabetes mellitus and provide a way to lower cost in comparison with insulin pumps and improve patient compliance. FROM THE CLINICAL EDITOR: The authors present a novel intraepidermal insulin delivery technology by combining skin pretreatment with short microneedles and iontophoresis transdermal patch to provide a continuous basal dose and on-demand bolus dosing. This new method is has the potentials to replace insulin pumps by offering a cost effective alternative with less inconvenience and improved compliance.

Enhanced transdermal delivery of 18ß-glycyrrhetic acid via elastic vesicles: in vitro and in vivo evaluation.

OBJECTIVE: The aim of this work was to develop an elastic vesicular formulation to enhance the skin permeation of a poorly water-soluble 18ß-glycyrrhetic acid (GA) and treat dermatitis. METHODS: Elastic vesicles of GA were prepared by the film method with high pressure homogenizer and characterized by storage stability. In vitro permeation studies were carried on rat skin using Franz diffusion cell. In vivo skin deposition of GA was studied using HPLC assay. Chronic allergic contact dermatitis model was built to evaluate pharmacodynamic of GA elastic vesicles. RESULTS: The GA elastic vesicles developed have high flexibility and the storage stability was at least for 6 months at 4°C and for 4 months at 25°C. In vitro cumulative penetration of GA from elastic vesicles within 8 hours was 5.3-fold and 23.2-fold higher than that of conventional liposomes and saturated solution, respectively. After non-occlusive application to mice ears in vivo, skin deposition of GA increased immediately and reached the C(max) at 3 h (1.95 ± 0.32 µg/cm²) and still detected, even after 16 hours GA removed. In vivo anti-inflammatory activity study, GA elastic vesicles showed significant reduction in ear thickness and mass (25.52% and 49.23%) (P < 0.05). The suppressive activity was comparable to that of positive control group (Triamcinolone Acetonide and Econazole Nitrate cream in market), while few side effects were observed in present model. CONCLUSION: The results suggested that of GA elastic vesicular was safe and effective in treatment of contact dermatitis by transdermal administration.

Two-Layer Sustained-Release Microneedles Encapsulating Exenatide for Type 2 Diabetes Treatment.

Daily administration of multiple injections can cause inconvenience and reduce compliance in diabetic patients; thus, microneedle (MN) administration is favored due to its various advantages. Accordingly, the two-layer sustained-release MNs (TS-MNs) were fabricated by encapsulating exenatide (EXT) in calcium alginate (CA) gel in this work. The TS-MNs were composed of a sodium alginate (SA) tip and a water-soluble matrix-containing calcium chloride (CaCl2). Subsequently, the calcium ion (Ca2+) contained in the matrix layer penetrated the tip layer for cross-linking, leaving the drug in the cross-linked network. The patches have adequate mechanical strength to pierce the skin; then, the matrix layer is dissolved, leaving the tip layer to achieve sustained release. Additionally, the TS-MNs encapsulating EXT retained high activity during long-term storage at room temperature. The pharmacokinetic results indicated that the plasma concentrations of EXT were sustained for 48 h in the EXT MN group, which agreed with the in vitro release test. Furthermore, they had high relative bioavailability (83.04%). Moreover, the hypoglycemic effect was observed to last for approximately 24 h after a single administration and remained effective after multiple administrations without drug resistance. These results suggest that the TS-MNs are a promising depot for the sustained delivery of encapsulated EXT.

Process optimization of Ca2+ cross-linked alginate-based swellable microneedles for enhanced transdermal permeability: More applicable to acidic drugs.

We describe a swellable microneedle (SMN) consisting of Ca2+ cross-linked alginate, which expands the types of natural polymers available for SMN fabrication. After investigation of different fabrication methods, the alginate in situ hydrogel-based SMN with a flat substrate was successfully constructed, whose gelation was triggered by ethylenediaminetetraacetic acid calcium disodium salt and D-(+)-glucono-1,5-lactone. With the addition of polyvinyl alcohol and trehalose, SMN possessed good mechanical properties. The biocompatibility of SMN was demonstrated through the tests of in vitro cytotoxicity and in vivo skin irritation. With the assistance of SMN, the in vitro transdermal delivery efficiencies of drugs were significantly improved throughout 16 h. 3-O-ethyl ascorbic acid (EAA, pH = 4.81) exhibited a cumulative release of up to 83.83 ± 6.30%, which was consistent with zero-order kinetics, while tranexamic acid (TA, pH = 6.90) showed the most significant increase in delivery efficiency, which was consistent with the Higuchi model and Ritger-Peppas model. The SMN remained intact after the 16 h of EAA transdermal delivery, indicating its better suitability for acidic drugs. We believe that this technology has the potential to expand the range of drugs available for transdermal administration as well as the breadth of patient care applications.

Preparation, characterization, and in vivo evaluation of levonorgestrel-loaded thermostable microneedles.

To facilitate the storage and use of poly (lactic-co-glycolic acid) (PLGA)-based microneedles (MNs) in hot seasons and regions, thermally stable MNs loaded with levonorgestrel (LNG) were developed. Due to its good biocompatibility and high glass transition temperature (Tg), Hydroxypropyl methylcellulose (HPMC) was added to the PLGA-based MNs to increase thermal stability. MNs with HPMC exhibited excellent thermal stability at high temperatures. After the MNs has been applied to the skin for 10 min, the backing layer of the MNs was dissolved by contact with the interstitial fluid of skin, which resulted in the separation of the MN tips from the backing layer. The MN tips were implanted intradermally and sustained-release LNG. Biodegradable polymers were used to encapsulate the LNG, providing long-acting contraception. The in vitro release rate of LNG from the MNs reached 72.78%-83.76% within 21 days. In rats, the MNs maintained plasma concentrations of LNG above the human contraceptive level for 8-12 days. In mice, the time required for complete degradation of the MN tips was 12-16 days. MNs have excellent medication adherence due to the advantages of painlessness, minimally invasive, and self-administered. MNs can make long-acting contraceptives more readily available to humans.

Design and Evaluation of Complex Polypeptide-Loaded Dissolving Microneedles for Improving Facial Wrinkles in Different Areas.

Wrinkles are one of the most intuitive manifestations of skin aging. Complex polypeptide-loaded dissolving microneedles (CP-DMNs) for facial wrinkles in different areas have been developed and evaluated for the first time. In optimizing formulations, we compared the differences in CP-DMNs heights on skin insertion depth and skin repair and healing. Furthermore, systemic safety experiments were carried out to provide a reference for clinical application. On this basis, an 84-day efficacy assessment based on the improvement of facial wrinkles in different areas and a comparison between CP-DMNs vs. placebo was performed on 30 healthy subjects. As a result, DMNs with a height of 300 µm presented sufficient strength to pierce the stratum corneum with minimized skin damage. In addition, CP-DMNs possessed excellent biological safety and skin compatibility for clinical application. Compared with placebo, CP-DMNs exhibited obvious improvements in wrinkles distributed in the corners of eyes, under-eyes, and nasolabial folds. Furthermore, after using CP-DMNs for 84 days, facial wrinkles in five different areas were smoothed. In short, the complex polypeptides showed apparent anti-wrinkle efficacy with the aid of DMNs technology, and CP-DMNs seemed to work better on deeper wrinkles, such as frown lines and nasolabial folds.

Enhanced bioavailability of L-carnitine after painless intradermal delivery vs. oral administration in rats.

PURPOSE: In vitro and in vivo permeation studies were conducted to evaluate the characteristic of percutaneous administration of high hydrophilic drug L-carnitine (LC) by Functional MicroArray (FMA) painless intradermal delivery system. METHODS: In vitro study was designed to assess the effects of various skins, donor concentration and hydrogels from different carbomer derivatives on the release of LC in a Franz-type diffusion cell. The LC gel patches with carbomer 980 P were prepared and successfully applied to pharmacokinetic study of SD rats with and without FMA. Intravenous injection and oral administration were performed to support pharmacokinetic calculations and comparison of bioavailability. RESULTS: Enhanced delivery of LC using FMA was achieved in skin of different species in vitro studies. The 750 mg LC gel patches were applied to rats over 6 h, and approximately 27% of loaded dose was transported into rat. A 2.8-fold enhancement of absolute bioavailability for LC with FMA intradermal delivery system was observed compared with oral LC administration in vivo study. CONCLUSIONS: Both in vitro and in vivo studies demonstrated that the FMA intradermal delivery system can enhance the delivery and bioavailability of LC.