losartan-potassium and Burns

Topics: Animals; Burns; Erythroid Precursor Cells; Erythropoietin; Hematopoiesis; Hematopoietic Stem Cells; Humans

Although it was initially assumed that erythropoietin (EPO) was a hormone that only affected erythropoiesis, it has now been proposed that EPO plays an additional key role in the regulation of acute and chronic tissue damage.. This is a large, prospective, randomized, double-blind, multi-center study, funded by the German Federal Ministry of Education and Research, and fully approved by the designated ethics committee. The trial, which is to investigate the effects of EPO in severely burned patients, is in its recruitment phase and is being carried out in 13 German burn care centers. A total of 150 patients are to be enrolled to receive study medication every other day for 21 days (EPO 150 IU/kg body weight or placebo). A follow-up of one year is planned. The primary endpoint of this study is the time until complete re-epithelialization of a defined skin graft donor site is reached. Furthermore, clinical parameters such as wound healing, scar formation (using the Vancouver scar scale), laboratory values, quality of life (SF-36), angiogenic effects, and gene- and protein-expression patterns are to be determined. The results will be carefully evaluated for gender differences.. We are seeking new insights into the mechanisms of wound healing in thermally injured patients and more detailed information about the role EPO plays, specifically in these complex interactions. We additionally expect that the biomimetic effects of EPO will be useful in the treatment of acute thermal dermal injuries.. EudraCT Number: 2006-002886-38, Protocol Number: 0506, ISRCT Number: http://controlled-trials.com/ISRCTN95777824/ISRCTN95777824.

Topics: Adolescent; Adult; Aged; Burns; Clinical Protocols; Double-Blind Method; Drug Administration Schedule; Erythropoietin; Female; Germany; Humans; Male; Middle Aged; Prospective Studies; Re-Epithelialization; Recombinant Proteins; Regeneration; Research Design; Severity of Illness Index; Skin; Skin Transplantation; Time Factors; Treatment Outcome; Young Adult

To evaluate the effects of recombinant human erythropoietin (r-HuEPO) in attempting to prevent anemia in acutely burned patients.. Prospective double-blind randomized study of 40 patients.. Patients with burns from 25% to 65% total body surface were enrolled. r-HuEPO or a placebo was begun within 72 hours of admission. Cell blood count, reticulocyte counts, transfusion requirements, and blood loss were measured. Comparison was carried out by the unpaired t test.. There was no statistically significant difference in hemoglobin, hematocrit, reticulocyte count, ferritin, serum iron, total iron blinding capacity, or transfusion requirements. In patients with burns from 25% to 35%, the reticulocyte counts were statistically significantly higher.. In our work the administration of r-HuEPO in acutely burned patients did not prevent the development of postburn anemia or decrease transfusion requirements. Increased erythropoiesis in smaller burns (25% to 35%) was observed and may indicate a reason for further study.

Topics: Adolescent; Adult; Anemia; Blood Transfusion; Burns; Double-Blind Method; Erythropoietin; Female; Humans; Male; Middle Aged; Prospective Studies; Recombinant Proteins; Reticulocyte Count

Topics: Burns; Erythropoietin; Humans; Recombinant Proteins; Skin; Wound Healing

Stem cells have shown promising potential to treat burn wounds. Erythropoietin was capable of promoting in vitro transdifferentiation of mesenchymal stem cells (MSCs). The aim of the study was to investigate possible role of erythropoietin-pretreated mesenchymal stem cells (EPOa/MSCs) in burn wounds healing and to evaluate its in vivo differentiation into keratinocytes.. Forty rats were utilised in this study divided into four groups (n = 10 for each). Control group (I), burn group (II), burn + MSCs, group (III), burn + EPOa/MSCs. 1 × 10⁶ cells were injected locally for each 1 cm² of burn areas. Burn areas were followed-up morphologically. After 21 days of the experiment, the rats were euthanised, skin specimens were assessed biochemically, histologically and immunohistochemically.. EPOa/MSCs enhanced significantly (p < 0.05) burn wound vimentin gene expression and level of interleukin (IL)-10 while decreased IL-1 and COX2 as compared to the burn group. Histologically, EPOa/MSCs improved epithelialisation despite stem cells' differentiation into keratinocytes was rarely detected by PKH26 red fluorescence. EPOa/MSCs promoted angiogenesis as detected by significant increase in VEGF and PDGF immunoexpression as compared to burn group.. EPOa/MSCs may improve burn wound healing, probably through anti-inflammatory, immunomodulatory and angiogenic action. However, in vivo transdifferentiation into keratinocytes was rarely detected.

Topics: Animals; Biomarkers; Burns; Cell Proliferation; Cell Transdifferentiation; Erythropoietin; Gene Expression Regulation; Inflammation Mediators; Keratinocytes; Male; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mice; Organic Chemicals; Platelet-Derived Growth Factor; Rats; Skin; Treatment Outcome; Vascular Endothelial Growth Factor A; Vimentin; Wound Healing

Burn-induced neuromuscular dysfunction may contribute to long-term morbidity; therefore, it is imperative to develop novel treatments. The present study investigated whether erythropoietin (EPO) administration attenuates burn-induced motor neuron apoptosis and neuroinflammatory response. To validate our hypothesis, a third-degree hind paw burn rat model was developed by bringing the paw into contact with a metal surface at 75°C for 10 s. A total of 24 male Sprague-Dawley rats were randomly assigned to four groups: Group A, sham-control; Group B, burn-induced; Group C, burn + single EPO dose (5000 IU/kg i.p. at D0); and Group D, burn + daily EPO dosage (3000 IU/kg/day i.p. at D0-D6). Two treatment regimens were used to evaluate single versus multiple doses treatment effects. Before sacrifice, blood samples were collected for hematological parameter examination. The histological analyses of microglia activation, iNOS, and COX-2 in the spinal cord ventral horn were performed at week 1 post-burn. In addition, we examined autophagy changes by biomarkers of LC3B and ATG5. The expression of BCL-2, BAX, cleaved caspase-3, phospho-AKT, and mTOR was assessed simultaneously through Western blotting. EPO administration after burn injury attenuated neuroinflammation through various mechanisms, including the reduction of microglia activity as well as iNOS and COX-2 expression in the spinal cord ventral horn. In addition, the expression of phospho-AKT, mTOR and apoptotic indicators, such as BAX, BCL-2, and cleaved caspase-3, was modulated. Furthermore, the activity of burn-induced autophagy in the spinal cord ventral horn characterized by the expression of autophagic biomarkers, LC3B and ATG5, was reduced after EPO administration. The present results indicate that EPO inhibits the AKT-mTOR pathway to attenuate burn-induced motor neuron programmed cell death and microglia activation. EPO can modulate neuroinflammation and programmed cell death and may be a therapeutic candidate for neuroprotection.

Topics: Animals; Apoptosis; Burns; Disease Models, Animal; Erythrocyte Count; Erythropoietin; Hematocrit; Immunohistochemistry; Male; Motor Neurons; Rats; Rats, Sprague-Dawley

We have previously reported that the topical application of erythropoietin (EPO) to cutaneous wounds in rats and mice with experimentally induced diabetes accelerates their healing by stimulating angiogenesis, reepithelialization, and collagen deposition, and by suppressing the inflammatory response and apoptosis. Aquaporins (AQPs) are integral membrane proteins whose function is to regulate intracellular fluid hemostasis by enabling the transport of water and glycerol. AQP3 is the AQP that is expressed in the skin where it facilitates cell migration and proliferation and re-epithelialization during wound healing. In this report, we provide the results of an investigation that examined the contribution of AQP3 to the mechanism of EPO action on the healing of burn wounds in the skin of pigs with experimentally induced type 1 diabetes. We found that topical EPO treatment of the burns accelerated their healing through an AQP3-dependent mechanism that activates angiogenesis, triggers collagen and hyaluronic acid synthesis and the formation of the extracellular matrix (ECM), and stimulates reepithelialization by keratinocytes. We also found that incorporating fibronectin, a crucial constituent of the ECM, into the topical EPO-containing gel, can potentiate the accelerating action of EPO on the healing of the burn injury.

Topics: Administration, Topical; Angiogenesis Inducing Agents; Animals; Aquaporin 3; Burns; Collagen; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Erythropoietin; Extracellular Matrix; Fibronectins; Hyaluronic Acid; Keratinocytes; Neovascularization, Physiologic; Re-Epithelialization; Skin; Swine; Wound Healing

Inflammation-mediated impairment of erythropoiesis plays a central role in the development of the anemia of critical illness (ACI). ACI develops despite elevation of endogenous erythropoietin (EPO), does not respond to exogenous erythropoietin (EPO) supplementation, and contributes significantly to transfusion requirements in burned patients. We have reported previously that the reduction of red blood cell mass in the bone marrow of a burn-injured ACI mouse model is granulocyte colony-stimulating factor (G-CSF) dependent. Given that elevated G-CSF levels also have been associated with lower hemoglobin levels and increased transfusion requirements in trauma victims, we postulated that G-CSF mediates postburn EPO resistance. In ACI mice, we found that bone marrow erythroid differentiation, viability, and proliferation are impaired after thermal injury of the skin. These changes in the marrow were associated with attenuated phosphorylation of known EPO-responsive signaling nodes, signal transducer and activator of transcription 5 (STAT5) Y694 and STAT3 S727, in bone marrow erythroid cells and developed despite highly elevated levels of endogenous EPO. Severely blunted STAT5 Y694 phosphorylation in bone marrow erythroid cells after exogenous EPO supplementation confirmed that EPO signaling was impaired in ACI mice. Importantly, parenteral administration of anti-G-CSF largely rescued postburn bone marrow erythroid differentiation arrest and EPO signaling in erythroid cells. Together, these data provide strong evidence for a role for G-CSF in the development of ACI after burn injury through suppression of EPO signaling in bone marrow erythroid cells.

Topics: Anemia; Animals; Burns; Cell Differentiation; Erythroid Cells; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Mice; Phosphorylation; Signal Transduction; Skin; STAT3 Transcription Factor; STAT5 Transcription Factor

Recent studies have suggested that pyruvate-enriched oral rehydration solution (Pyr-ORS) may be superior to the standard bicarbonate-based ORS in the protection of intestine from ischemic injury. The aim of this study was to compare the effects of Pyr-ORS with citrate-enriched ORS (Cit-ORS) on the intestinal hypoxia-inducible factor-1 (HIF-1)-erythropoietin (EPO) signaling pathway for enteral rehydration in a rat model of burn injury.. Rats were randomly assigned to 4 groups (N = 20, 2 subgroups each: n = 10): scald sham (group SS), scald with no fluid resuscitation (group SN), scald and resuscitation with enteral Cit-ORS (group SC), and scald and resuscitation with enteral Pyr-ORS (group SP). At 2.5 and 4.5 hours after a 35% total body surface area (TBSA) scald, intestinal mucosal blood flow (IMBF), contents of HIF-1, EPO, endothelial nitric oxide synthase (eNOS), nitric oxide (NO), barrier protein (ZO-1), levels of serum diamine oxidase (DAO), and intestinal mucosal histology injury score were determined.. Serum DAO activities in the scalded groups were significantly elevated, but less raised in group SP than in group SC, at 2.5 hours and at 4.5 hours after the scald. Further, group SP more profoundly preserved intestinal HIF-1 expression compared with group SC at the 2 time points. Compared with group SC, group SP had markedly elevated intestinal EPO, eNOS, and NO levels at the same time points, respectively (P < .05). Similarly, IMBF and ZO-1 levels were significantly higher in group SP than in group SC. Intestinal mucosal histopathological scores were statistically higher at 2.5 hours and 4.5 hours after scalding but were more attenuated in group SP than in group SC (P < .05). Immunofluorescence expression of intestinal mucosal ZO-1 was consistent with the above changes. The above parameters were also significantly different between groups SC and SN (all P < .05).. Pyr-ORS provides a superior option to Cit-ORS for the preservation of intestinal blood flow and barrier function and the attenuation of histopathological alterations in enteral resuscitation of rats with burn injury. Its underlying mechanism may be closely related to the pyruvate in activation of intestinal HIF-1-EPO signaling cascades.

Topics: Amine Oxidase (Copper-Containing); Animals; Bicarbonates; Body Surface Area; Burns; Citric Acid; Erythropoietin; Fluid Therapy; Glucose; Hypoxia-Inducible Factor 1; Intestinal Mucosa; Intestines; Male; Nitric Oxide Synthase Type III; Potassium Chloride; Pyruvic Acid; Rats; Rats, Sprague-Dawley; Resuscitation; Signal Transduction; Sodium Chloride; Zonula Occludens-1 Protein

Cutaneous burns are often exacerbated by poor perfusion and subsequent necrosis of the microvasculature surrounding the primary injury. Preservation of these vessels can reduce necrotic tissue expansion and increase success rates of skin graft procedures. Recent work has identified a peptide derived from erythropoietin, ARA290, with the ability to mediate tissue protection in a variety of cell types. Here we demonstrate the advantages of fusing ARA290 to an elastin-like polypeptide (ELP) to salvage microvascular endothelial cells in harsh proteolytic conditions following thermal shock. These fusion proteins were expressed recombinantly in bacterial hosts and rapidly purified by inverse transition cycling. They were shown to spontaneously aggregate into particles at subphysiological temperatures. The bifunctional submicron particles were resistant to digestion in enzymes upregulated after burn injury. Furthermore, the data strongly suggest these ARA290-functionalized particles were superior to treatment with the peptide alone in preventing microvascular cell death in these conditions. The results bring to light an efficient and cost-effective strategy for the delivery therapeutic peptides to proteolytically active wound sites.

Topics: Biopolymers; Burns; Cell Line; Cell Survival; Elastin; Erythropoietin; Escherichia coli; Hot Temperature; Humans; Microvessels; Oligopeptides; Proteolysis; Recombinant Fusion Proteins; Skin; Wound Healing

Large doses of recombinant growth factors formulated in solution form directly injected into the body is usual clinical practice in treating second-degree scald injuries, with promising results, but this approach creates side effects; furthermore, it may not allow appropriate levels of the factor to be sensed by the target injured tissue/organ in the specific time frame, owing to complications arising from regeneration. In this research, two delivery methods (infusion pumping and local topical application) were applied to deliver recombinant human erythropoietin (rHuEPO) for skin regeneration. First, rHuEPO was given in deep second-degree scald injury sites in mice by infusion pump. Vascularization was remarkably higher in the rHuEPO pumping group than in controls. Second, local topical application of rHuEPO gel was given in deep second-degree scald injury sites in rats. Histological analysis showed that epithelialization rate was significantly higher in the rHuEPO gel-treated group than in controls. Immunohistochemical studies showed that the rHuEPO gel-treated group showed remarkably higher expression of skin regeneration makers than the control group. An accurate method for visualization and quantification of blood vessel networks in target areas has still not been developed up to this point, because of technical difficulties in detecting such thin blood vessels. A method which utilizes a series of steps to enhance the image, removes noise from image background, and tracks the vessels edges for vessel segmentation and quantification has been used in this study. Using image analysis methods, we were able to detect the microvascular networks of newly formed blood vessels (less than 500 μm thickness), which participate in the healing process, providing not only nutrition and oxygen to grow tissues but also necessary growth factors to grow tissue cells for complete skin regeneration. The rHuEPO-treated group showed higher expression of stem cell markers (CD 31, CD 90, CD 71, and nestin), which actively contribute to in-wound-healing processes for new hair follicle generation as well as skin regeneration. Collectively, both rHuEPO group pumping into the systemic circulation system, and injection into the local injury area, prompted mice and rats to form new blood vessel networks in scald injury sites, which significantly participate in the scald healing process. These results may lead to the development of novel treatments for scald wounds.

Topics: Administration, Topical; Animals; Blood Vessels; Burns; Epithelium; Erythropoietin; Female; Gels; Humans; Infusion Pumps; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Neovascularization, Physiologic; Rats; Recombinant Proteins; Regeneration; Skin Physiological Phenomena; Wound Healing

Deep second-degree burns are characterized by delayed formation of granulation tissue and impaired angiogenesis. Erythropoietin (EPO) is able to stimulate angiogenesis and mitosis, activating vascularization and cell cycle. The aim of our study was to investigate whether two biosimilar recombinant human erythropoietins, EPO-α and EPO-Z, may promote these processes in an experimental model of burn injury. A total of 84 mice were used and a scald burn was produced on the back after shaving, in 80°C water for 10 seconds. Mice were then randomized to receive EPO-α (400 units/kg/day/sc) or EPO-Z (400 units/kg/day/sc) or their vehicle (100 μL/day/sc 0.9% NaCl solution). After 12 days, both EPO-α and EPO-Z increased VEGF protein expression. EPO-α caused an increased cyclin D1/CDK6 and cyclin E/CDK2 expression compared with vehicle and EPO-Z (p<0.001). Our study showed that EPO-α and EPO-Z accelerated wound closure and angiogenesis; however EPO-α resulted more effectively in achieving complete skin regeneration. Our data suggest that EPO-α and EPO-Z are not biosimilars for the wound healing effects. The higher efficacy of EPO-α might be likely due to its different conformational structure leading to a more efficient cell proliferation and skin remodelling.

Topics: Animals; Brain Injuries; Burns; Epoetin Alfa; Erythropoietin; Gene Expression Regulation; Humans; Mice; Models, Theoretical; Neovascularization, Physiologic; Recombinant Proteins; Vascular Endothelial Growth Factor A; Wound Healing

Topics: Animals; Burns; Erythropoietin; Male; Neovascularization, Physiologic; Wound Healing

To investigate whether systemic erythropoietin administration can prevent secondary burn progression in an experimental model and to elucidate the underlying mechanisms.. Prospective study.. University-based laboratory research.. Twenty-one male Wistar rats.. The burn comb model creates four rectangular burned surfaces that are intercalated by three unburned zones (interspaces) prone to secondary necrosis. Twenty-one animals were randomized to three experimental groups: 1) Local cooling with water for 20 minutes (control, 17°C); 2) and 3) local cooling with water and intraperitoneal erythropoietin once a day for five days starting 45 minutes after burn injury (500 IU/kg body weight: EPO 500 or 2500 IU/kg body weight: EPO 2500).. Secondary burn progression-both in depth (histology) and in surface (planimetry)-as well as interspace perfusion (laser Doppler flowmetry) and hematocrit were analyzed. Further, dilatory response (inducible nitric oxide synthase expression), inflammation (leukocyte count), and angiogenesis (CD31 expression) were assessed. Finally, wound healing time and contracture rate were reported. Burn progression resulted in complete dermal destruction as well as in important interspace necrosis in control animals, whereas burn progression was significantly reduced in a dose-dependent manner in animals treated with erythropoietin. Tissue protection was associated with an increased interspace perfusion with EPO 500, but not with EPO 2500, and was paralleled by a significant increase in inducible nitric oxide synthase expression and decreased inflammation, independent of the erythropoietin dosage. EPO 2500 led to a significant increase of hematocrit at day 4. Finally, faster wound healing and less contracture were observed in animals treated with EPO 500 only.. Erythropoietin represents an easy-to-use therapeutic approach to prevent secondary burn progression, i.e., to control damage after burn injury. It preserves microcirculatory perfusion within the endangered areas in a dose-dependent manner.

Topics: Animals; Burns; Disease Models, Animal; Disease Progression; Erythropoietin; Male; Necrosis; Neovascularization, Physiologic; Oxidative Stress; Prospective Studies; Random Allocation; Rats; Rats, Wistar; Wound Healing

Alternate erythropoietin (EPO)-mediated signaling via the heteromeric receptor composed of the EPO receptor and the β-common receptor (CD131) exerts the tissue-protective actions of EPO in various types of injuries. Herein we investigated the effects of the EPO derivative helix beta surface peptide (synonym: ARA290), which specifically triggers alternate EPO-mediated signaling, but does not bind the erythropoietic EPO receptor homodimer, on the progression of secondary tissue damage following cutaneous burns. For this purpose, a deep partial thickness cutaneous burn injury was applied on the back of mice, followed by systemic administration of vehicle or ARA290 at 1, 12, and 24 h postburn. With vehicle-only treatment, wounds exhibited secondary microvascular thrombosis within 24 h postburn, and subsequent necrosis of the surrounding tissue, thus converting to a full-thickness injury within 48 h. On the other hand, when ARA290 was systemically administered, patency of the microvasculature was maintained. Furthermore, ARA290 mitigated the innate inflammatory response, most notably tumor necrosis factor-alpha-mediated signaling. These findings correlated with long-term recovery of initially injured yet viable tissue components. In conclusion, ARA290 may be a promising therapeutic approach to prevent the conversion of partial- to full-thickness burn injuries. In a clinical setting, the decrease in burn depth and area would likely reduce the necessity for extensive surgical debridement as well as secondary wound closure by means of skin grafting. This use of ARA290 is consistent with its tissue-protective properties previously reported in other models of injury, such as myocardial infarction and hemorrhagic shock.

Topics: Animals; Burns; Cell Line; Erythropoietin; Inflammation; Male; Mice; Mice, Inbred C57BL; Microvessels; Signal Transduction; Skin; Thrombosis; Tumor Necrosis Factor-alpha; Wound Healing

Acceleration of skin regeneration is still an unsolved problem in the clinical treatment of patients suffering from deep burns and scalds. Although erythropoietin (EPO) has a protective role in a wide range of organs and cells during ischemia and after trauma, it has been recently discovered that EPO is not tissue-protective in the common β subunit receptor (βCR) knockout mouse. The protective capacity of EPO in tissue is mediated via a heteroreceptor complex comprising both the erythropoietin receptor (EPOR) and βCR. However, proof of coexpression of these heterogenic receptors in regenerating skin after burns is still lacking.. To understand the role of nanosized recombinant human erythropoietin (rhEPO) in wound healing, we investigated the effects of subcutaneous injections of EPO on skin regeneration after deep second-degree scalding injuries. Our aim was to determine if joint expression of EPOR and βCR is a prerequisite for the tissue-protective effect of rhEPO. The efficiency in wound regeneration in a skin scalding injury mouse model was examined. A deep second-degree dermal scald injury was produced on the backs of 20 female Balb/c mice which were subsequently randomized to four experimental groups, two of which received daily subcutaneous injections of rhEPO. At days 7 and 14, the mice were sacrificed and the effects of rhEPO were analyzed with respect to grade of re-epithelialization (wound closure) and stage of epidermal maturation. This was investigated using different histological parameters of epithelial covering, such as depth of the epidermal layer, epidermal stratification, and presence of conical and hair follicle structures.. Expression of EPOR, βCR, and growth hormone receptor at the mRNA and protein levels was demonstrated with reverse transcriptase polymerase chain reaction and Western blot analysis. After rhEPO treatment, the rate of re-epithelialization of the scalding injury was increased and the time to final wound closure was reduced. In addition, the quality of regenerated skin was improved. In this investigation, for the first time, we demonstrated coexpression of EPOR and βCR at the RNA and protein levels in vivo using a deep second-degree scalding injury mouse model. These results highlight the potential role of rhEPO in the improved treatment of burns patients, which might be crucial for the development of innovative new therapy regimes.. Local injection of nanosized rhEPO directly to the injury site rather than systemic administration for deep second-degree scalding injuries achieved complete skin regeneration with conical and hair follicle structure via combined expression of EPOR and βCR.

Topics: Animals; Burns; Epidermis; Erythropoietin; Female; Gene Expression Profiling; Hair Follicle; Histocytochemistry; Humans; Injections, Subcutaneous; Mice; Mice, Inbred BALB C; Nanoparticles; Protein Subunits; Receptors, Erythropoietin; Receptors, Somatotropin; Recombinant Proteins; Reverse Transcriptase Polymerase Chain Reaction; Skin; Wound Healing

Damage control is essential in first aid of burn lesions. The aim of the present study was to investigate whether systemic erythropoietin (EPO) administration could prevent secondary burn progression in an experimental model.. The burn comb model creates four rectangular burn surfaces intercalated by three unburned zones prone to progression. Twenty-one Wistar rats were randomized to a control group or to receive intraperitoneal EPO (500 units per kg) once a day for 5 days starting 45 min (EPO45min) or 6 h (EPO6h) after burn injury. Histological analyses assessing burn depth, inflammation and neoangiogenesis, planimetric evaluation of burn progression, and laser Doppler flowmetry to assess perfusion were performed after 1, 4 and 7 days. Final scarring time and contracture rate were assessed once a week.. Burn progression was decreased significantly with EPO45min but not EPO6h; progression of burn depth stopped in the intermediate dermis (mean(s.e.m.) burn depth score 3·3(0·6) for EPO45min versus 4·7(0·3) and 5·0(0·0) for EPO6h and control respectively on day 7; P = 0·026) and the surface extension was significantly reduced (45(8), 65(4) and 78(4) respectively on day 7; P = 0·017). This was paralleled by faster re-establishment of perfusion with EPO45min (114(5) per cent on day 4 versus 85(6) and 91(3) per cent for EPO6h and control respectively; P = 0·096). The reduction in progression resulted in a decreased healing time (7·3(0·7) weeks for EPO45min versus 11·5(1·0) and 10·8(0·5) weeks for EPO6h and control; P = 0·020) and contracture rate (P = 0·024).. Early EPO prevented burn progression, mainly by improved vascular perfusion.

Topics: Animals; Blood Circulation; Burns; Contracture; Disease Progression; Erythropoietin; Hematinics; Hematocrit; Leukocyte Count; Male; Necrosis; Neovascularization, Physiologic; Nitric Oxide Synthase Type II; Random Allocation; Rats; Rats, Wistar; Recombinant Proteins; Wound Healing

Recent data demonstrate a possible mortality benefit in traumatically injured patients when given subcutaneous recombinant human erythropoietin (rhEPO). The purpose of this report is to examine the effect of rhEPO on mortality and transfusion in burn patients. We conducted a review of burn patients (greater than 30% total body surface area, intensive care unit [ICU] days greater than 15) treated with 40,000 u rhEPO over an 18-month period (January 2007 to July 2008). Matched historical controls were identified and a contemporaneous cohort of subjects not administered rhEPO was used for comparison (NrhEPO). Mortality, transfusions, ICU and hospital length of stay were assessed. A total of 105 patients were treated (25 rhEPO, 53 historical control group, 27 NrhEPO). Hospital transfusions (mean 13,704 +/- mL vs. 13,308 +/- mL; P = 0.42) and mortality (29.6 vs. 32.0%; P = 0.64) were similar. NrhEPO required more blood transfusions (13,308 +/- mL vs. 6827 +/- mL; P = 0.004). No difference in mortality for the rhEPO and NrhEPO (32.0 vs. 22.2%; P = 0.43) was found. Thromboembolic complications were similar in all three groups. No effect was seen for rhEPO treatment on mortality or blood transfusion requirements in the severely burned.

Topics: Adolescent; Adult; Anemia; Blood Transfusion; Burns; Critical Illness; Erythropoietin; Female; Humans; Injury Severity Score; Length of Stay; Male; Middle Aged; Recombinant Proteins; Young Adult

Erythropoietin interacts with vascular endothelial growth factor (VEGF) and stimulates endothelial cell mitosis and motility; thus it may be of importance in the complex phenomenon of wound healing. The purpose of this study was to investigate the effect of recombinant human erythropoietin (rHuEPO) on experimental burn wounds.. Randomized experiment.. Research laboratory.. C57BL/6 male mice weighing 25-30 g.. Mice were immersed in 80 degrees C water for 10 secs to achieve a deep-dermal second degree burn. Animals were randomized to receive either rHuEPO (400 units/kg/day for 14 days in 100 microL subcutaneously) or its vehicle alone (100 microl/day distilled water for 14 days subcutaneously). On day 14 the animals were killed. Burn areas were used for histologic examination, evaluation of neoangiogenesis by immunohistochemistry, and expression (Western blot) of the specific endothelial marker CD31 as well as quantification of microvessel density, measurement of VEGF wound content (enzyme-linked immunosorbent assay), expression (Western blot) of endothelial and inducible nitric oxide synthases, and determination of wound nitric oxide (NO) products.. rHuEPO increased burn wound reepithelialization and reduced the time to final wound closure. These effects were completely abated by a passive immunization with specific antibodies against erythropoietin. rHuEPO improved healing of burn wound through increased epithelial proliferation, maturation of the extracellular matrix, and angiogenesis. The hematopoietic factor augmented neoangiogenesis as suggested by the marked increase in microvessel density and by the robust expression of the specific endothelial marker CD31. Furthermore, rHuEPO enhanced the wound content of VEGF caused a marked expression of endothelial and inducible nitric oxide synthases and increased wound content of nitric oxide products.. Our study suggests that rHuEPO may be an effective therapeutic approach to improve clinical outcomes after thermal injury.

Topics: Animals; Burns; Erythropoietin; Male; Mice; Mice, Inbred C57BL; Neovascularization, Physiologic; Recombinant Proteins; Skin; Wound Healing

Topics: Animals; Burns; Erythropoietin; Recombinant Proteins; Wound Healing

To observe the effects of burn serum on the erythropoiesis and granulopoiesis in bone marrow in mice, and to explore the possible underlying mechanism.. Murine bone marrow cell (BMC) strain was prepared routinely and was employed in the establishment of the culture system of colony forming units of erythrocytes, or granulocytes and monocytes. To both sets of culture system normal murine serum (N group) and burn serum, which was collected from the mice with 15% full thickness burn at 12 postburn hours (PBH) and 1, 3, 5, 7 and 10 postburn days (PBD), (burn serum group) was added. In addition, positive control and blank control groups were set accordingly. The stimulating activity of all kinds of sera on the BMCs in the two sets of culture system was determined. The changes in the burn serum concentrations of EPO and GM-CSF were detected by radioimmunoassay, and the data were analyzed by logarithmic linear fitting correlation with the former influence of burn sera on the erythrocytes and granulocytes.. (1) Burn sera exhibited obvious stimulation promoting activity on the erythropoiesis and granulopoiesis in BMC, and the activity peaked (384 +/- 60 and 127 +/- 16 CFU) on 1 PBD and decreased thereafter to approach the values found in normal sera group (125 +/- 14 and 34 +/- 20 CFU) on 7 PBD. (2) The EPO content in burn serum was evidently higher than the normal value (P < 0.01) during 12 PBH to 7PBD period. The GM-CSF concentration was obviously higher than the normal value (P < 0.05) at 12 PBH and on 1 PBD. (3) The EPO concentration in burn serum was significantly and logarithmically correlated with the stimulation promoting activity of burn serum on erythropoiesis (r = 0.8570, P = 0.0137). But the GM-CSF concentration in culture with burn serum was not correlated with the stimulation promoting activity of burn serum on granulopoiesis (r = 0.7049, P > 0.05).. The sera harvested from burned mice during early postburn stage exhibited strong stimulation promoting activity on the erythropoiesis and granulopoiesis in bone marrow. The increased EPO level in burn serum might be the important factor contributing strong stimulation action on erythropoiesis, while increased GM-CSF level was not.

Topics: Animals; Bone Marrow; Burns; Erythropoiesis; Erythropoietin; Granulocyte-Macrophage Colony-Stimulating Factor; Granulocytes; Hematopoietic Stem Cell Mobilization; Mice; Mice, Inbred Strains; Serum

A 3.5-year-old girl suffered from a thermal injury affecting 37% of the body surface area. The parents, being Jehovah's witnesses, refused permission for their child to receive blood transfusions. As the haemoglobin level was only 7.5% and a necrectomy was planned, the patient was likely to need blood transfusions. Indications for transfusion were defined as clinical signs of hypoxia and/or cardiovascular instability. A judicial declaration was proposed. Hb decreased during the therapy. To stimulate the erythropoiesis erythropoietin and iron were administered. During the clinical course the anaemia worsened. First, a conservative treatment with polyvidoniodine ointment for tanning was started, to avoid an operation during the acute phase after the injury, as in this case it was thought a blood transfusion would definitely be necessary. On the 19th day after the injury a necrectomy of 10% of the body surface was necessary because of fever and leucocytosis not responding to antibiotics. The most likely cause of the symptoms was an infection of the burned area. Hb was 4.6 g/dl%. General anaesthesia was performed with midazolam, ketamine and vecuronium and mechanical ventilation. No blood transfusion was required during the operation. Vital signs were stable during the preoperative period, during anaesthesia and following the operation. There were no signs of tissue hypoxia. Peripheral oxygen saturation ranged between 98% and 100%, lactate and arterial blood gas samples were normal, and the child was awake and cooperative before and after anaesthesia. The lowest Hb was 3.3 g/dl on the 22th day after injury (3rd postoperative day). In this phase the patient was still playing and riding a tricycle. On the 45th day after injury the child was discharged home with Hb of 10.9 g/dl and reticulocytosis of 33%.

Topics: Anemia; Anesthesia; Blood Cell Count; Blood Transfusion; Burns; Child, Preschool; Christianity; Critical Care; Erythropoietin; Female; Hemoglobins; Humans; Iron

The treatment rationale of a burn victim (35% TBSA) who was child of Jehova's witnesses is described. Following a combined approach including erythropoetin and blood saving surgical techniques we were able to excise and graft the burn areas without blood transfusion. An extremely low hemoglobin of 3.4 g/dl was tolerated postoperatively and showed an increase to 10.9 g/dl 25 days later when the child was dismissed from the burn unit in stable condition. Possibilities to minimize blood loss and to avoid blood transfusions are discussed.

Topics: Blood Loss, Surgical; Blood Transfusion; Burns; Child, Preschool; Christianity; Debridement; Erythropoietin; Female; Hemoglobinometry; Humans; Infusions, Intravenous; Iron-Dextran Complex; Skin Transplantation; Treatment Refusal

Topics: Blood Loss, Surgical; Blood Transfusion; Burns; Child, Preschool; Christianity; Erythropoietin; Female; Humans; Recombinant Proteins

Haemostatic debridement, recombinant-human erythropoietin and cultured epithelial autografts have been used successfully in a Jehovah's Witness with a major burn injury. Tourniquet ischaemia complemented by a topical haemostatic agent minimized excisional blood loss, while recombinant-human erythropoietin accelerated erythropoiesis, thereby correcting postburn anaemia. Cultured epithelial autografts provided coverage of the granulating wounds without creating donor sites.

Topics: Adult; Anemia; Burns; Cells, Cultured; Christianity; Epithelium; Erythropoietin; Humans; Male; Recombinant Proteins; Religion and Medicine; Skin; Skin Transplantation; Transplantation, Autologous

Since controversy exists over whether erythropoietin levels are increased or decreased after thermal injury, a prospective study was performed to answer this question as well as to characterize the erythropoietic response to thermal injury.. The concept of using erythropoietin to reduce the need for blood transfusions after thermal injury is attractive. However, since the etiology of burn anemia is both unclear and multifocal, prior to initiating a trial of erythropoietin therapy, it will be necessary to better define the erythropoietic response to thermal injury.. Twenty-four burn patients with a mean burn size of 31 +/- 18% had serial measurements of serum iron, total iron binding capacity (TIBC), ferritin, erythropoietin, transferrin saturation, hemoglobin, and reticulocyte counts performed on burn days 1, 3, 5, 7, 10, 14, and then weekly.. The erythropoietic response was characterized by a decrease in hemoglobin levels as well as serum iron, TIBC, and transferrin saturation (p < 0.05). Ferritin and erythropoietin levels increased as did the reticulocyte count. The erythropoietin response to anemia appeared to be at least grossly intact, since there was an appropriate inverse relationship between the degree of anemia and the magnitude of the erythropoietin response (r2 = .61, p < 0.00001).. Since the erythropoietin levels of these anemic burn victims reached supranormal levels and they manifested a moderate reticulocytosis, the role of replacement erythropoietin therapy after thermal injury requires further study.

Topics: Burns; Erythropoiesis; Erythropoietin; Hemoglobins; Humans; Iron; Prospective Studies; Regression Analysis; Transferrin

Surgical procedures result in blood loss that can require replacement transfusions. Such therapy may result in multiple adverse sequelae, including transmission of infectious diseases and immune impairment. Alternative therapies are therefore desirable.. We evaluated the ability of recombinant human erythropoietin (rEPO) to increase red blood cell production in both normal healthy volunteers and patients with burn injuries. The effect of rEPO on immune function in the volunteers was also evaluated. The volunteers received 150 units/kg rEPO daily for 7 days, with immune function and hematopoiesis assayed on days 0, 7, and 14. The patients with burn injuries received either 500 units/kg/day rEPO with iron supplementation or merely the iron.. rEPO increased erythropoiesis in both the volunteers and the patients with burn injuries. Failure to provide iron supplementation to the volunteers resulted in significant depletion of iron stores with a concomitant impairment in immune function that paralleled the iron depletion.. rEPO therapy offers the potential to increase red blood cell production in surgical patients. Failure to provide iron supplementation in patients receiving rEPO can lead to a rapid depletion of iron stores and may contribute to an immune dysfunction.

Topics: Adult; Anemia; Burns; Child; Erythrocytes; Erythropoietin; Female; Hematopoiesis; Humans; Immune System; Iron; Iron Deficiencies; Male; Middle Aged; Recombinant Proteins; Reference Values; Retrospective Studies

We report the use of recombinant human erythropoietin (rh-Ep) as an alternative to transfusion therapy in a burn victim who was a Jehovah's Witness and refused blood transfusion. Despite endogenous elevated erythropoietin levels, the patient was reticulocytopenic and administration of rh-Ep was accompanied by a 10-fold increase in reticulocyte response and a rise in hemoglobin from 7.4 to 10.4 g/dl over a 12-day period. We suggest that the exogenously administered erythropoietin overcame a clinically inadequate endogenous erythropoietin response.

Topics: Anemia; Blood Transfusion; Burns; Christianity; Erythropoietin; Hemoglobins; Humans; Male; Middle Aged; Recombinant Proteins; Reticulocytes; Treatment Refusal

Recombinant-human erythropoietin was given to two burn patients who are Jehovah's Witnesses and hence refused transfusion. Anaemia developing postburn was corrected in both patients. Serum erythropoietin levels were found to be elevated prior to initiation of therapy in both patients.

Topics: Adolescent; Adult; Anemia; Burns; Christianity; Erythropoietin; Female; Humans; Skin Transplantation

Topics: Anemia; Animals; Burns; Erythropoietin; Female; Hematopoietic Stem Cells; Humans; Male; Mice

The anemia of thermal injury is a multifactorial process and includes hemorrhage and hemolysis. Much evidence suggests that a reduced rate of erythropoiesis contributes to this anemia. Prior studies show that this anemia is temporally related to the appearance in burn patients sera of a substance(s) capable of inhibiting erythropoiesis in vitro. Four experiments were done to elucidate the mechanism of action of this inhibitor. In all experiments sera from burn patients previously shown to be inhibitory to erythropoiesis in vitro were studied. In the first, inhibitory sera were exposed to erythropoietin solutions without loss of erythropoietic activity. Second, mouse marrow cells were preincubated with serum without loss of their ability to form erythroid colonies. Third, the inhibitory effect could not be overcome with increasing amounts of erythropoietin. Finally, erythroid colony formation was effected only if the inhibitory serum was present during the first 8 to 12 hr of culture. The data suggest that the erythropoietic inhibitor in these sera acts directly on erythroid stem cells in vitro and not by inactivating or interference with erythropoietin.

Topics: Anemia; Animals; Burns; Dose-Response Relationship, Drug; Erythropoiesis; Erythropoietin; Humans; Mice; Receptors, Cell Surface; Receptors, Erythropoietin

Anemia is invariably seen in patients who have been severely burned, and a number of factors have been implicated in its etiology. Prior studies have suggested that a depressed rate of erythropoiesis is involved. In order to study this, we evaluated the effect of serum from burned patients on red cell and white cell colony growth in vitro. We found that these sera were capable of inhibiting red cell, but not white cell, colony growth. Additional experiments indicated that this was related to the presence of some substance in the burned serum rather than the absence of a factor required for colony formation. Further studies, including review of clinical data, suggested that this effect was not due to topical medications nor to episodes of bacterial sepsis. Serial studies showed that inhibition was often not present in the immediate postburn period but developed gradually, reaching maximum intensity approximately 20 to 30 days following the burn and then returning toward normal as patients healed their injury. Our studies permit the hypothesis that inhibition of erythropoiesis plays a role in the pathogenesis of the anemia of thermal injury.

Topics: Adult; Aged; Anemia; Animals; Bacterial Infections; Burns; Cells, Cultured; Erythropoiesis; Erythropoietin; Female; Humans; Mice

Topics: Anemia; Biological Transport; Blood Preservation; Blood Transfusion; Burns; Diphosphoglyceric Acids; Erythropoiesis; Erythropoietin; Hemoglobins; Humans; Hydrogen-Ion Concentration; Oxygen; Oxyhemoglobins

The relationship between metabolism, oxygen transport, and anemia was assessed in burn patients. A significant negative correlation was found between erythrocyte 2,3 DPG, the major modulator of oxygen transport, and erythropoietin synthesis. Simultaneous bioassay and radioimmunoassay for erythropoietin revealed elevated values in the anemic burn patients. Elevated 2,3 DPG values during convalescence from thermal injury may remove the "anemic hypoxia" stimulus to erythropoieitn production, resulting in persistence of the anemia.

Topics: Adult; Aged; Anemia; Burns; Diphosphoglyceric Acids; Energy Intake; Erythropoietin; Female; Humans; Male; Middle Aged; Nitrogen; Oxygen Consumption; Phosphates; Radioimmunoassay

Erythropoietin excretion was persistently increased following major thermal injury in 4 of 5 patients. A good correlation was found between erythropoietin excretion and red cell mass but not between erythropoietin excretion and hematocrit. In spite of the increased erythropoietin, erythropoiesis in these thermally injured patients was inadequate to compensate for erythrocyte deficits as judged by bone marrow morphology, reticulocyte counts, and transfusion requirements.

Topics: Adolescent; Adult; Anemia; Biological Assay; Bone Marrow Examination; Burns; Erythropoietin; Hematocrit; Humans; Male

Topics: Anemia; Animals; Burns; Erythropoietin; Mice

Topics: Adolescent; Adult; Aged; Anemia; Animals; Burns; Erythropoietin; Female; Humans; Iron Radioisotopes; Male; Mice; Middle Aged; Reticulocytes