Full-thickness Skin Biopsies Research Articles

Analysis of Enhanced Wound Closure Potential of ADSC-Derived Secretome Compared to Aquacel Extra in Diabetic Rats

Background: Diabetes Mellitus (DM) can lead to serious complications, such as Diabetic Foot Ulcers (DFUs). Currently, Aquacel Extra is one of the standard therapies for DFUs, but its efficacy has some limitations that may be overcome by alternative treatments, such as the secretome derived from Adipose-Derived Stem Cells (ADSCs). ADSC secretome has shown promising results in promoting wound healing by enhancing cell proliferation, collagen synthesis, and migration of fibroblasts. Aims: This study aims to compare the effect of the adipose-derived stem cell (ADSC) secretome to Aquacel Extra, a hydrofiber dressing containing gel-forming agents, on wound healing in a diabetic rat model. Aquacel Extra has sodium carboxymethylcellulose fibers, which contribute to its unique properties. Objective: The objective of this study is to compare the wound healing outcome of ADSC secretome and Aquacel Extra in streptozotocin-induced diabetic rats. Methods: Sprague Dawley rats received a single intravenous administration of streptozotocin to induce diabetes mellitus (DM). A full-thickness excisional circular wound was created at the dorsum aspect of the rats two weeks after diabetic induction. Rats were randomly divided into two groups: the Aquacel Extra group, which received the hydrofiber dressing Aquacel Extra containing gel-forming agents, and the secretome group, which received the ADSC secretome on days 0, 3, and 7. The gross examination was performed on days 0, 3, 7, and 14. On day 14, the rats were euthanized, and full-thickness skin biopsies of the wounds were taken for histological and immunohistochemical analysis. Results: The secretome group demonstrated accelerated cutaneous wound healing compared to the Aquacel Extra group, as indicated by faster wound closure (p < 0.05). Hematoxylin and eosin staining revealed a thicker epidermal layer in the secretome group than in the Aquacel Extra group (p < 0.05). The mean histological score of the secretome group was higher at 10 ± 1.73 compared to the score of 8.33 ± 1.52 in the Aquacel Extra group, indicating a more mature tissue architecture. Immunohistochemical analysis showed lower expression of α-smooth muscle actin (α-sma) in the secretome group than in the Aquacel Extra group, which correlated with reduced collagen secretion and deposition in the secretome group, as indicated by Masson’s trichrome staining. Conclusion: The ADSC secretome accelerated the healing of diabetic wounds compared to Aquacel Extra. However, as this study is preliminary, future studies should be conducted similar to this study with a larger sample and wound size. In addition, the optimal dosage of ADSC secretome and the components of the secretome that promoted wound healing should be determined to fully elucidate the clinical potential of ADSC secretome.

Exploring the Influence of Cold Plasma on Epidermal Melanogenesis In Situ and In Vitro.

Epidermal melanin synthesis determines an individual's skin color. In humans, melanin is formed by melanocytes within the epidermis. The process of melanin synthesis strongly depends on a range of cellular factors, including the fine-tuned interplay with reactive oxygen species (ROS). In this context, a role of cold atmospheric plasma (CAP) on melanin synthesis was proposed due to its tunable ROS generation. Herein, the argon-driven plasma jet kINPen® MED was employed, and its impact on melanin synthesis was evaluated by comparison with known stimulants such as the phosphodiesterase inhibitor IBMX and UV radiation. Different available model systems were employed, and the melanin content of both cultured human melanocytes (in vitro) and full-thickness human skin biopsies (in situ) were analyzed. A histochemical method detected melanin in skin tissue. Cellular melanin was measured by NIR autofluorescence using flow cytometry, and a highly sensitive HPLC-MS method was applied, which enabled the differentiation of eu- and pheomelanin by their degradation products. The melanin content in full-thickness human skin biopsies increased after repeated CAP exposure, while there were only minor effects in cultured melanocytes compared to UV radiation and IBMX treatment. Based on these findings, CAP does not appear to be a useful option for treating skin pigmentation disorders. On the other hand, the risk of hyperpigmentation as an adverse effect of CAP application for wound healing or other dermatological diseases seems to be neglectable.

Overview on Lumpy Disease: Pathogenesis, Sign and Symptoms, Risk Factor, and Treatment

The lumpy diseases is an infectious disease. It is caused by viruses (LSDV) of the family Capripoxvirus.An investigation on the risk variables linked to the proliferation of LSD in Ethiopia has been conducted through a cross-sectional questionnaire-based study LSD virus,virus has been detected in stomoxy calcitrans and Anopheles stephens. virus can survive in necrotic skin for about 33 days and remains viable for at least 18 days in air-dried skin lesions at room temperature. The virus has been reported to persist in necrotic skin nodules for up to 33 days or longer, in dried crusts for up to 35 days and at least 18 days in air-dried hides.LSD has gross lesions that are well-described. Skin nodules are typically elevated, hard, and spherical in shape, however some might fuse together to form enormous, uneven plaques with defined boundaries. Biphasic fever, a clinical symptom of LSD, is characterised by two episodes of fever that develop after a varying incubation period of 4–12 days (typically 7 days). The affected animals' body temperature rises to 40–41.5°C, and this can last for at least 6–72 hours and, in exceptional cases, up to 10 days.To guarantee sufficient defence against LSD, susceptible adult cattle should receive a yearly vaccination.To reduce the possibility of the disease spreading by vectors, cattle should get routine treatments with insect repellents. While not completely effective, this method can lower the likelihood of transmission..The first report of LSD's clinical symptoms was published in Zambia (Morris 1931 ) The affected animals displayed symptoms of nasal discharge, emaciation, necrotic plaques in the mouth and nose mucous membranes, and excessive salivation. The most popular technique for diagnosis is full-thickness skin biopsies or scabs with electron microscopy showing the presence of characteristic capripox virions.To identify the antigens, techniques such as ELISA, PCR, and viral neutralization test procedures are helpful. Doctors may administer vitamin injections, anti-inflammatory drugs, or antibiotics to treat secondary bacterial infection fever and inflammation.needed a full recuperation period of six months A27L, L1R, and a combination of the two LSDV antigens were included in the development of three vaccines. Insect repellents, disinfectant cleaning, proper disposal of contaminated areas, and awareness-raising are examples of preventive approaches.The economic impact of LSD was caused by both direct consequences, like control expenses, and indirect ones, including milk loss, mortality loss, and draft power loss.

When pink is a question: Comparative gross and microscopic skin structure analyses reveal the histological basis of skin colour in Galápagos pink land iguanas (Conolophus marthae)

AbstractOne of the rarest and most unusual iguanas on the planet is the Galápagos pink land iguana (Conolophus marthae). There have been a number of hypotheses on the source of their pink coloration, including that the colour is from blood and a relative lack of dermal pigmentation. We obtained full thickness skin biopsies of three species and compared tissue from darkly pigmented areas and lightly pigmented surfaces. “Pink” areas of pink iguanas are devoid of pigment cells (e.g. melanophores) and the dermal tissue is rich with aggregates of confluent capillaries. This was in sharp contrast to the minimally vascular (only capillaries were observed) dermal areas of the marine and yellow iguanas. The dermal stratum laxum of every biopsy site contained melanophores except for the pink skin of pink iguanas. Interestingly, marine iguanas have a much thicker epidermal stratum germinativum/granulosum, between 2 and 10 cells thick depending on location, compared to the thinner epidermal stratum germinativum/granulosum of land iguanas (one to three cells thick with most areas possessing just one or two cell layers). These microscopic differences might reflect differences in habitat and ecology of the three species.

OA15 Missing the wood(-y induration) for the trees

OA15 Missing the wood(-y induration) for the trees

Spatial transcriptomics combined with single-cell RNA-sequencing unravels the complex inflammatory cell network in atopic dermatitis.

Atopic dermatitis (AD) is the most common chronic inflammatory skin disease with complex pathogenesis for which the cellular and molecular crosstalk in AD skin has not been fully understood. Skin tissues examined for spatial gene expression were derived from the upper arm of 6 healthy control (HC) donors and 7 AD patients (lesion and nonlesion). We performed spatial transcriptomics sequencing to characterize the cellular infiltrate in lesional skin. For single-cell analysis, we analyzed the single-cell data from suction blister material from AD lesions and HC skin at the antecubital fossa skin (4 ADs and 5 HCs) and full-thickness skin biopsies (4 ADs and 2 HCs). The multiple proximity extension assays were performed in the serum samples from 36 AD patients and 28 HCs. The single-cell analysis identified unique clusters of fibroblasts, dendritic cells, and macrophages in the lesional AD skin. Spatial transcriptomics analysis showed the upregulation of COL6A5, COL4A1, TNC, and CCL19 in COL18A1-expressing fibroblasts in the leukocyte-infiltrated areas in AD skin. CCR7-expressing dendritic cells (DCs) showed a similar distribution in the lesions. Additionally, M2 macrophages expressed CCL13 and CCL18 in this area. Ligand-receptor interaction analysis of the spatial transcriptome identified neighboring infiltration and interaction between activated COL18A1-expressing fibroblasts, CCL13- and CCL18-expressing M2 macrophages, CCR7- and LAMP3-expressing DCs, and T cells. As observed in skin lesions, serum levels of TNC and CCL18 were significantly elevated in AD, and correlated with clinical disease severity. In this study, we show the unknown cellular crosstalk in leukocyte-infiltrated area in lesional skin. Our findings provide a comprehensive in-depth knowledge of the nature of AD skin lesions to guide the development of better treatments.

Acellular Dermal Matrix Cover Improves Skin Growth during Tissue Expansion by Affecting Distribution of Mechanical Forces.

Biological cover over tissue expander prostheses has been introduced to provide soft-tissue support for tissue expanders during breast reconstruction. However, its impact on mechanically induced skin growth remains unknown. This study investigates the hypothesis that covering the tissue expander with acellular dermal matrix (ADM) affects mechanotransduction without compromising the efficacy of tissue expansion. Tissue expansion, with and without use of ADM, was performed on a porcine model. The tissue expanders were inflated twice with 45 mL of saline, and the full-thickness skin biopsy specimens were harvested from expanded and control unexpanded skin 1 week and 8 weeks after the final inflation. Histologic evaluation, immunohistochemistry staining, and gene expression analysis were performed. Skin growth and total deformation were evaluated using isogeometric analysis. The authors' results demonstrate that use of ADM as a biological cover during tissue expansion does not impede mechanotransduction that leads to skin growth and blood vessel formation. Isogeometric analysis revealed similar total deformation and growth of expanded skin with and without a biological cover, confirming that its use does not inhibit mechanically induced skin growth. In addition, the authors found that use of an ADM cover results in more uniform distribution of mechanical forces applied by the tissue expander. These results suggest that ADM improves mechanically induced skin growth during tissue expansion by facilitating a more uniform distribution of mechanical forces applied by the tissue expander. Therefore, the use of a biological cover has potential to improve outcomes in tissue expansion-based reconstruction.

Anagen hair follicles transplanted into mature human scars remodel fibrotic tissue

Despite the substantial impact of skin scarring on patients and the healthcare system, there is a lack of strategies to prevent scar formation, let alone methods to remodel mature scars. Here, we took a unique approach inspired by how healthy hairbearing skin undergoes physiological remodelling during the regular cycling of hair follicles. In this pilot clinical study, we tested if hair follicles transplanted into human scars can facilitate tissue regeneration and actively remodel fibrotic tissue, similar to how they remodel the healthy skin. We collected full-thickness skin biopsies and compared the morphology and transcriptional signature of fibrotic tissue before and after transplantation. We found that hair follicle tranplantation induced an increase in the epidermal thickness, interdigitation of the epidermal-dermal junction, dermal cell density, and blood vessel density. Remodelling of collagen type I fibres reduced the total collagen fraction, the proportion of thick fibres, and their alignment. Consistent with these morphological changes, we found a shift in the cytokine milieu of scars with a long-lasting inhibition of pro-fibrotic factors TGFβ1, IL13, and IL-6. Our results show that anagen hair follicles can attenuate the fibrotic phenotype, providing new insights for developing regenerative approaches to remodel mature scars.

581 Spontaneous, Intrinsic Aging in Organ-Cultured Human Skin as Novel Ex Vivo Assay to Test Candidate Anti-Aging Actives

Since in vitro methods (2D and 3D) used for the preclinical testing of candidate anti-aging compounds mimic the complexity of human skin aging unsatisfactorily, we wished to develop an ex vivo assay for interrogating key aging-read outs in healthy human skin. Healthy, full-thickness skin biopsies (2 males, 4x females; 22 - 77 years) were organ-cultured in serum-free, supplemented medium for 3 days in the presence/absence of recognized anti-aging compounds (tretinoin (5μM), resveratrol (10μM), or caffeine (200μM), and key skin aging biomarkers were analyzed by quantitative (immuno-)histomorphometry and/or qRT-PCR.

OA16 A great imitator of scleroderma

Introduction/BackgroundEosinophilic fasciitis is a rare disease of idiopathic aetiology, which can often mimic scleroderma in presentation and has been associated with haematological diseases. We present a case of eosinophilic fasciitis where the diagnosis was delayed due to the attribution of skin manifestations to scleroderma. This case highlights the importance of careful history and examination to recognize these differential diagnoses, prevent delay and improve outcome.Description/MethodA 69-year-old gentleman was referred to the Rheumatology clinic with suspected diffuse cutaneous systemic sclerosis. He presented with a 9-month history of skin tightening, pain and swelling affecting his legs, finding it difficult to fully extend at the knees, to drive or walk. His symptoms progressed overtime with skin changes spreading to all four extremities, abdomen and back. He reported weight loss of 12Kg associated with early satiety. He also experienced intermittent numbness of his hands. There were no respiratory symptoms, dysphagia or Raynaud’s phenomenon reported. His medical history includes atrial fibrillation and asbestos exposure. Currently on Apixaban 5mg/daily and Bisoprolol 2.5mg/daily.Physical examination revealed diffuse brown hyperpigmentation, erythema and thickened skin on his upper and lower limbs, abdomen and back, it was difficult to crease the skin and there was ‘peau d’orange’ appearance in his forearms. His face and hands were spared. Phalen's and Tinel’s tests were negative.Prior to clinical review he was investigated for paraneoplastic disease: a CT chest, abdomen and pelvis showed an incidental cyst in the thymus and emphysema with no underlying malignancy. Laboratory tests showed normocytic normochromic anaemia with haemoglobin 112 g/L and eosinophilia 1.5 x 109/L, elevated erythrocyte sedimentation rate 62mm/h, CRP 57mg/L, polyclonal hypergammaglobulinaemia IgA 4.3g/L and IgG 24.85g/L. The myeloma screen were negative.The absence of Raynaud’s, and the distribution of skin changes sparing his face, hands and feet, eosinophilia and raised inflammatory markers, made the diagnosis of eosinophilic fasciitis. Histological analysis of a full thickness skin biopsy confirmed patchy lymphoplasmacytic inflammatory cell infiltrate within the fascia and spilling into subcutis, with eosinophils.Subsequently, he was commenced on prednisolone 40mg/daily reducing regimen and Methotrexate 20mg/weekly, with a good response and regression of the swelling and pain. He is now able to walk without difficulty.Discussion/ResultsEosinophilic fasciitis is rare. Symptoms can be similar to those of scleroderma, sometimes considered a variant. The exact aetiology is unknown; however, the literature mentions a potential paraneoplastic, inflammatory, infectious or physical trigger, leading to an abnormal response with accumulation of eosinophils in different tissues. Consequently, this may release transforming growth factor beta to activate fibroblasts, increasing expression of other factors in the connective tissue, resulting in fibrosis.This case highlights that eosinophilic fasciitis present with hardening and thickening of the skin and surrounding tissue similar to that of scleroderma, however, lacks Raynaud’s phenomenon, sclerodactyly. Another striking features from his examination which helped us distinguishing from scleroderma was that his skin changes spare his face, hands and feet.Carpal tunnel syndrome is described in the literature which can progress if untreated.Typical blood tests findings documented in the literature include eosinophilia in 80% of the cases, elevated ESR and C-reactive protein (CRP), polyclonal hypergammaglobulinaemia, are usually present, however, eosinophilia may be transient, therefore normal eosinophil count does not exclude the disease. ANA and Rheumatoid factor can be positive in up to 10 % of the cases.MRI of the extremities to look for thickening of the fascia has been used in other cases, however, it was not required in our case because our Orthopaedics colleagues helped us with performing an urgent full thickness biopsy.Recommended treatment includes high dose steroids and immunosuppressive medication such Methotrexate is most often used for severe or resistant cases.The prognosis is good, however if treatment is delayed patients can develop joint contractures in 85% of patients and skin hardness may persist.In conclusion, careful history and examination is essential in differentiating scleroderma from the scleroderma-mimics especially when there is an atypical distribution, no sclerodactyly and no autoantibodies.Key learning points/Conclusion1. This case suggest that clinicians should be aware that eosinophilic fasciitis typically spare the hands, feet and face.2. Eosinophilia may be transient, therefore normal count does not exclude the disease.3. Recognizing this rare but great imitator of scleroderma set the stage for prompt therapy in either condition and reduce morbidity.

34982 Transcriptome analysis reveals intrinsic proinflammatory signaling in healthy African American skin

Background: The disparities in prevalence and clinical features for many inflammatory skin diseases including atopic dermatitis and psoriasis are well recognized; however, the underlying mechanisms are largely unknown. Methods: Using bulk RNA-seq analysis, we compared the baseline gene expression in upper arm full thickness skin biopsies from 17 healthy individuals self-reporting as African American (AA) or White non-Hispanic (WNH) and in 3D human skin equivalents (HSE) made from AA and WNH primary skin keratinocytes. Results: Extensively validated RNA-Seq analysis identified 570 differentially expressed genes (DEG) in AA skin including immunoglobulins and their receptors such as FCER1G; proinflammatory genes such as TNFα, IL-32; EDC (epidermal differentiation cluster) and keratin genes. DEGs were functionally enriched for inflammatory responses, keratinization, cornified envelope formation. RNA-seq analysis of gene expression in AA versus WNH HSEs revealed 360 DEGs (some shared with skin) which were enriched by similar to skin functions. In addition, AA HSE appeared more responsive to TNFα proinflammatory effects. Finally, AA-specific DEGs in skin and HSE significantly overlapped with molecular signatures of lesional skin in AD and psoriasis patients. Interestingly, AA skin DEGs overlapping with AD molecular signature in skin were enriched by IL13 and IL4 signaling central for AD. Conclusions: Overall, these findings suggest the existence of intrinsic proinflammatory circuits in AA keratinocytes/skin that may account for disease disparities and will help to build a foundation for the development of targeted skin disease prevention and treatment.

413 Quantitative morphometric analysis of intrinsic and extrinsic ageing in Caucasian individuals

Introduction: The field of skin ageing is important due to the world ageing population. Due to its interface with the external environment, skin ageing is attributed to both intrinsic and extrinsic factors. The aim of this study was to define histomorphic disparities between intrinsically and extrinsically aged skin, and increase understanding of how skin structures change with age and photoexposure. Methods: Full-thickness skin biopsies were obtained from the photoexposed dorsal forearm and photoprotected buttock of young (21-24 years, n=10 females) and aged Caucasian volunteers (61-65 years, n=10 females). Biometrics analysis of epidermal thickness (Emin), epidermal proliferation, basal keratinocyte morphology, interdigitation index and morphology of rête ridges and dermal papilla was performed on blinded histological and immunofluorescence images (n=40 biopsies; n=160-240 images). Results: Our study found that Emin and epidermal proliferation do not change with age or photoexposure (P>0.05). The size and height of basal keratinocytes is increased by photoexposure in young and ageing individuals, but no differences were identified with age (P<0.0001). The dermal-epidermal junction is affected by both age and photoexposure with regards to decreased interdigitation and decreased height of rête ridges and dermal papilla (P<0.0001). Discussion: Our data suggest that basal keratinocyte morphology is affected by photoexposure, and dermal-epidermal junction characteristics are altered by age and photoexposure. Interestingly, flattening of the dermal-epidermal junction was most apparent in photoexposed areas, and we propose that there is an accelerated ageing phenotype due to the contribution of extrinsic factors. These data are important for both academic and industrial scientists to validate in vitro findings, and identify targets for cosmetic interventions.

Disease surveillance in England and Wales, June 2022.

Disease surveillance in England and Wales, June 2022.

A Microcurrent Dressing Reduces Cutibacterium Acnes Colonization in Patients Undergoing Shoulder Arthroplasty or Arthroscopy: A Prospective Case Series.

Background: Cutibacterium acnes infections in the shoulder remain a significant concern in the setting of shoulder arthroplasty. Purpose: We sought to evaluate the efficacy of a microcurrent dressing in reducing C. acnes skin colonization and thereby reducing the risk of periprosthetic joint infection of the shoulder. Methods: This study was designed as a prospective case series. From October 2017 to February 2019, patients undergoing elective shoulder arthroplasty or arthroscopic shoulder surgery at a major academic medical center were offered enrollment; they signed an informed consent to participate. Patients under the age of 18, scheduled for revision shoulder arthroplasty, or with sensitivity or allergy to silver, zinc, or latex were excluded. Subjects underwent skin culture swab of the shoulder in the mid-point of the planned deltopectoral incision. The JumpStart (Arthrex; Naples, FL) microcurrent dressing was then placed over the area of the planned incision, and a full-thickness skin biopsy was harvested from the incision at the initiation of the surgical procedure. All specimens were cultured for C. acnes by the hospital's clinical microbiology laboratory with standard anaerobic technique. Results: Thirty-one subjects were enrolled in the study. Those who demonstrated no growth at baseline for the control specimen were excluded from further analysis (N = 11), given the absence of preoperative C. acnes colonization. Culture results from the 20 remaining subjects revealed significantly diminished C. acnes skin growth at the time of surgery compared to baseline. Sixty percent (12 of 20) of the subjects with positive skin swabs at baseline demonstrated no growth in the skin biopsy specimens at the time of surgery. There were no adverse events associated with the application of the microcurrent dressing. Conclusion: This prospective case series found that preoperative application of a microcurrent dressing resulted in significantly diminished C. acnes skin burden at the time of surgery in patients undergoing elective shoulder arthroplasty or arthroscopic shoulder surgery. Further study is warranted to investigate whether this preoperative intervention may contribute to a reduction in perioperative infections, including prosthetic joint infection.

Interrogation of RDEB Epidermal Allografts after BMT Reveals Coexpression of Collagen VII and Keratin 15 with Proinflammatory Immune Cells and Fibroblasts

Recessive dystrophic epidermolysis bullosa (RDEB) is a devastating genodermatosis characterized by dysfunctional collagen VII protein resulting in epithelial blistering of the skin, mucosa, and gastrointestinal tract. There is no cure for RDEB, but improvement of clinical phenotype has been achieved with bone marrow transplantation and subsequent epidermal allografting from the bone marrow transplant donor. Epidermal allografting of these patients has decreased wound surface area for up to 3 years after treatment. This study aimed to determine the phenotype of the epidermal allograft cells responsible for durable persistence of wound healing and skin integrity. We found that epidermal allografts provide basal keratinocytes coexpressing collagen VII and basal stem cell marker keratin 15. Characterization of RDEB full-thickness skin biopsies with single-cell RNA sequencing uncovered proinflammatory immune and fibroblast phenotypes potentially driven by the local environment of RDEB skin. This is further highlighted by the presence of a myofibroblast population, which has not been described in healthy control human skin. Finally, we found inflammatory fibroblasts expressing profibrotic gene POSTN, which may have implications in the development of squamous cell carcinoma, a common, lethal complication of RDEB that lacks curative treatment. In conclusion, this study provides insights into and targets for future RDEB studies and treatments.

Preparation of shikonin liposome and evaluation of its in vitro antibacterial and in vivo infected wound healing activity

BackgroundThe emergence of antibiotic resistance over the past decade has made the treatment of Staphylococcus aureus infection difficult. Burn wounds infected with methicillin-resistant S. aureus (MRSA) can cause mortality in animals. Shikonin (SH) has been reported to possess antimicrobial and anti-inflammatory properties, and is also responsible for the process of wound healing. However, the pharmacological mechanism of its wound healing process remains poorly comprehended, hence the probable mechanism deserves further investigation. PurposeThe current study was designed to develop a novel SH-liposome with improved anti-MRSA effect and to detect its beneficial wound healing effects. Study DesignIn vitro antibacterial tests and in vivo infected wound healing test were conducted. MethodsSH-liposome was produced by the film formation method, and the characteristics were measured using a laser particle size analyzer, transmission electron microscopy, and the dialysis method. Additionally, in vitro antibacterial tests were conducted to investigate the antibacterial effects and the relative mechanism of SH-liposome. Furthermore, the therapeutic effects and bioactivity of SH-liposome in MRSA infected burn wounds were investigated in rats. Sixty-four male Sprague Dawley rats (250 ± 10 g) were randomly divided into four groups, including Group I (control group); Group II (model group); Group III (SH-liposome group) and Group IV (Arnebia oil® group), and the drug treatments were applied topically twice daily for 21 days. Further, full thickness skin biopsies at different periods were collected aseptically to evaluate tissue cytokines, recognize flora, observe histopathological changes, and determine the mechanism underlying the wound healing effects of SH-liposome. The data were analyzed via one-way analysis of variance (ANOVA) and Duncan's multiple range test. ResultsThe results showed that SH-liposome was successful with a drug load of 4.6 ± 0.17%. Moreover, SH-liposome showed a sustained-release behavior and improved antibacterial ability in a dose-dependent manner. For the possible antibacterial mechanism, we observed that SH-liposome achieved antibacterial activity by damaging the integrity of bacterial cell wall and membrane to further disturb the physiological activities of S. aureus. In addition, SH-liposome facilitated wound healing by inhibiting bacterial activities to control infection, regulating the I-κBα/NFκB-p65 pathway to alleviate inflammation, and directly promoting repair in burn wounds. ConclusionIn conclusion, SH-liposome showed an antibacterial effect against S. aureus, promoted effective healing of infected burn wounds; hence, it could be used as an alternative therapy for drug-resistant infections.

Oral 11β-HSD1 inhibitor AZD4017 improves wound healing and skin integrity in adults with type 2 diabetes mellitus: a pilot randomized controlled trial.

BackgroundChronic wounds (e.g. diabetic foot ulcers) reduce the quality of life, yet treatments remain limited. Glucocorticoids (activated by the enzyme 11β-hydroxysteroid dehydrogenase type 1, 11β-HSD1) impair wound healing.ObjectivesEfficacy, safety, and feasibility of 11β-HSD1 inhibition for skin function and wound healing.DesignInvestigator-initiated, double-blind, randomized, placebo-controlled, parallel-group phase 2b pilot trial.MethodsSingle-center secondary care setting. Adults with type 2 diabetes mellitus without foot ulcers were administered 400 mg oral 11β-HSD1 inhibitor AZD4017 (n = 14) or placebo (n = 14) bi-daily for 35 days. Participants underwent 3-mm full-thickness punch skin biopsies at baseline and on day 28; wound healing was monitored after 2 and 7 days. Computer-generated 1:1 randomization was pharmacy-administered. Analysis was descriptive and focused on CI estimation. Of the 36 participants screened, 28 were randomized.ResultsExploratory proof-of-concept efficacy analysis suggested AZD4017 did not inhibit 24-h ex vivoskin 11β-HSD1 activity (primary outcome; difference in percentage conversion per 24 h 1.1% (90% CI: −3.4 to 5.5) but reduced systemic 11β-HSD1 activity by 87% (69–104%). Wound diameter was 34% (7–63%) smaller with AZD4017 at day 2, and 48% (12–85%) smaller after repeat wounding at day 30. AZD4017 improved epidermal integrity but modestly impaired barrier function. Minimal adverse events were comparable to placebo. Recruitment rate, retention, and data completeness were 2.9/month, 27/28, and 95.3%, respectively.ConclusionA phase 2 trial is feasible, and preliminary proof-of-concept data suggests AZD4017 warrants further investigation in conditions of delayed healing, for example in diabetic foot ulcers.Significance statementStress hormone activation by the enzyme 11β-HSD type 1 impairs skin function (e.g. integrity) and delays wound healing in animal models of diabetes, but effects in human skin were previously unknown. Skin function was evaluated in response to treatment with a 11β-HSD type 1 inhibitor (AZD4017), or placebo, in people with type 2 diabetes. Importantly, AZD4017 was safe and well tolerated. This first-in-human randomized, controlled, clinical trial found novel evidence that 11β-HSD type 1 regulates skin function in humans, including improved wound healing, epidermal integrity, and increased water loss. Results warrant further studies in conditions of impaired wound healing, for example, diabetic foot ulcers to evaluate 11β-HSD type 1 as a novel therapeutic target forchronic wounds.

Noninvasive Tape-Stripping with High-Resolution RNA Profiling Effectively Captures a Preinflammatory State in Nonlesional Psoriatic Skin

Tape stripping is a minimally invasive, nonscarring method that can be utilized to assess gene expression in the skin but is infrequently used given technical constraints. By comparing different tape stripping technologies and full-thickness skin biopsy results of lesional and nonlesional psoriatic skin from the same patients, we demonstrate that tape stripping with optimized high-resolution transcriptomic profiling can be used to effectively assess and characterize inflammatory responses in the skin. Upon comparison with single-cell RNA-sequencing data from psoriatic full-thickness skin biopsies, we illustrate that tape-stripping efficiently captures the transcriptome of the upper layers of the epidermis with sufficient resolution to assess the molecular components of the feed-forward immune amplification pathway in psoriasis. Notably, nonlesional psoriatic skin sampled by tape stripping demonstrates activated, proinflammatory changes when compared to healthy control skin, suggesting a prepsoriatic state, which is not captured on full-thickness skin biopsy transcriptome profiling. This work illustrates an approach to assess inflammatory response in the epidermis by combining noninvasive sampling with high throughput RNA-sequencing, providing a foundation for biomarker discoveries and mechanism of action studies for inflammatory skin conditions.

Transcriptome Analysis Reveals Intrinsic Proinflammatory Signaling in Healthy African American Skin

Differences in the morphology and physiology of darkly pigmented skin compared with those of lightly pigmented skin are well-recognized. There are also disparities in the prevalence and clinical features for many inflammatory skin diseases, including atopic dermatitis and psoriasis; however, the underlying mechanisms are largely unknown. We compared the baseline gene expression in full-thickness skin biopsies from healthy individuals self-reporting as African American (AA) or as White non-Hispanic (WNH). Extensively validated RNA-sequencing analysis identified 570 differentially expressed genes in AA skin, including Igs and their receptors such as FCER1G; proinflammatory genes such as TNFα and IL32; and epidermal differentiation cluster and keratin genes. Differentially expressed genes were functionally enriched for inflammatory responses, keratinization, and cornified envelope formation. RNA-sequencing analysis of three-dimensional human skin equivalents made from AA and WNH primary keratinocytes revealed 360 differentially expressed genes (some shared with skin) that were enriched by similar functions. AA human skin equivalents appeared more responsive to TNF-α proinflammatory effects. Finally, AA-specific differentially expressed genes in the skin and human skin equivalents significantly overlapped with molecular signatures of skin in patients with atopic dermatitis and psoriasis. Overall, these findings suggest the existence of intrinsic proinflammatory circuits in AA keratinocytes/skin that may account for disease disparities and will help to build a foundation for the development of targeted skin disease prevention.

4: ACTA2+ Cells Activation and Dermal Changes During Skin Adaptation to Mechanical Forces

Purpose:Tissue expansion (TE) is based on the skin’s exceptional ability to regenerate under mechanical stress and is widely used to repair skin defects. However, knowledge about molecular mechanisms involved in maintaining skin integrity and homeostasis is limited. The present study aims 1) to elucidate the role of myofibroblasts in the mechanism of adaptation to mechanical stress exerted by an inflated tissue expander, and 2) to describe morphological changes in collagen structure that could lead to the re-establishment of dermal tension.Methods:TE was performed on a porcine model. Each expander was placed subcutaneously over the ribs and two weeks later inflated with 30 cc of saline to induce subtle tension. After 1 day (acute stretch) and 7 days (sub-acute stretch) of expansion, the full-thickness skin biopsies were collected from the apex of the expander and control unexpanded skin (contralateral sites). Skin samples were fixed in formalin and embedded in paraffin for histological evaluation (Russell-Movat Pentachrome staining) or fixed in 4% PFA and embedded in OCT for immunohistochemistry staining (IF) of α-smooth muscle actin (α-SMA), a marker of myofibroblast. Area of fluorescent signal from α-SMA was calculated using ImageJ while collagen morphology was evaluated optically after staining.Results:We compared the presence of α-SMA fluorescence between control biopsies and expanded biopsies after 1-day and 7-days of stretch. The immunofluorescence staining revealed 2.32 times more α-SMA fluorescent staining in skin expanded for 1 day (p = 0.0065) and 2.19 times more α-SMA fluorescence in skin expanded for 7 days (p = 0.0047). The increase in number of α-SMA positive cells were mostly observed in the outermost 400µm of papillary dermis. Histological staining showed minimal collagen morphological changes in both the papillary and reticular dermis after 1-day of expansion. However, shortening of fibril length, increased density, and increased disorder were observed in the papillary dermis collagen after 7-days of subtle expansion.