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SUMMARY: Keloid scar is a unique benign fibroproliferative tumor of the human skin. Previously, it was reported that early growth response 1 (EGR1), a transcription factor, promotes keloid fibrosis; however, the mechanism by which EGR1 modulates keloid formation was not elaborated. In this research, the specific function and the microRNA (miRNA) regulatory network of EGR1 in keloids was examined. Keloid fibroblasts (KFs) were transfected with EGR1-small interfering RNA (siEGR1), EGR1-overexpression plasmid (pcDNA3.1-EGR1), and microRNA (miR-183-5p)-mimics to regulate the expression of EGR1 and miR-183-5p. The study employed dual-luciferase reporter assays to explore the targeting regulation of miR-183-5p on EGR1. Additionally, Western blotting, flow cytometry, qRT-PCR, cell count kit-8 (CCK-8), transwell, and wound healing assays, and RNA sequencing were conducted. EGR1 was upregulated in KFs, and EGR1 silencing diminished proliferation, fibrosis, migration, invasion, and apoptosis of cells. In KFs, the expression of miR- 183-5p was reduced, leading to the inhibition of cell proliferation, migration, and invasion. Conversely, it enhanced apoptosis. By targeting EGR1, miR-183-5p partially counteracted the impact of EGR1 on migration, invasion, and fibrosis in KFs. The findings imply that miR-183-5p suppresses keloid formation by targeting EGR1. As a result, EGR1 holds promise as a potential therapeutic target for preventing and treating keloids.
La cicatriz queloide es un tumor fibroproliferativo benigno único de la piel humana. Anteriormente, se informó que la respuesta de crecimiento temprano 1 (EGR1), un factor de transcripción, promueve la fibrosis queloide; sin embargo, no se explicó el mecanismo por el cual EGR1 modula la formación de queloides. En esta investigación, se examinó la función específica y la red reguladora de microARN (miARN) de EGR1 en queloides. Se transfectaron fibroblastos queloides (KF) con ARN de interferencia pequeño de EGR1 (siEGR1), plásmido de sobreexpresión de EGR1 (pcDNA3.1-EGR1) y miméticos de microARN (miR-183-5p) para regular la expresión de EGR1 y miR-183. -5p. El estudio empleó ensayos de indicador de luciferasa dual para explorar la regulación dirigida de miR-183-5p en EGR1. Además, se realizaron pruebas de transferencia Western, citometría de flujo, qRT-PCR, kit de recuento celular-8 (CCK-8), transwell y curación de heridas, y secuenciación de ARN. EGR1 estaba regulado positivamente en KF, y el silenciamiento de EGR1 disminuyó la proliferación, fibrosis, migración, invasión y apoptosis de las células. En KF, la expresión de miR- 183-5p se redujo, lo que llevó a la inhibición de la proliferación, migración e invasión celular. Por el contrario, mejoró la apoptosis. Al apuntar a EGR1, miR-183-5p contrarrestó parcialmente el impacto de EGR1 en la migración, invasión y fibrosis en KF. Los hallazgos implican que miR-183-5p suprime la formación de queloides al apuntar a EGR1. Como resultado, EGR1 es prometedor como objetivo terapéutico potencial para prevenir y tratar los queloides.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Young Adult , Early Growth Response Protein 1 , Fibroblasts , Keloid/genetics , Keloid/pathology , Wound Healing , Transfection , Down-Regulation , Cell Movement , Blotting, Western , Sequence Analysis, RNA , Apoptosis , MicroRNAs/physiology , Cell Proliferation , Real-Time Polymerase Chain ReactionABSTRACT
Queloides e cicatrizes hipertróficas são lesões formadas a partir da resposta fibroproliferativa anormal ao processo de cicatrização de feridas, gerando uma proliferação excessiva do colágeno nas lesões. Geralmente, predominam em pacientes do sexo feminino e em indivíduos com tons de pele mais escuros. A abordagem terapêutica dessas cicatrizes pode ser indicada de acordo com alguns critérios, como déficit funcional, tamanho e tempo de cicatrização da ferida. Nesse sentido, o presente estudo objetivou realizar uma revisão descritiva da literatura, buscando as evidências de tratamento dos últimos cinco anos neste tema. A revisão foi realizada com base no guideline PRISMA, utilizando as bases de dados PubMed, LILACS, Cochrane Library, SCOPUS, Web of Science e Grey Literature, entre os anos de 2018 e 2022. Foram encontrados 740 artigos, dos quais 16 ensaios clínicos randomizados foram selecionados. Foi evidenciado que manejo do queloide apresenta abordagem multimodal, não havendo um padrão-ouro de tratamento, com taxa de recorrência baixa. Além disso, a terapia combinada de diferentes agentes pareceu ser superior ao uso isolado de métodos terapêuticos no tratamento dessas lesões.
Keloids and hypertrophic scars are lesions formed from the abnormal fibroproliferative response to the wound healing process, generating excessive collagen proliferation in the lesions. They generally predominate in female patients and individuals with darker skin tones. The therapeutic approach to these scars can be indicated according to criteria such as functional deficit, size, and wound healing time. In this sense, the present study aimed to conduct a descriptive review of the literature, seeking evidence of treatment over the last five years. The review was carried out based on the PRISMA guideline, using the databases PubMed, LILACS, Cochrane Library, SCOPUS, Web of Science, and Grey Literature between 2018 and 2022. Seven hundred forty articles were found, of which 16 randomized clinical trials were selected. It was demonstrated that keloid management presents a multimodal approach, with no gold standard of treatment with a low recurrence rate. Furthermore, combined therapy with different agents appeared superior to the isolated therapeutic methods in treating these injuries.
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Introduction: Keloids are one of the most aggressive spectrums of healing disorders. They have a unique pathophysiology and multiple specific genetic and cellular factors, which have not yet been fully elucidated. So far, literature reviews have found the influence of genetics, injury site, and ethnicity on the incidence and rate of recurrence. Furthermore, the need to associate an adjuvant method with surgical excision has already been verified, but the best therapy has yet to be defined. Method: A retrospective analysis of medical records was carried out to assess the profile of patients who underwent postoperative radiotherapy with an electron beam to treat keloids at the Hospital das Clínicas da Faculdade de Medicina de Botucatu between 2015 and 2019. Results: Data from 131 patients were evaluated, with 269 keloid scars treated. The average duration of treatment was 51 days, and the number of sessions was 17. White patients were predominant (78%) and women (70%), with surgical incision being the main cause of formation (49%) and ear the most identified location (33%). Women were more likely to complete the proposed treatment (p=0.04), while non-literates completed less than those who had at least completed primary or secondary education (p<0.0001). Conclusion: Postoperative radiotherapy for keloid scars is a well-established treatment in the literature and an important tool for plastic surgeons. Knowing the profile of patients who need this therapy is essential to create methods that improve adherence and results.
Introdução: Os queloides correspondem a um dos espectros mais agressivos dos distúrbios da cicatrização. Possuem fisiopatologia ímpar e múltiplos fatores genéticos e celulares específicos, ainda não totalmente elucidados. Até o momento, revisões literárias encontraram influência da genética, local da lesão e etnia sobre a incidência e taxa de recorrência. Ademais, já foi constatada a necessidade de associação de um método adjuvante com a excisão cirúrgica, mas ainda sem definição da melhor terapia. Método: Realizada uma análise retrospectiva de prontuários para avaliação do perfil dos pacientes submetidos a radioterapia pós-operatória com feixe de elétrons para o tratamento de queloides no Hospital das Clínicas da Faculdade de Medicina de Botucatu, entre 2015 e 2019. Resultados: Foram avaliados os dados de 131 pacientes, com um total de 269 cicatrizes queloideanas tratadas. A média da duração do tratamento foi de 51 dias e do número de sessões, de 17. Houve predominância de pacientes brancos (78%) e de mulheres (70%), sendo incisão cirúrgica a principal causa de formação (49%) e a orelha a localização mais identificada (33%). As mulheres tiveram mais chance de completar o tratamento proposto (p=0,04), enquanto os não alfabetizados completaram menos do que aqueles que tinham pelo menos ensino fundamental ou médio completo (p<0,0001). Conclusão: A radioterapia pós-operatória em cicatrizes queloideanas é um tratamento consagrado na literatura e uma importante ferramenta do cirurgião plástico. Conhecer o perfil dos pacientes que necessitam desta terapia é fundamental para criar métodos que melhorem a adesão e o resultado da mesma.
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Resumen El acné queloide nucal es una forma crónica de foliculitis cicatricial, caracterizada por el desarrollo de papulo-pústulas que se convierten en queloides cuyo proceso fisiopatológico no está bien dilucidado. Se presenta un paciente de 19 años, con antecedente familiar de acné queloideo nucal, cuya aparición es posterior al ingreso de una bacteria en el cuero cabelludo. El caso se reporta por la rara frecuencia de la enfermedad y como conocimiento de una posible activación de la enfermedad por el ingreso de una bacteria al organismo del paciente. En este caso, se realiza una revisión breve del tema y resalta la exacerbación del proceso inflamatorio y empeoramiento de la enfermedad con medicina alternativa usada deliberadamente por familiares del paciente.
Abstract Keloid acne nuchae is a chronic form of cicatricial folliculitis, characterized by the development of papules, pustules, and keloid-like plaques which pathophysiological process is not well elucidated. We report a 19-year-old patient, with a family backgroundof keloid acne nuchae, whose appearance is after the entry of a bacterium into the scalp. The case is also reported due to the rare frequency of the disease and as knowledge of a possible activation of the disease by the entry of a bacterium into the patient's organism. In this case, a brief review of the subject is made and the exacerbation of the inflammatory process and worsening of the disease with alternative medicine deliberately used by the patient's relatives is emphasized.
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Objective:To verify that hypoxia induces epithelial mesenchymal transition in HaCaT cells, and to observe the effect of LncRNA HOTAIR on epithelial mesenchymal transition in HaCaT cells.Methods:From Jan. 2018 to Dec. 2018 in Chinese Academy of Medical Science, HaCaT cells were divided into four groups: group A cultured under normoxia, group B cultured under hypoxia, group C transfected with sh-control cultured under hypoxia, and group D transfected with sh-LncRNA HOTAIR cultured under hypoxia. After 36 hours of culturing, the expression of E-cadherin and vimentin were detected using qPCR and Western blot. The migration of HaCaT cells was evaluated by Transwell assay.Results:The relative quantity of E-cadherin mRNA in the four groups were 3.076±0.271, 1.000±0.089, 1.024±0.222, and 2.595±0.085, while vimentin mRNAs were 1.002±0.183, 4.170±0.279, 4.111±0.477, and 2.412±0.134, respectively. In addition, the Transwll invasion assay showed that numbers of cell migration in the four groups were 32.70±3.93, 125.40±6.26, 120.10±6.79, and 58.24±7.06, respectively.Conclusions:The study suggests that hypoxia promotes epithelial mesenchymal transition in keratinocytes. Furthermore, downregulation of HOTAIR is noted to inhibit epithelial mesenchymal transition of keratinocytes under hypoxia condition.
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Objective:To explore the staged treatment of keloid scars and to optimise treatment options.Methods:From June 2005 to June 2019, 629 keloid patients, 253 males and 376 females, aged 9 to 78 years, with a mean age of 31.3±16.8 years, were admitted to the Department of Aesthetic Plastic Surgery of Weifang People＇s Hospital. Three-stage comprehensive treatment was administered according to the stage of the disease from low to high, and the treatment effect was observed regularly, and the recurrence rate was counted.Results:In stage Ⅰ, 251 patients had local recurrence in 14 cases (5.5%) and 4 cases (1.6%) at 2 years 6 months after treatment, all of whom were cured after re-injection; in stage Ⅱ, 302 patients had local recurrence in 56 cases (18.6%) at 6 months after treatment, 49 patients (87.5%) were cured after re-treatment and 35 patients (11.6%) at 2 years; in stage Ⅲ, at 6 months after treatment, 36 patients (47.3%) had recurrence and 19 patients (25%) had recurrence at 2 years after re-treatment.Conclusions:The results and recurrence rate of keloids after comprehensive treatment are related to their severity, and a more satisfactory outcome can be achieved by staging the treatment according to the stage of the disease.
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Objective:To determine the expression of transmembrane protein 45A (TMEM45A) in keloid tissues and fibroblasts, and to evaluate its effect on extracellular matrix (ECM) synthesis by keloid-derived fibroblasts (KFs) .Methods:Samples of surgically excised keloid and normal foreskin tissues were collected from the Department of Dermatology and Department of Urology of Yanbian University Hospital from January 2019 to December 2020, and TMEM45A protein expression was determined in keloid tissues and KFs by Western blot analysis. KFs were divided into TMEM45A-specific small interfering RNA (siRNA) group and control siRNA group to be transfected with the TMEM45A-specific siRNA and control siRNA respectively. Then, Western blot analysis was performed to evaluate the effects of down-regulation of the TMEM45A gene on the expression of myofibroblast marker protein (α-smooth muscle actin) and ECM-related proteins.Results:Compared with normal skin tissues (1.00 ± 0.11) and fibroblasts (1.00 ± 0.20), TMEM45A expression levels significantly decreased in keloid tissues (0.26 ± 0.05) and KFs (0.41 ± 0.09), respectively ( t = 10.76, 4.75, P < 0.001, = 0.009, respectively). The expression levels of α-smooth muscle actin, ECM-related type Ⅰ collagen, type Ⅲ collagen, and fibronectin were significantly higher in the TMEM45A-specific siRNA group than in the control siRNA group ( t = -5.98, -4.57, -4.90, -7.19, P = 0.004, 0.010, 0.008, 0.002, respectively) . Conclusion:Lowly expressed TMEM45A in keloids may play an important role in the pathogenesis of keloids by promoting ECM synthesis.
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Objective:To investigate effects of the ROCK1 gene on proliferation and migration of and related molecular expression in keloid fibroblasts.Methods:Immunohistochemical technique was used to detect ROCK1 protein expression in human keloids and normal skin tissues, and Western blot analysis was performed to detect the expression of ROCK1, transforming growth factor β1 (TGF-β1) and E-cadherin in keloid tissues. In vitro cultured human keloid fibroblasts (HKFs) were divided into 4 groups: ROCK1 gene overexpression control group (ROCK1 NC group) transfected with ROCK1 gene overexpression control vectors, ROCK1 gene overexpression group (ROCK1 OE group) transfected with ROCK1 gene overexpression vectors, ROCK1 gene knockdown control group (sh NC group) transfected with ROCK1 gene knockdown control vectors, and ROCK1 gene knockdown group (shROCK1 group) transfected with ROCK1 gene knockdown vectors. Cell counting kit-8 (CCK8) assay was performed to evaluate the effect of ROCK1 gene on the survival rate of HKFs, Transwell assay to evaluate the effect on the migration of HKFs, and real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were conducted to determine the mRNA and protein expression of ROCK1, TGF-β1 and E-cadherin, respectively. Results:Immunohistochemical study showed that ROCK1 protein expression decreased significantly in the human keloid tissues compared with the normal tissues ( t = 6.47, P = 0.003) ; Western blot analysis showed that the expression levels of ROCK1 and E-cadherin significantly decreased ( t = 14.02, 162.20, respectively, both P < 0.001), while TGF-β1 expression significantly increased ( t = 76.01, P < 0.001) in the keloid tissues compared with the expression levels of corresponding proteins in the normal tissues. CCK8 assay showed that the cell activity was significantly lower in the ROCK1 OE group than in the ROCK1 NC group after 24-hour transfection ( t = 3.25, 3.78, P = 0.031, 0.019, respectively), and significantly higher in the shROCK1 group than in the sh NC group ( t = 3.12, 2.79, P = 0.036, 0.049, respectively). Transwell assay showed that the number of migratory cells was significantly lower in the ROCK1 OE group than in the ROCK1 NC group ( t = 5.17, P = 0.004), and significantly higher in the shROCK1 group than in the sh NC group ( t = 9.28, P < 0.001). Compared with the ROCK1 NC group, the ROCK1 OE group showed significantly increased mRNA and protein expression levels of ROCK1 and E-cadherin ( P < 0.05 or < 0.001), but decreased mRNA and protein expression levels of TGF-β1 (both P < 0.001) ; compared with the sh NC group, the shROCK1 group showed significantly decreased mRNA and protein expression levels of ROCK1 and E-cadherin ( P < 0.05 or < 0.001), but significantly increased mRNA and protein expression levels of TGF-β1 ( P = 0.005 or < 0.001) . Conclusions:The ROCK1 gene can inhibit the proliferation and migration of HKFs. Overexpression of the ROCK1 gene can down-regulate the TGF-β1 gene expression and up-regulate the E-cadherin gene expression in HKFs.
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Introdução: Calcinose cutânea é uma doença rara caracterizada por precipitações de cristais de cálcio no tecido cutâneo. Pode ser localizada ou generalizada, distrófica, metastática, iatrogênica ou idiopática. Relato do Caso: Paciente feminina, 66 anos, vítima de queimaduras de segundo e terceiro graus por fogo em abdome inferior e coxas aos 8 anos de idade atingindo 25% de superfície corpórea. Após 58 anos, recebeu o diagnóstico de calcinose distrófica na cicatriz da queimadura, contemplado através de biópsia e análise histopatológica. Submetida a exérese cirúrgica associada a rotação de retalho dermogorduroso de abdome superior e enxertia de pele total para correção de cicatrizes. Conclusão: Embora a melhor escolha terapêutica ainda não seja clara, o tratamento de complicações que podem culminar em incapacidade funcional é fundamental para reduzir a morbidade e aumentar a qualidade de vida do paciente.
Introduction: Cutaneous calcinosis is a rare disease characterized by the precipitation of calcium crystals in the skin tissue. It can be localized, generalized, dystrophic, metastatic, iatrogenic, or idiopathic. Case Report: Female patient, 66 years old, victim of second and third-degree burns by fire in the lower abdomen and thighs at 8 years old, reaching 25% of the body surface. After 58 years, she was diagnosed with dystrophic calcinosis in the burn scar, which was confirmed through biopsy and histopathological analysis. She underwent surgical excision associated with rotation of the upper abdomen dermal-fat flap and total skin graft for scar correction. Conclusion: Although the best therapeutic choice is still unclear, treating complications leading to functional disability is essential to reduce morbidity and increase the patient's quality of life.
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Queloides e cicatrizes hipertróficas são muito prevalentes, mas seu tratamento ainda é desafiador. Vários tratamentos se mostraram benéficos, mas ainda não há um protocolo ou algoritmo para abordar cicatrizes, que siga um passo-a-passo organizado e que justifique o uso de cada tipo de tratamento em uma fase da cicatriz. O objetivo deste estudo é apresentar uma proposta de algoritmo para tratar cicatrizes, usado em nosso serviço por alguns anos. Ele ajuda a organizar as diversas terapias empregadas no tratamento de cicatrizes. Combinadas, estas sequências de tratamentos levam a uma melhora progressiva em queloides e cicatrizes hipertróficas.
Keloids (K) and hypertrophic scars (HS) are highly prevalent, but their treatment is still challenging. Several treatments are beneficial to treat K/HS, but there is still no protocol or algorithm to address scars following an organized step-by-step and justifying each type of treatment in the scar phases. This study aims to present an algorithm to treat keloids/scars. It has been used for several years in our clinic and helps organize several therapies to treat scars. Combined, these sequences of treatments lead to progressive improvement in K and HS.
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As cicatrizes queloidianas afetam diversas populações, comprometendo a qualidade de vida dos pacientes. Vários tratamentos são apresentados na literatura. O presente estudo visou a realização de uma revisão integrativa dos artigos de revisões sistemáticas e/ou metanálises que abordam o seu tratamento nas bases de dados PubMed, LILACS, MEDLINE e Cochrane no período de 2015 a 2021. Após a identificação, e seguindo os critérios de seleção e elegibilidade, foram incluídos 24 artigos para revisão qualitativa. Observamos que as diferentes modalidades de tratamento empregadas para os queloides são afetadas pela dificuldade de avaliar recorrência, ainda mantendo muitas taxas de insucesso e necessidade de novos estudos.
Keloid scars affect different populations, compromising patients' quality of life. The literature presents several treatments. The study aimed to conduct an integrative review of systematic review articles and/or meta-analyses addressing keloid treatment in the PubMed, LILACS, MEDLINE, and Cochrane databases from 2015 to 2021. After identification and following the selection and eligibility criteria, 24 articles were included for qualitative review. We observed that the difficulty in evaluating recurrence affected different keloids treatment modalities, still presenting many failure rates and the need for further studies.
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@#A 44-years-old woman with hard and thick keloids after chronic mercury burn injury underwent a several times of reconstructive surgery. The patient had excised surgery and injected Umbilical Cord-derived Mesenchymal Stem Cells (UC-MSCs) and Umbilical Cord Mesenchymal Stem Cells-derived Condtioned Medium (UCMSC-CM) 9 times within 9 months. Seven times injection of UC-MSCs of 5x106 cells and UCMSC-CM of 20 cc 2 times at months 4 and 9. The results of this therapy showed gradual improvement without any side effects and the size of the keloid reduced from 25x16 cm to 23x14 cm after 9 months. Keloid thickness from 12 mm to 4 mm. Keloid color becomes lighter and skin texture is more supple and soft. The cases showed that UC-MSCs and UCMSC-CM can be a potential treatment for keloids after chronic burn either alone or in combination with surgery.
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Objective:To investigate the clinical effect of 5-aminolevulinic acid (ALA) photodynamic therapy (PDT) combined with CO 2 fractional laser on hypertrophic scars. Methods:From 2017 to 2019, 20 patients [17 males, 3 females, aged (24.4±4.24) years] with mandibular acne keloid in the Department of Dermatology of Air Force Medical Center, were treated with ALA-PDT combined with CO 2 fractional laser treatment, once a month, 3 times in total, and were followed up for 6 months. The efficacy, safety, and recurrence were evaluated by the Vancouver scar scale (VSS) score. Results:The effective rate was 100% after three treatments. The VSS score decreased significantly after the first treatment, and as the number of treatments increased, the VSS score further decreased. The first improvement after treatment was thickness and hardness. There was no recurrence during the 6-month follow-up. The main adverse reactions were pain, erythema and pigmentation.Conclusions:ALA-PDT combined with CO 2 fractional laser can be used as a new treatment for hypertrophic scars.
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Objective:To investigate the effect of keloid fibroblasts on the polarization and expression of inflammatory factors of M0 macrophages and possible mechanisms, and provide theoretical basis for new targets for keloid therapy.Methods:Keloids, normal skin tissues and paraffin specimens from patients undergoing plastic surgery in the First Affiliated Hospital of Sun Yat-sen University from November 2020 to September 2021 were collected, and fibroblasts of keloids and normal skins were isolated and co-cultured with M0 cells formed form THP-1 by phorbol ester (PMA)-stimulation to detect the expression of macrophage polarization markers and cytokines. Besides, keloid fibroblasts were treated with exogenous tumor necrosis factor-α(TNF-α) to detect its effect on the proliferation and extracellular matrix expression.Results:Macrophages were dominated by CD163 + (M2) in keloid tissues. Moreover, M0 cells expressed more TNF-α when co-cultured with keloid fibroblasts, compared with those with normal skin fibroblasts, in which, the positive staining rates of TNF-α were 19.32% and 29.52% respectively by flow cytometry. Furthermore, the proliferation was promoted and the expression of extracellular matrix proteins (COL3A1 and FN1)and Vimentin were upregulated in keloid fibroblasts under TNF-α stimulation. However, there was no significant difference in the expression of polarization surface markers CD86 and CD163 in macrophages, when co-cultured with keloid fibroblasts or normal skin fibroblasts. Conclusions:Keloid fibroblasts promote the expression of TNF-α in macrophages, which in turn promotes the proliferation and extracellular matrix secretion of keloid fibroblasts.
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Objective:To observe the effect of continuous shaping and compression in the comprehensive treatment of auricle keloids.Methods:From June 2018 to April 2020, 40 patients with auricular keloids (53 ears) admitted to the Plastic Surgery Department of Henan Provincial People＇s Hospital were divided into surgical injection group (20 cases, 27 ears) and surgical injection compression group (20 cases, 26 ears) according to the treatment method. The patients were followed up for more than 18 months continuously; the clinical efficacy, satisfaction with auricle morphology, and the occurrence of infection and recurrence were observed in the two groups.Results:The effective rate of surgical injection compression group (24 ears, 92.31%) was higher than that of surgical injection group (16 ears, 59.26%), and the difference was statistically significant (χ 2=5.121, P<0.05). The Vancouver scar scale (VSS) scores of the two groups at 6 months and 12 months after treatment were significantly lower than that before treatment, and the VSS score of the surgical injection compression group was lower than that of the surgical injection group, the difference was also statistically significant ( P<0.05). The satisfactory rate of the surgical injection compression group (22 ears, 84.62%) was higher than that of the surgical injection group (13 ears, 48.15%), and the difference was statistically significant (χ 2=4.048, P<0.05). The recurrence rate of the surgical injection compression group was significantly lower than that of the surgical injection group (χ 2=5.779, P<0.05); there was no significant difference in the infection rate between the two groups of patients after treatment (χ 2=0.001, P>0.05). Conclusions:Surgical resection of auricle keloids combined with local injection of corticosteroids and local shaping and compression can significantly improve the clinical efficacy, maintain the shape of the auricle, and the therapeutic effect is satisfactory.
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Objective:To observe the effect of LncRNA HOTAIR on the expression of HIF-1α in HaCat cell under low oxygen condition, and to explore the role of LncRNA HOTAIR in the pathogenesis and development of keloid.Methods:From Jan. 2018 to Dec. 2018 in Chinese Academy of Medical Science, recombinant plasmids were designed and constructed by specific shRNA-HOTAIR. After transfected HaCat cells with sh-LncRNA HOTAIR, RT-PCR was used to detect the expression level of HOTAIR. HaCat cells were cultured in different conditions and divided into four groups: group A cultured under normoxia condition, the other three groups cultured under hypoxia condition. Group C was transfected with sh-control before hypoxic culture, while group D were transfected with sh-LncRNA HOTAIR. After 24 hours′ culture, dual luciferase reporter gene assay was used to verify the relationship of LncRNA HOTAIR and HIF-1α. Expression of HIF-1α in four groups of HaCat cells were determined by Western blot analysis.Results:Recombinant plasmids of shRNA-HOTAIR were successfully constructed, and the HOTAIR expression was significantly decreased. The relative luciferase activity was 0.94±0.30 in group A, 20.39±1.15 in group B, 18.09±0.80 in group C and 3.04±1.15 in group D. The relative luciferase activity of group B was higher than those of group A ( t=29.03, P<0.05), and the difference was statistically significant ( P<0.05). According to the results of Western blotting, group B (1.19±0.07) and group D (1.15±0.06) had higher expression of HIF-1α than group A (0.56±0.29) and group C (0.37±0.38). Down-regulation of HOTAIR significantly inhibited the protein level of HIF-1α. Conclusions:LncRNA HOTAIR plays a positive role in upregulation of HIF-1α in HaCat cells under hypoxia condition. Thus LncRNA HOTAIR may take part in the pathogenesis and development of keloid through HIF-1α pathway.
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Objective:To investigate the effect of ultrasound combined with 4-hydroxyphenyl-retinamide (4-HPR) lipid microbubbles on type Ⅰ collagen α1 chain (COL1A1) protein expression in keloid-derived fibroblasts.Methods:In vitro cultured keloid-derived fibroblasts were divided into 3 groups: control group receiving conventional culture with incomplete Dulbecco′s modified Eagle′s medium (DMEM) , 4-HPR lipid microbubble group cultured with incomplete DMEM containing 15 mg/L 4-HPR lipid microbubbles, and ultrasound + 4-HPR lipid microbubble group cultured with incomplete DMEM containing 15 mg/L 4-HPR lipid microbubbles under ultrasound treatment. After 24-hour treatment, reverse transcription (RT) -PCR and Western blot analysis were performed to determine the mRNA and protein expression of COL1A1 in keloid-derived fibroblasts in each group. Intergroup comparison was carried out by using t test. Results:The mRNA relative expression level of COL1A1 was 1.00 ± 0.18, 0.69 ± 0.15 and 0.35 ± 0.18 in the control group, 4-HPR lipid microbubble group and ultrasound + 4-HPR lipid microbubble group respectively, and the protein relative expression level of COL1A1 was 0.93 ± 0.03, 0.74 ± 0.07 and 0.44 ± 0.06 in the above 3 groups respectively. Moreover, the mRNA and protein expression of COL1A1 was significantly lower in the 4-HPR lipid microbubble group and ultrasound + 4-HPR lipid microbubble group than in the control group ( P < 0.05 or 0.001) , and lower in the ultrasound + 4-HPR lipid microbubble group than in the 4-HPR lipid microbubble group ( P < 0.05) . Conclusion:Ultrasound combined with 4-HPR lipid microbubbles could markedly inhibit the mRNA and protein expression of COL1A1 in keloid-derived fibroblasts.
ABSTRACT
Abstract Background: Tacrolimus is used to prevent unaesthetic scars due to its action on fibroblast activity and collagen production modulation. Objectives: To evaluate the action pathways, from the histopathological point of view and in cytokine control, of tacrolimus ointment in the prevention of hypertrophic scars. Methods: Twenty-two rabbits were submitted to the excision of two 1-cm fragments in each ear, including the perichondrium. The right ear received 0.1% and 0.03% tacrolimus in ointment base twice a day in the upper wound and in the lower wound respectively. The left ear, used as the control, was treated with petrolatum. After 30 days, collagen fibers were evaluated using special staining, and immunohistochemistry analyses for smooth muscle actin, TGF-β and VEGF were performed. Results: The wounds treated with 0.1% tacrolimus showed weak labeling and a lower percentage of labeling for smooth muscle actin, a higher proportion of mucin absence, weak staining, fine and organized fibers for Gomori's Trichrome, strong staining and organized fibers for Verhoeff when compared to controls. The wounds treated with 0.03% tacrolimus showed weak labeling for smooth muscle actin, a higher proportion of mucin absence, strong staining for Verhoeff when compared to the controls. There was absence of TGF-β and low VEGF expression. Study limitations: The analysis was performed by a single pathologist. Second-harmonic imaging microscopy was performed in 2 sample areas of the scar. Conclusions: Both drug concentrations were effective in suppressing TGF-β and smooth muscle actin, reducing mucin, improving the quality of collagen fibers, and the density of elastic fibers, but only the higher concentration influenced elastic fiber organization.
Subject(s)
Animals , Cicatrix, Hypertrophic/pathology , Cicatrix, Hypertrophic/prevention & control , Cicatrix, Hypertrophic/drug therapy , Ointment Bases , Rabbits , Wound Healing , Tacrolimus , Ear/pathologyABSTRACT
SUMMARY: The establishment of primary keloid fibroblast culture has always been a fundamental measure for studying mechanisms of keloid disease. The quality of the primary cell culture can directly affect the results of further experiments. This study was performed to investigate the optimal growth conditions, including the optimal storage time and collagenase treatment time, for in vitro cell culture models and the suitable methods for epidermis-dermis separation in different tissues. Keloid tissues, keloid-surrounding tissues, and normal skin tissues were collected from patients, for primary fibroblast culture. Two methods, tissue explant and collagenase digestion, were deployed and compared. Expression levels of the keloid-related genes α -SMA, Col1, and Col3 were assessed in cells cultured using both methods, to verify the qualities of the primary cells. A comparative analysis was conducted between the two methods and among the three different tissues used. Bacterial and lipid contamination was immediately minimized after the samples were processed. Different methods of epidermis removal and different durations of collagenase digestion were required in different tissues to generate optimal results. Real-time PCR results showed that the mRNA expression levels of keloid-related genes in cultured fibroblasts correlated to their in vivo expression profile, as previously reported in other studies. The results of this study have revealed several key points in the culture of primary keloid fibroblasts and demonstrated the correlation in gene expression between in vivo keloid fibroblasts and in vitro primary keloid fibroblasts.
RESUMEN: La identificación de un cultivo de fibroblastos queloides primarios, siempre ha sido una medida fundamental para estudiar los mecanismos de la enfermedad queloide. La calidad del cultivo de células primarias puede afectar directamente los resultados de otros experimentos. Este estudio se realizó para investigar las condiciones óptimas de crecimiento, incluido el tiempo óptimo de almacenamiento y el tiempo de tratamiento con colagenasa, para modelos de cultivo celular in vitro y los métodos adecuados para la separación epidermis-dermis en diferentes tejidos. Se recogieron de los pacientes tejidos queloides, tejidos circundantes queloides y tejidos cutáneos normales, para cultivo primario de fibroblastos. Se implementaron y compararon dos métodos, explante de tejido y digestión con colagenasa. Los niveles de expresión de los genes relacionados con queloides α -SMA, Col1 y Col3 se evaluaron en células cultivadas usando ambos métodos, para verificar las cualidades de las células primarias. Se realizó un análisis comparativo entre los dos métodos y entre los tres tejidos diferentes utilizados. La contaminación de bacterias y lípidos se minimizó inmediatamente después de que se procesaron las muestras. Se requirieron varios métodos de eliminación de la epidermis y diferentes tiempos de digestión con colagenasa en los tejidos para generar resultados óptimos. Los resultados de la PCR en tiempo real mostraron que los niveles de expresión de ARNm de genes relacionados con queloides en fibroblastos cultivados se correlacionaban con su perfil de expresión in vivo, como se informó en estudios anteriores. Los resultados de este studio indicaron varios puntos clave en el cultivo de fibroblastos queloides primarios y han demostrado la correlación en la expresión génica entre fibroblastos queloides in vivo y fibroblastos queloides primarios in vitro.
Subject(s)
Humans , Adolescent , Adult , Young Adult , Skin , Primary Cell Culture/methods , Fibroblasts , Keloid , Fluorescent Antibody Technique , Actins , Collagen , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
El acné queloideo de la nuca (AKN) o foliculitis esclerosante es un proceso inflamatorio crónico del folículo piloso en la región de la nuca. Su incidencia es baja y su etiología desconocida. Afecta con mayor frecuencia a hombres de mediana edad y de raza negra. A lo largo del tiempo se han utilizado diferentes modalidades terapéuticas con resultados variables. Presentamos el caso de un paciente con AKN que mostró una excelente respuesta al uso diario de imiquimod 5% tópico durante ocho semanas
Keloid acne of the neck (AKN) or sclerosing folliculitis of the nape of the neck is a chronic inflammatory process of the nape region. Its incidence is low and its etiology is unknown. It mainly affects brown-black males in middle age. Different treatment modalities have been used with different responses. We present the case of a patient with AKN who presented an excellent response to the daily use of topical imiquimod 5% for eight weeks