The Effect of Boric Acid and Sodium Pentaborate Pentahydrate-Treated Foreskin Derived Mesenchymal Stem Cells on Liver Fibrosis.

Liver fibrosis is a worldwide public health problem due to its life-threatening complications, including portal hypertension, liver failure, cirrhosis, and hepatocellular carcinoma (HCC). Liver fibrosis is the net result of a complex excessive accumulation of extracellular matrix (ECM). Activation of hepatic stellate cells (HSCs) are the cause of deposition of ECM and are commonly recognized as a key step in liver fibrosis. The aim of this study was to investigate the effect of foreskin-derived mesenchymal stem cells treated with boron compounds on liver fibrosis. Rats were injected intraperitoneally with thioacetamide (TAA) at a dose of 150 mg/kg except sham and control groups' rats. Thioacetamide (TAA), foreskin-derived mesenchymal stem cells (TAA + FSDMSC), FSDMSC treated with boric acid (TAA + FSDMSC + BA), FSDMSC treated with sodium pentaborate pentahydrate (TAA + FSDMSC + NaB), control and sham groups were studied. Boron compound treated foreskin-derived mesenchymal stem cells were injected into the tail vein, and evaluations were conducted after 4 weeks and liver tissues were obtained for structural, immunohistochemical, and western blot studies and blood samples were taken for biochemical analysis. FSDMSC (BA) alleviates TAA-induced rats liver fibrosis, and BA showed a positive effect on foreskin-derived mesenchymal stem cells viability. After using BA-treated mesenchymal stem cells, we observed that there was regression in the fibrotic areas at TAA-induced liver fibrosis. The result demonstrates that the contribution of TAA + FSDMSC and TAA + FSDMSC (NaB) at the level of structure is not effective in regression of fibrosis in TAA-generated liver fibrosis. We concluded that FSDMSC treated with BA may be a factor in the regression of fibrosis.

Foreskin leiomyosarcoma: a case report. / [Leiomiosarcoma de prepucio: reporte de un caso]

Introduction: Primary skin leiomyosarcomas are infrequent neoplasms. They correspond to 2-3% of skin sarcomas and are most frequently located on the lower extremities, trunk and genitals. Methods: We present a case of a 73-year-old man with a 4-month evolution of foreskin leiomyosarcoma. The lesion was biopsied for histopathological study with HE and immunohistochemistry with smooth muscle actin, specific muscle actin, CD34, p63 and S-100 (-). Results: We observed a leiomyosarcoma of high histological grade and mitotic count. It was positive by immunohistochemistry for smooth muscle actin, while the other markers were negative. Surgical limits were compromised so a reoperation with wide margins of healthy tissue was necessary. Conclusion: The skin lesions should be removed all, without exception, since they can be neoplasms of variable biological behavior. The histological study must be complemented with immunohistochemistry to differentiate them from other neoplasms. For the prognosis, the histological grade, size, location and the possibility of resection with wide margins must be taken into account.

Introducción: Los leiomiosarcomas primarios de piel son neoplasias infrecuentes. Corresponden al 2-3 % de los sarcomas cutáneos y se localizan con mayor frecuencia en las extremidades inferiores, tronco y genitales. Método: Presentamos un caso de un varón de 73 años con un leiomiosarcoma en prepucio de 4 meses de evolución. Se le practicó biopsia excisional de la lesión para estudio histopatológico con HE e inmunohistoquímica con actina de músculo liso, actina muscular específica, CD34, p63 y S-100 (-). Resultados: Observamos un leiomiosarcoma de alto grado histológico y recuento mitótico. Presentó positividad por inmunohistoquímica para actina de músculo liso, en tanto que los otros marcadores fueron negativos. Los límites quirúrgicos estuvieron comprometidos por lo que fue necesaria una reintervención con amplios márgenes de tejido sano. Conclusión: Las lesiones de piel deben extirparse todas, sin excepción, ya que pueden tratarse de neoplasias de conducta biológica variable. El estudio histológico debe complementarse con inmunohistoquímica para diferenciarlas de otras neoplasias. Para el pronóstico se debe tener en cuenta el grado histológico, el tamaño, la localización y la posibilidad de resección con amplios márgenes.

Folliculosebaceous units are a frequent finding in early lichen sclerosus of the foreskin.

BACKGROUND: Folliculosebaceous units (FSU) has been considered an early target of inflammation in vulvar lichen sclerosus (VLS). This diagnostic clue is not reported in lichen sclerosus (LS) of the foreskin (FLS) that is normally hairless. We evaluated the presence and inflammation of FSU and sebaceous glands (SG) in LS of the foreskin. METHODS: Histological specimens from therapeutic circumcision were assessed in order to evaluate the frequency and inflammation of FSU and SG in LS. RESULTS: Ninety-eight cases, grouped into 46 early (group 1) and 52 overt (group 2) FLS were included in the study. SG-FSU were found in 95.7% of group 1, and 65.4% of group 2 cases. Their density was inversely correlated with patient age (P=0.0014). We observed perifollicular inflammation in all cases with visible SG-FSU and frequent FSU abnormalities. CONCLUSIONS: SG and FSU were frequent in early FLS and decreased in advanced disease and adults. We hypothesize that SG and FSU are involved in the inflammatory process leading to FLS. These data, which need further investigation, could help to better understand the pathogenesis of FLS.

The role of occlusion and micro-incontinence in the pathogenesis of penile lichen sclerosus: an observational study of pro-inflammatory cytokines' gene expression.

PURPOSE: To assess the expression of selected cytokines in penile lichen sclerosus (PLS) and associate them with the occurrence of micro-incontinence (MI) in different stages of PLS. METHODS: The skin biopsies from 49 PLS affected, and 13 from nonlesional foreskins (healthy control adult males undergoing circumcision due to phimosis caused by short frenulum) were obtained. All specimens were used for RNA extraction and RT-qPCR. Quantitative assessment of the gene expression of interleukin 1-A (IL-1A), interleukin 1-B (IL-1B), interleukin 1 receptor antagonist (IL-1RN), interleukin 6 (IL-6), transforming growth factor ß1 (TGF-ß1), and interferon-gamma (INF-γ) was performed. To determinate the presence of MI, the patients were asked about voiding patterns, especially leaking tiny drops of urine from the urethral meatus after urination. RESULTS: IL-1A, IL-6, and INF-γ mRNA levels were approximately 150, 16, and 59 times higher in PLS than in control samples, respectively. The highest IL-1A mRNA levels were observed in early PLS (n = 13), INF-γ in moderate PLS (n = 32), while IL-6 in severe PLS (n = 4). MI was noted in 45 PLS patients vs. 0 in control (p < 0.0001). IL-1A and IL-6 vs control ratios were concentration (ca.) 400 and 30 times higher, respectively, in MI PLS samples than in PLS without MI. CONCLUSION: Occlusion and irritating urine effect are associated with the clinical progression of penile LS with increased mRNA expression of IL-1A, INF-γ, and IL-6 pro-inflammatory cytokines in the foreskin.

Long-Chain and Very Long-Chain Ceramides Mediate Doxorubicin-Induced Toxicity and Fibrosis.

Doxorubicin (Dox) is a chemotherapeutic agent with cardiotoxicity associated with profibrotic effects. Dox increases ceramide levels with pro-inflammatory effects, cell death, and fibrosis. The purpose of our study was to identify the underlying ceramide signaling pathways. We aimed to characterize the downstream effects on cell survival, metabolism, and fibrosis. Human fibroblasts (hFSF) were treated with 0.7 µM of Dox or transgenically overexpressed ceramide synthase 2 (FLAG-CerS2). Furthermore, cells were pre-treated with MitoTempo (MT) (2 h, 20 µM) or Fumonisin B1 (FuB) (4 h, 100 µM). Protein expression was measured by Western blot or immunofluorescence (IF). Ceramide levels were determined with mass spectroscopy (MS). Visualizations were conducted using laser scanning microscopy (LSM) or electron microscopy. Mitochondrial activity was measured using seahorse analysis. Dox and CerS2 overexpression increased CerS2 protein expression. Coherently, ceramides were elevated with the highest peak for C24:0. Ceramide- induced mitochondrial ROS production was reduced with MT or FuB preincubation. Mitochondrial homeostasis was reduced and accompanied by reduced ATP production. Our data show that the increase in pro-inflammatory ceramides is an essential contributor to Dox side-effects. The accumulation of ceramides resulted in a lipotoxic shift and subsequently mitochondrial structural and functional damage, which was partially reversible following inhibition of ceramide synthesis.

Phototoxic versus photoprotective effects of tattoo pigments in reconstructed human skin models: In vitro phototoxicity testing of tattoo pigments: 3D versus 2D.

The increasing number of tattooed persons urges the development of reliable test systems to assess tattoo associated risks. The alarming prevalence of 60 % phototoxic reactions in tattoos ask for a more comprehensive investigation of phototoxic reactions in tattooed skin. Here, we aimed to compare the cellular responses of human skin cells to ultraviolet (UV)A and UVB irradiation in doses of short to intermitted sun exposure (3-48 J/cm² and 0.05-5 J/cm², respectively) in the presence of tattoo pigments. Therefore, we used fibroblast monolayer culture (2D), our recently developed three dimensional full-thickness skin model with dermal-located tattoo pigments (TatSFT) and its dermal equivalents (TatSDE) that lack keratinocytes. We tested the most frequently used tattoo pigments carbon black, titanium dioxide (TiO2) anatase and rutile as well as Pigment Orange (P.O.)13 in ranges from 0.067 to 2.7 ng/cell in 2D. For TatSDE and TatSFT, concentrations were 1.3 ng/cell for TiO2, 0.67 ng/cell for P.O.13 and 0.067 ng/cell for carbon black. We assessed cell viability and cytokine release in all systems, and cyclobutane pyrimidine dimer (CPD) formation in TatSFT. Phototoxicity of tattoo pigments was exclusively observed in 2D, where especially TiO2 anatase induced phototoxic effects in all concentrations (0.067-2.7 ng/cell). In contrast, fibroblasts were protected from UV irradiation in TatSDE by TiO2 and carbon black. Neither toxic nor protective effects were recorded in TatSFT. P.O.13 showed altered cytokine secretion in 2D (0.067-1.3 ng/cell) and TatSDE, despite the absence of significant effects on viability in all systems. All pigments reduced the number of CPDs in TatSFT compared to the pigment-free controls. In conclusion, our study shows that within a 3D arrangement, intradermal tattoo pigments may act photoprotective despite intrinsic phototoxic properties in 2D. Thus, dermal 3D equivalents should be considered to evaluate acute tattoo pigment toxicology.

A competitive nature-derived multilayered scaffold based on chitosan and alginate, for full-thickness wound healing.

The aim of this study was to evaluate a bioactive multilayer wound dressing, based on chitosan and alginate. To enhance healing potential, Dracaena Cinnabari and Aloe Vera were loaded as separate layers into the scaffold. The bare and bioactive multilayered scaffolds were fabricated by an iterative layering freeze-drying technique. Following of topographical, chemical, and physical assessment, the performance of the scaffolds was evaluated in vitro and in vivo. The results revealed adequate attachment, and proliferation of human foreskin fibroblasts, indicating excellent biocompatibility of the bioactive scaffold. In vivo, the performance of the multi-layered scaffold loaded with the bioactive materials was comparable with Comfeel plus®. The wounds treated with the bioactive scaffold exhibited superior hypergranulation, fibroblast maturation, epithelization, and collagen deposition, with minimal inflammation, and crust formation. It is concluded that the synergism of extracellular matrix-mimicking multi-layered scaffolding with Aloe Vera and Dracaena Cinnabari could be considered as a supportive wound dressing.

Biobanked human foreskin epithelial cell sheets reduce inflammation and promote wound healing in a nude mouse model.

BACKGROUND: Human epithelial cell sheets (ECSs) are used to clinically treat epithelial conditions such as burns, corneal blindness, middle ear cholesteatoma and vitiligo. As a widely used material in clinic, there is little information on the biobanking of ECSs and its repair effect after storage. RESULTS: Two methods for biobanking foreskin ECSs were compared in a short term (7 days): 4-degree storage and programmed cryopreservation. Cell sheet integrity, viability, apoptosis, immunogenicity, mechanical properties and function were evaluated. In vivo, ECSs were directly transplanted to skin defect models and histological examination was performed at 1 week postoperatively. We successfully extracted human foreskin-derived primary epithelial cells and fabricated them into ECSs. Compared with 4-degree storage, programmed cryopreservation preserved the ECS structural integrity, enhanced the mechanical properties, decreased HLA-I expression, and increased cell viability and survival. An increased proportion of melanocytes with proliferative capacity remained in the cryopreserved sheets, and the undifferentiated epithelial cells were comparable to those of the fresh sheets. In vivo, cryopreserved ECSs could reduce inflammatory cell infiltration and promote connective tissue remodeling, epithelial cell proliferation and vascular regeneration. CONCLUSIONS: Programmed cryopreservation of ECSs was superior and more feasible than 4-degree storage and the cryopreserved ECSs achieved satisfying skin wound healing in vivo. We anticipate that the off-the-shelf ECSs could be quickly used, such as, to repair human epithelial defect in future.

Impact of chemokine C-C ligand 27, foreskin anatomy and sexually transmitted infections on HIV-1 target cell availability in adolescent South African males.

We compared outer and inner foreskin tissue from adolescent males undergoing medical male circumcision to better understand signals that increase HIV target cell availability in the foreskin. We measured chemokine gene expression and the impact of sexually transmitted infections (STIs) on the density and location of T and Langerhans cells. Chemokine C-C ligand 27 (CCL27) was expressed 6.94-fold higher in the inner foreskin when compared with the outer foreskin. We show that the density of CD4+CCR5+ cells/mm2 was higher in the epithelium of the inner foreskin, regardless of STI status, in parallel with higher CCL27 gene expression. In the presence of STIs, there were higher numbers of CD4+CCR5+ cells/mm2 cells in the sub-stratum of the outer and inner foreskin with concurrently higher number of CD207+ Langerhans cells (LC) in both tissues, with the latter cells being closer to the keratin surface of the outer FS in the presence of an STI. When we tested the ability of exogenous CCL27 to induce T-cell migration in foreskin tissue, CD4 + T cells were able to relocate to the inner foreskin epithelium in response. We provide novel insight into the impact CCL27 and STIs on immune and HIV-1 target cell changes in the foreskin.

Topical estradiol increases epidermal thickness and dermal collagen of foreskin prior to hypospadia surgery - Randomized double blinded controlled trial.

INTRODUCTION: The use of preoperative topical testosterone stimulation prior to hypospadias correction aims to increase penile size and achieve better surgical results. Topical estradiol has been shown to improve the quality of skin in other sites, but its use in boys with hypospadia has not yet been elucidated. OBJECTIVE: This study aims to evaluate the primary effects in epidermal thickness and collagen distribution of estradiol compared to testosterone and placebo in skin of prepuce before hypospadia surgery. MATHERIALS AND METHODS: Patients were randomized into three groups according to the topical hormone used: TG: Testosterone ointment; EG: Estradiol ointment; CG: Neutral base ointment. Fragments of foreskin were excised, fixed and then sectioned for histology. For each sample, epidermal thickness and dermal collagen expression was measured by specific computer analysis, P-values of <0.05. RESULTS: Thirty-three patients with a mean age of 4.01 ± 2.92 years were included. Hypospadia classification was similar in all three groups. Mean epidermal thickness and collagen type I expression in EG were greater than those of the other groups. Collagen type III expression was similar in all groups. DISCUSSION: Foreskin has a fundamental role in many techniques of hypospadias surgery and can be used either as a graft or a flap in the correction of the penile defect. Increase of epidermal thickness and dermal collagen observed in the present study has already been related to use of estradiol in other skin sites, but not yet in foreskin. Further studies are needed to evaluate the real significance of these findings in boys with hypospadias. CONCLUSION: Use of topical estradiol before hypospadias surgery lead to greater epidermal thickness and increases dermal collagen expression in foreskin.

Zebras in Foreskin Dermatopathology: A Review.

OBJECTIVES: The aim of the study was to review uncommon foreskin dermatopathology conditions clinically and pathologically. METHODS: A database search of PubMed and Google Scholar were extracted between March 1, 2009, and March 1, 2019, using the search terms "foreskin," "prepuce," "penis," "pathology," "dermatology," and "rare." The search was limited to "humans" and "dermatopathology." Full article texts were reviewed. Reference lists were screened for additional articles. Patient details (diagnosis, dermatopathology, treatment, and follow-up if available) were extracted. We excluded articles written in the non-English language, unusual variants of common conditions, and cases of common dermatologic conditions. RESULTS: A list of 369 articles was identified and another screening identified 30 articles for rare foreskin pathologies. Those are divided into categories based on the following etiologies: (a) benign, including congenital (e.g., aposthia), infectious (graft versus host disease and histoplasma), autoimmune (Crohn's disease and pyoderma gangrenosum), and benign neoplasms (neurofibroma, apocrine hidrocystoma, verruciform xanthoma, porokeratosis, penile cutaneous horn, localized amyloidosis) and (b) malignancies, including primary (myeloid sarcoma, basal cell carcinoma, Kaposi's sarcoma, mucosal-associated lymphoid tissue lymphoma), and metastasis. CONCLUSIONS: We reviewed and discussed unusual benign and malignant dermatopathology conditions that can affect the foreskin.

Foreskin reconstruction at the time of single-stage hypospadias repair: is it a safe procedure?

INTRODUCTION: Foreskin reconstruction (FR) is a recognised, yet debated, option for patients undergoing single-stage hypospadias repair (HR). METHODS: We evaluated the incidence of complications after single-stage HR in our institution. This is a retrospective review of all single-stage HR. Patients were classified into group 1 (circumcision) and group 2 foreskin reconstruction (FR). Urethroplasty and foreskin complications were recorded. Statistics used are as follows: Mann-Whitney test to compare age at operation and length of follow-up (FU); Chi-Square test to analyse the incidence of urethral complications and need for reoperation; Log rank test to compare the survival curves; p statistically significant < 0.05. Data are presented as median (range). RESULTS: 304 patients were identified, operated between January 2010 and December 2016, and 20 were excluded: 6 already circumcised at the time of the surgery, 3 with megameatus intact prepuce, 11 lost at FU. 284 patients were included: 161 circumcised and 123 FR. Median age at the operation was 17 months (8-179) (group 1) and 17 months (8-148) (group 2) (p = 0.71). Length of FU was 19 months (8-91) (group 1) and 17 months (4-87) (group 2) (p = 0.45). The survival curve was homogeneous (p = 0.28). Urethroplasty complications occurred in 32/161 (20%) (group 1) and in 21/123 (17%) (group 2) (p = 0.55). Foreskin complications occurred in 18/123 (15%). A second operation was required in 33 boys in each group, (20% group 1 and 27% group 2) (p = 0.21). CONCLUSION: FR does not increase the complication rate or the need for a reoperation after single-stage HR. Parents should be offered the option between the two procedures according to their personal preference.