Antitumor efficacy and reduced toxicity using an anti-CD137 Probody therapeutic.

Costimulation via CD137 (4-1BB) enhances antitumor immunity mediated by cytotoxic T lymphocytes. Anti-CD137 agonist antibodies elicit mild liver inflammation in mice, and the maximum tolerated dose of Urelumab, an anti-human CD137 agonist monoclonal antibody, in the clinic was defined by liver inflammation-related side effects. A protease-activated prodrug form of the anti-mouse CD137 agonist antibody 1D8 (1D8 Probody therapeutic, Pb-Tx) was constructed and found to be selectively activated in the tumor microenvironment. This construct, which encompasses a protease-cleavable linker holding in place a peptide that masks the antigen binding site, exerted antitumor effects comparable to the unmodified antibody but did not result in liver inflammation. Moreover, it efficaciously synergized with both PD-1 blockade and adoptive T-cell therapy. Surprisingly, minimal active Pb-Tx reached tumor-draining lymph nodes, and regional lymphadenectomy did not abrogate antitumor efficacy. By contrast, S1P receptor-dependent recirculation of T cells was absolutely required for efficacy. The preferential cleavage of the anti-CD137 Pb-Tx by tumor proteases offers multiple therapeutic opportunities, including neoadjuvant therapy, as shown by experiments in which the Pb-Tx is given prior to surgery to avoid spontaneous metastases.

Radioprotective Effects from Propolis: A Review.

Propolis is a natural bee-produced substance with antimicrobial, anti-inflammatory, and wound-healing properties, containing some components from the leaves, buds and resins of plants. It has been used for centuries for various health benefits. In this manuscript, our group reviewed the radioprotective effect of propolis using PubMed and Embase, and our review was conducted according to the PRISMA statement. Finally, 27 articles were included in this review, which includes the radioprotective effect of propolis from cell-based studies (n = 8), animal models (n = 14), and human trials (n = 5). Results reflected that the dosage forms of propolis extracted in the scientific literature were ethanolic extracts of propolis, a water-soluble derivate of propolis, or capsules. The efficacy of the radioprotective properties from propolis is extracted from the bibliography, as several compounds of this resinous mixture individually or synergistically are possible candidates that have the radioprotective effect. In fact, studies prior to 2011 lacked a comprehensive characterization of propolis due to the variability in active compounds among different batches of propolis and were limited to analytical techniques. Furthermore, in this manuscript, we have selected studies to include primarily propolis types from Brazil, Croatia, Egypt, European countries, and those commercialized in Spain. They all contained ethanolic extract of propolis (EEP) and were influenced by different dosage forms. EEP showed a significant presence of lipophilic bioactive compounds like flavones, flavonols, and flavanones.

Epicardial cell shape and maturation are regulated by Wt1 via transcriptional control of Bmp4.

The epicardium plays a crucial role in embryonic heart development and adult heart repair; however, the molecular events underlying its maturation remain unknown. Wt1, one of the main markers of the embryonic epicardium, is essential for epicardial development and function. Here, we analyse the transcriptomic profile of epicardial-enriched cells at different stages of development and from control and epicardial-specific Wt1 knockout (Wt1KO) mice. Transcriptomic and cell morphology analyses of epicardial cells from epicardial-specific Wt1KO mice revealed a defect in the maturation process of the mutant epicardium, including sustained upregulation of Bmp4 expression and the inability of mutant epicardial cells to transition into a mature squamous phenotype. We identified Bmp4 as a transcriptional target of Wt1, thus providing a molecular basis for the retention of the cuboidal cell shape observed in the Wt1KO epicardium. Accordingly, inhibition of the Bmp4 signalling pathway both ex vivo and in vivo rescued the cuboidal phenotype of the mutant epicardium. Our findings indicate the importance of the cuboidal-to-squamous transition in epicardial maturation, a process regulated by Wt1.

Cholinergic Senescence in the Ts65Dn Mouse Model for Down Syndrome.

Down syndrome (DS) induces a variable phenotype including intellectual disabilities and early development of Alzheimer's disease (AD). Moreover, individuals with DS display accelerated aging that affects diverse organs, among them the brain. The Ts65Dn mouse is the most widely used model to study DS. Progressive loss of cholinergic neurons is one of the hallmarks of AD present in DS and in the Ts65Dn model. In this study, we quantify the number of cholinergic neurons in control and Ts65Dn mice, observing a general reduction in their number with age but in particular, a greater loss in old Ts65Dn mice. Increased expression of the m1 muscarinic receptor in the hippocampus counteracts this loss. Cholinergic neurons in the Ts65Dn mice display overexpression of the early expression gene c-fos and an increase in the expression of ß-galactosidase, a marker of senescence. A possible mechanism for senescence induction could be phosphorylation of the transcription factor FOXO1 and its retention in the cytoplasm, which we are able to confirm in the Ts65Dn model. In our study, using Ts65Dn mice, we observe increased cholinergic activity, which induces a process of early senescence that culminates in the loss of these neurons.

Annexin A6 modulates TBC1D15/Rab7/StARD3 axis to control endosomal cholesterol export in NPC1 cells.

Cholesterol accumulation in late endosomes is a prevailing phenotype of Niemann-Pick type C1 (NPC1) mutant cells. Likewise, annexin A6 (AnxA6) overexpression induces a phenotype reminiscent of NPC1 mutant cells. Here, we demonstrate that this cellular cholesterol imbalance is due to AnxA6 promoting Rab7 inactivation via TBC1D15, a Rab7-GAP. In NPC1 mutant cells, AnxA6 depletion and eventual Rab7 activation was associated with peripheral distribution and increased mobility of late endosomes. This was accompanied by an enhanced lipid accumulation in lipid droplets in an acyl-CoA:cholesterol acyltransferase (ACAT)-dependent manner. Moreover, in AnxA6-deficient NPC1 mutant cells, Rab7-mediated rescue of late endosome-cholesterol export required the StAR-related lipid transfer domain-3 (StARD3) protein. Electron microscopy revealed a significant increase of membrane contact sites (MCS) between late endosomes and ER in NPC1 mutant cells lacking AnxA6, suggesting late endosome-cholesterol transfer to the ER via Rab7 and StARD3-dependent MCS formation. This study identifies AnxA6 as a novel gatekeeper that controls cellular distribution of late endosome-cholesterol via regulation of a Rab7-GAP and MCS formation.

Caffeic acid skin absorption: Delivery of microparticles to hair follicles.

Caffeic acid (CA) is a polyphenol that can be found in a wide range of vegetal dietary sources. It presents a remarkable antioxidant potential, but what is more interesting from the therapeutic point of view is, that it has demonstrated in vitro antimicrobial properties. Folliculitis is a common skin condition, usually caused by a bacterial or fungal infection, in which hair follicles become inflamed. A typical challenge in dermal application when the actives diffuse passively through the skin in a quick manner, as it is the case of CA, is to provide the effective concentration of the compound at the target site for the sufficient time to finalize the treatment adequately and reduce the possibility to trigger systemic side effects. To achieve this goal, it is necessary to appropriately design the drug delivery system. In this case, we leverage the ability of microparticles to accumulate into the hair follicles to design O/W-emulsions containing CA-loaded controlled-release microparticles. Two different emulsion types containing CA were prepared, one containing free CA and the other containing microencapsulated CA. Traditional and differential tape stripping techniques were performed to investigate drug distribution within the different skin layers and into the hair follicles. The Tape stripping results demonstrated that the tapes S3-S5 and S6-S10 presented a higher total amount of CA. The strips are collected and extracted in groups to assure the extraction of quantifiable amounts of drug. Samples S11-15 and S16-20 show a decrease in the amount of quantified CA, as it was expected. Thus, it can be seen that the amount of active decreases while the stratum corneum depth increases. The retention studies demonstrated that, the microparticles tend to produce a more homogeneous distribution of CA, within the stratum corneum and a higher retention into the hair follicle, which can be attributed to their size and uniformity. Besides, MPs present an additional advantage because they guarantee a continuous release of CA in the target for a prolonged period, allowing the treatment of folliculitis with a single dose until the MPs are removed from the hair follicle by its natural regeneration process or particle depletion of CA.

Effect of freezing and storage temperature on stability and antimicrobial activity of an antibiotic mixture used for decontamination of tissue allografts.

One of the most important risks to be controlled in tissue banking is the infection associated with the clinical use of auto- and allografts. Thus, tissue disinfection protocols are used, in addition to processing in controlled environments. For this purpose, combinations of antibiotics are designed to ensure a broad spectrum of antimicrobial activity. This type of protocol is usually validated by testing its antimicrobial efficacy. In this work, we have studied the effect of several factors on the potential of an antibiotic mixture: container, freezing, storage at 4 °C, storage at - 30 °C and storage at - 80 °C. The molecular stability of the compounds has also been tested, additionally to their efficacy. Our findings show that storage conditions affect the molecular stability of Fungizone and Tobramycin (only in case of frozen storage for the last one). Nevertheless, the solution retains its antimicrobial activity for several weeks. The availability of stored aliquots of disinfectant solution and defining expiry dates for different storage conditions can help to schedule tissue bank tasks.

Annexin A6 and Late Endosomal Cholesterol Modulate Integrin Recycling and Cell Migration.

Annexins are a family of proteins that bind to phospholipids in a calcium-dependent manner. Earlier studies implicated annexin A6 (AnxA6) to inhibit secretion and participate in the organization of the extracellular matrix. We recently showed that elevated AnxA6 levels significantly reduced secretion of the extracellular matrix protein fibronectin (FN). Because FN is directly linked to the ability of cells to migrate, this prompted us to investigate the role of AnxA6 in cell migration. Up-regulation of AnxA6 in several cell models was associated with reduced cell migration in wound healing, individual cell tracking and three-dimensional migration/invasion assays. The reduced ability of AnxA6-expressing cells to migrate was associated with decreased cell surface expression of αVß3 and α5ß1 integrins, both FN receptors. Mechanistically, we found that elevated AnxA6 levels interfered with syntaxin-6 (Stx6)-dependent recycling of integrins to the cell surface. AnxA6 overexpression caused mislocalization and accumulation of Stx6 and integrins in recycling endosomes, whereas siRNA-mediated AnxA6 knockdown did not modify the trafficking of integrins. Given our recent findings that inhibition of cholesterol export from late endosomes (LEs) inhibits Stx6-dependent integrin recycling and that elevated AnxA6 levels cause LE cholesterol accumulation, we propose that AnxA6 and blockage of LE cholesterol transport are critical for endosomal function required for Stx6-mediated recycling of integrins in cell migration.

Nanoencapsulation in lipid-core nanocapsules controls mometasone furoate skin permeability rate and its penetration to the deeper skin layers.

AIMS: The influence of nanoencapsulation of mometasone furoate (MF) in poly(ε-caprolactone) lipid-core nanocapsules (LNC) on its in vitro human skin permeation and penetration was evaluated. METHODS: Semisolid formulations were prepared by increasing the viscosity of LNC using a carbomer (Carbopol(®) Ultrez at 0.5% w/v). Two complementary techniques (the static Franz diffusion cell model and the Saarbrücken penetration model) were used to evaluate skin permeation/penetration. RESULTS: The drug release rate was decreased by nanoencapsulation. The skin permeability of MF was controlled by the nanoencapsulation as well as by increasing the viscosity. Furthermore, the formulation containing the nanoencapsulated MF controlled the amount of drug reaching the deeper skin layers without changing its accumulation in the stratum corneum. CONCLUSION: This formulation is suitable for prolonged treatment of skin disorders which should avoid systemic absorption.

Latest advances in glucose-responsive microneedle-based systems for transdermal insulin delivery.

The development of a self-regulated minimally invasive system for insulin delivery can be considered as the holy grail in the field of diabetes mellitus. A delivery system capable of releasing insulin in response to blood glucose levels would significantly improve the quality of life of diabetic patients, eliminating the need for frequent finger-prick tests and providing better glycaemic control with lower risk of hypoglycaemia. In this context, the latest advances in glucose-responsive microneedle-based transdermal insulin delivery are here compiled with a thorough analysis of the delivery mechanisms and challenges lying ahead in their clinical translation. Two main groups of microneedle-based systems have been developed so far: glucose oxidase-containing and phenylboronic acid-containing systems. Both strategies in combination have also been tested and two other novel strategies are under development, namely electronic closed-loop and glucose transporter-based systems. Results from preclinical studies conducted using these different types of glucose-triggered release systems are comprehensively discussed. Altogether, this analysis from both a mechanistic and translational perspective will provide rationale and/or guidance for future trends in the research hotspot of glucose-responsive microneedle-based insulin delivery systems.

Cyclosporin A-loaded dissolving microneedles for dermatitis therapy: Development, characterisation and efficacy in a delayed-type hypersensitivity in vivo model.

Several drugs can be used for treating inflammatory skin pathologies like dermatitis and psoriasis. However, for the management of chronic and long-term cases, topical administration is preferred over oral delivery since it prevents certain issues due to systemic side effects from occurring. Cyclosporin A (CsA) has been used for this purpose; however, its high molecular weight (1202 Da) restricts the diffusion through the skin structure. Here, we developed a nano-in-micro device combining lipid vesicles (LVs) and dissolving microneedle array patches (DMAPs) for targeted skin delivery. CsA-LVs allowed the effective incorporation of CsA in the hydrophilic DMAP matrix despite the hydrophobicity of the drug. Polymeric matrix composed of poly (vinyl alcohol) (5% w/v), poly (vinyl pyrrolidine) (15% w/v) and CsA-LV dispersion (10% v/v) led to the formation of CsA-LVs@DMAPs with adequate mechanical properties to penetrate the stratum corneum barrier. The safety and biocompatibility were ensured in an in vitro viability test using HaCaT keratinocytes and L929 fibroblast cell lines. Ex vivo permeability studies in a Franz-diffusion cell setup showed effective drug retention in the skin structure. Finally, CsA-LVs@DMAPs were challenged in an in vivo murine model of delayed-type hypersensitivity to corroborate their potential to ameliorate skin inflammatory conditions. Different findings like photon emission reduction in bioluminescence study, normalisation of histological damage and decrease of inflammatory cytokines point out the effectivity of CsA-LVs@DMAPs to treat these conditions. Overall, our study demonstrates that CsA-LVs@DMAPs can downregulate the skin inflammatory environment which paves the way for their clinical translation and their use as an alternative to corticosteroid-based therapies.

Antibiotic resistance and tolerance: What can drug delivery do against this global threat?

Antimicrobial resistance and tolerance (AMR&T) are urgent global health concerns, with alarmingly increasing numbers of antimicrobial drugs failing and a corresponding rise in related deaths. Several reasons for this situation can be cited, such as the misuse of traditional antibiotics, the massive use of sanitizing measures, and the overuse of antibiotics in agriculture, fisheries, and cattle. AMR&T management requires a multifaceted approach involving various strategies at different levels, such as increasing the patient's awareness of the situation and measures to reduce new resistances, reduction of current misuse or abuse, and improvement of selectivity of treatments. Also, the identification of new antibiotics, including small molecules and more complex approaches, is a key factor. Among these, novel DNA- or RNA-based approaches, the use of phages, or CRISPR technologies are some potent strategies under development. In this perspective article, emerging and experienced leaders in drug delivery discuss the most important biological barriers for drugs to reach infectious bacteria (bacterial bioavailability). They explore how overcoming these barriers is crucial for producing the desired effects and discuss the ways in which drug delivery systems can facilitate this process.

Skin drug delivery using lipid vesicles: A starting guideline for their development.

Lipid vesicles can provide a cost-effective enhancement of skin drug absorption when vesicle production process is optimised. It is an important challenge to design the ideal vesicle, since their properties and features are related, as changes in one affect the others. Here, we review the main components, preparation and characterization methods commonly used, and the key properties that lead to highly efficient vesicles for transdermal drug delivery purposes. We stand by size, deformability degree and drug loading, as the most important vesicle features that determine the further transdermal drug absorption. The interest in this technology is increasing, as demonstrated by the exponential growth of publications on the topic. Although long-term preservation and scalability issues have limited the commercialization of lipid vesicle products, freeze-drying and modern escalation methods overcome these difficulties, thus predicting a higher use of these technologies in the market and clinical practice.

Microneedle-assisted transdermal delivery of nanoparticles: Recent insights and prospects.

Transdermal delivery of drugs offers an interesting alternative for the administration of molecules that present certain troubles when delivered by the oral route. It can produce systemic effects or perform a local action when the formulation exerts an optimal controlled drug release or a targeted delivery to the specific cell type or site. It also avoids several inconveniences of the oral administration such as the hepatic first pass effect, gastric pH-induced hydrolysis, drug malabsorption because of certain diseases or surgeries, and unpleasant organoleptic properties. Nanomedicine and microneedle array patches (MAPs) are two of the trendiest delivery systems applied to transdermal research nowadays. However, the skin is a protective barrier and nanoparticles (NPs) cannot pass through the intact stratum corneum. The association of NPs and MAPs (NPs@MAPs) work synergistically, since MAPs assist NPs to bypass the outer skin layers, and NPs contribute to the system providing controlled drug release and targeted delivery. Vaccination and tailored therapies have been proposed as fields where both NPs and MAPs have great potential due to inherent characteristics. MAPs conception and easy use could allow self-administration and therefore facilitate mass vaccination campaigns in undeveloped areas with weak healthcare services. Additionally, nanomedicine is being explored as a platform to personalize therapies in such an important field as oncology. In this work we show recent insights that prove the benefits of NPs@MAPs association and analyze the prospects and the discrete interest of the industry in NPs@MAPs, evaluating different limiting steps that restricts NPs@MAPs translation to the clinical practice. This article is categorized under: Nanotechnology Approaches to Biology > NA Therapeutic Approaches and Drug Discovery > NA.

A Narrative Review of the Herbal Preparation of Ayurvedic, Traditional Chinese, and Kampo Medicines Applied as Radioprotectors.

In recent years, there has been growing scientific interest in the search for natural radioprotectors that can be used to mitigate the effects of radiation on patients, healthcare personnel, and even for space travel. This narrative review covers the past fifty years and focuses on herbal preparations of Ayurvedic, Traditional Chinese, and Kampo Medicines that have the potential to reduce or eliminate the harmful effects of radiation. Our findings highlight ten herbal preparations, namely Abana, Amalakyadi Churna, Amritaprasham, Brahma, Bu-zhong-yi-qi-tang (BZYQT), Chyavanaprasha, Cystone, Geriforte, Mentat, and Triphala, which have demonstrated potential radioprotective effects. This review examines their composition, properties, and possible mechanisms of action in relation to their radioprotective properties. Exploring the ethnobotany of traditional Asian medicine is particularly interesting as it may lead to the discovery of new active compounds with radioprotective properties.

Nanotechnology-based techniques for hair follicle regeneration.

The hair follicle (HF) is a multicellular complex structure of the skin that contains a reservoir of multipotent stem cells. Traditional hair repair methods such as drug therapies, hair transplantation, and stem cell therapy have limitations. Advances in nanotechnology offer new approaches for HF regeneration, including controlled drug release and HF-specific targeting. Until recently, embryogenesis was thought to be the only mechanism for forming hair follicles. However, in recent years, the phenomenon of wound-induced hair neogenesis (WIHN) or de novo HF regeneration has gained attention as it can occur under certain conditions in wound beds. This review covers HF-specific targeting strategies, with particular emphasis on currently used nanotechnology-based strategies for both hair loss-related diseases and HF regeneration. HF regeneration is discussed in several modalities: modulation of the hair cycle, stimulation of progenitor cells and signaling pathways, tissue engineering, WIHN, and gene therapy. The HF has been identified as an ideal target for nanotechnology-based strategies for hair regeneration. However, some regulatory challenges may delay the development of HF regeneration nanotechnology based-strategies, which will be lastly discussed.

Vitamins and Radioprotective Effect: A Review.

The radioprotective effect ex vivo, in vitro and in vivo of vitamins was reviewed using PubMed and Embase and conducted according to the PRISMA statement. A total of 38 articles were included in this review, which includes the radioprotective effect of vitamins from ex vivo, in vitro and in vivo studies. Vitamins A, C, D and E were used alone, in combination or with other nutritional and non-nutritional compounds. The use of vitamins in natural form or supplementation can be useful to reduce the radiation effect in the body, organs and/or cells. Only four (A, C, D and E) out of thirteen vitamins have been detected with radioprotective properties being mainly vitamin E followed by vitamin C, A and D.

Nursing capacity building in health coaching with hospitalised chronic heart failure patients: a quasi-experimental study.

BACKGROUND: Difficulty in adherence to treatment and self-care behaviours is a leading cause of preventable readmission in people with chronic heart failure (CHF). Although there is evidence of benefits of health coaching for the management of this situation, few interventions have been tested in the hospital setting. AIM: To evaluate a coaching programme (H-Coaching) designed to develop nursing capacity in health coaching for chronic heart failure inpatients. METHODS: A quasi-experimental pre-post study including all nurses in a single centre cardiology ward (N = 19). The intervention consisted of two training packages: (1) five theoretical-practical sessions on health-coaching competencies, emotional intelligence, communication and support of chronic heart failure patients in their illness in the hospital setting; and (2) training sessions seven months after the first training package to reinforce the theoretical and practical knowledge. On four occasions, the Competence Instrument of Health Education for the Nursing professional was used to measure nurses' knowledge, skills and attitudes in health coaching for chronic heart failure patients. RESULTS: The difference between the preintervention and postintervention scores were statistically significant for knowledge [mean difference = 1.00 (95% CI -1.45 to -0.51; p = 0.000)], skills in general [mean difference = 0.50 (95% CI -1.41 to -0.21; p = 0.015)] and personal/social skills [mean difference = 1.00 (95% CI -1.10 to -0.01; p = 0.048)]. While attitudinal and affective domains did not differ, there were differences in knowledge and skills. CONCLUSION: The H-Coaching programme proved to be effective for building nursing capacity in health coaching CHF inpatients. Similar programmes designed to improve knowledge in verbal and nonverbal communication techniques, and skills for coaching interventions adapted to meet the needs of individual patients, should be tested in future interventional experimental studies. CLINICAL TRIAL REGISTRATION NUMBER: NCT05300880. IMPACT STATEMENT: To our knowledge, this is the first nursing training intervention in health coaching for chronic heart failure the inpatient setting. This study has demonstrate improvements in both the knowledge and personal and social skills of cardiology nurses with regard to the development of health coaching in a hospital setting. Given the study design, further research is warranted. PLAIN LANGUAGE SUMMARY: Many patients with chronic heart failure have problems in adhering to the treatment and self-care behaviours and this is one of the main causes of preventable readmission. To promote self-care, patients need to be empowered to integrate these habits into their daily lives and we should implement innovative strategies to achieve this. Health coaching is an ideal alternative to this but very few nurses in the hospital cardiology setting are experienced in health coaching. Our study has shown preliminary results demonstrating that a structured theoretical and practical training programme for nurses can improve nurses' knowledge and skills in health coaching for inpatient patients with chronic heart failure. This study provides an opportunity for future research to demonstrate whether nurses with this training have a positive impact on the health outcomes of chronic heart failure patients and, more specifically, on their levels of self-care and empowerment.

α-Enolase, a multifunctional protein: its role on pathophysiological situations.

α-Enolase is a key glycolytic enzyme in the cytoplasm of prokaryotic and eukaryotic cells and is considered a multifunctional protein. α-enolase is expressed on the surface of several cell types, where it acts as a plasminogen receptor, concentrating proteolytic plasmin activity on the cell surface. In addition to glycolytic enzyme and plasminogen receptor functions, α-Enolase appears to have other cellular functions and subcellular localizations that are distinct from its well-established function in glycolysis. Furthermore, differential expression of α-enolase has been related to several pathologies, such as cancer, Alzheimer's disease, and rheumatoid arthritis, among others. We have identified α-enolase as a plasminogen receptor in several cell types. In particular, we have analyzed its role in myogenesis, as an example of extracellular remodelling process. We have shown that α-enolase is expressed on the cell surface of differentiating myocytes, and that inhibitors of α-enolase/plasminogen binding block myogenic fusion in vitro and skeletal muscle regeneration in mice. α-Enolase could be considered as a marker of pathological stress in a high number of diseases, performing several of its multiple functions, mainly as plasminogen receptor. This paper is focused on the multiple roles of the α-enolase/plasminogen axis, related to several pathologies.

Exploration of Microneedle-assisted skin delivery of cyanocobalamin formulated in ultraflexible lipid vesicles.

Vitamin B12 (cyanocobalamin) deficiency is a widespread condition because of its different aetiologies, like malabsorption syndrome or lifestyles as strict veganism that is increasing its incidence and prevalence in developed countries. It has important haematological consequences that require pharmacological treatment. Current therapy consists of oral or parenteral supplements of cyanocobalamin; however, the oral route is discarded for malabsorption syndrome patients and the parenteral route is not well accepted generally. Topical treatments have been suggested as an alternative, but the molecular weight and hydrophilicity of cyanocobalamin limits its diffusion through the skin. Lipid vesicles can allow the transdermal absorption of molecules > 500 Da. The aim of this work was to use different ultraflexible lipid vesicles (transfersomes and ethosomes) to enhance cyanocobalamin transdermal delivery. Vesicles were characterized and lyophilised for long-term stability. The ability to deliver cyanocobalamin through the skin was assessed in vitro using full-thickness porcine skin in Franz diffusion cells. As expected, the best transdermal fluxes were provided by ultraflexible vesicles, in comparison to a drug solution. Moreover, the pre-treatment of the skin with a solid microneedle array boosts the amount of drug that could potentially reach the systemic circulation.