Activity and Effectiveness of Recombinant hEGF Excreted by Escherichia coli BL21 on Wound Healing in Induced Diabetic Mice

Sriwidodo Sriwidodo,1 Iman Permana Maksum,2 Toto Subroto,2 Nasrul Wathoni,1 Anas Subarnas,3 Abd Kakhar Umar1 1Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Universitas Padjadjaran, Jatinangor 45363, Indonesia; 2Department of Chemistry, Faculty of Mathematics and Nat...

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Main Authors: Sriwidodo S, Maksum IP, Subroto T, Wathoni N, Subarnas A, Umar AK
Format: Article
Language:English
Published: Dove Medical Press 2020-09-01
Series:Journal of Experimental Pharmacology
Subjects:
Online Access:https://www.dovepress.com/activity-and-effectiveness-of-recombinant-hegf-excreted-by-escherichia-peer-reviewed-fulltext-article-JEP
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spelling doaj-ea0989ed796e48a08c960fb0215e2ef82021-09-02T21:46:51ZengDove Medical PressJournal of Experimental Pharmacology1179-14542020-09-01Volume 1233934857492Activity and Effectiveness of Recombinant hEGF Excreted by Escherichia coli BL21 on Wound Healing in Induced Diabetic MiceSriwidodo SMaksum IPSubroto TWathoni NSubarnas AUmar AKSriwidodo Sriwidodo,1 Iman Permana Maksum,2 Toto Subroto,2 Nasrul Wathoni,1 Anas Subarnas,3 Abd Kakhar Umar1 1Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Universitas Padjadjaran, Jatinangor 45363, Indonesia; 2Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jatinangor 45363, Indonesia; 3Department of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Universitas Padjadjaran, Jatinangor 45363, IndonesiaCorrespondence: Sriwidodo SriwidodoDepartment of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Universitas Padjadjaran, Jatinangor 45363, IndonesiaTel +6222 842 888888Email sriwidodo@unpad.ac.idContext: Human epidermal growth factor (hEGF) has biological activities and can be used in medicines and cosmetics. A high level of effectiveness of hEGF can be obtained when three disulfide bonds fold perfectly. Extracellular secretion from E. coli BL21 using the PelB signal peptide is a new way to obtain hEGF with a structure that folds appropriately.Object: This study aimed to determine the activity and effectiveness of recombinant hEGF excreted by E. coli BL21 on wound healing in induced diabetic mice.Methods: Cell proliferation and migration tests were performed on NIH3T3 cells, followed by wound healing tests in induced diabetic mice, along with histological and endotoxin test at various hEGF concentrations (25, 50, and 75 μg/mL).Results: Based on the results, hEGF at a level of 50 μg/mL showed optimal proliferation and migration activities. Wound healing in induced diabetic mice showed faster-wound closure within 12 days at hEGF 50 and 75 μg/mL with a percentage wound closure of 95% and 98.5%, respectively, which was significant versus control. In the histology test, the number of fibroblasts showed an increase and was significant at hEGF 75 μg/mL compared to the control group. The single test vial (STV) showed that hEGF solution was free of endotoxin.Conclusion: Recombinant hEGF produced by extracellular secretion using E. coli BL21 has optimal diabetic wound healing activity through increased fibroblast proliferation.Keywords: recombinant hEGF, fibroblast proliferation, diabetic wound healinghttps://www.dovepress.com/activity-and-effectiveness-of-recombinant-hegf-excreted-by-escherichia-peer-reviewed-fulltext-article-JEPecombinant hegffibroblast proliferationdiabetic wound healing
collection DOAJ
language English
format Article
sources DOAJ
author Sriwidodo S
Maksum IP
Subroto T
Wathoni N
Subarnas A
Umar AK
spellingShingle Sriwidodo S
Maksum IP
Subroto T
Wathoni N
Subarnas A
Umar AK
Activity and Effectiveness of Recombinant hEGF Excreted by Escherichia coli BL21 on Wound Healing in Induced Diabetic Mice
Journal of Experimental Pharmacology
ecombinant hegf
fibroblast proliferation
diabetic wound healing
author_facet Sriwidodo S
Maksum IP
Subroto T
Wathoni N
Subarnas A
Umar AK
author_sort Sriwidodo S
title Activity and Effectiveness of Recombinant hEGF Excreted by Escherichia coli BL21 on Wound Healing in Induced Diabetic Mice
title_short Activity and Effectiveness of Recombinant hEGF Excreted by Escherichia coli BL21 on Wound Healing in Induced Diabetic Mice
title_full Activity and Effectiveness of Recombinant hEGF Excreted by Escherichia coli BL21 on Wound Healing in Induced Diabetic Mice
title_fullStr Activity and Effectiveness of Recombinant hEGF Excreted by Escherichia coli BL21 on Wound Healing in Induced Diabetic Mice
title_full_unstemmed Activity and Effectiveness of Recombinant hEGF Excreted by Escherichia coli BL21 on Wound Healing in Induced Diabetic Mice
title_sort activity and effectiveness of recombinant hegf excreted by escherichia coli bl21 on wound healing in induced diabetic mice
publisher Dove Medical Press
series Journal of Experimental Pharmacology
issn 1179-1454
publishDate 2020-09-01
description Sriwidodo Sriwidodo,1 Iman Permana Maksum,2 Toto Subroto,2 Nasrul Wathoni,1 Anas Subarnas,3 Abd Kakhar Umar1 1Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Universitas Padjadjaran, Jatinangor 45363, Indonesia; 2Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jatinangor 45363, Indonesia; 3Department of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Universitas Padjadjaran, Jatinangor 45363, IndonesiaCorrespondence: Sriwidodo SriwidodoDepartment of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Universitas Padjadjaran, Jatinangor 45363, IndonesiaTel +6222 842 888888Email sriwidodo@unpad.ac.idContext: Human epidermal growth factor (hEGF) has biological activities and can be used in medicines and cosmetics. A high level of effectiveness of hEGF can be obtained when three disulfide bonds fold perfectly. Extracellular secretion from E. coli BL21 using the PelB signal peptide is a new way to obtain hEGF with a structure that folds appropriately.Object: This study aimed to determine the activity and effectiveness of recombinant hEGF excreted by E. coli BL21 on wound healing in induced diabetic mice.Methods: Cell proliferation and migration tests were performed on NIH3T3 cells, followed by wound healing tests in induced diabetic mice, along with histological and endotoxin test at various hEGF concentrations (25, 50, and 75 μg/mL).Results: Based on the results, hEGF at a level of 50 μg/mL showed optimal proliferation and migration activities. Wound healing in induced diabetic mice showed faster-wound closure within 12 days at hEGF 50 and 75 μg/mL with a percentage wound closure of 95% and 98.5%, respectively, which was significant versus control. In the histology test, the number of fibroblasts showed an increase and was significant at hEGF 75 μg/mL compared to the control group. The single test vial (STV) showed that hEGF solution was free of endotoxin.Conclusion: Recombinant hEGF produced by extracellular secretion using E. coli BL21 has optimal diabetic wound healing activity through increased fibroblast proliferation.Keywords: recombinant hEGF, fibroblast proliferation, diabetic wound healing
topic ecombinant hegf
fibroblast proliferation
diabetic wound healing
url https://www.dovepress.com/activity-and-effectiveness-of-recombinant-hegf-excreted-by-escherichia-peer-reviewed-fulltext-article-JEP
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