Mouse Skin Research Articles

Impact of Hyaluronic Acid on the Cutaneous T-Cell Lymphoma Microenvironment: A Novel Anti-Tumor Mechanism of Bexarotene.

Cutaneous T-cell lymphoma (CTCL) is a type of non-Hodgkin's lymphoma that primarily affects the skin, rich in hyaluronic acid (HA). HA is a component of the extracellular matrix in the dermis and likely affects the development of CTCL, but the mechanism is poorly understood. Here we show that low-molecular-weight HA (LMWHA) possibly exacerbates CTCL, and bexarotene, already used in CTCL treatment, decreases HA production. We conducted immunohistochemistry, qRT-PCR, immunoblotting, and HA quantification using both mouse and human specimens to evaluate the impact of HA on CTCL. Additionally, we assessed the effect of bexarotene, which is already used for CTCL treatment, on HA metabolism. HA expression was higher in patients' serum and skin sections than in healthy controls. HA extracted from the skin of mice inoculated with tumors showed an increase in LMWHA. LMWHA increased lymphoma cell proliferation in vitro and accelerated tumor formation in mice in vivo. LMWHA also created a favorable environment for tumor cells by affecting fibroblasts, vascular endothelial cells, and tumor-associated macrophages. Thus, increased levels of HA, mainly LMWHA, exacerbate CTCL progression by affecting tumor cells and their microenvironment. Bexarotene treatment reduced the amount of total HA in murine tumor-inoculated skin, as well as the supernatant of cultured normal human dermal fibroblasts (NHDFs) and HuT78 cells. Detailed in vitro analyses showed that bexarotene treatment decreased HA synthase (HAS)1 and HAS2 expression in NHDFs and HAS1 and HAS3, and CEMIP expression in HuT78 cells. Chromatin immunoprecipitation assays revealed that bexarotene reduced retinoid X receptor-α binding to the HAS1 and HAS2 promoters in NHDFs. Bexarotene potentially exerts its anti-tumor effect by reducing HA levels through decreased expression of HAS. These findings provide new insights into the process of CTCL development and additional insights regarding bexarotene treatment.

D-mannose promotes diabetic wound healing through inhibiting advanced glycation end products formation in keratinocytes

BackgroundDiabetic chronic foot ulcers pose a significant therapeutic challenge around the world, resulting in adverse effects and complications in patients. D-mannose is enriched in cirtus peel and exerts beneficial effects among various diseases, especially against inflammation-related disorders.MethodsHere, we examined the potential effect of D-mannose during wound healing process in streptozotocin (STZ)-induced diabetes mice in vivo and by culturing keratinocytes under high glucose condition in vitro. The skin lesion healing was recorded in photos and evaluated by histochemical staining. What’s more, the advanced glycation end products (AGEs) concentration in blood and mice skin was quantified. Apoptotic cells were assessed by flow cytometry and Western blotting. Inflammatory cytokines and cellular differential gene expression levels were measured by real-time PCR. The expression of the AMPK/Nrf2/HO-1 signaling-related molecules was determined by Western blotting.ResultsWe first found that topical supplementation of D-mannose remarkably improved skin wound healing in diabetes mice. Furthermore, both in vivo and in vitro experiments demonstrated that D-mannose reduced the AGEs generation. Mechanistically, D-mannose inhibited AGEs, then upregulated AMPK/Nrf2/HO-1 signaling in the context of high glucose to maintain keratinocyte normal functions, which positively influenced macrophage and fibroblast to accelerate diabetic wound healing. Noteworthily, these protective effects of D-mannose were abolished by the pretreatment with inhibitors of AGEs or AMPK.ConclusionAs far as we know, this is the first study exploring the protective role of D-mannose on diabetic wound healing via topical supplementation. We find that D-mannose protects keratinocytes from high glucose stimulation via inhibition of AGEs formation as well as orchestrates inflammatory microenvironment in diabetic wounded skin, suggesting its supplementation as a potential therapy to promote refractory wound healing in diabetic patients.

Preparation of Ethosome Gel with Total Flavonoids from Vernonia anthelmintica (L.) Willd. for the Treatment of Vitiligo.

Vernonia anthelmintica (L.) Willd. is a traditional medicinal herb in Chinese medicine, extensively used by various ethnic groups due to the numerous advantages derived from its total flavonoids. These benefits encompass anti-inflammatory and antioxidant effects, and the promotion of melanin production, showcasing its significant efficacy in addressing vitiligo. To improve transdermal absorption and enhance the antioxidant effectiveness of the treatment, ethosome containing total flavonoids were prepared utilizing the ultrasound injection technique. The resulting ethosome was then carefully mixed with 0.7% Carbomer 934 gel in equal parts, yielding a gel concentration of 0.302 mg/g. This formulation produced small, consistent ethosome that exhibited high encapsulation efficiency and notable stability. In vitro analyses demonstrated sustained release characteristics of the gel and considerable therapeutic effectiveness against vitiligo resulting from hydroquinone exposure. Histological examinations performed through hematoxylin and eosin (H&E) staining of mouse skin revealed increased melanin production and increased activities of tyrosinase (TYR), cholinesterase (CHE), and mouse monoamine oxidase (MAO), while levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were reduced. These findings underscore the promising effectiveness of this treatment strategy and validate the efficacy of the dosage form.

Casticin reduces rosacea-related inflammation by inhibiting mast cell activation via Mas-related G protein-coupled receptor X2.

Rosacea is a chronic inflammatory disease characterized by persistent erythema, papules, and pustules, mainly on the skin of the face. Rosacea is difficult to treat; therefore, identifying new treatments is crucial. Mas-related G protein-coupled receptor X2 (MRGPRX2)-mediated mast cell (MC) activation is essential in the pathogenesis of rosacea. Casticin has been shown to exert anti-inflammatory effects; however, it remains unclear whether it can inhibit MRGPRX2 in treating rosacea. This study determined the therapeutic efficacy of casticin against rosacea by inhibiting MRGPRX2-mediated MC activation. A mouse model of LL37-induced rosacea-like dermatitis was employed. The pathological changes were evaluated using hematoxylin and eosin (H&E) staining, and MCs and CD4+ T cells were observed. Inflammatory mediators were analyzed using ELISA. Mouse skin lesions were collected for transcriptomic sequencing. We used an MRGPRX2-mediated MC degranulation model to evaluate the inhibitory effects of casticin in vitro. Molecular docking analysis, molecular dynamics simulations, and surface plasmon resonance evaluated the binding between casticin and MRGPRX2. Casticin attenuated the LL37-induced inflammatory phenotype and reactions in rosacea-like dermatitis. RNA-seq data showed that casticin inhibited MC activation in a mouse model of rosacea. Furthermore, casticin significantly reduced CD4 + T-cell infiltration. Moreover, casticin inhibited MC activation as an MRGPRX2 antagonist in vitro and in vivo by influencing the NF-κB signaling pathway. Our study demonstrated that casticin exhibits therapeutic efficacy against rosacea by inhibiting MC activation via MRGPRX2.

Bactericidal Hemostatic Sponge: A Point of Care Solution to Combat Traumatic Injury.

Uncontrollable haemorrhage and associated microbial contamination in the battlefield and civilian injuries pose a tremendous threat to healthcare professionals. Such traumatic wounds often necessitate an effective point-of-care solution to prevent the consequent morbidity owing to blood loss or haemorrhage. However, developing superior hemostatic materials with anti-infective properties remains a challenge. To address this, an injectable, cationic dextran-mesoporous silica nanoparticle-based bactericidal hemostatic sponge (BACSTAT) has been developed. A dual crosslinking approach is adopted through in situ covalent cross-linking through photo polymerization and silica nanoparticle-induced non-covalent interactions. This interconnected macroporous BACSTAT sponge has superior fluid absorption properties and fluid-induced rapid shape recovery of the sponge helps to seal the irregularly shaped wound. Furthermore, this sponge can stimulate a coagulation cascade for rapid blood clotting in mice femoral vein incision and liver puncture model. The optimum sponge exhibited potent antibacterial activity against wide-spectrum Gram-positive and Gram-negative pathogens. Notably, it is completely biocompatible with mammalian cells and mice skin. Significantly, this sponge reduces Pseudomonas aeruginosa burden >99% in mice subcutaneous infection model with substantially lessening inflammatory responses in infected tissues. Collectively, the optimized sponge bears immense potential to be developed as point-of-care solution for military and civilian traumatic injury.

Microfluidic Approach for Enhanced Paeoniflorin Transdermal Delivery: A Comparative Study on Different Chips and Mixing Dynamics

Paeoniflorin is a natural pharmaceutical ingredient with a widely biological activity. However, as a hydrophilic drug, the problem of low transdermal rate limits its clinical application. To overcome this shortage, LUVs were used as biocompatible carriers of paeoniflorin in this study. We prepared paeoniflorin-loaded large unilamellar vesicles (LUVs) with W/O/W structure by microfluidics. We used four kinds of chips to prepare paeoniflorin LUVs and explored the effects of the chip structures on LUVs properties applying both experiments and numerical simulations. The difference of fluid mixing mechanisms was analyzed among four different channels, including straight and curved structures. Then we evaluated the differences in skin permeability among the three groups, paeoniflorin aqueous solution group, drug-loaded liposome group and blank liposome & drug mixture group, using the abdominal skin of male mice. The results showed that the structure of the microfluidic channel was a key factor affecting the flow rate and mixing efficiency. The mixing efficiency further affected the liposome size. The mixing efficiency of curved channel was not better than that of a straight channel due to the low flow rate and long mixing time. By the results of transdermal experiments, LUVs could reduce the transdermal time and increase the total transdermal amount. LUVs effectively improved the transdermal absorption efficiency of paeoniflorin. In conclusion, paeoniflorin LUVs with highly efficient transdermal were successfully prepared by using microfluidics. We explored the underlying fluid dynamics that lead to variations in the preparation with different chip structures. The transdermal effect of the LUVs was verified.Graphical

Characterization of Cutaneous Radiation Syndrome in a Mouse Model Using [18F]F- Fluorodeoxyglucose Positron Emission Tomography.

Ionizing radiation on the skin has the potential to cause various sequelae affecting quality of life and even leading to death due to multi-system failure. The development of radiation dermatitis is attributed to oxidative damage to the skin's basal layer and alterations in immune response, leading to inflammation. Past studies have shown that [18F]F-2-fluoro-2-deoxyglucose positron emission tomography-computed tomography ([18F]F-FDG PET/CT) can be used effectively for the detection of inflammatory activity, especially in conditions like hidradenitis suppurativa, psoriasis, and early atherosclerosis. Since currently there are no specific tests for radiation dermatitis, our study aimed to validate whether radiation dermatitis induced in mice can be accurately visualized and measured using [18F]F-FDG PET/CT. We induced cutaneous radiation syndrome in BALB/c mice with different radiation absorbed doses and monitored symptom development through photography, PET imaging, and histopathology, marking the first attempt at non-invasively quantifying radiation dermatitis effects at the molecular level using PET imaging. Our results showed that there were progressive changes in the dorsal skin of irradiated mice, with notable differences between those exposed to varying doses of radiation. Erythema, epilation, and desquamation were more pronounced in mice exposed to lower doses (25 Gy and 35 Gy) than at 45 Gy; however, by the third week, severe skin deterioration, including ulceration and dermal atrophy, was evident in mice irradiated with 35 Gy and 45 Gy. PET/CT imaging revealed increased [18F]F-FDG uptake in the irradiated dorsal skin area of all mice compared to controls, with more pronounced avidity for the lesion in the 25 Gy and 35 Gy than the 45 Gy. Comparison of tissue-normalized SUVMax values showed that both the 25 Gy and 35 Gy mice exhibited fourfold [18F]F-FDG uptake in the dorsal skin compared to controls, while a twofold uptake was seen at 45 Gy, thus indicating substantial metabolic changes in the dorsal skin induced by radiation exposure. Histopathological analyses correlated with the above findings, demonstrating generalized hypertrophy and epidermal thickening in all irradiated mice compared to controls, with thicker epidermis observed with higher radiation doses and increased destruction of microvasculature. In conclusion, PET/CT emerges as a successful tool for imaging cutaneous radiation syndrome, with the observed dermal changes in irradiated mice closely aligning with metabolic alterations of the affected area.

Study on the Effect of Bee Venom and Its Main Component Melittin in Delaying Skin Aging in Mice.

Bee venom (BV) and its main compound melittin (MLT) have antioxidant, anti-inflammatory, and anti-aging activities; however, very little research has been conducted on their effects on skin aging. In this study, a mouse skin aging model induced by D-galactose was constructed via subcutaneous injection into the scruff of the neck, and different doses of BV and MLT were used as interventions. The anti-aging effects and mechanisms of BV and MLT were explored by detecting the skin morphology and structure, and anti-aging-related factors and performing non-targeted metabolomics of mice. BV and MLT improved dermal and epidermal thickness, boosted the collagen fiber content, increased hydroxyproline and hyaluronic acid levels, and enhanced transcript-level expression of IL-10, Col1a1, and Col3a1, while decreasing that of IL-1β. Metabolomic analysis showed that BV and MLT regulated the levels of some metabolites (compared to those in the skin aging control). BV effectively alleviated skin aging by regulating the pentose phosphate pathway, and pathways associated with carbon, galactose, and β-alanine metabolism, whereas MLT regulated pathways related to lipid metabolism, cholesterol metabolism, and atherosclerosis. This study highlights the potential applicability of BV and MLT in skin aging treatments and cosmetic products.

Subcytotoxic transepidermal delivery using low intensity cold atmospheric plasma

Cold atmospheric plasma (CAP) has been utilized in various medical devices using its oxidative nature. Recent studies have provided evidence that CAP can facilitate the delivery of large, hydrophilic molecules through the epidermis to the dermis. On the other hand, a new approach called low-intensity CAP (LICAP) has been developed, allowing the plasma level to be controlled within a subtoxic range, thereby demonstrating various biological benefits without tissue damage. However, the ability of LICAP to enhance transepidermal delivery in sub-cytotoxic conditions has not been fully investigated. This study aims to determine the sub-cytotoxic range of exposure time for LICAP and, within the range, to investigate the effects of LICAP treatment on transepidermal drug delivery (TED) and mechanisms using human keratinocytes and a mouse model. For the in vitro studies, LICAP treatment was evaluated in human keratinocyte (HaCaT) cells by assessing reactive species production, DNA damage, and cytotoxicity profiles. Within the determined safety range, mechanistic analyses were conducted to examine LICAP-enhanced delivery pathways. mRNA expression and protein levels of tight and adherens junction genes were quantified, and changes in ultramicroscopic morphology of HaCaT monolayers were investigated. Intracellular delivery of fluorescein isothiocyanate (FITC)-dextran was also assessed. For the in vivo studies, E-cadherin expression and the transepidermal delivery (TED) of human epidermal growth factor (hEGF) were analyzed in LICAP-treated mouse dorsal skin. The upper safety range of LICAP exposure time, reducing cell viability by 70% (IC70 or LD30), was estimated at 34.3 s. Within the safety range, LICAP treatment downregulated multiple tight and adherens junction genes in HaCaT cells. Consistent with the in vitro results, the epidermal E-cadherin expression was reduced, and human epidermal growth factor (hEGF) was infiltrated in the dermis of the LICAP-treated mouse skin. Intercellular clefts were detected in the HaCaT cell monolayer immediately following LICAP treatment and intracellular delivery of FITC-dextran was confirmed after LICAP exposure. This study demonstrated that LICAP treatment enhances transepidermal permeation of hEGF, apparently via both paracellular and transcellular routes. Under our study conditions, LICAP treatment seems to be a novel approach to facilitate TED with low safety concerns in vitro. Further translational studies are needed for clinical evaluation.

UVB Induces Sympathetic Nervous System Activation and Norepinephrine Secretion to Regulate TheSkin Color of Mice Through the β2-AR/AP-1 Pathway in Epidermal Keratinocytes.

The aim of this study was to investigate how ultraviolet B (UVB) light regulates AP-1 expression via the β2-adrenergic receptor (β2-AR) in epidermal keratinocytes, which in turn regulates melanin synthesis in melanocytes, thereby modulating downstream melanin production in skin hair follicles and altering mouse skin color. We established a UV-irradiated mouse model to investigate the effects of UV radiation on changes in skin color. By measuring changes in the expression of genes related to cutaneous sympathetic nerves, norepinephrine synthesis and melanin synthesis, we investigated the relationship between β2-AR expression and cutaneous melanogenesis and determined the localization of β2-AR in cells. The results of the siRNA-mediated transfection of keratinized cells with downregulated β2-AR expression were further verified in vitro. Our results suggest that UVB alters the color of the dorsal skin in mice by activating the AP-1/IL-6 pathway, which triggers the sympathetic release of norepinephrine, thereby increasing β2-AR expression in keratinocytes. Overall, our study improves the current understanding of how UVB light influences skin color changes and highlights the complex interplay between ultraviolet radiation and skin physiology.

Canthaxanthin ameliorates atopic dermatitis in mice by suppressing Th2 immune response.

Atopic dermatitis (AD) is a prevalent chronic inflammatory skin disorder characterized by intense pruritus and complex immunopathogenic mechanisms. Recent evidence has highlighted the critical link between dysregulated intestinal microecology and altered immune responses in AD progression. As essential components of the intestinal microenvironment, metabolites play pivotal roles in various physiological processes. Through metabolomic profiling in an AD mouse model, we identified a significant reduction in canthaxanthin (CTX), a bacterial-derived metabolite naturally present in many foods, in AD mice compared to healthy controls. To investigate the therapeutic potential of CTX, we established an AD model by repeatedly applying 2,4-dinitrochlorobenzene (DNCB) to the ears and dorsal skin of mice, successfully inducing AD-like symptoms and lesions. Notably, oral administration of CTX significantly attenuated skin inflammation and reduced serum IgE levels in this DNCB-induced AD model. Both in vivo and in vitro studies demonstrated that CTX treatment effectively suppressed Th2 immune responses. Mechanistically, we found that CTX significantly inhibited the activation of the JAK2-STAT6 signaling pathway in Th2-polarized T cells. Our findings not only demonstrate the therapeutic efficacy of CTX in AD but also elucidate its molecular mechanism in modulating T helper cell subset balance. These insights suggest that CTX could serve as a promising therapeutic agent for AD and potentially other Th2 response-mediated immune disorders.

Targeted delivery of cytotoxic proteins via lipid-based nanoparticles to primary Langerhans cells.

Targeted delivery has emerged as a critical strategy in the development of novel therapeutics. The advancement of nanomedicine hinges on the safe and precise cell-specific delivery of protein-based therapeutics to the immune system. However, major challenges remain, such as developing an efficient delivery system, ensuring specificity, minimizing off-target effects, and attaining effective intracellular localization. Our strategy utilizes lipid-based nanoparticles conjugated with a glycomimetic ligand. These nanoparticles selectively bind to langerin, a C-type lectin receptor expressed on Langerhans cells in the skin. We opted for cytotoxic proteins, namely cytochrome c and saporin, as model proteins to showcase the potential of delivering intact proteins to Langerhans cells. These proteins are recognized for their ability to induce apoptosis upon entry into the cytosol. We observed specific killing of cells expressing langerin in vitro, and in primary Langerhans cells isolated from mouse and human skin ex vivo with minimal off target effects. By delivering functional proteins within lipid nanoparticles to Langerhans cells, our approach offers new potential to deliver effective therapeutics with minimal side effects.

Fatty acid chain modification enhances the serum stability of antimicrobial peptide B1 and activities against Staphylococcus aureus and Klebsiella pneumoniae

Antimicrobial peptides (AMPs) possess broad-spectrum antibacterial properties and low resistance development, making them promising candidates for new antibacterial drugs. Incorporating fatty acid chains into AMPs can increase their hydrophobicity, strengthen membrane affinity, and improve their antibacterial effectiveness and stability. This study introduces fatty acid chains of varying lengths into the naturally derived antimicrobial peptide B1. These modified peptides were evaluated for their antibacterial activity, stability, and biocompatibility to identify the optimal chain length for analogues. The analogues B1-C6 and B1-C8 exhibited significantly enhanced antimicrobial activities against Staphylococcus aureus (S. aureus) and Klebsiella pneumoniae (K. pneumonia), demonstrating better stability and biocompatibility. Following acute toxicity and skin irritation tests on mice, further in vivo tests using a mouse skin inflammation model showed that these peptides significantly restrain bacterial growth and promote wound healing. The skin healing rate in the high-concentration group reached 95.92%, 97.35% 98.42% and 98.17%, respectively. These findings indicated that optimizing the hydrophobic-hydrophilic balance in AMPs is crucial for maximizing their therapeutic potential. This research offers a promising approach for designing effective AMPs to treat infections caused by S. aureus and K. pneumoniae.

α-arbutin prevents UVA-induced skin photodamage via alleviating DNA damage and collagen degradation in NIH-3T3 cells.

Ultraviolet radiation (UV) causes certain side effects to the skin, and their accumulation to a certain extent can lead to accelerated aging of the skin. Recent studies suggest that α-arbutin may be useful in various disorders such as hyperpigmentation disorders, wound healing, and antioxidant activity. However, the role of α-arbutin in skin photodamage is unclear. In this study, under UVA-induced photodamage conditions, α-arbutin treated mouse skin fibroblasts (NIH-3T3) can repair DNA damage and resist apoptosis by reducing the production of reactive oxygen species (ROS) and increasing the phosphorylation of glycogen synthase kinase 3 beta (GSK3β) to orchestra AKT/GSK3β pathway. Meanwhile, α-arbutin can also regulate collagen metabolism and facilitate the replenishment of collagen by targeting the phosphorylation of SMAD3 to mediate the TGFβ/SMAD pathway in NIH-3T3. In conclusion, we found that α-arbutin can mitigate the detrimental effects of skin photodamage induced by UVA irradiation, and provides a theoretical basis for the use of α-arbutin in the treatment of skin photodamage.

Receptor activity-modifying protein 1 regulates the differentiation of mouse skin fibroblasts by downregulating α-SMA expression via suppression of high mobility group AT-hook 1 to promote skin wound repair.

Skin innervation is very important for normal wound healing, and receptor activity-modifying protein 1 (RAMP1) has been reported to modulate calcitonin gene-related peptide (CGRP) receptor function and thus be a potential treatment target. This study aimed to elucidate the intricate regulatory effect of RAMP1 on skin fibroblast function, thereby addressing the existing knowledge gap in this area. Immunohistochemical staining and immunofluorescence (IF) staining were used to measure the dynamic changes in the expression of RAMP1 and α-smooth muscle actin (α-SMA) in skin wound tissue in mice. Mouse skin fibroblasts (MSFs) stably transfected with Tet-on-Flag-RAMP1 overexpression (OE) and Tet-on-Flag control (Ctrl) lentiviruses were constructed for in vitro experiments. High mobility group AT-hook 1 (HMGA1) plasmids and α-SMA plasmids were used to overexpress HMGA1 and α-SMA, respectively. An α-SMA siRNA was used to silence α-SMA. Quantitative real-time polymerase chain reaction (qPCR), western blot and IF staining analyses were used to determine the mRNA and protein levels in the cells in different groups. A scratch wound healing assay was used to evaluate the cell migration ability of different groups. Cleavage under targets and release using nuclease (CUT & RUN) assays and dual-luciferase reporter assays were used to predict and verify the interaction between HMGA1 and the α-SMA promoter. RAMP1 and α-SMA protein expression levels in the dermis changed dynamically and were negatively correlated during dorsal skin wound healing in mice. RAMP1 OE in vitro inhibited the differentiation and promoted the migration of MSFs by decreasing α-SMA expression via the suppression of HMGA1, which was shown for the first time to bind to the α-SMA promoter and increase α-SMA transcription. RAMP1 OE also modulated extracellular matrix (ECM) synthesis and remodeling by promoting collagen III and MMP9 expression and decreasing collagen I, MMP2, and tissue inhibitor of metalloproteinases 1 expression. Our findings suggest that RAMP1 OE decreases differentiation and promotes migration in MSFs by downregulating α-SMA expression via the suppression of HMGA1 and modulates ECM synthesis and remodeling, revealing a novel mechanism regulating α-SMA transcription, providing new insights into the RAMP1-mediated regulation of fibroblast function, and identifying effective nerve-related targets for skin wound repair.

Berberine nanostructured lipid carriers by ultrasonication technique: initial study for skin and oral administration

Abstract Berberine (BBR), a natural bioactive ingredient from Eastern nations, has low solubility and permeability that limits its applications. Hence, berberine nanostructured lipid carriers (BBR-NLCs) were fabricated to improve the drug therapeutic effectiveness. Glycerol monostearate, stearic acid and oleic acid were chosen for lipid base whereas Pluronic F127, Span 80, and Transcutol-P were used as the surfactant and co-surfactant. BBR-NLCs had an average particle size of 82 nm, zeta potential of −32 mV, and narrow size distribution (PDI approximately 0.2), prepared with probe ultrasonication at 490 W in 15 min. In addition, BBR-NLCs prepared at optimized conditions showed around 92% of encapsulation efficiency with drug loading over 5.5%. NLCs presented sustained released through mouse skin, dialysis membrane in Franz cell model, and oral dissolution test, compared with free drug over 24 h. Moreover, blank NLCs even increased the cell viability of HaCaT and HEK293T cells at lower concentrations. NLCs significantly enabled higher interactive BBR quantity with investigated cells. Therefore, BBR-NLCs could be considered as a potential nanocarrier for improved bioavailability of this therapeutic agent.

Beneficial effects of ultrafine bubble shower on a mouse model of atopic dermatitis.

Atopic dermatitis (AD) is a common and relapsing skin disease characterized by skin barrier dysfunction, inflammation, and chronic pruritus. Both cutaneous barrier dysfunction and immune dysregulation are critical etiologies of the pathology of AD. Although various anti-inflammatory pharmacological agents, including cytokine inhibitors and signaling pathway blockers, have been developed recently, keeping the skin clean is of utmost importance in maintaining physiological cutaneous barrier function and avoiding an AD flare. Ultrafine bubbles (UFBs) are less than 1 μm in diameter and usually used to clean medical equipment. A UFB shower is expected to keep skin clean with attention to the temperature and strength of the shower. We examined the effects of a UFB shower on two mouse models of AD: Dermatophagoides farinae body (Dfb)- induced AD in NC/Nga mice and interleukin (IL)-33 transgenic (tg) mice. Each model comprised three groups: UFB shower-treated, normal shower-treated, and untreated. We evaluated the mice using a dermatitis score, scratching counts, histology, and the expression of inflammatory cytokines and skin barrier-related proteins. In the Dfb-induced AD mouse model, clinical features improved markedly in the UFB shower-treated mice compared to other groups. IL-4 and IL-13 levels decreased in the skin of normal and UFB shower-treated mice. In addition, in the skin of UFB shower-treated mice, the expression levels of skin barrier-related proteins were increased compared to normal showertreated mice. However, we found no significant differences in IL33tg mice. These results suggest that UFB shower can recover the skin barrier function and improve skin inflammation, especially in conditions such as extrinsic AD.

Indole-3-carbinol alleviates allergic skin inflammation via periostin/thymic stromal lymphopoietin suppression in atopic dermatitis.

Atopicdermatitis(AD) is a chronic multifactorial inflammatory skin disorder with a complex etiology. Despite its increasing prevalence, treatment of AD is still limited.Indole-3-carbinol (I3C) is found in cruciferous vegetables and is formed when these vegetables are cut, chewed, or cooked; it exerts diverse pharmacological activities. HaCaT keratinocytes stimulated with tumor necrosis factor-α and interferon-γ mixture and NC/Nga mice stimulated with 2,4-dinitrochlorobenzen (DNCB) were used for AD models, in vitro and in vivo, respectively. The results showed that I3C reduced the expression of pro-inflammatory cytokines, thymic stromal lymphopoietin (TSLP), and periostin in in vitro model. Oral administration of I3C alleviated AD-like skin inflammatory symptoms, including serum IgE levels, epidermal thickening, inflammatory cell infiltration, transepidermal water loss, and scratching behavior. Moreover, I3C decreased the expression of TSLP and periostin and recovered the expression of skin barrier proteins by regulating Aryl Hydrocarbon Receptor and inhibiting the mitogen-activated protein kinase and nuclear factor-κB pathways in the skin of DNCB-induced AD mice. I3C is suggested as a potential therapeutic alternative for the treatment of AD by repressing allergic inflammatory pathways.

Effect of non-invasive fractional photothermolysis on the efficacy of transdermal photosensitization in the experiment in vivo

In an in vivo pilot study, the efficiency of noninvasive fractional laser photothermolysis (NFLP) as a transdermal system for application photosensitization of mouse skin before photodynamic therapy (PDT) was studied. For NFLP, a laser (λ = 970 nm) with an average power of 4 W and a pulse frequency of 50 Hz was used. An area of the skin of the anterior abdominal wall of mice was irradiated. After NFLP, a photosensitizer (PS) based on chlorin e6 in the form of a gel (0.5%) was applied to the skin with an application time of 30 min. Then, laser PDT (λ = 662 nm) was performed with a power of 2 W in a scanning pulse-periodic mode with a frequency of 5 Hz and a light spot area on the skin of 1.2 mm². The results of histological examination, confocal and electron microscopy showed the features of transdermal distribution of chlorin e6 after NFLP. PS fluoresces in all skin layers and the subcutaneous fat layer, indicating its deep penetration into the hypodermis after NFLP compared to conventional cutaneous application. The advantages of NFLP as a transport system for successful penetration of the gel form of chlorin e6 through all skin layers are demonstrated. Electron microscopy showed transdermal transport of PS in the form of nanosized microspheres and particles absorbed by macrophages and fibroblasts. It was also shown for the first time that pulsed PDT after NFLP leads to the formation of nanosized foci of photodestruction up to the border of the reticular layer of the skin and the hypodermis.

Gasdermin D-mediated neutrophil pyroptosis drives inflammation in psoriasis

Psoriasis is a multifactorial immune-mediated inflammatory disease. Its pathogenesis involves abnormal accumulation of neutrophils and T-cell-related abnormalities. Pyroptosis is a type of regulated cell death associated with innate immunity, but its role in psoriasis is unclear. In this study, we found that gasdermin D (GSDMD) is higher in human psoriatic skin than that in normal skin, and in imiquimod-induced psoriasis-like mouse skin, the expression of Gsdmd was most significantly altered in neutrophils and Il1b was also mainly expressed in neutrophils. Immunohistochemical staining of serial sections of skin lesions from psoriasis patients and healthy control also showed that GSDMD expression is higher in psoriasis lesion, especially in neutrophils. Gsdmd deficiency mitigates psoriasis-like inflammation in mice. GSDMD in neutrophils contributes to psoriasis-like inflammation, while Gsdmd depletion in neutrophils attenuates the development of skin inflammation in psoriasis and reduces the release of the inflammatory cytokines. We found that neutrophil pyroptosis is involved in and contributes to psoriasis inflammation, which provides new insights into the treatment of psoriasis by targeting neutrophil pyroptosis.