Skin Architecture Research Articles

Ribbontail Stingray Skin Employs a Core–Shell Photonic Glass Ultrastructure to Make Blue Structural Color

AbstractStructural blue colors are common in animals, with the tissue nanostructures and material systems that produce them—especially bright blues—typically based on highly ordered nano‐architectures. In this study, we describe an unusually bright and angle‐independent structural blue from the skin of ribbontail stingray, arising from a more disordered array of scattering elements with a previously undescribed core–shell ultrastructure, involving nano‐vesicles enclosing guanine nano‐platelets. We show that this skin architecture functions as an intracellular photonic glass, coherently scattering blue, while broadband absorption from closely associated melanophores obviates the low color saturation typical for photonic glasses. Our characterization of skin ultrastructure and color in a stingray demonstrates how disordered systems can be harnessed to produce brilliant hues while illustrating that the capacity for guanine‐based colors likely arose extremely early in vertebrate evolution. Moreover, the material‐structure‐function associations underlying ribbontail stingray coloration, employing two distinct photonic phenomena, illustrate how the evolution of nanoscale architectures can have profound effects at much larger size scales (e.g., in visual ecology and communication), and provide fundamental guidelines for color‐saturated manmade photonic glasses.

In Vivo Wound Healing Potential and Molecular Pathways of Amniotic Fluid and Moringa Olifera-Loaded Nanoclay Films.

Cutaneous wounds pose a significant health burden, affecting millions of individuals annually and placing strain on healthcare systems and society. Nanofilm biomaterials have emerged as promising interfaces between materials and biology, offering potential for various biomedical applications. To explore this potential, our study aimed to assess the wound healing efficacy of amniotic fluid and Moringa olifera-loaded nanoclay films by using in vivo models. Additionally, we investigated the antioxidant and antibacterial properties of these films. Using a burn wound healing model on rabbits, both infected and non-infected wounds were treated with the nanoclay films for a duration of twenty-one days on by following protocols approved by the Animal Ethics Committee. We evaluated wound contraction, proinflammatory mediators, and growth factors levels by analyzing blood samples. Histopathological changes and skin integrity were assessed through H&E staining. Statistical analysis was performed using SPSS software (version 2; Chicago, IL, USA) with significance set at p < 0.05. Our findings demonstrated a significant dose-dependent increase in wound contraction in the 2%, 4%, and 8% AMF-Me.mo treatment groups throughout the study (p < 0.001). Moreover, macroscopic analysis revealed comparable effects (p > 0.05) between the 8% AMF-Me.mo treatment group and the standard treatment. Histopathological examination confirmed the preservation of skin architecture and complete epidermal closure in both infected and non-infected wounds treated with AMF-Me.mo-loaded nanofilms. RT-PCR analysis revealed elevated concentrations of matrix metalloproteinases (MMPs) and vascular endothelial growth factor (VEGF), along with decreased levels of tumor necrosis factor-alpha (TNF-α) in AMF-Me.mo-loaded nanofilm treatment groups. Additionally, the antimicrobial activity of AMF-Me.mo-loaded nanofilms contributed to the decontamination of the wound site, positioning them as potential candidates for effective wound healing. However, further extensive clinical trials-based studies are necessary to confirm these findings.

Ultrafast readout, crosstalk suppression iontronic array enabled by frequency-coding architecture

The development of iontronic skin (I-skin) capable of ultrafast sensing in a wide pressure range, comparable to human skin, is of paramount importance for intelligent robotics. However, this remains a major challenge due to the lack of iontronic array architectures that can achieve ultrafast readout and crosstalk-free under large capacitance response generated within a wide pressure range. Here, we report a frequency-coding architecture of artificial ion mechanoreceptor skin (AIM-skin) that can provide a universal mode of iontronic array sensing and bypass the dependence of complex integrated back-end interface electronics. Notably, the successful implementation of orthogonal frequency coding in the AIM-skin with high sensitivity and ultrawide pressure range achieve ultrafast parallel readout for the spatiotemporal mechanical stimuli. Furthermore, the parallel zero-potential mechanism (PZPM) of the architecture effectively mitigates electrical crosstalk between sensing units. We have demonstrated that combhination of proposed device and deep learning has a broad application prospect in intelligent human-machine interaction and real-time dynamic robotic manipulation.

An optimized protocol for generating appendage-bearing skin organoids from human-induced pluripotent stem cells.

Organoid generation from pluripotent stem cells is a cutting-edge technique that has created new possibilities for modelling human organs in vitro, as well as opening avenues for regenerative medicine. Here, we present a protocol for generating skin organoids (SKOs) from human-induced pluripotent stem cells (hiPSCs) via direct embryoid body formation. This method provides a consistent start point for hiPSC differentiation, resulting in SKOs with complex skin architecture and appendages (e.g. hair follicles, sebaceous glands, etc.) across hiPSC lines from two different somatic sources.

Exploring the potential of an Aloe vera and honey extract loaded bi-layered nanofibrous scaffold of PCL-Col and PCL-SBMA mimicking the skin architecture for the treatment of diabetic wounds.

Diabetic wounds are often chronic in nature, and issues like elevated blood sugar, bacterial infections, oxidative stress and persistent inflammation impede the healing process. To ensure the appropriate healing of wounds, scaffolds should promote complete tissue regeneration in wounds, both functionally and structurally. However, the available scaffolds lack the explicit architecture and functionality that could match those of native skin, thus failing to carry out the scar-free skin regeneration in diabetic wounds. This study deals with the synthesis of a bi-layered nanofibrous scaffold mimicking the native skin architecture in terms of porosity and hydrophobic-hydrophilic gradients. In addition, herbal extracts of Aloe vera and litchi honey were added in consecutive layers to manage the high blood glucose level, inflammation, and increased ROS level associated with diabetic wounds. In vitro studies confirmed that the prepared scaffold with herbal extracts showed enhanced proliferation of skin cells with good mechanical strength, degradability, anti-bacterial and anti-diabetic properties. The scaffold also demonstrated superior wound healing in vivo with quicker scar-free wound recovery and appropriate skin regeneration, compared to conventional treatment. Altogether, the synthesized herbal extract loaded bi-layered nanofibrous scaffold can be used as a regenerative template for hard-to-heal diabetic wounds, offering a new strategy for the management of chronic wounds.

Design and Bioprinting of Decellularized Extracellular Matrix-Based Bioinks for Skin Tissue Engineering

3D bioprinting is a powerful enabling technology for the automated fabrication of biomimetic constructs for skin modelling and repair with high resolution and reproducibility. Bioinks, which often comprise the combination of cells and printable biomaterials, are essential for bioprinting 3D cell-laden constructs emulating the architecture, composition and function of native skin. Tissue- and organ-specific decellularized extracellular matrix (dECM) materials are assuming a significant role in bioink design owing their capacity to provide native biophysical and biochemical signals that can elicit distinct cellular responses toward tissue development. Herein, we discuss the rational design of dECM-based bioinks, focusing on processing methods to obtain dECM as well as on bioprinting strategies to engineer 3D constructs for applications in wound healing and in vitro skin modelling.

Molecular Organization of the Skin Barrier.

Cryo-electron microscopy of vitreous sections allows for investigation directly in situ of the molecular architecture of skin. Recently, this technique has contributed to the elucidation of the molecular organization of the skin's permeability barrier and its stepwise formation process. The aim of this review is to provide an overview of the procedure for cryo-electron microscopy of vitreous sections, its analysis using atomic detail molecular dynamics modelling and electron microscopy simulation, and its application in the investigation of the barrier structure and formation process of the skin.

Chitosan-sodium percarbonate-based hydrogels with sustained oxygen release potential stimulated angiogenesis and accelerated wound healing.

The prolonged hypoxic conditions hinder chronic wounds from healing and lead to severe conditions such as delayed re-epithelialization and enhanced risk of infection. Multifunctional wound dressings are highly required to address the challenges of chronic wounds. Herein, we report polyurethane-coated sodium per carbonate-loaded chitosan hydrogel (CSPUO2 ) as a multifunctional dressing. The hydrogels (Control, CSPU, and CSPUO2 ) were prepared by freeze gelation method and the developed hydrogels showed high porosity, good absorption capacity, and adequate biodegradability. The release of oxygen from the CSPUO2 hydrogel was confirmed by the increase in pH and a sustained oxygen release was observed over the period of 21 days, due to polyurethane (CSPU) coating. The CSPUO2 hydrogel exhibited around 2-fold increased angiogenic potential in CAM assay when compared with Control and CSPU dressing. CSPUO2 also showed good level of antibacterial efficacy against E. coli and S. aureus. In a full-thickness rat wound model, CSPUO2 hydrogel considerably accelerated wound healing with exceptional re-epithelialization granulation tissue formation less inflammatory cells and improved skin architecture highlighting the tremendous therapeutic potential of this hydrogel when compared with control and CSPU to treat chronic diabetic and burn wounds.

Smart battery-free and wireless bioelectronic platform based on a nature-skin-derived organohydrogel for chronic wound diagnosis, assessment, and accelerated healing

Using smart electroactive hydrogels with emerging bioelectronics to monitor wound status wirelessly in real-time is a promising approach as an unremitting therapy for refractory chronic wounds. Based on a natural skin-derived organohydrogel (SGC@MA-Gel) triboelectric nanogenerator, a bioresorbable, battery-free, and wireless smart wound treatment bioelectronic system for pharmacological treatment, nanogenerator-driven electrostimulation, and visual wound condition assessment was engineered in this study. The inherent hierarchical architecture and origins of natural skin, in tandem with the substantial loading capabilities of collagen-based biomaterials, endowed SGC@MA-Gel with favorable characteristics. These included mechanical and adhesive robustness, electrical conductivity, antibacterial properties, controlled on-demand drug release capabilities, and human physiological signal monitoring performance. The customizable bioelectronic system, developed in-house, employs a single-electrode triboelectric nanogenerator, which facilitates skin wound healing via electrostimulation. Consequently, the system's biosensors can simultaneously measure glucose, uric acid, and pH at wound sites, thereby enabling accurate wound diagnosis and real-time assessment. This system accelerates cell proliferation and wound healing in vitro and in vivo, which validates its effectiveness in chronic wound treatment. The results demonstrated that the smart wound treatment bioelectronic system could provide a facile therapeutic strategy to assist with integrated wound monitoring, diagnosis, and therapy for hospitalized patients and those recovering at home.

Mitigation of Nitrogen Mustard-Induced Skin Injury by the β-Blocker Carvedilol and Its Enantiomers.

The chemical warfare agent sulfur mustard and its structural analog nitrogen mustard (NM) cause severe vesicating skin injuries. The pathologic mechanisms for the skin injury following mustard exposure are poorly understood; therefore, no effective countermeasure is available. Previous reports demonstrated the protective activity of carvedilol, a US Food and Drug Administration (FDA)-approved β-blocker, against UV radiation-induced skin damage. Thus, the current study evaluated the effects of carvedilol on NM-induced skin injuries in vitro and in vivo. In the murine epidermal cell line JB6 Cl 41-5a, β-blockers with different receptor subtype selectivity were examined. Carvedilol and both of its enantiomers, R- and S-carvedilol, were the only tested ligands statistically reducing NM-induced cytotoxicity. Carvedilol also reduced NM-induced apoptosis and p53 expression. In SKH-1 mice, NM increased epidermal thickness, damaged skin architecture, and induced nuclear factor κB (NF-κB)-related proinflammatory genes as assessed by RT2 Profiler PCR (polymerase chain reaction) Arrays. To model chemical warfare scenario, 30 minutes after exposure to NM, 10 μM carvedilol was applied topically. Twenty-four hours after NM exposure, carvedilol attenuated NM-induced epidermal thickening, Ki-67 expression, a marker of cellular proliferation, and multiple proinflammatory genes. Supporting the in vitro data, the non-β-blocking R-enantiomer of carvedilol had similar effects as racemic carvedilol, and there was no difference between carvedilol and R-carvedilol in the PCR array data, suggesting that the skin protective effects are independent of the β-adrenergic receptors. These data suggest that the β-blocker carvedilol and its enantiomers can be repurposed as countermeasures against mustard-induced skin injuries. SIGNIFICANCE STATEMENT: The chemical warfare agent sulfur mustard and its structural analog nitrogen mustard cause severe vesicating skin injuries for which no effective countermeasure is available. This study evaluated the effects of US Food and Drug Administration (FDA)-approved β-blocker carvedilol on nitrogen mustard-induced skin injuries to repurpose this cardiovascular drug as a medical countermeasure.

Multicellular bioprinted skin facilitates human-like skin architecture in vivo.

Bioprinting is a promising alternative method to generate skin substitutes because it can replicate the structural organization of the skin into biomimetic layers in vitro. In this study, six primary human skin cell types were used to bioprint a trilayer skin construct consisting of epidermis, dermis, and hypodermis. Transplantation of the bioprinted skin with human cells onto full-thickness wounds of nu/nu mice promoted rapid vascularization and formation of epidermal rete ridges analogous to the native human epidermis, with a normal-looking extracellular matrix. Cell-specific staining confirmed the integration of the implanted cells into the regenerated skin. Using a similar approach, a 5 centimeter-by-5 centimeter bioprinted autologous porcine skin graft was transplanted onto full-thickness wounds in a porcine excisional wound model. The bioprinted skin graft improved epithelialization, reduced skin contraction, and supported normal collagen organization with reduced fibrosis. Differential gene expression demonstrated pro-remodeling protease activity in wounds transplanted with bioprinted autologous skin grafts. These results demonstrate that bioprinted skin can support skin regeneration to allow for nonfibrotic wound healing and suggest that the skin bioprinting technology may be applicable for human clinical use.

Method for high-plex analysis of immune cells in human skin using the GeoMx system.

Specific T cell populations in the skin have been demonstrated as important disease drivers in several dermatoses. Due to the unique skin architecture, these cells are not grouped together in structures but dispersedly spread out throughout the epidermis. Following tissue disruption and isolation, only about 10% of skin T cells are recovered and any in vitro expansion may alter their bona fide phenotype. The Nanostring GeoMx system was developed to address cellular phenotype and protein expression in a tissue spatial context. To do so, regions of interest (ROI) must exceed a certain area threshold (usually 100 μm in diameter) to generate a sufficient signal-to-noise ratio. Here, we present an approach that allows for the pooling of numerous smaller ROIs within the skin, enabling T cell and melanocyte phenotyping. Skin samples from healthy individuals and vitiligo patients were analysed using the GeoMx system and several immune profiling panels. A sufficient signal-to-noise ratio was achieved by pooling smaller ROIs and analysing them as a single group. While this prevents spatial analysis, this method allows for detailed analysis of cells as a population in the context of their physiological environment, making it possible to investigate in situ phenotype of rare cells in different tissue compartments.

Fibrinogen-Based Bioink for Application in Skin Equivalent 3D Bioprinting.

Three-dimensional bioprinting has emerged as an attractive technology due to its ability to mimic native tissue architecture using different cell types and biomaterials. Nowadays, cell-laden bioink development or skin tissue equivalents are still at an early stage. The aim of the study is to propose a bioink to be used in skin bioprinting based on a blend of fibrinogen and alginate to form a hydrogel by enzymatic polymerization with thrombin and by ionic crosslinking with divalent calcium ions. The biomaterial ink formulation, composed of 30 mg/mL of fibrinogen, 6% of alginate, and 25 mM of CaCl2, was characterized in terms of homogeneity, rheological properties, printability, mechanical properties, degradation rate, water uptake, and biocompatibility by the indirect method using L929 mouse fibroblasts. The proposed bioink is a homogeneous blend with a shear thinning behavior, excellent printability, adequate mechanical stiffness, porosity, biodegradability, and water uptake, and it is in vitro biocompatible. The fibrinogen-based bioink was used for the 3D bioprinting of the dermal layer of the skin equivalent. Three different normal human dermal fibroblast (NHDF) densities were tested, and better results in terms of viability, spreading, and proliferation were obtained with 4 × 106 cell/mL. The skin equivalent was bioprinted, adding human keratinocytes (HaCaT) through bioprinting on the top surface of the dermal layer. A skin equivalent stained by live/dead and histological analysis immediately after printing and at days 7 and 14 of culture showed a tissuelike structure with two distinct layers characterized by the presence of viable and proliferating cells. This bioprinted skin equivalent showed a similar native skin architecture, paving the way for its use as a skin substitute for wound healing applications.

Roles of bovine sialoglycoproteins for anti-skin aging and accelerating skin wound healing.

Fibroblasts are the most predominant cell subpopulation in the dermal layer of human skin, they play an important role in maintaining skin architecture and function. The senescence of fibroblasts is one of major causes of skin aging and chronic wound in the elderly, which is accompanied with a reduction of α2,6-sialylation on the cell surface. In this study, we investigated the effects of the bovine sialoglycoproteins on normal human dermal fibroblasts (NHDF). The results showed that bovine sialoglycoproteins could promote the proliferation and migration of NHDF cells, and accelerate the contraction of fibroblast-populated collagen lattice (FPCL). The average doubling time of NHDF cells treated with bovine sialoglycoproteins (0.5 mg/mL) was 31.1 ± 1.0 h whereas that was 37.9 ± 2.7 h for the control (p ˂ 0.05). Moreover, the expression of basic fibroblast growth factor (FGF-2) was upregulated, while that of transforming growth factor-beta 1 (TGF-β1) and human type I collagen (COL-I) were downregulated in treated NHDF cells. Furthermore, bovine sialoglycoproteins treatment significantly enhanced the α2,6-sialylation on the cell surfaces, which was consistent with the upregulation of α2,6-sialyltransferase I (ST6GAL1) expression. These results indicated that the bovine sialoglycoproteins might be developed as a reagent against skin aging in the cosmetic industry, or as a new candidate for accelerating skin wound healing and inhibiting scar formation.

In vivo Performance of a Hydrogel of Tribulus terrestris in Alloxan induced Diabetic Rats

Abstract: Background: Diabetes is one of the most important factors in chronic injuries. The gold standard for treating diabetic foot ulcers includes debridement of wounds, management of all infections, revascularization when indicated, and non-loading of ulcers. Traditionally, the aerial parts of Tribulus terrestris L. (Zygophyllaceae) are used in the folklore for the treatment of various kinds of wounds. Materials and Methods: Based on the evaluation of phytochemicals, a hydrogel of the hydroalcoholic and the aqueous of T. terrestris were prepared in this study. It was subjected to evaluation of the different parameters. The hydrogel of hydroalcoholic extract and aqueous extract was brown in color, uniform and non-irritant. It was further evaluated for pH, spreadability and viscosity. Furthermore, the in vivo performance of the hydrogel was evaluated in alloxan monohydrate induced diabetic rats. Results: In excision wound model and incision wound model topical application of 5% hydroalcoholic extract hydrogel in diabetic Wistar albino rat showed remarkable repair effects. It significantly (***p<0.001) enhanced the rate of % wound contraction. It also exhibited significant (***p<0.001) content of hydroxyproline and extremely high tensile strength (***p<0.001) as compared to the diabetic control group. It may be due to enhanced collagen synthesis. The histopathological examination of 5% hydroalcoholic extract hydrogel of T. terrestris L. treated rats showed skin architecture which was comparable to the normal group. The diabetes-induced group rats’ wounds failed to heal even 25 days post the infliction of the wounds. Our model precisely reproduced the pathophysiology of diabetic ulcers and can serve as a valid alternative model for diabetic foot ulcer and to explore new therapies. Conclusion: The results strongly support that the hydrogel of the hydroalcoholic extract of T. terrestris seems to be a novel and promising biomaterial used for wound healing applications. Keywords: Diabetes, Tribulus terrestris, Wound healing, Alloxan, Hydroxyproline, Hydrogel.

Modulation of Piezo1 influences human skin architecture and oxytocin expression.

Throughout our existence, the skin senses and analyses the mechanical forces imposed by the environment. In response to these environmental forces, skin can deform itself and achieve a biological response. The subsequent cutaneous plasticity emerges from mechanical properties arising from the collective action of the skin cells, particularly keratinocytes, that govern the tensile strength via cell-to-cell adhesions and via cell-matrix adhesion structures. In addition to serving as force-bearing entities, keratinocytes respond to forces by activating signalling pathways to control their own fate and function. To detect and adapt to mechanical signals, keratinocytes possess a panel of sensory receptors and junctional intercellular structures. Mechanically activated ion channel Piezo1 has been described as a force sensor and as being involved in pleasant touch perception. In this study, relationships between Piezo1 modulation and oxytocin synthesis were investigated. The expression of Piezo1 in the skin was studied and compared with the expression of TRPV1. Dooku1 antagonist and Jedi1 agonist were used to modulate Piezo1. The level of E-cadherin and oxytocin was monitored in ex vivo skin biopsies by immunodetection. Taken together, our results illustrate the major role of mechanosensitive ion channel Piezo1 in skin barrier integrity, and in peripheral oxytocin synthesis in the skin. In conclusion, this study highlights the relationships between pleasant touch, soft touch and local oxytocin synthesis.

17. Mechanical Offloading Prevents and Rescues Microtrauma Induced Fibrosis in Large Animal Model

PURPOSE: Repetitive subdermal injections, as seen with insulin in diabetes mellitus, cause microtrauma leading to skin fibrosis and consequential pain. We hypothesized that microtrauma induced fibrosis could be attenuated by injecting through a mechanical offloading dressing (MOD). We developed a large animal porcine model to evaluate injection associated fibrosis. METHODS: Two, Red-Duroc (3-month-old) pigs were subjected to four treatment conditions over a 16-week period, in two 8-week phases. All conditions received daily bleomycin injections in Phase 1 to induce microtrauma induced fibrosis. Group 1 (NN) received no MOD. Group 2 (EE) received the MOD through Phases 1, 2. Group 3 (EN) received the MOD in Phase 1 only. Lastly group 4 (NE) received the MOD in Phase 2 only. Biopsies from each group, with their respective adjacent unwounded skin (UW), were acquired at the 16-week timepoint for both histology and transcriptional analysis using single-cell-RNA sequencing. RESULTS: Skin histology by H&E and Masson trichome demonstrated that EE-treated skin significantly reduced dermal thickness and restored hair follicle formation (*p<0.05). Picrosirius red staining confirmed that EE-treatment resembled the architecture of unwounded skin (*p<0.05). Mechanical testing of EE, EN, and NE treated skin exhibited significantly decreased stiffness compared to NN skin (*p<0.05). Finally, skin deformation, was significantly greater for EE, EN, and NE treated skin (*p<0.05). scRNA-seq analysis identified specific profibrotic notch pathways responsible for fibrosis in NN treated skin, which were downregulated following MOD application. CONCLUSION: To date there is no effective treatment to prevent injection induced fibrosis, which is a huge clinical problem in diseases such as diabetes. This pig study demonstrates for the first time that mechanical offloading may prevent and treat microtrauma induced fibrosis.

PANX3 Channels Regulate Architecture, Adhesion, Barrier Function, and Inflammation in the Skin

The channel-forming glycoprotein PANX3 functions in cutaneous wound healing and keratinocyte differentiation, but its role in maintaining skin homeostasis through aging is not yet understood. We found that PANX3 is absent in newborn skin but becomes upregulated with age. We characterized the skin of global Panx3-knockout (KO) mice and found that KO dorsal skin showed sex differences at different ages but generally had reduced dermal and hypodermal areas compared with age-matched controls. Transcriptomic analysis of the KO epidermis revealed reduced E-cadherin stabilization and Wnt signaling compared with that of wild-type, consistent with the inability of primary KO keratinocytes to adhere in culture and diminished epidermal barrier function in KO mice. We also observed increased inflammatory signaling in the KO epidermis and a higher incidence of dermatitis in aged KO mice compared with that in wild-type controls. These findings suggest that during skin aging, PANX3 is critical in the maintenance of dorsal skin architecture, keratinocyte cell-cell and cell-matrix adhesion, and inflammatory skin responses.

Cytosine Deaminase Base Editing to Restore COL7A1 in Dystrophic Epidermolysis Bullosa Human: Murine Skin Model

Recessive dystrophic epidermolysis bullosa is a debilitating blistering skin disorder caused by loss-of-function mutations in COL7A1, which encodes type VII collagen, the main component of anchoring fibrils at the dermal-epidermal junction. Although conventional gene therapy approaches through viral vectors have been tested in preclinical and clinical trials, they are limited by transgene size constraints and only support unregulated gene expression. Genome editing could potentially overcome some of these limitations, and CRISPR/Cas9 has already been applied in research studies to restore COL7A1 expression. The delivery of suitable repair templates for the repair of DNA cleaved by Cas9 is still a major challenge, and alternative base editing strategies may offer corrective solutions for certain mutations. We show highly targeted and efficient cytidine deamination and molecular correction of a defined recessive dystrophic epidermolysis bullosa mutation (c.425A>G), leading to restoration of full-length type VII collagen protein expression in primary human fibroblasts and induced pluripotent stem cells. Type VII collagen basement membrane expression and skin architecture were restored with de novo anchoring fibrils identified by electron microscopy in base-edited human recessive dystrophic epidermolysis bullosa grafts recovered from immunodeficient mice. The results show the potential and promise of emerging base editing technologies in tackling inherited disorders with well-defined single nucleotide mutations.

Planar cell polarity proteins determine basal cell height in the later stage embryonic mouse epidermis'.

Background: Complex organ formation requires the coordinated morphogenesis of adjacent tissue layers. Here, we report a role for the planar cell polarity (PCP) proteins Fz6 and Celsr1 in generating squamous basal cells in the later stage embryonic epidermis of the mouse is reported, which may impact upon the shape of overlying suprabasal cells. Methods: The depth of the epidermis and basal layer as well as cell proliferation index was scored from immunostained wax sections taken from different mouse embryos mutant in planar cell polarity signalling and their wild-type littermates. Orientation of epidermal cell division in Celsr1 Crash/Crash mutants was determined from thick frozen immunostained sections. Immunostained wax sections of wild-type skin explants cultured using the Lumox method enabled any changes in epidermal and basal layer depth to be measured following the release of surface tension upon dissection of skin away from the whole embryo. Results: Increased numbers of columnar and cuboidal basal epidermal cells were observed in fz6-/- mutant and Celsr1 mouse mutant Crash/Crash which correlated with visibly more rounded suprabasal cells and a thicker epidermis. Conclusions: Altogether these data support tissue intrinsic roles for PCP proteins in 'outside-in' (radial) skin architecture.