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Comparison of EGF with VEGF Non-Viral Gene Therapy for Cutaneous Wound Healing of Streptozotocin Diabetic Mice
Junghae Ko, Haejung Jun, Hyesook Chung, Changshin Yoon, Taekyoon Kim, Minjeong Kwon, Soonhee Lee, Soojin Jung, Mikyung Kim, Jeong Hyun Park
Diabetes Metab J. 2011;35(3):226-235.   Published online June 30, 2011
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  • 35 Download
  • 20 Crossref
AbstractAbstract PDFPubReader   

To accelerate the healing of diabetic wounds, various kinds of growth factors have been employed. It is the short half-life of administered growth factors in hostile wound beds that have limited wide-spread clinical usage. To overcome this limitation, growth factor gene therapy could be an attractive alternative rather than direct application of factors onto the wound beds. We administered two growth factor DNAs, epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) into a cutaneous wound on diabetic mice. We compared the different characteristics of the healing wounds.


Streptozotocin was injected intraperitoneally to induce diabetes into C57BL/6J mice. The ultrasound micro-bubble destruction method with SonoVue as a bubbling agent was used for non-viral gene delivery of EGF828 and VEGF165 DNAs. Each gene was modified for increasing efficacy as FRM-EGF828 or minicircle VEGF165. The degree of neoangiogenesis was assessed using qualitative laser Doppler flowmetry. We compared wound size and histological findings of the skin wounds in each group.


In both groups, accelerated wound closure was observed in the mice receiving gene therapy compared with non treated diabetic control mice. Blood flow detected by laser doppler flowmetry was better in the VEGF group than in the EGF group. Wound healing rates and histological findings were more accelerated in the EGF gene therapy group than the VEGF group, but were not statistically significant.


Both non-viral EGF and VEGF gene therapy administrations could improve the speed and quality of skin wound healing. However, the detailed histological characteristics of the healing wounds were different.


Citations to this article as recorded by  
  • Gene therapy to enhance angiogenesis in chronic wounds
    Elnaz Shaabani, Maryam Sharifiaghdam, Reza Faridi-Majidi, Stefaan C. De Smedt, Kevin Braeckmans, Juan C. Fraire
    Molecular Therapy - Nucleic Acids.2022; 29: 871.     CrossRef
  • Approaches to Modulate the Chronic Wound Environment Using Localized Nucleic Acid Delivery
    Adam G. Berger, Jonathan J. Chou, Paula T. Hammond
    Advances in Wound Care.2021; 10(9): 503.     CrossRef
  • Limited Treatment Options for Diabetic Wounds: Barriers to Clinical Translation Despite Therapeutic Success in Murine Models
    May Barakat, Luisa A. DiPietro, Lin Chen
    Advances in Wound Care.2021; 10(8): 436.     CrossRef
  • Minicircle‐based expression of vascular endothelial growth factor in mesenchymal stromal cells from diverse human tissues
    Joana Serra, Cláudia P. A. Alves, Joaquim M. S. Cabral, Gabriel A. Monteiro, Cláudia L. da Silva, Duarte Miguel F. Prazeres
    The Journal of Gene Medicine.2021;[Epub]     CrossRef
  • Mathematical Model Predicts that Acceleration of Diabetic Wound Healing is Dependent on Spatial Distribution of VEGF-A mRNA (AZD8601)
    S. Michaela Rikard, Paul J. Myers, Joachim Almquist, Peter Gennemark, Anthony C. Bruce, Maria Wågberg, Regina Fritsche-Danielson, Kenny M. Hansson, Matthew J. Lazzara, Shayn M. Peirce
    Cellular and Molecular Bioengineering.2021; 14(4): 321.     CrossRef
  • β-Neoendorphin Enhances Wound Healing by Promoting Cell Migration in Keratinocyte
    Dong Joo Yang, Sang Hyun Moh, Yun-Hee Choi, Ki Woo Kim
    Molecules.2020; 25(20): 4640.     CrossRef
  • Orf Virus IL-10 and VEGF-E Act Synergistically to Enhance Healing of Cutaneous Wounds in Mice
    Lyn M. Wise, Gabriella S. Stuart, Nicola C. Jones, Stephen B. Fleming, Andrew A. Mercer
    Journal of Clinical Medicine.2020; 9(4): 1085.     CrossRef
  • Engineering of Human Mesenchymal Stem/Stromal Cells with Vascular Endothelial Growth Factor–Encoding Minicircles for AngiogenicEx VivoGene Therapy
    Joana Serra, Cláudia P.A. Alves, Liliana Brito, Gabriel A. Monteiro, Joaquim M.S. Cabral, Duarte Miguel F. Prazeres, Cláudia L. da Silva
    Human Gene Therapy.2019; 30(3): 316.     CrossRef
  • Therapeutic strategies for enhancing angiogenesis in wound healing
    Austin P. Veith, Kayla Henderson, Adrianne Spencer, Andrew D. Sligar, Aaron B. Baker
    Advanced Drug Delivery Reviews.2019; 146: 97.     CrossRef
  • Nanomedicines and gene therapy for the delivery of growth factors to improve perfusion and oxygenation in wound healing
    Céline M. Desmet, Véronique Préat, Bernard Gallez
    Advanced Drug Delivery Reviews.2018; 129: 262.     CrossRef
  • Electrospun Fibers as a Dressing Material for Drug and Biological Agent Delivery in Wound Healing Applications
    Mulugeta Gizaw, Jeffrey Thompson, Addison Faglie, Shih-Yu Lee, Pierre Neuenschwander, Shih-Feng Chou
    Bioengineering.2018; 5(1): 9.     CrossRef
  • Non-viral gene therapy: Gains and challenges of non-invasive administration methods
    Marianna Foldvari, Ding Wen Chen, Nafiseh Nafissi, Daniella Calderon, Lokesh Narsineni, Amirreza Rafiee
    Journal of Controlled Release.2016; 240: 165.     CrossRef
  • RETRACTED ARTICLE: Decellularized scaffolds containing hyaluronic acid and EGF for promoting the recovery of skin wounds
    Zhengzheng Wu, Yan Tang, Hongdou Fang, Zhongchun Su, Bin Xu, Yongliang Lin, Peng Zhang, Xing Wei
    Journal of Materials Science: Materials in Medicine.2015;[Epub]     CrossRef
  • Acute and chronic wound fluids inversely influence adipose‐derived stem cell function: molecular insights into impaired wound healing
    Paola Koenen, Timo A Spanholtz, Marc Maegele, Ewa Stürmer, Thomas Brockamp, Edmund Neugebauer, Oliver C Thamm
    International Wound Journal.2015; 12(1): 10.     CrossRef
  • Adipose‐derived stem cells and keratinocytes in a chronic wound cell culture model: the role of hydroxyectoine
    Oliver C Thamm, Panagiotis Theodorou, Ewa Stuermer, Max J Zinser, Edmund A Neugebauer, Paul C Fuchs, Paola Koenen
    International Wound Journal.2015; 12(4): 387.     CrossRef
  • Genetic and cellular techniques emerge as promising modalities for the treatment of diabetic foot syndrome
    Vladimir Iosifovich Konenkov, Vadim Valerievich Klimontov
    Diabetes mellitus.2014; 17(1): 63.     CrossRef
  • Analysis of blood flow and local expression of angiogenesis-associated growth factors in infected wounds treated with negative pressure wound therapy
    Molecular Medicine Reports.2014; 9(5): 1749.     CrossRef
  • Alteration of Skin Properties with Autologous Dermal Fibroblasts
    Rajesh Thangapazham, Thomas Darling, Jon Meyerle
    International Journal of Molecular Sciences.2014; 15(5): 8407.     CrossRef
  • Angiopoietin-like 4 Stimulates STAT3-mediated iNOS Expression and Enhances Angiogenesis to Accelerate Wound Healing in Diabetic Mice
    Han Chung Chong, Jeremy Soon Kiat Chan, Chi Qin Goh, Natalia V Gounko, Baiwen Luo, Xiaoling Wang, Selin Foo, Marcus Thien Chong Wong, Cleo Choong, Sander Kersten, Nguan Soon Tan
    Molecular Therapy.2014; 22(9): 1593.     CrossRef
  • Genome Editing of Mouse Fibroblasts by Homologous Recombination for Sustained Secretion of PDGF-B and Augmentation of Wound Healing
    Jenny C. Barker, Adam D. Barker, Jessica Bills, Jiying Huang, Mary Wight-Carter, Imelda Delgado, Debby L. Noble, Lily J. Huang, Matthew H. Porteus, Kathryn E. Davis
    Plastic and Reconstructive Surgery.2014; 134(3): 389e.     CrossRef
Treatment of Type 1 Diabetes through Genetically Engineered K-cell Transplantation in a Mouse Model.
Ju Yeon Sim, Ju Hee Kim, Yu Bae Ahn, Ki Ho Song, Je Ho Han, Bong Yun Cha, Sook Kyung Lee, Sung Dae Moon
Korean Diabetes J. 2009;33(6):466-474.   Published online December 1, 2009
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AbstractAbstract PDF
K-cells function as targets for insulin gene therapy. In a previous study, we constructed EBV-based plasmids expressing rat preproinsulin controlled by glucose-dependent insulinotropic polypeptide promoters. In the present study, we attempted to correct hyperglycemia in vivo using genetically engineered K-cells in a mouse model of type 1 diabetes. METHODS: K-cells expressing insulin were transplanted under the kidney capsules of STZ-induced diabetic mice. The blood glucose levels and body weights of the experimental animals were measured daily. After four weeks, the mice were injected intra-peritoneally with 2 g/kg glucose following a 6 hr fast. Blood glucose levels were measured immediately following glucose injections. All animals were sacrificed at the end of the glucose tolerance study, and pancreas and graft-bearing kidney tissue samples were stained with antibodies against insulin, glucagon, and C-peptide. RESULTS: The body weights of K-cell-transplanted diabetic mice increased after transplantation, whereas those of untreated diabetic control mice continued to decline. The blood glucose levels of K-cell-transplanted diabetic mice decreased gradually during the two weeks following transplantation. After intra-peritoneal injection of glucose into K-cell-transplanted diabetic mice, blood glucose levels increased at 30 minutes, and were restored to the normal range between 60 and 90 minutes, while untreated control diabetic mice continued to experience hyperglycemia. Kidney capsules containing transplanted K-cells were removed, and sections were stained with anti-insulin antibodies. We detected insulin-positive cells in the kidney capsules of K-cell-transplanted diabetic mice, but not in untreated control mice. CONCLUSION: We detected glucose-dependent insulin secretion in genetically engineered K-cells in a mouse model of type 1 diabetes. Our results suggest that genetically modified insulin producing K-cells may act as surrogate beta-cells to effectively treat type 1 diabetes.
Cloning of Novel Epidermal Growth Factor (EGF) Plasmid for Gene Therapy on Diabetic Foot Ulcer.
Hye Sook Chung, Chang Shin Yoon, Min Jeong Kwon, Mi Kyung Kim, Soon Hee Lee, Kyung Soo Ko, Byung Doo Rhee, Jeong Hyun Park
Korean Diabetes J. 2008;32(2):131-140.   Published online April 1, 2008
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  • 44 Download
  • 1 Crossref
AbstractAbstract PDF
Epidermal Growth Factor (EGF) is one of the important growth factors involved in the epithelialization during cutaneous wound healing. Peptide EGF has been used for the treatment of diabetic foot ulcer. But the inferiority of cost-effectiveness and the inconvenience of daily application might have restricted its wide clinical usage. EGF gene therapy could dramatically improve the efficacy and inconvenience through long-term expression and bypassing the EGF degradation by hostile non-specific proteinases expressed in the wound bed. METHODS: EGF DNAs were amplified via PCR. For the more effective secretion from the transfected cell, we inserted furin cleavage site into EGF plasmids. The efficacy of novel plasmid pbeta-EGF was verified by transfection into the various animal cell lines, and the biologic potency of expressed EGF was confirmed via phosphorylation of PI3K and GSK3beta by Western blotting. RESULTS: We tested various kinds of human EGFs. One of the human EGF isoforms, EGF(828) including a membrane-anchoring domain was successfully released as the mature EGF protein in the cell culture media. Also EGF plasmid including furin cleavage site showed more than 2-fold increased EGF expression compared with the sequence without furin cleavage site. CONCLUSION: In conclusion, these findings suggest that mature EGF could be released easily out of cells by modifying EGF DNA sequence. Our novel EGF plasmid DNA could markedly increase the efficiency of non-viral gene therapy for diabetic foot ulcer.


Citations to this article as recorded by  
  • Effective healing of diabetic skin wounds by using nonviral gene therapy based on minicircle vascular endothelial growth factor DNA and a cationic dendrimer
    Min J. Kwon, Songhie An, Sunghyun Choi, Kihoon Nam, Hye S. Jung, Chang S. Yoon, Jung H. Ko, Hye J. Jun, Tae K. Kim, Soo J. Jung, Jeong H. Park, Yan Lee, Jong‐Sang Park
    The Journal of Gene Medicine.2012; 14(4): 272.     CrossRef
Comparison of Minicircle with Conventional Plasmid for the Non-viral Vascular Endothelial Growth Factor (VEGF) Gene Therapy.
Minjeong Kwon, Soonhee Lee, Heysook Chung, Changshin Yoon, Mikyung Kim, Jeonghyun Park
Korean Diabetes J. 2007;31(6):465-471.   Published online November 1, 2007
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  • 22 Download
AbstractAbstract PDF
Delayed wound healings in diabetic patients are related with the impairment of the expressions of various growth factors. Treatments using growth factors have been attempted on diabetic foot ulcer. VEGF (vascular endothelial growth factor) accelerates neo-angiogenesis on the early phase of the wound healing and exerts chemo-attractive effect for the other growth factors and cytokines. Non-viral gene transfer strategies are attractive tool for the gene therapy due to the safety and the versatility, but the low efficiency has been the serious problem. METHODS: We performed the VEGF gene therapy using reconstructed minicircle MINI-pbetaVEGF DNA with a polymeric carrier, polyethylenimine (PEI, 25 kDa) in HEK293, CHO, and NIH3T3 cell lines, and compared its efficiency with the conventional VEGF plasmid pbetaVEGF. RESULTS: The levels of expressed VEGF were higher in the groups using BPEI (branched PEI) as a gene carrier than naked plasmid transfer in all cell lines (P < 0.05). The minicircle MINI-pbetaVEGF DNA showed much higher VEGF expression than conventional plasmid pbetaVEGF (P < 0.05). CONCLUSION: Minicircle DNA MINI-pbetaVEGF showed much higher transfection efficiency than conventional plasmid pbetaVEGF. It might be used in actual human clinical trial due to its higher efficiency and possible safety for the treatment of diabetic foot ulcer.
Glucose-dependent Insulin Secretion from Genetically Engineered K-cells Using EBV-based Episomal Vector.
Ju Hee Kim, Sung Dae Moon, Seung Hyun Ko, Yu Bai Ahn, Ki Ho Song, Hyang Sook Lim, Sook Kyung Lee, Soon Jip Yoo, Hyun Shik Son, Kun Ho Yoon, Bong Yun Cha, Ho Young Son, Sung Joo Kim, Je Ho Han
Korean Diabetes J. 2007;31(1):9-21.   Published online January 1, 2007
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  • 17 Download
  • 2 Crossref
AbstractAbstract PDF
Type 1 diabetes mellitus is an autoimmune disease resulting in destruction of the pancreatic beta cells. Insulin gene therapy for these patients has been vigorously researched. The strategy for achieving glucose-dependent insulin secretion in gene therapy relies on glucose-responsive transcription of insulin mRNA and the constitutive secretory pathway of target non-beta cells. We observed that genetically engineered K-cells using Epstein-Barr virus (EBV)-based episomal vector can produce glucose-regulated insulin production. METHODS: Green fluorescent protein (GFP) or rat-preproinsulin (PPI) expression cassette transcriptionally controlled by the promoter of glucose dependent insulinotropic peptide (GIPP) is fused to pCEP4 containing the origin of replication (oriP) and Epstein-Barr virus nuclear antigen 1 (EBNA-1). CMV promoter was replaced by subcloning the GIPP into pCEP4 to generate pGIPP/CEP4. Two recombinant EBV-based episomal vectors, pGIPP/GFP/CEP4 and pGIPP/PPI/CEP4, were constructed. pGIPP/GFP/CEP4 and pGIPP/PPI/CEP4 containing K-cell specific GIPP were co-transfected into STC-1. K-cell was isolated from the clonal expansion of the fluorescent cells selected by hygromycin treatment in STC-1, and were analyzed for the expression of glucokinase (GK) or transcription factors involved in pancreas development. K-cells concurrently transfected with pGIPP/PPI/CEP4 and pGIPP/GFP/CEP4 were analyzed for the transcripts of PPI by RT-PCR, and for the glucose dependent insulin expression by immunocytochemistry or insulin assay using ultra-sensitive rat-specific insulin ELISA kit. RESULT: STC-1 was stably-transfected with pGIPP/GFP/CEP4 along with pGIPP/PPI/CEP4. Genetically selected fluorescent K-cells expressed GK and transcription factors involved in pancreas development. And K-cells transfected with pGIPP/PPI/CEP4 contained detectable levels of PPI transcripts and showed glucose-dependent immunoreactive insulin secretion. CONCLUSION: We identified genetically engineered K-cells which exert a glucose-dependent insulin expression using EBV-based episomal vector. The similarities between K-cells and pancreatic beta cells support that K-cells may make effective and ideal targeting cells for insulin gene therapy or alternative cell therapy.


Citations to this article as recorded by  
  • Relationship of traditional and nontraditional cardiovascular risk factors to coronary artery calcium in type 2 diabetes
    Ju-Yeon Sim, Ju-Hee Kim, Yu-Bae Ahn, Ki-Ho Song, Je-Ho Han, Bong-Yun Cha, Sook-Kyung Lee, Sung-Dae Moon
    Korean Diabetes Journal.2009; 33(6): 466.     CrossRef
  • Transdifferentiation of Enteroendocrine K-cells into Insulin-expressing Cells
    Esder Lee, Jun Mo Yu, Min Kyung Lee, Gyeong Ryul Ryu, Seung-Hyun Ko, Yu-Bae Ahn, Sung-Dae Moon, Ki-Ho Song
    Korean Diabetes Journal.2009; 33(6): 475.     CrossRef
Insulin Gene Therapy Using HVJ-liposome and Epstein-Barr Virus Plasmid in Murine Streptozotocin Induced Diabetes.
Yong Deuk Kim, Keun Gyu Park, Seong Wook Han, Jong Doek Ahn, Hyo Jung Lee, Mi Jung Kim, Hye Soon Kim, Nam Hee Park, In Kyu Lee
Korean Diabetes J. 2003;27(5):405-413.   Published online October 1, 2003
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  • 17 Download
AbstractAbstract PDF
Despite improvement in insulin preparation and delivery, the use of insulin therapy alone to maintain normal glucose concentration and prevent the development of diabetic complication is not easy. Therefore, there has been considerable interest in developing gene therapy to supply insulin. We investigated that the administration of hemagglutinating virus of Japan (HVJ)- liposome complex, containing human insulin construct into the portal vein to control the blood glucose level in murine streptozotocin (STZ)-induced diabetes. METHODS: Human insulin gene was delivered to STZ-induced diabetic rats through the portal vein using HVJ-liposome containing Epstein-Barr virus (EBV) replicon-based plasmid (pEB). Blood glucose and body weight were measured after insulin gene delivery. The animals were sacrificed 28 days later and the livers were collected for immuno-histochemical staining of insulin. In addition plasma insulin and C-peptide levels were measured. RESULTS: Significant decrease in blood glucose levels and an increase in insulin and C-peptide levels were observed in the insulin gene transfection group as compared to the control group. Immunohistochemical staining of insulin also showed significant differences between these two groups. CONCLUSION: This study demonstrated the possibility of insulin gene therapy through the portal vein using pEB and HVJ-liposome method to produce a sustained improvement of diabetic glucose metabolism.
Insulin Gene Therapy using Vascular Smooth Muscle Cells in Diabetic Rats.
Tae Geun Oh, Mi Ja Lee, Young Ku Kim, Seung Tak Kim
Korean Diabetes J. 2002;26(1):32-46.   Published online February 1, 2002
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AbstractAbstract PDF
Type 1 diabetes mellitus is caused by a lack of insulin. The purpose of this study was to test whether blood glucose control in severe diabetic animals can be achieved by transplanting of rat vascular smooth muscle cells which are transduced with the insulin gene using a retroviral vector system. METHODS: After cloning the recombinant retroviral plasmid including human mutated proinsulin cDNA which contains furin endopeptidase cleavage site, the resulting plasmid, LInABCSN, was transfected into the retroviral packaging cell line (PA317/LhInABCSN). The resulting retrovirus in the supernatant of PA317/ LhInABCSN infected the F344 rat vascular smooth muscle cell (SMC) and produced the SMC/LhInABCSN cells. After transplanting SMC/LInABCSN cells into the internal carotid artery of the rat, diabetes was induced by an intraperitoneal streptozotocin (STZ) injection (50 mg/kg) 2 week later. The blood glucose and insulin levels, percent weight change and the survival rates between the control group (SMC/LNFZ) and the treatment group (SMC/LInABCSN) were compared. RESULTS: The insulin concentrations in the supernatant of the SMC/LhInABCSN mice were 160.2 IU/mL in 24 hours, 243.6 IU/mL in 48 hours and 350.2 IU/mL in 72 hours, but the proinsulin concentrations in 24, 48 and 72 hours were all lower than 1 pmol/L. After 1 day and 3 days of the STZ injection, there were no differences in glucose concentrations between treatment group (n=10) and control group (n=10). There were no statistical differences in the percent weight change between the control and treatment group but the treated rats showed bad a lower weight loss than control rats. After 3 days of the STZ injection, serum insulin concentration of treatment group showed slightly higher levels than the control group (2.7+/-.5 IU/mL vs. 1.6+/-.1 IU/mL, p=0.077). The survival showed a significant increase in treatment group (median survival: 29 days, 9-104 days) compared to the control group (median survival: 6 days, 3-49 days, p < 0.05). CONCLUSION: Although this study did not show a normal glucose concentration in treated rats, it did show significantly higher survival compared to control rats. It is believed that gene therapy using rat vascular smooth muscle cells which transduced the insulin gene may be a new insulin delivery method.

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