The use of human acellular dermal matrices in advanced wound healing and surgical procedures: State of the art.

Wound closure after post-traumatic injuries and/or localized at peculiar body sites (head-and-neck, oral cavity, legs) are particularly challenging and can often be delayed due to local and systemic factors. In case of deep wounds and/or hard-to-heal wounds, grafting of dermal acellular matrices (ADM) is often needed. Though a great variety of synthetic and semisynthetic dermal and skin equivalents are available, viable human dermis, is still considered the most physiological alternative to replace the loss of autologous dermis, by acting as a physiological scaffold that add structural support to soft tissues. To date, human ADMs (hADMs) have been employed in the reconstruction of skin defects affecting almost all body sites, ranging from visceral sites to the skin and subcutaneous tissues. This review aims to investigate the use of hADM at different body sites and their peculiar advantages. A literature search was using the search terms "acellular dermal matrices", "dermal regeneration", "advances wound healing", "human acellular dermal matrices surgery". A total of 50 out of 150 papers was included. Based on the current body if evidence, hADMs appear to bring several advantages, such as: protection of deep structures (eg, tendons, bones, cartilage and nerves); stimulation of a functional new dermis (rather than a scar); reduction of wound closure time; control of pain and exudate. Finally, hADMs may represent the best treatment option for hard-to-heal wound not only in terms of efficacy and patient satisfaction bout also in terms of sanitary costs, especially across Europe, where hADMs cannot be commercialized as medical devices.

A new clinical and dermoscopic monitoring of infantile hemangiomas treated with oral propranolol.

Oral propranolol (OP) demonstrated high efficacy and safety profile for treatment of critical infantile hemangiomas (IHs). Our aim was to assess the morphologic changes of IHs with standard and high-resolution video dermoscopy (HRVD) from baseline to 18 months either in presence or absence of OP therapy; to investigate if extended anamnestic perinatal data and clinical-dermoscopic characteristics of the IHs can correlate with therapeutic outcome. We enrolled 94 patients (112 IHs): 58 were treated with OP, 35 (42 IHs) for 6 months (group 1), and 23 (25 IHs) for 12-months (group 2); 36 (45 IHs) were followed-up. Clinical-dermoscopic examinations were performed every 3 months during therapy and follow-up. Among 67 treated IHs, superficial and deep IHs with homogenous clinical-dermoscopic aspect developed after the 2 weeks of life achieved the better outcome, stable at 9-month follow-up, independently form treatment duration. Under HRVD, glomerular vessels were prevalent at baseline; corckscrew, comma, and linear-irregular vessels were the prevalent pattern at 1, 3, and 6 months of therapy, respectively. At 12-month follow-up, adequate healing was achieved by 96% of IHs in group 2 and by 78% in group 1, showing dotted vessels. Persistent IHs displayed a reticulated aspect and linear irregular vessels, while arborizing vessels characterized relapsed IHs. A 12-month OP therapy can be considered for newborns presenting with nonhomogenous mixed IHs >3 cm on the perineal area/lower extremities. In conclusion, HRVD allows a real time monitoring of vascular changes in IHs treated with OP and can support physicians in identifying relapses before they become clinically evident.

Long-term therapy of multiple basal cell carcinomas: Clinicodermoscopic score for monitoring of intermittent vismodegib treatment.

Vismodegib treatment of multiple basal cell carcinomas (BCCs) is limited by adverse effects and high relapse rates: intermittent regimens are therefore preferred for long-term administration. The objective of this study was to investigate clinical and dermoscopic changes in BCCs during long-term intermittent treatment and to identify those most indicative of tumor persistence/clearing. Clinical and dermoscopic images (n = 380 each) of 38 BCCs were acquired at 10 observation times (t0-t9). Biopsies were performed at baseline (t0) and after 72 weeks of treatment (t9). All images were evaluated retrospectively by experts who assessed the presence/absence of 12 clinical and 14 dermoscopic features: clinical scores (CScs) and dermoscopic scores (DScs) were then calculated.

Line-Field Optical Coherence Tomography: Usefulness in the Non-Invasive Differential Diagnosis of Congenital Alopecia of Infancy.

INTRODUCTION: Soon after birth, the clinical differential diagnosis between sebaceous of Jadassohn (NSJ), congenital triangular alopecia (CTA) and aplasia cutis congenita (ACC) may be challenging. A certain overlap of standard dermoscopic features can occur, especially in atypical cases, depending on scalp skin morphology and maturation age. The recently developed line-field confocal optical coherence tomography (LC-OCT) can provide morphological skin details with cellular resolution trough a rapid non-invasive examination. OBJECTIVES: To assess the LC-OCT features of 6 cases of congenital alopecia of different aetiologies, with both typical and atypical clinical appearance. METHODS: A non-invasive imaging examination combining standard dermoscopy, high-resolution videodermoscopy (HRVD) and LC-OCT was realized in 7 babies presenting for congenital alopecia with overlapping features, aged between 5 months and 5 years. RESULTS: Based on the specific LC-OCT features, and supported by HRVD features, a diagnosis of NSJ, congenital triangular alopecia (CTA) and AC) were made in 4, 2 and 1 case, respectively. CONCLUSIONS: The combined LC-OCT plus HRVD non-invasive imaging bring the advantage to have a real time diagnosis, to set the proper management and allows to avoid a skin biopsy in the perinatal age/first years of life at delicate skin site.

Importance of good manufacturing practices in microbiological monitoring in processing human tissues for transplant.

Skin allografts represent an important therapeutic resource in the treatment of severe skin loss. The risk associated with application of processed tissues in humans is very low, however, human material always carries the risk of disease transmission. To minimise the risk of contamination of grafts, processing is carried out in clean rooms where air quality is monitored. Procedures and quality control tests are performed to standardise the production process and to guarantee the final product for human use. Since we only validate and distribute aseptic tissues, we conducted a study to determine what type of quality controls for skin processing are the most suitable for detecting processing errors and intercurrent contamination, and for faithfully mapping the process without unduly increasing production costs. Two different methods for quality control were statistically compared using the Fisher exact test. On the basis of the current study we selected our quality control procedure based on pre- and post-processing tissue controls, operator and environmental controls. Evaluation of the predictability of our control methods showed that tissue control was the most reliable method of revealing microbial contamination of grafts. We obtained 100 % sensitivity by doubling tissue controls, while maintaining high specificity (77 %).

Retraction: SNHG5 promotes proliferation and induces apoptosis in melanoma by sponging miR-155.

[This retracts the article DOI: 10.1039/C7RA12520H.].

Line field confocal optical coherence tomography: An adjunctive tool in the diagnosis of autoimmune bullous diseases.

Autoimmune bullous diseases (AIBDs) still represent a considerable a source of morbidity and mortality: early identification of a specific AIBD is often difficult due to overlapping clinical and/or laboratory features and time-consuming invasive laboratory tests. We aimed to investigate the potential role of a new imaging technology, line-field confocal optical coherence tomography (LC-OCT), in the non-invasive diagnosis of AIBDs. LC-OCT was performed at lesional, perilesional and contralateral healthy sites in 30 patients, before histology and direct immunofluorescence. LC-OCT examination was able to identify the level of split (subcorneal/suprabasal/subepidermal/sublamina densa), to provide detailed images of the bulla roof morphology and content (eg, erythrocytes/acantholytic cells/polymorphonucleates). Areas of intra/subepidermal detachment were also detected also at clinically normal perilesional skin sites. LC-OCT can support physicians, real time and at bed-site, in the differential diagnosis of various AIBDs and their mimickers. Moreover, it can be used for the identification of subclinical lesions and therapy tapering.

SNHG5 promotes proliferation and induces apoptosis in melanoma by sponging miR-155.

Background: Melanoma is the most common malignancy of skin cancer. Small nucleolar RNA host gene 5 (SNHG5), a long non-coding RNA (lncRNA), has been demonstrated to be abnormally expressed in multiple malignances. However, the roles and molecular mechanisms of SNHG5 in melanoma progression have not been well identified. Methods: RT-qPCR assays were used to detect the expression patterns of SNHG5 and microRNA-155 (miR-155). Cell proliferation was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and colony formation assays. Cell apoptosis rate was measured by flow cytometry via double-staining of fluorescein isothiocyanate (FITC)-labeled annexin V (Annexin V-FITC) and propidium iodide (PI). The interaction between SNHG5 and miR-155 was validated using bioinformatics analysis, subcellular fraction assay, luciferase assay and RNA immunoprecipitation (RIP) assay. A mouse model of melanoma was established to further verify the effect of SNHG5 on tumor growth in vivo. Results: SNHG5 expression was upregulated in melanoma tumor tissues and cell lines. Moreover, higher SNHG5 expression was associated with advanced pathogenic status and poor prognosis. Functional analysis showed that SNHG5 knockdown suppressed proliferation and facilitated apoptosis in melanoma cells. Mechanical exploration revealed that SNHG5 acted as a molecular sponge of miR-155 in melanoma cells. Restoration experiments delineated that miR-155 down-regulation partly abrogated SNHG5-knockdown-mediated anti-proliferation and pro-apoptosis effect in melanoma cells. In vivo assays further demonstrated that SNHG5 depletion hindered tumor growth through up-regulating miR-155 expression. Conclusion: SNHG5 promoted the development of melanoma by sponging miR-155 in vitro and in vivo, implying the important implication of lncRNAs in melanoma progression and providing a potential therapeutic target for melanoma.

Prevalence of skin allograft discards as a result of serological and molecular microbiological screening in a regional skin bank in Italy.

BACKGROUND: Postmortem skin is widely used in the treatment of patients with severe burns. Skin specimens must be screened for transmissible agents including human immunodeficiency virus (HIV), hepatitis B (HBV) and C (HCV) virus, human T-cell lymphotropic virus (HTLV), cytomegalovirus (CMV) and Treponema pallidum. METHODS: Four hundred and sixty-one cadaveric donors underwent serological and molecular microbiological (polymerase chain reaction, PCR) screening at Siena Skin Bank between 2000 and 2004. RESULTS: 74/461 donors (16.1%) were found ineligible under current regulations. CONCLUSIONS: These results are interesting in a local context and underline the importance of screening involving both routine serological procedures and molecular microbiological investigation. The latter has not been uniformly introduced in many countries and very limited data is available to assess its cost-benefit ratio in the field of skin donor screening.

Autografts and cultured epidermis in the treatment of vitiligo.

Skin transplants can be a useful and efficacious method to treat vitiligo. The aim is to repopulate areas lacking melanocytes with functional cells taken from normally pigmented areas. Several procedures have been devised and tested: some consist in the simple transfer of epidermis sampled and implanted as is, whereas others are based on the transplantation of disaggregated and manipulated cells. The therapeutic success of the former methods is partly determined by the ability and experience of the surgeon performing the operation, whereas the results of the latter methods mainly depend on the laboratory facilities and abilities of the personnel who manipulate the cells to be transplanted. The transplantation of cultured cells is the most fascinating and promising procedure but requires the observance of still not completely predictable procedures. The use of biological material of animal origin and the use of factors to stimulate cell proliferation, such as growth factors and promoting agents, are other points that require attention.

Skin bank organization.

Skin allografts were first used at the end of the last century by Girdner [Girdner JH. Skin grafting with graft taken from the dead subject. Med Rec (NY) 1881;20:119-20]; however, routine storage of human tissue developed only in the 1930s to 1940s [Webster JP. Refrigerated skin grafts. Ann Surg 1944;120:431-49] when reliable preservation methods became available. The first proper skin bank was the US Navy Skin Bank, set up in 1949 [McCauley RL. The skin Bank. In: Herndon DN, editor. Total burn care. 1st ed. Philadelphia: Saunders; 1996. p. 159-63]. Several skin banks were subsequently established in the United States and Europe, and in most cases they were organized as multitissue banks. Nowadays, it is estimated that 30 to 50 tissue banks are active in the United States, working according to the American Association of Tissue Banking (AATB) standards (AATB. Standards for tissue banking; 1984) and federal regulations (Real E S and regulations. Fed Regist. 1993).

Other uses of homologous skin grafts and skin bank bioproducts.

The main use of homologous skin grafts or grafts of related bioproducts is in the treatment of severe burns. However, various new clinical and experimental sectors, in which this type of skin substitute can be useful, have recently emerged. The main new clinical indications for skin allografts include: skin loss, surgical wounds and bullous diseases. In these fields donor skin can be used for different purposes: as a physiological biological dressing to control pain and protect deep structures such as tendons, bones, cartilage and nerves, and to promote reepithelization with a significant reduction in healing time, and as skin substitute with dermal tissue to guide repair and make it as physiological as possible. In particular, skin bank bioproducts are currently used in the treatment of several conditions such venous and arterial leg ulcers, pressure ulcers, diabetic foot ulcers, pyoderma gangrenosum, post traumatic lesions, Mohs surgery, reconstructive surgery, wound cover in critical areas, aesthetic surgery, congenital epidermolysis bullosa and Lyell's syndrome. Skin bank bioproducts have also been used for experimental indications, to study in vitro toxicology and in vitro skin biology. Recently the demonstration that de-epidermized dermis (DED) has all the characteristics of an excellent dermal substitute into which various types of cells can be introduced and made to develop, opens exciting new possibilities of research in the field of wound healing and tissue engineering. Our preliminary observations seems to indicate that CD 34+ stem cells from umbilical cord blood can survive in DED and in a few weeks populate collagen bundles. The observation of tubular structures without lumina close to collagen bundles as well as clusters of epithelioid or fibroblast-shaped cells may represent aspects of differentiation of CD 34+ stem cells. More detailed and sophisticated studies are clearly needed to answer all the questions that these initial observations pose. Anyway the 3-dimensional model proposed seems to be suitable for the study of the behaviour of peripheral CD 34+ and perhaps also other types of stem cells in 3-dimensional dermal matrix.

Identification of a novel, alpha2-fucosylation-dependent uptake system in highly proliferative cells.

In this paper we describe a new structure present in highly proliferative cells and absent in cells with normal growth potential. We used cultured bovine venular endothelial cells (CVEC) as examples of high proliferation, and dermal fibroblasts of a primary culture as examples of normal proliferation. The structure, consisting of tubules radiating from the nuclear region to the tips of cell protrusions, was revealed by its strong positivity to the fucose-binding lectin from Lotus (LTL) that prefers glycans with alpha-1,2-linked fucose. Another fucose-binding lectin that prefers glycans with alpha-1,6-linked fucose was instead found to localize glycans exclusively in Golgi complexes. LTL binding sites were also found at the surface of CVEC in a restricted region close to the nucleus. The role of alpha-1,2-linked fucose in forming or maintaining the tubules was confirmed by the fact that down-regulation of the fucosyltransferases FUT1 and FUT2 resulted in disappearance of the tubular structure. LTL also proved able to penetrate the cells through the tubular structures up to the nuclear region and to inhibit proliferation. Endostatin was also found to massively penetrate the cells in the tubular structures in control cells but not in FUT1/2 depleted cells. In cells of a first passage primary culture of dermal fibroblasts the tubular LTL-positive structure was absent as well as the LTL-positive sites at the external surface, and both fucose-binding lectins were found to exclusively localize glycans in Golgi complexes. Tubules were again found progressively in fibroblasts derived from repeated passages, where faster growing cells predominate. Disappearance of LTL-positivity in Golgi complexes paralleled appearance of LTL-positive tubules. The role of Golgi complexes in forming the tubules is discussed.

Congenital subcutaneous granuloma annulare.

Granuloma annulare is a palisading granulomatous skin disease which may be generalized, localized, perforating, or subcutaneous. Subcutaneous granuloma annulare is self-limiting, affecting infants and children, with typical postnatal onset. Here we report a patient with congenital clinical manifestations.

Autologous epidermal cultures and narrow-band ultraviolet B in the surgical treatment of vitiligo.

BACKGROUND: Vitiligo is an acquired skin disorder with a great social impact. It can be successfully treated with autologous epidermal grafting. OBJECTIVE: To evaluate the possibility of treating vitiligo by autologous grafting of epidermal cells and narrow-band ultraviolet B (UVB). METHODS: Autologous epidermal cultures were prepared starting from small biopsies of normally pigmented skin. Cells were cultured on hyaluronic acid membranes using medium supplemented with patient's serum. Cell cultures were grafted onto laser-abraded depigmented areas. Patients underwent narrow-band UVB therapy 3 weeks after grafting. RESULTS: Repigmentation of the grafted areas started 1 month after transplant and continued until 4 months after grafting. All patients were evaluated 3, 6, 12, and 18 months after grafting. At the 18-month follow-up, repigmentation was observed in 75% of patients with focal and segmental vitiligo and in 30% of patients with generalized vitiligo. CONCLUSIONS: This therapy can be considered for the treatment of stable vitiligo (especially focal and segmental) resistant to standard therapies. Their results are encouraging from the clinical and esthetic point of view, although the treatment is costly and highly specialized.

Skin allograft in the treatment of toxic epidermal necrolysis (TEN).

BACKGROUND: TEN is a severe form of exfoliative dermatitis. Its course is acute and its outcome fatal in 40% of cases. Wound cover to prevent fluid/protein loss and infections and to control pain, is the first step, as for burns. Skin allograft can be successfully used for this purpose. OBJECTIVE: We report two cases of TEN with de-epithelialization of 50 and 70% of the total body surface area. The patients were given support therapy and treated with human glycerol-preserved skin allografts for wound cover. METHODS: Patients were grafted with glycerol-preserved donor skin, obtained from a skin bank. RESULTS: Re-epithelization of treated areas was complete in 8 days; pain relief was obtained soon after the graft. CONCLUSIONS: Glycerol-preserved skin allograft is an effective treatment in extensive skin loss, for its barrier and analgesic effect. Quality standards of this product ensure safety and simplicity of use at limited cost.