Effect of topical application of monocerin on surgical wounds in murine model

Detalhes bibliográficos
Autor(a) principal: Anwar, Usama Bilal
Data de Publicação: 2020
Tipo de documento: Tese
Idioma: eng
Título da fonte: Biblioteca Digital de Teses e Dissertações da USP
Texto Completo: https://www.teses.usp.br/teses/disponiveis/10/10132/tde-23022021-113548/
Resumo: In some cases, the wound healing process demands treatment for a long time, especially in complex situation such as skin burn with severe degree that requires tissue remodeling without scars or keloids on the patient’s skin. Microorganism’s secondary metabolites are being explored as a new source of molecules that could be applied on pharmaceutical area for several purposes and previous studies showed that the secondary metabolite monocerin obtained from the fungus Exserohilum rostratum induces proliferation in human umbilical vein endothelial cells (HUVECs) and in normal human fibroblasts (FN1). In this study, the monocerin compound was obtained from the culture of the fungus E. rostratum and after purification by high performance liquid chromatography (HPLC), it was incorporated into a cream formulation at concentrations of 0.0006 and 0.005%. The effect of the formulations was evaluated on surgical wounds induced on the dorsal part of male Balb/c mice, and as control groups animals received no treatment (Untreated), or were treated with the commercial Kollagenase ointment plus chloramphenicol (collagenase 0.6 U / g + chloramphenicol 0.01 g / g, Cristália), or with the vehicle of the cream formulation. The animals were treated for 1, 3, 7 and 14 days and the wound size was measured daily with paquimeter and X-ray analysis. At the end of the treatments, the animal’s blood was collected for hematological and cytokine / chemokine analysis (IL-1β, IL-6, IL-10, TNF-α, TGF-β, FGF-2, VEGF). The wounded tissue was collected for morphological analysis hydroxyproline, histological (hematoxylin & eosin, masson trichrome and picrosirius red staining), scanning electron microscopy (SEM), and immunohistochemistry (MMP-3, MMP-9, TIMP-2, VEGF). Together with the hematological, cytokine/chemokine, SEM and Immunohistochemistry parameters, the results indicate that the treatment with monocerin was able to induce wound healing with tissue regeneration and intense collagen formation. The treatment was effective and no toxicity was observed on the animals; the wound healing process was effective without scar on the regenerated skin.
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spelling Effect of topical application of monocerin on surgical wounds in murine modelEfeito da aplicação tópica da monocerina em feridas cirúrgicas em modelo murinoCicatrizaçãoFerida cirúrgicaMonocerinMonocerinaSurgical woundWound healingIn some cases, the wound healing process demands treatment for a long time, especially in complex situation such as skin burn with severe degree that requires tissue remodeling without scars or keloids on the patient’s skin. Microorganism’s secondary metabolites are being explored as a new source of molecules that could be applied on pharmaceutical area for several purposes and previous studies showed that the secondary metabolite monocerin obtained from the fungus Exserohilum rostratum induces proliferation in human umbilical vein endothelial cells (HUVECs) and in normal human fibroblasts (FN1). In this study, the monocerin compound was obtained from the culture of the fungus E. rostratum and after purification by high performance liquid chromatography (HPLC), it was incorporated into a cream formulation at concentrations of 0.0006 and 0.005%. The effect of the formulations was evaluated on surgical wounds induced on the dorsal part of male Balb/c mice, and as control groups animals received no treatment (Untreated), or were treated with the commercial Kollagenase ointment plus chloramphenicol (collagenase 0.6 U / g + chloramphenicol 0.01 g / g, Cristália), or with the vehicle of the cream formulation. The animals were treated for 1, 3, 7 and 14 days and the wound size was measured daily with paquimeter and X-ray analysis. At the end of the treatments, the animal’s blood was collected for hematological and cytokine / chemokine analysis (IL-1β, IL-6, IL-10, TNF-α, TGF-β, FGF-2, VEGF). The wounded tissue was collected for morphological analysis hydroxyproline, histological (hematoxylin & eosin, masson trichrome and picrosirius red staining), scanning electron microscopy (SEM), and immunohistochemistry (MMP-3, MMP-9, TIMP-2, VEGF). Together with the hematological, cytokine/chemokine, SEM and Immunohistochemistry parameters, the results indicate that the treatment with monocerin was able to induce wound healing with tissue regeneration and intense collagen formation. The treatment was effective and no toxicity was observed on the animals; the wound healing process was effective without scar on the regenerated skin.Em alguns casos, o processo de cicatrização de feridas exige tratamento por muito tempo, principalmente em situações complexas como queimaduras na pele de grau severo que requerem remodelação do tecido sem formação de cicatrizes ou queloides na pele do paciente. Metabólitos secundários produzidos por micro-organismos têm sido explorados como uma fonte de moléculas que podem ser aplicadas na área farmacêutica para diversas aplicações, e estudos anteriores mostraram que o metabólito secundário monocerina produzido pelo fungo Exserohilum rostratum induz proliferação em células endoteliais da veia umbilical humana (HUVECs) e em fibroblastos humanos normais (FN1). Neste estudo, o composto monocerina foi obtido a partir da cultura do fungo E. rostratum e após processo de purificação por cromatografia líquida de alta eficiência (CLAE) foi incorporado em formulação em creme nas concentrações de 0,0006 e 0,005%. O efeito das formulações foi avaliado no tratamento de ferida cirúrgica induzida no dorso de camundongos Balb/c machos, e como grupos controles foram empregados animais sem tratamento, animais tratados com a pomada comercial Kollagenase mais cloranfenicol (colagenase 0,6 U/ g + cloranfenicol 0,01 g/g, Cristália), e animais tratados com o veículo da formulação em creme. Os animais foram tratados por 1, 3, 7 e 14 dias e o tamanho da ferida foi medido diariamente com paquímetro digital e análise por raio-X. Ao final dos tratamentos, o sangue dos animais foi coletado para análise hematológica e de citocinas/quimiocinas (IL-1β, IL-6, IL-10, TNF-α, TGF-β, FGF-2, VEGF). O tecido da área cicatricial foi coletado para dosagem de hidroxiprolina, análises histológicas (hematoxilina/eosina, tricrômio de masson e picrosirius red) e morfológicas por microscopia eletrônica de varredura (MEV) e imunoistoquímica (MMP-3, MMP-9, TIMP-2, VEGF). Juntamente com os parâmetros hematológicos, dosagens de citocinas/quimiocinas, análise por MEV e imunoistoquímica, os resultados indicam que o tratamento com monocerina a 0,005% foi capaz de induzir a cicatrização da ferida com regeneração tecidual e intensa formação de colágeno de forma mais efetiva que os grupos controles. O tratamento foi eficaz e nenhuma toxicidade foi observada nos animais; o processo de cicatrização foi eficiente e sem cicatriz na pele regenerada.Biblioteca Digitais de Teses e Dissertações da USPSouza, Ana Olívia deAnwar, Usama Bilal2020-12-16info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/10/10132/tde-23022021-113548/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPReter o conteúdo por motivos de patente, publicação e/ou direitos autoriais.info:eu-repo/semantics/openAccesseng2023-02-14T19:24:37Zoai:teses.usp.br:tde-23022021-113548Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212023-02-14T19:24:37Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false
dc.title.none.fl_str_mv Effect of topical application of monocerin on surgical wounds in murine model
Efeito da aplicação tópica da monocerina em feridas cirúrgicas em modelo murino
title Effect of topical application of monocerin on surgical wounds in murine model
spellingShingle Effect of topical application of monocerin on surgical wounds in murine model
Anwar, Usama Bilal
Cicatrização
Ferida cirúrgica
Monocerin
Monocerina
Surgical wound
Wound healing
title_short Effect of topical application of monocerin on surgical wounds in murine model
title_full Effect of topical application of monocerin on surgical wounds in murine model
title_fullStr Effect of topical application of monocerin on surgical wounds in murine model
title_full_unstemmed Effect of topical application of monocerin on surgical wounds in murine model
title_sort Effect of topical application of monocerin on surgical wounds in murine model
author Anwar, Usama Bilal
author_facet Anwar, Usama Bilal
author_role author
dc.contributor.none.fl_str_mv Souza, Ana Olívia de
dc.contributor.author.fl_str_mv Anwar, Usama Bilal
dc.subject.por.fl_str_mv Cicatrização
Ferida cirúrgica
Monocerin
Monocerina
Surgical wound
Wound healing
topic Cicatrização
Ferida cirúrgica
Monocerin
Monocerina
Surgical wound
Wound healing
description In some cases, the wound healing process demands treatment for a long time, especially in complex situation such as skin burn with severe degree that requires tissue remodeling without scars or keloids on the patient’s skin. Microorganism’s secondary metabolites are being explored as a new source of molecules that could be applied on pharmaceutical area for several purposes and previous studies showed that the secondary metabolite monocerin obtained from the fungus Exserohilum rostratum induces proliferation in human umbilical vein endothelial cells (HUVECs) and in normal human fibroblasts (FN1). In this study, the monocerin compound was obtained from the culture of the fungus E. rostratum and after purification by high performance liquid chromatography (HPLC), it was incorporated into a cream formulation at concentrations of 0.0006 and 0.005%. The effect of the formulations was evaluated on surgical wounds induced on the dorsal part of male Balb/c mice, and as control groups animals received no treatment (Untreated), or were treated with the commercial Kollagenase ointment plus chloramphenicol (collagenase 0.6 U / g + chloramphenicol 0.01 g / g, Cristália), or with the vehicle of the cream formulation. The animals were treated for 1, 3, 7 and 14 days and the wound size was measured daily with paquimeter and X-ray analysis. At the end of the treatments, the animal’s blood was collected for hematological and cytokine / chemokine analysis (IL-1β, IL-6, IL-10, TNF-α, TGF-β, FGF-2, VEGF). The wounded tissue was collected for morphological analysis hydroxyproline, histological (hematoxylin & eosin, masson trichrome and picrosirius red staining), scanning electron microscopy (SEM), and immunohistochemistry (MMP-3, MMP-9, TIMP-2, VEGF). Together with the hematological, cytokine/chemokine, SEM and Immunohistochemistry parameters, the results indicate that the treatment with monocerin was able to induce wound healing with tissue regeneration and intense collagen formation. The treatment was effective and no toxicity was observed on the animals; the wound healing process was effective without scar on the regenerated skin.
publishDate 2020
dc.date.none.fl_str_mv 2020-12-16
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