epidermal-growth-factor and Wounds--Penetrating

Diabetic wound is a chronic wound in which normal process of wound healing is interrupted. Lack of blood supply, infection and lack of functional growth factors are assumed as some of the conditions that lead to non-healing environment. Epidermal growth factor (EGF) acts primarily to stimulate epithelial cell growth across wound. Erythropoietin (EPO) is a haematopoietic factor, which stimulates the production, differentiation and maturation of erythroid precursor cells. This study hypothesised combining these two factors, non-healing process of diabetic wound will be compensated and eventually lead to acceleration of wound healing compared with single growth factor treatment. A total of 30 diabetic Sprague-Dawley rats were divided into three treatment groups (single treatment of rh-EPO or rh-EGF or combined treatment on a full-thickness skin wound). To assess the wound healing effects of the components, the wound size and the healing time were measured in each treatment groups. The skin histology was examined by light microscopy and immunohistochemical analysis of proliferating markers was performed. The combined treatment with rh-EPO and rh-EGF improved full-thickness wound significantly (P < 0·05) accelerating 50% healing time with higher expression of Ki-67 compared with single growth factor-treated groups. The combined treatment failed to accelerate the total healing time when compared with single growth factor treatments. However, the significant improvement were found in wound size reduction in the combined treatment group on day 4 against single growth factor-treated groups (P < 0·05). This study demonstrated that the combined treatment of rh-EPO and rh-EGF improved the wound healing possibly through a synergistic action of each growth factor. This application provides further insight into combined growth factor therapy on non-healing diabetic wounds.

Topics: Animals; Bandages, Hydrocolloid; Diabetes Mellitus, Experimental; Drug Therapy, Combination; Epidermal Growth Factor; Erythropoietin; Follow-Up Studies; Humans; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Skin; Treatment Outcome; Wound Healing; Wounds, Penetrating

Materials able to deliver topically bioactive molecules represent a new generation of biomaterials. In this article, we describe the use of silk mats, made of electrospun nanoscale silk fibers containing epidermal growth factor (EGF), for the promotion of wound healing processes. In our experiments, we demonstrated that EGF is incorporated into the silk mats and slowly released in a time-dependent manner (25% EGF release in 170h). We tested these materials using a new model of wounded human skin-equivalents displaying the same structure as human skin and able to heal using the same molecular and cellular mechanisms found in vivo. This human three-dimensional model allows us to demonstrate that the biofunctionalized silk mats, when placed on the wounds as a dressing, aid the healing by increasing the time of wound closure by the epidermal tongue by 90%. The preservation of the structure of the mats during the healing period as demonstrated by electronic microscopy, the biological action of the dressing, as well as the biocompatibility of the silk demonstrate that this biomaterial is a new and very promising material for medical applications, especially for patients suffering from chronic wounds.

Topics: Absorption; Administration, Topical; Bandages; Diffusion; Drug Carriers; Electrochemistry; Epidermal Growth Factor; Materials Testing; Rotation; Silk; Skin; Wound Healing; Wounds, Penetrating

This study was designed to elucidate the in vivo efficacy of epidermal growth factor (EGF) on wound healing in non diabetic and diabetic rats.. Ninety-six male Wistar-Albino rats were randomly divided into six groups. Saline-moistened gauze, pure gelatin or EGF in gelatin-microsphere dressings were used in a dermal excision model in both normal and streptomycin-induced diabetic rats. Wound healing was evaluated on day 7 and 14. Reduction in wound area, hydroxypyroline content and tensile strength of the wound were evaluated in each rat. Tissue samples taken from the wounds were examined histopathologically for reepithelialisation, cellular infiltration, number of fibroblasts, granulation and neovascularisation.. On day 7, the use of EGF-containing dressing was observed to reduce the wound area better when compared with the other dressings tested. This effect was significant in normal rats rather than diabetic rats. The difference in reduction of wound area did not persist on day 14. No significant effect on hydroxyproline content of the wound was found with EGF-containing dressing in either normal or diabetic rats. There was a statistically significant increase in tensile strength values of EGF-applied non diabetic rats over the 14 day period. An increase in tensile strength was prominent in also EGF-applied diabetic rats on day 14. Histological examination revealed higher histopathologic scores in EGF-applied diabetic and non diabetic rats.. These findings implicate that use of EGF in gelatin-microsphere dressings improves wound healing both in normal and diabetic rats.

Topics: Administration, Topical; Animals; Diabetes Mellitus, Experimental; Epidermal Growth Factor; Gelatin Sponge, Absorbable; Male; Microspheres; Rats; Rats, Wistar; Streptozocin; Wound Healing; Wounds, Penetrating

Topics: Animals; Chitosan; Delayed-Action Preparations; Disease Models, Animal; Epidermal Growth Factor; Humans; Random Allocation; Recombinant Proteins; Swine; Wound Healing; Wounds and Injuries; Wounds, Penetrating

The purpose of this article is to evaluate the effect of continuously released recombinant human epidermal growth factor (rh-EGF) in chitosan film in full thickness porcine wounds. A total of 10 domestic pigs (Yorkshire species) weighing 18 to 22 kg between the ages of 50 to 60 days were used. The wounds were divided into 3 groups and treated selectively with rh-EGF in chitosan film (EGF 20 ug/wound/d), chitosan film without rh-EGF, or remained as the control group. One hundred percent healing time was observed, and hematoxylin and eosin and Anti Ki-67 antibody immunohistochemical staining were performed. The 100% healing time and Anti Ki-67 antibody immunohistochemical staining showed statistical significance of the rh-EGF chitosan film-treated group against the control group (P < 0.05). But it did not reveal any statistical significance over the chitosan film-treated group. In this preliminary study, although continuous release of rh-EGF in chitosan film accelerates epithelialization, the benefit of the combination of rh-EGF in chitosan cannot be determined over the use of chitosan alone. Further analysis using complex wound models such as diabetes or infection, which may have different pathology in healing, will be needed to evaluate the potential benefit/synergistic effectiveness.

Topics: Animals; Bandages, Hydrocolloid; Chitosan; Delayed-Action Preparations; Drug Carriers; Epidermal Growth Factor; Humans; Recombinant Proteins; Skin; Swine; Wound Healing; Wounds, Penetrating

We examined in vivo efficiency of a gelatin film sheet with epidermal growth factor (EGF) for a novel therapeutic device for cutaneous wound repair. NIH3T3 fibroblasts and PAM212 keratinocytes proliferated when the cells were incubated with the homogenate of EGF containing gelatin sheets, indicating that the gelatin sheet retained biologic activity of EGF in the process of sheet formation. To evaluate therapeutic effects, the EGF containing gelatin sheets or control sheets were applied onto partial-thickness skin wounds made on dorsa of hairless dogs. Wound closure in wounds treated with EGF containing gelatin sheets was accelerated when compared to that of wounds treated with control sheets, exhibiting earlier reepithelialization of the epidermis and highly regulated repair of extracellular matrix in the dermis. Therefore, we concluded that the gelatin film is a useful material to keep EGF stable and the EGF containing sheet has the ability to become an efficient therapeutic agent for superficial or deep partial-thickness wounds in the skin.

Topics: Animals; Bandages; Dogs; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Evaluation Studies as Topic; Gelatin; Histological Techniques; Male; Mice; NIH 3T3 Cells; Skin; Wound Healing; Wounds, Penetrating

Growth factors have proved to be an effective therapeutic strategy. However, some controversies have arisen concerning their efficacy in topical wound treatments. Stabilization of epidermal growth factors at the wound site and long-lasting receptor occupancy are important factors for wound repair. This study evaluated the cumulative profiles of two jellies containing 10 or 20 microg of 125I-rhEGF per gram of jelly, in a rat full-thickness skin lesion model. The prolonged time-courses at the wound sites for both strengths compared with saline solutions previously evaluated using a similar skin lesion model are reported. It seems that these two topical formulations that provide more sustained amounts of 125I-rhEGF over the period of sampling, would probably achieve the required wound healing response in terms of cell proliferation, collagen deposition and protein synthesis. Further studies need to be developed in order to elucidate whether such an in vivo disposition pattern is consistent with an earlier and stronger promotion of wound healing events.

Topics: Administration, Topical; Animals; Chemistry, Pharmaceutical; Epidermal Growth Factor; Gels; Humans; Iodine Radioisotopes; Male; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Wounds, Penetrating

Epidermal growth factor (EGF) and its receptor (EGF-R) are attractive targets for cancer immunotherapy. Tolerance has been broken with an EGF-vaccine and antibodies against EGF have been produced in animals and in cancer patients. EGF also plays an important role in the inflammation stage of wound healing. Because this therapeutic approach may be of importance after surgery procedures in cancer patients, we decided to investigate the possible role of the EGF-vaccine in the croton-oil-induced ear edema and in the wound healing experimental animal models.. Mice were immunized with an EGF-vaccine by intramuscular injections and serum titers against EGF were measured through ELISA techniques. Control animals received saline.. Immunized mice produced antibodies against EGF while no antibody titers could be measured in control animals. Croton oil applied to the inner ear surface of EGF-vaccine treated mice caused a 61.3% lower ear punch weight and a 60.2% lower myeloperoxidase activity than control mice. In the EGF-vaccine treated animals, planimetry measurements and histological analysis did not led to significant impairment in tissue repair.. The EGF-vaccination in mice decreased the normal croton-oil-induced inflammation response, without apparent impairment in tissue healing.

Topics: Animals; Antibody Formation; Autoantibodies; Cancer Vaccines; Croton Oil; Dermatologic Agents; Drug Combinations; Ear Diseases; Edema; Epidermal Growth Factor; Image Processing, Computer-Assisted; Immunization; Inflammation; Injections, Intramuscular; Mice; Mice, Inbred BALB C; Skin; Wound Healing; Wounds, Penetrating

Human corneal epithelial cells (HCECs) were functionally depleted of erbB2 to elucidate its role in epidermal growth factor (EGF) receptor (EGFR) activation-dependent cell migration.. The retrovirus pBabe-5R, which encodes an erbB2 single-chain antibody with an endoplasmic reticulum (ER)-targeting sequence, and control pBabe-puro were used to infect THCE cells (an SV40-immortalized HCEC line). Several cell lines expressing 5R were selected along with a pBabe-puro control line. The depletion of erbB2 was verified by cell surface biotinylation of proteins, followed by streptavidin precipitation and subsequent detection of erbB2 by immunoblot analysis. Activation of erbBs was analyzed by immunoprecipitation using the phosphotyrosine antibody pY20, followed by Western blot analysis with erbB1 or erbB2 antibodies. Phosphorylation of extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3'-kinase (PI3K) was analyzed by Western blot with antibodies specific to phosphorylated proteins. Effects of erbB2 depletion on heparin-binding EGF-like growth factor (HB-EGF)-induced cell migration were determined by Boyden chamber migration assay and by scratch wound assay.. Wounding induced erbB2 tyrosine phosphorylation. Expression of 5R encoding an erbB2 single-chain antibody with an endoplasmic reticulum-targeting sequence depleted the cell surface expression of erbB2 in HCECs. Wounding resulted in a rapid increase in the phosphorylation of erbB1 in both 5R-expressing and control cells, whereas wound-induced erbB2 phosphorylation in 5R-expressing cells was not detectable. Depletion of functional erbB2 attenuated the healing of scratch wounds in the presence of HB-EGF and impaired both chemotactic migration stimulated by HB-EGF and haptotactic migration toward a fibronectin-collagen I (3:1; FNC) coating mix. Expression of 5R affected both the intensity and the duration of wound-induced, EGFR-elicited ERK and PI3K activation. Inhibition of ERK and PI3K pathways in cultured porcine corneas impaired ex vivo epithelial wound healing.. ErbB2 serves as a critical component that couples erbB receptor tyrosine kinase to the migration machinery of corneal epithelial cells.

Topics: Cell Line, Transformed; Cell Membrane; Cell Movement; Coated Materials, Biocompatible; Collagen Type I; Enzyme Activation; Enzyme Inhibitors; Epidermal Growth Factor; Epithelium, Corneal; ErbB Receptors; Extracellular Signal-Regulated MAP Kinases; Eye Injuries; Fibronectins; Heparin-binding EGF-like Growth Factor; Humans; Intercellular Signaling Peptides and Proteins; Membranes, Artificial; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Receptor, ErbB-2; Tyrosine; Wound Healing; Wounds, Penetrating

Epidermal growth factor (EGF) and zinc promote re-epithelization and reparative tissue strength by enhancing deposition of collagen at the site of the wound. In this study two EGF dosage forms were chosen to assess the effect of zinc levels on wound healing and for comparison with wound tear strengths. A solution of EGF in 0.9% w/v NaCl and an EGF gel in 0.2% Carbopol 940 polymer (5 microL) were applied to full-thickness skin wounds of mice twice a day for 7 and 15 days. Wound zinc levels were higher on day 7 than on day 15, especially in wounds treated with EGF. The wound zinc levels of the gel + EGF group on day 15 were similar to those of normal control skin. These results imply that there is a close connection, but no direct relationship, between EGF application in both dosage forms and wound zinc levels during healing.

Topics: Animals; Chemistry, Pharmaceutical; Dermatologic Agents; Epidermal Growth Factor; Female; Male; Mice; Mice, Inbred C57BL; Skin; Wound Healing; Wounds, Penetrating; Zinc

Dehiscence of colonic anastomoses is a multifactorial phenomenon. One mechanism by which this can occur is a deficiency of colonic submucosal collagen. Peptide growth factors (PGFs) have been shown to play a role in the synthesis, deposition, and maturation of collagen. Specifically, in tissues other than the colon, the transforming growth factor-beta (TGF-beta1) gene has been shown to be temporally associated with expression of the procollagen gene. This study examines the temporal expression of the TGF-beta1, epidermal growth factor (EGF), and platelet-derived growth factor B (PDGF-BB) genes and their temporal relationship to the expression of the procollagen type 1 (PROC I) gene.. Forty-eight Sprague-Dawley rats underwent transection of the descending colon with primary anastomosis. Perianastomotic colonic tissue was harvested on Day 0 and postoperative days 3, 5, 6, 7, and 14. Colonic tissue was analyzed using semiquantitative reverse transcriptase-polymerase chain reaction and primers specific for the TGF-beta1, EGF, and PDGF-B growth factors. Relative expression ratios of PGFs and PROC I genes were calculated versus a constitutive gene.. The data show that although all three of the PGFs genes were expressed in healing postoperative colonic tissue, only TGF-beta1 showed a significant increase in its level of expression versus a constitutive gene from a mean ratio of 0.4 +/- 0. 08 on Day 0 to a mean ratio of 1.9 +/- 0.27 on Day 7 (P < 0.0001 by ANOVA). The PROC I gene also showed a significant increase in expression (P < 0.001 by ANOVA) in the postoperative period which temporally correlated with the increase in the expression of the TGF-beta1 gene (r = 0.89, P < 0.05).. The temporal correlation between an increase in the gene expression of TGF-beta1 and PROC I is initial evidence that that TGF-beta1 plays a significant role in collagen metabolism in a healing colonic anastomosis.

Topics: Animals; Becaplermin; Colon; Epidermal Growth Factor; Gene Expression Regulation; Male; Platelet-Derived Growth Factor; Procollagen; Proto-Oncogene Proteins c-sis; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; Time Factors; Transforming Growth Factor beta; Wound Healing; Wounds, Penetrating

Using hamster as an oral wound healing model, we examined eosinophils and their expression of transforming growth factor-alpha (TGF-alpha) and transforming growth factor-beta 1 (TGF-beta 1). Oral wounds healed approximately two times faster than their cutaneous counterparts. Eosinophils infiltrated prominently into oral wounds; however, unlike the dual expression of TGF-alpha and TGF-beta 1 in skin wounds, oral wound-associated eosinophils expressed TGF-beta 1, but not TGF-alpha. Because saliva is present in oral environments and contains epidermal growth factor (EGF) and TGF-alpha, sialoadenectomy was performed in this model to determine whether the lack of TGF-alpha expression by eosinophils in oral wounds is due to the presence of salivary EGF and/or TGF-alpha. We found that eosinophils in sialoadenectomized hamsters did express TGF-alpha during oral wound healing but that such expression was suppressed when EGF was added to their drinking water. Taken together, our findings suggest that eosinophil-derived TGF-alpha and salivary TGF-alpha/ EGF may have complementary roles in contributing to TGF-alpha in oral wound healing.

Topics: Animals; Cricetinae; Eosinophils; Epidermal Growth Factor; Male; Mouth Mucosa; RNA, Messenger; Saliva; Salivary Glands; Time Factors; Transforming Growth Factor alpha; Wound Healing; Wounds, Penetrating

Epidermal growth factor (EGF) has been shown to stimulate connective tissue repair in the perforated mesentery of rats. The aim of the present investigation was to study the effect of EGF on the formation of healing tissue and angiogenesis in such repair. After laparotomy standardised perforations were made in the centre of the mesenteric "windows" with a scalpel. The rats were given intraperitoneal injections of either 10 micrograms EGF dissolved in phosphate-buffered saline (PBS), or PBS alone, twice daily for four consecutive days beginning on the day of operation. In the first experiment, healing tissue formation and angiogenesis was quantified morphometrically in perpendicularly cut mesenteric windows on days 1 to 10 after operation. Treatment with EGF caused the formation of significantly more healing tissue on days 2 to 7, but no stimulation of angiogenesis. In the second experiment, angiogenesis was quantified morphometrically on days 14 and 21. Mesenteric windows were spread out on objective slides after the capillary bed had been visualised by perfusion of carbon ink. Perforation caused a significant increase of microvascular density in the centre of the mesenteric windows on days 14 and 21. Treatment with EGF did not stimulate angiogenesis at any observation point. In conclusion, treatment with EGF significantly increased the formation of healing tissue in connective tissue repair in the perforated mesentery of rats, but did not affect angiogenesis.

Topics: Animals; Connective Tissue; Epidermal Growth Factor; Male; Mesentery; Neovascularization, Pathologic; Rats; Rats, Sprague-Dawley; Wound Healing; Wounds, Penetrating

Impaired wound healing results in significant morbidity for the surgical patient. The genetically diabetic (C57BL/KsJ-db/db) mouse is obese, hyperglycemic, insulin-resistant, and exhibits markedly impaired wound healing. Previous studies have demonstrated that the fibroblast mitogens, BB homodimer of platelet-derived growth factor (PDGF-BB) or basic fibroblast growth factor, plus insulin-like growth factor, act synergistically to enhance wound closure in the genetically diabetic mouse. The purpose of this study was to determine whether the keratinocyte mitogens, epidermal growth factor (EGF) or transforming growth factor-alpha (TGF-alpha), in combination with the fibroblast mitogen, PDGF-BB, would produce a similar synergistic enhancement in tissue repair. Full-thickness skin wounds created on the backs of diabetic mice received topical applications of vehicle (5% polyethylene glycol), PDGF-BB (10 micrograms), EGF (1 microgram), TGF-alpha (1 microgram), or the combination of PDGF (10 micrograms) and EGF (1 microgram) or TGF-alpha (1 microgram) for 5 consecutive days starting at wounding. Application of PDGF-BB or TGF-alpha alone to wounds in diabetic animals improved wound closure when compared to vehicle treatment. EGF did not affect healing and did not have any additive effects when combined with PDGF-BB. Significant improvements in wound closure were observed with the combination of PDGF-BB and TGF-alpha when compared to treatment with the individual growth factors. The PDGF-BB/TGF-alpha combination accelerated healing in the diabetic animals to a rate that was closer to that seen in nondiabetic mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Becaplermin; Diabetes Mellitus; Drug Synergism; Epidermal Growth Factor; Female; Mice; Platelet-Derived Growth Factor; Proto-Oncogene Proteins c-sis; Recombinant Proteins; Skin; Transforming Growth Factor alpha; Wound Healing; Wounds, Penetrating

A significant percentage of tympanic membrane (TM) perforations require some form of therapy to heal. Topical application of epidermal growth factor (EGF), a potent mitogen for epidermal and mesodermal cells, promotes healing of injuries in tissues histologically similar to TM, such as cornea and skin. We evaluated TM as a target tissue for EGF action. Specific, high affinity receptors for EGF were detected in TM (Kd = 3.1 nM, 150 fmol receptor/mg protein). Autoradiography of iodine 125-EGF binding to intact TMs revealed that EGF receptors were present on cells in the stratified squamous epithelial layer and in the stromal/mucoepithelial layer. Repetitive treatment of perforated cat TMs with 20 micrograms of EGF formulated in saline induced substantial hyperplasia of epithelial and stromal layers compared to paired TMs treated with saline. A single treatment of perforated cat TMs with 50 micrograms of EGF formulated in a hydrogel or in shredded Gelfoam produced significantly (p less than .05) smaller perforations at 6 days following the operation compared to paired TMs treated with vehicles. These results demonstrate that TM is a target tissue for EGF and that topical treatment with EGF stimulates healing of TM perforations.

Topics: Animals; Autoradiography; Binding, Competitive; Cats; Epidermal Growth Factor; ErbB Receptors; Swine; Tympanic Membrane; Wound Healing; Wounds, Penetrating