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BACKGROUND: Debridement is key to removing devitalised tissue, debris and biofilm as part of wound-bed preparation. Unlike many other methods of debridement, mechanical debridement with a pad is effective enough to be used independently without an adjunctive method of debridement, while being more accessible than other standalone options. OBJECTIVE: To explore the clinical performance and safety of a debridement pad with both abrasive and non-abrasive surfaces in daily clinical practice. METHODS: This was a prospective, non-controlled, non-randomised, single-arm, open-label, multicentred observational evaluation. Inclusion criteria were wounds >4 cm2 covered with at least 30% debris, necrotic tissue or slough in patients aged ≥18 years. The treatment protocol comprised a single application of the debridement pad. The primary outcome measure was the amount of necrotic tissue, slough or debris in the wound bed. Secondary outcomes included the appearance of the wound bed, edges and periwound skin; self-reported pain scores; foreseeable negative impacts; and clinician satisfaction. RESULTS: A total of 62 participants with a variety of wound types were included in the analysis. Most wounds (87%) had been present for over 3 months and had high or moderate exudate levels (90%). A significant reduction was observed in all three parameters: necrotic tissue (p=0.043), slough (p<0.001) and debris (p<0.001). Necrotic tissue, slough and debris showed mean relative reductions of 40%, 72% and 40%, respectively. Of participants, 84% did not experience an increase in pain during the debridement procedure. CONCLUSION: This clinical real-world data shows the debridement pad to be an effective and well-tolerated device for debridement and wound bed preparation.
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Desbridamento , Humanos , Desbridamento/métodos , Masculino , Estudos Prospectivos , Feminino , Pessoa de Meia-Idade , Idoso , Adulto , Cicatrização , Ferimentos e Lesões/terapia , Idoso de 80 Anos ou mais , NecroseRESUMO
Hypertrophic scarring in burn wounds is caused by overactive fibroblasts and myofibroblasts. Blue light reveals wavelength- and dose-dependent antibacterial and antiproliferative effects and may serve as a therapeutic option against wound infection and fibrotic conditions. Therefore, we evaluated in this study the effects of single and multiple irradiations with blue light at 420 nm (BL420) on the intracellular ATP concentration, and on the viability and proliferation of the human skin fibroblast (HDFs). In addition, possible BL420-induced effects on the catalase expression and differentiation were assessed by immunocytochemical staining and western blot analyses. Furthermore, we used RNA-seq analyses to identify BL420-affected genes. We found that BL420 induced toxicity in HDFs (up to 83%; 180 J/cm2). A low dose of 20 J/cm2 reduced the ATP concentration by ~50%. Multiple irradiations (4 × 20 J/cm2) inhibited proliferation without visible toxicity and reduced catalase protein expression by ~37% without affecting differentiation. The expression of about 300 genes was significantly altered. Many downregulated genes have functions in cell division/mitosis. BL420 can strongly influence the fibroblast physiology and has potential in wound therapy. However, it is important to consider the possible toxic and antiproliferative effects, which could potentially lead to impaired wound healing and reduced scar breaking strength.
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The field of reconstructive surgery encompasses a wide range of surgical procedures and regenerative approaches to treat various tissue types. Every surgical procedure is associated with the risk of surgical site infections, which are not only a financial burden but also increase patient morbidity. The surgical armamentarium in this area are biomaterials, particularly natural, biodegradable, biocompatible polymers, including the silk proteins fibroin (SF) and sericin (SS). Silk is known to be derived from silkworms and is mainly composed of 60-80% fibroin, which provides the structural form, and 15-35% sericin, which acts as a glue-like substance for the SF threads. Silk proteins possess most of the desired properties for biomedical applications, including biocompatibility, biodegradability, minimal immunogenicity, and tunable biomechanical behaviour. In an effort to alleviate or even prevent infections associated with the use of biomaterials in surgery, antibacterial/antimicrobial properties have been investigated in numerous studies. In this systematic review, the following question was addressed: Do silk proteins, SF and SS, possess an intrinsic antibacterial property and how could these materials be tailored to achieve such a property?
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Antibacterianos , Fibroínas , Sericinas , Cirurgia Plástica , Antibacterianos/farmacologia , Materiais Biocompatíveis/farmacologia , Fibroínas/química , Fibroínas/farmacologia , Sericinas/farmacologia , Sericinas/química , CicatrizaçãoRESUMO
To facilitate the recovery process of chronic and hard-to-heal wounds novel pro-resolving treatment options are urgently needed. We investigated the pro-regenerative properties of soluble CD83 (sCD83) on cutaneous wound healing, where sCD83 accelerated wound healing not only after systemic but also after topical application, which is of high therapeutic interest. Cytokine profile analyses revealed an initial upregulation of inflammatory mediators such as TNFα and IL-1ß, followed by a switch towards pro-resolving factors, including YM-1 and IL-10, both expressed by tissue repair macrophages. These cells are known to mediate resolution of inflammation and stimulate wound healing processes by secretion of growth factors such as epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF), which promote vascularization as well as fibroblast and keratinocyte differentiation. In conclusion, we have found strong wound healing capacities of sCD83 beyond the previously described role in transplantation and autoimmunity. This makes sCD83 a promising candidate for the treatment of chronic- and hard-to-heal wounds.
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Interleucina-10 , Fator de Necrose Tumoral alfa , Fator de Crescimento Epidérmico , Mediadores da Inflamação/metabolismo , Interleucina-10/metabolismo , Macrófagos , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Cicatrização/fisiologiaRESUMO
Oral wounds are among the most troublesome injuries which easily affect the patients' quality of life. To date, the development of functional antibacterial dressings for oral wound healing remains a challenge. In this regard, we investigated antibacterial silk protein-based membranes for the application as wound dressings in oral and maxillofacial surgery. The present study includes five variants of casted membranes, i.e., i) membranes-silver nanoparticles (CM-Ag), ii) membranes-gentamicin (CM-G), iii) membranes-control (without functionalization) (CM-C), iv) membranes-silk sericin control (CM-SSC), and v) membranes-silk fibroin/silk sericin (CM-SF/SS), and three variants of nonwovens, i.e., i) silver nanoparticles (NW-Ag), ii) gentamicin (NW-G), iii) control (without functionalization) (NW-C). The surface structure of the samples was visualized with scanning electron microscopy. In addition, antibacterial testing was accomplished using agar diffusion assay, colony forming unit (CFU) analysis, and qrt-PCR. Following antibacterial assays, biocompatibility was evaluated by cell proliferation assay (XTT), cytotoxicity assay (LDH), and live-dead assay on L929 mouse fibroblasts. Findings indicated significantly lower bacterial colony growth and DNA counts for CM-Ag with a reduction of bacterial counts by 3log levels (99.9% reduction) in CFU and qrt-PCR assay compared to untreated control membranes (CM-C and CM-SSC) and membranes functionalized with gentamicin (CM-G and NW-G) (p < 0.001). Similarly, NW-G yielded significantly lower DNA and colony growth counts compared to NW-Ag and NW-C (p < 0.001). In conclusion, CM-Ag represented 1log level better antibacterial activity compared to NW-G, whereas NW-G showed better cytocompatibility for L929 cells. As data suggest, these two membranes have the potential of application in the field of bacteria-free oral wound healing. However, provided that loading strategy and cytocompatibility are adjusted according to the antibacterial agents' characteristic and fabrication technique of the membranes.
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Fibroínas , Nanopartículas Metálicas , Sericinas , Cirurgia Bucal , Animais , Antibacterianos/farmacologia , Fibroínas/farmacologia , Gentamicinas/farmacologia , Nanopartículas Metálicas/uso terapêutico , Camundongos , Qualidade de Vida , Sericinas/farmacologia , Seda/química , Prata/farmacologia , CicatrizaçãoRESUMO
Biomaterials of natural origin have recently gained increasing attention in the field of dental implantology. The requirements for such materials, however, are very high. In addition to high clinical efficiency in tissue regeneration, wound healing should be demonstrably positively influenced. The translational division for regenerative orofacial medicine of the Clinic and Polyclinic for Oral and Maxillofacial Surgery of the University Medical Center Hamburg-Eppendorf (UKE) is examining this research topic by investigating which innovative treatment methods for the reconstruction of bone defects or for augmentative procedures can be applied in the future or are already being applied in the field of oral and maxillofacial surgery.
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Tissue adhesives have been successfully used in various kind of surgeries such as oral and maxillofacial surgery for some time. They serve as a substitute for suturing of tissues and shorten treatment time. Besides synthetic-based adhesives, a number of biological-based formulations are finding their way into research and clinical application. In natural adhesives, proteins play a crucial role, mediating adhesion and cohesion at the same time. Silk fibroin, as a natural biomaterial, represents an interesting alternative to conventional medical adhesives. Here, the most commonly used bioadhesives as well as the potential of silk fibroin as natural adhesives will be discussed.
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Fibroínas , Cirurgia Plástica , Adesivos Teciduais , Materiais Biocompatíveis/uso terapêutico , Fibroínas/uso terapêutico , Seda , Adesivos Teciduais/uso terapêutico , Engenharia Tecidual , Alicerces TeciduaisRESUMO
Marine long-chain omega-3 polyunsaturated fatty acids (ω3 FA) are involved in numerous cell responses and therefore vital for the mammal organism. Because of the attribution of immunomodulatory effects, a favourable impact on the inflammatory response in chronic wounds and cells involved in wound healing can be suspected. In the experimental setup, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were investigated regarding their impact on metabolic activity, cell proliferation and migration of human keratinocytes (HaCaT) and newborn foreskin fibroblasts (CRL-2522). For simulation of the microenvironment of a chronic wound, human chronic wound fluid (CWF) was used in the experimental setup addressing the in vitro influence of DHA, EPA and CWF on regenerative processes. The results showed a significant increase in the metabolic activity of keratinocytes and fibroblasts after 72 h treatment with DHA and EPA. In contrast, treatment with ω3 FA had no significant positive effect on skin cell proliferation. Both ω3 FA had no influence on in vitro wound closure. CWF demonstrated significantly adverse effects, which ω3 FA were unable to mitigate. It can be concluded that CWF exhibited the expected adverse effect on both skin cell types, especially inhibiting in vitro wound closure. ω3 FAs showed a slightly positive, yet rarely significant effect on human skin cells. Overall, the addition of DHA or EPA showed no relevant benefit for skin cells challenged with human CWF, merely in combination with DHA an initial significant increase in cell metabolism (fibroblasts) and cell proliferation (keratinocytes) could be observed.
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Ácidos Graxos Ômega-3 , Animais , Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/farmacologia , Ácidos Graxos/metabolismo , Ácidos Graxos Ômega-3/farmacologia , Humanos , Recém-Nascido , Mamíferos , Pele/metabolismo , CicatrizaçãoRESUMO
Objective: To determine effective treatment strategies against bacterial infections of chronic wounds, we tested different blue light (BL)-emitting light-emitting diode arrays (420, 455, and 480 nm) against wound pathogens and investigated in parallel BL-induced toxic effects on human dermal fibroblasts. Background: Wound infection is a major factor for delayed healing. Infections with Pseudomonas aeruginosa and Staphylococcus aureus are clinically relevant caused by their ability of biofilm formation and their quickly growing antibiotics resistance. BL has demonstrated antimicrobial properties against various microbes. Methods: Determination of antibacterial and cell toxic effects by colony-forming units (CFUs)/biofilm/cell viability assays, and live cell imaging. Results: A single BL irradiation (180 J/cm2), of P. aeruginosa at both 420 and 455 nm resulted in a bacterial reduction (>5 log10 CFU), whereas 480 nm revealed subantimicrobial effects (2 log10). All tested wavelengths of BL also revealed bacteria reducing effects on Staphylococcus epidermidis and Escherichia coli (maximum 1-2 log10 CFU) but not on S. aureus. Dealing with biofilms, all wavelengths using 180 J/cm2 were able to reduce significantly the number of P. aeruginosa, E. coli, and S. epidermidis. Here, BL420nm achieved reductions up to 99%, whereas BL455nm and BL480nm were less effective (60-83%). Biofilm-growing S. aureus was more BL sensitive than in the planktonic phase showing a reduction by 63-75%. A significant number of cell toxic events (>40%) could be found after applying doses (>30 J/cm2) of BL420nm. BL455nm showed only slight cell toxicity (180 J/cm2), whereas BL480nm was nontoxic at any dose. Conclusions: BL treatment can be effective against bacterial infections of chronic wounds. Nevertheless, using longer wavelengths >455 nm should be preferred to avoid possible toxic effects on skin and skin cells. To establish BL therapy for infected chronic wounds, further studies concerning biofilm formation and tissue compatibility are necessary.
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Anti-Infecciosos , Infecção dos Ferimentos , Antibacterianos/farmacologia , Escherichia coli , Humanos , Staphylococcus aureus , Infecção dos Ferimentos/tratamento farmacológicoRESUMO
OBJECTIVES: A major problem for wound healing is contamination with bacteria, often resulting in biofilm formation and wound infection, which, in turn, needs immediate intervention such as surgical debridement and through irrigation. A topical treatment with cold atmospheric pressure plasma (CAP) for wound disinfection may present an alternative and less painful approach. METHODS: This study investigated the antibacterial effects of a cold atmospheric pressure argon plasma jet (kINPen® MED) as a CAP source, using the three-dimensional Staphylococcus aureus immunocompetent biofilm system hpBIOM in addition to a standard planktonic test. Furthermore, skin cell compatibility was evaluated using a keratinocyte (HaCat) model. RESULTS: CAP treatment (0-240 s) followed by incubation (15, 120 min) within the CAP-treated media showed slight bactericidal efficacy under planktonic conditions but no effect on biofilms. However, indirect CAP treatment of keratinocytes performed under the same conditions resulted in a significant decrease in metabolic activity. Short CAP treatment and exposure time (30 s; 15 min) induced a slight increase in the metabolic activity; however, longer treatments and/or exposure times led to pronounced reductions up to 100%. These effects could partially be reversed by addition of catalase, indicating a dominant role of CAP-generated hydrogen peroxide. CONCLUSIONS: These results indicate that plasma treatment does not lead to the desired disinfection or significant reduction in the bacterial burden of Staphylococcus aureus in a wet milieu or in biofilms. Thus, treatment with CAP could not be recommended as a single anti-bacterial therapy for wounds but could be used to support standard treatments.
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Antibacterianos/farmacologia , Argônio/farmacologia , Queratinócitos/efeitos dos fármacos , Gases em Plasma/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Pressão Atmosférica , Biofilmes/efeitos dos fármacos , Células HaCaT , Humanos , Viabilidade Microbiana , Pele/efeitos dos fármacos , Staphylococcus aureus/citologia , Infecção dos Ferimentos/microbiologia , Infecção dos Ferimentos/terapiaRESUMO
Studies have demonstrated that blue light induces biological effects, such as cell death, and inhibition of proliferation and differentiation. Since blue light at longer wavelength (>440 nm) exerts less injurious effects on cells than at shorter wavelengths, (400-440 nm), we have investigated the impact of non-toxic (LED) blue light at 453 nm wavelength on human skin fibroblasts (hsFBs). We found that besides its decreasing effects on the proliferation rate, repeated blue light irradiations (80 J/cm2) also significantly reduced TGF-ß1-induced myofibrogenesis as shown by diminished α-SMA and EDA-FN expression accompanied by reduced protein expression and phosphorylation of ERK 1/2, SMAD 2/3, and p38-key players of TGF-ß1-induced myofibrogenesis. In parallel, catalase protein expression, intracellular FAD concentrations as well as NADP+/NADPH ratio were reduced, whereas intracellular reactive oxygen species (ROS) were increased. We postulate that as a molecular mechanism downregulation of catalase and photoreduction of FAD induce intracellular oxidative stress which, in turn, affects the signaling factors of myofibrogenesis leading to a lower rate of α-SMA and EDA-FN expression and, therefore, myofibroblast formation. In conclusion, blue light even at longer wavelengths shows antifibrotic activity and may represent a suitable and safe approach in the treatment of fibrotic skin diseases including hypertrophic scarring and scleroderma.
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Antioxidantes/metabolismo , Luz , Transdução de Sinais/efeitos da radiação , Fator de Crescimento Transformador beta1/metabolismo , Proliferação de Células/efeitos da radiação , Humanos , Miofibroblastos/citologia , Miofibroblastos/efeitos da radiação , Estresse OxidativoRESUMO
BACKGROUND: After severe polytrauma the dynamic process of coagulation may deteriorate towards a trauma-induced coagulopathy (TIC) promoting a dramatic increase in morbidity and mortality. Recent evidence suggests that microparticles (MPs) play a pivotal role at the interface between cellular and plasmatic coagulation systems. However, the impact of MPs on functional coagulation has not been clarified yet in the setting of traumatic injuries. We assessed the temporal patterns of circulating MP concentrations including their cellular origin in the context of clinical presentation and global coagulation assays. METHODS: Blood samples from 22 consecutive polytrauma patients (ISS ≥16) from 2015 were collected at hospital admission, after 24 and 72 h and compared to those from healthy individuals and minor injured patients with isolated extremity fractures. Flow cytometry (BD Accuri C6; Heidelberg/Germany) was used to determine MP concentrations and cellular origin using cell-specific markers (platelet derived (PDMP): CD42b+, CD61+, CD62p+; endothelial cell derived (EDMP): CD144+, CD62e+, CD144+/62e+). Results were correlated with clinical data and results from viscoelastic testing (ROTEM). RESULTS: Twenty two polytrauma patients (17 males, agemedian 60 yrs) with a median ISS 26.5 (IQR 14.5) were assessed. PDMP and EDMP concentrations increased significantly in polytrauma patients as compared to healthy individuals and minor injured patients. MP concentrations correlated with injury severity (CD144+: ρsp = 0.79, p < 0.001; CD42b+: ρsp = 0.61, p < 0.001). EDMP displayed a negative correlation with aPTT (CD144/62e+, ρsp = - 0.55, p < 0.05), INR (CD144/62e+, ρsp = - 0.61, p < 0.05) and ROTEM-INTEM CT (CD144/62e+, ρsp = - 0.68, p < 0.05) reflecting increased dynamics of clot formation and an overall procoagulative effect. Additionally, EDMP showed a negative association with FIBTEM values (10 min amplitude, maximum clot firmness) indicating a fibrinolytic potential. DISCUSSION: In a small cohort, analysing most severly injured patients, the association of increased MP levels and altered coagulation parameters could be demonstrated. However, these findings are based on correlation analysis, which do not enable causel evidence. Therefore, further in-vitro studies are needed analysing the underlying pathomechanisms. CONCLUSION: In conclusion, this study could demonstrate that PDMP and EDMP levels increase significantly following polytrauma correlating with injury severity. Although severe coagulopathy was not observed, EDMP levels were associated with improved coagulation parameters suggesting their essential role for regulating blood coagulation after trauma.
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Transtornos da Coagulação Sanguínea/sangue , Coagulação Sanguínea/fisiologia , Micropartículas Derivadas de Células/metabolismo , Traumatismo Múltiplo/complicações , Biomarcadores/sangue , Transtornos da Coagulação Sanguínea/diagnóstico , Transtornos da Coagulação Sanguínea/etiologia , Testes de Coagulação Sanguínea , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Traumatismo Múltiplo/sangue , Tempo de Tromboplastina Parcial , Estudos Prospectivos , Tromboelastografia/métodosRESUMO
Wound healing is a complex and dynamic process with different distinct and overlapping phases from homeostasis, inflammation and proliferation to remodelling. Monitoring the healing response of injured tissue is of high importance for basic research and clinical practice. In traditional application, biological markers characterize normal and abnormal wound healing. Understanding functional relationships of these biological processes is essential for developing new treatment strategies. However, most of the present techniques (in vitro or in vivo) include invasive microscopic or analytical tissue sampling. In the present study, a non-invasive alternative for monitoring processes during wound healing is introduced. Within this context, hyperspectral imaging (HSI) is an emerging and innovative non-invasive imaging technique with different opportunities in medical applications. HSI acquires the spectral reflectance of an object, depending on its biochemical and structural characteristics. An in-vitro 3-dimensional (3-D) wound model was established and incubated without and with acute and chronic wound fluid (AWF, CWF), respectively. Hyperspectral images of each individual specimen of this 3-D wound model were assessed at day 0/5/10 in vitro, and reflectance spectra were evaluated. For analysing the complex hyperspectral data, an efficient unsupervised approach for clustering massive hyperspectral data was designed, based on efficient hierarchical decomposition of spectral information according to archetypal data points. It represents, to the best of our knowledge, the first application of an advanced Data Mining approach in context of non-invasive analysis of wounds using hyperspectral imagery. By this, temporal and spatial pattern of hyperspectral clusters were determined within the tissue discs and among the different treatments. Results from non-invasive imaging were compared to the number of cells in the various clusters, assessed by Hematoxylin/Eosin (H/E) staining. It was possible to correlate cell quantity and spectral reflectance during wound closure in a 3-D wound model in vitro.
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Modelos Biológicos , Análise Espectral/métodos , Cicatrização , Automação , Células Cultivadas , Análise por Conglomerados , HumanosRESUMO
BACKGROUND: Various studies have shown the deleterious effect of high volume resuscitation following severe trauma promoting coagulopathy by haemodilution, acidosis and hypothermia. As the optimal resuscitation strategy during prehospital trauma care is still discussed, we raised the question if the amount and kind of fluids administered changed over the recent years. Further, if less volume was administered, fewer patients should have arrived in coagulopathic depletion in the Emergency Department resulting in less blood product transfusions. METHODS: A data analysis of the 100 489 patients entered into the TraumaRegister DGU® (TR-DGU) between 2002 and 2012 was performed of which a total of 23512 patients (23.3%) matched the inclusion criteria. Volume and type of fluids administered as well as outcome parameter were analysed. RESULTS: Between 2002 and 2012, the amount of volume administered during prehospital trauma care decreased from 1790 ml in 2002 to 1039 ml in 2012. At the same time higher haemoglobin mean values, higher Quick's mean values and reduced mean aPTT can be observed. Simultaneously, more patients received catecholamines (2002: 9.2 to 2012: 13.0%). Interestingly, the amount of volume administered decreased steadily regardless of the presence of shock. Fewer patients were in the need of blood products and the number of massive transfusions (≥10 pRBC) more than halved. DISCUSSION: The changes in volume therapy might have reduced haemodilution potentially resulting in an increase of the Hb value. During the period observed transfusion strategies have become more restrictiveand ratio based; the percentage of patients receiving MT halved as blood products may imply negative secondary effects. Furthermore, preventing administration of high blood product ratios result in less impairment of coagulation factors and inhibitors and an therfore improved coagulation. CONCLUSION: The volume administered in severely injured patients decreased considerably during the last decade possibly supporting beneficial effects such as minimizing the risk of coagulopathy and avoiding potential harmful effects caused by blood product transfusions. Despite outstanding questions in trauma resuscitation, principle evidence merges quickly into clinical practice and algorithms.
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Soluções Isotônicas/administração & dosagem , Sistema de Registros , Ressuscitação/métodos , Adulto , Soluções Cristaloides , Serviços Médicos de Emergência , Feminino , Hidratação , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Índices de Gravidade do Trauma , Ferimentos e Lesões/fisiopatologiaRESUMO
BACKGROUND: Uncontrolled haemorrhage is still the leading cause of preventable death after trauma and the primary focus of any treatment strategy should be related to early detection and control of blood loss including haemostasis. METHODS: For assessing management practices across six European level I trauma centres with academic interest and research in the field of coagulopathy an online survey was conducted addressing local management practice for bleeding trauma patients including algorithms for detection, management and monitoring coagulation disorders and immediate interventions. Each centre provided their locally applied massive transfusion protocol. RESULTS: All participating trauma centres have developed and implemented a local algorithm and protocol for the bleeding trauma patient. These are uniformly activated by clinical triggers and deactivated once the bleeding has stopped according to clinical assessment in combination with laboratory signs of achieved haemostasis. The severity of coagulopathy and shock is mostly assessed via standard coagulation tests and partially used extended viscoelastic tests. All centres have implemented the immediate use of tranexamic acid. Initial resuscitation is started either pre-hospital or after hospital admission by using transfusion packages with pre-fixed universal blood product combinations and ratios following the concept of "damage control resuscitation" at which applied ratios substantially vary. Two centres initially start with transfusion packages but with viscoelastic tests running in parallel to quickly allow a shift towards a viscoelastic test-guided therapy. CONCLUSION: Diversity in the management of bleeding trauma patients such as pre-hospital blood administration and routinely performed viscoelastic tests exists even among level I trauma centres. The paucity of consensus among these centres highlights the need for further primary research followed by clinical trials to improve the evidence for sophisticated guidelines and strategies.
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Protocolos Clínicos , Hemorragia/diagnóstico , Hemorragia/terapia , Hemostasia , Padrões de Prática Médica/estatística & dados numéricos , Centros de Traumatologia/normas , Algoritmos , Transtornos da Coagulação Sanguínea/etiologia , Transtornos da Coagulação Sanguínea/terapia , Transfusão de Sangue/normas , Europa (Continente) , Humanos , Ressuscitação/normas , Inquéritos e QuestionáriosRESUMO
Wound healing is a complex biological process that requires a well-orchestrated interaction of mediators as well as resident and infiltrating cells. In this context, mesenchymal stem cells play a crucial role as they are attracted to the wound site and influence tissue regeneration by various mechanisms. In chronic wounds, these processes are disturbed. In a comparative approach, adipose-derived stem cells (ASC) were treated with acute and chronic wound fluids (AWF and CWF, respectively). Proliferation and migration were investigated using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test and transwell migration assay. Gene expression changes were analysed using quantitative real time-polymerase chain reaction. AWF had a significantly stronger chemotactic impact on ASC than CWF (77·5% versus 59·8% migrated cells). While proliferation was stimulated by AWF up to 136·3%, CWF had a negative effect on proliferation over time (80·3%). Expression of b-FGF, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 was strongly induced by CWF compared with a mild induction by AWF. These results give an insight into impaired ASC function in chronic wounds. The detected effect of CWF on proliferation and migration of ASC might be one reason for an insufficient healing process in chronic wounds.
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Tecido Adiposo/citologia , Exsudatos e Transudatos/fisiologia , Células-Tronco Mesenquimais/fisiologia , Cicatrização/fisiologia , Ferimentos e Lesões/metabolismo , Doença Aguda , Técnicas de Cultura de Células , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Doença Crônica , Humanos , Metaloproteinase 9 da Matriz/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Ferimentos e Lesões/patologiaRESUMO
OBJECTIVE: Long-lasting reconstruction of joint surface by using an osteochondral transfer procedure (OCT). Reduction of donor site morbidity by using a minimally invasive approach to the dorsal medial femoral condyle. INDICATIONS: Grade 3 and 4 cartilage lesions (according to ICRS [International Cartilage Repair Society]), osteochondral lesions, and osteochondrosis dissecans. CONTRAINDICATIONS: Grade 2 or higher-graded cartilage lesions at the dorsal medial femoral condyle, infection, axis deviation of more than 5 degrees in the frontal plane, advanced osteoarthritis. SURGICAL TECHNIQUE: Cylinders at recipient site are removed first, thereby determining number and diameter of donor cylinders. Supine position, skin incision over the dorsal medial femoral condyle. After dissection of soft tissue and superficial fascia, semitendinosus tendon and medial gastrocnemius muscle are retracted to the lateral side, followed by arthrotomy, introduction of two Hohmann retractors medial and lateral of the condyle, and harvesting of the donor cylinders with a tubular chisel. Advantages of the described approach: reduction of soft-tissue trauma, easy surgical technique, additional donor site area besides femoral trochlea and intercondylar notch. POSTOPERATIVE MANAGEMENT: Partial weight bearing of 10-20 kg for 4-6 weeks. Limitation of knee flexion to 90 degrees for 6 weeks. RESULTS: Between 01/2006 and 04/2007, the dorsal medial femoral condyle was used as a donor site in 16 patients. All patients were evaluated preoperatively and after 1 year using the American Knee Society Score (KSS), the Western Ontario and McMaster Universities (WOMAC) Score, the Tegner Score, and the visual analog scale (VAS) pain. The mean follow- up was 13.9 (+/-4.3) months. The mean defect area was 4.6 (+/-2.2) cm(2). The mean KSS, Tegner Score, and WOMAC Score improved from 123.1 (+/-41.5), 2.8 (+/-0.9), and 73.3 (+/-50.2) points preoperatively to 171.3 (+/-16.9), 3.4 (+/-0.6), and 26.1 (+/-17.6) points after 13.9 months (p < 0.05). The VAS pain improved from 5.3 (+/-2.7) to 2.4 (+/-1.8) points (p < 0.05). One patient with an osteochondral defect of 8 cm(2) at the medial femoral condyle (Ahlbäck's disease) still complains of pain during deep squatting. The dorsal medial femoral condyle can be recommended as donor site for OCT. The minimally invasive approach has proven to be safe and simple with a low complication rate.
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Transplante Ósseo/métodos , Cartilagem/transplante , Fêmur/cirurgia , Procedimentos Cirúrgicos Minimamente Invasivos/métodos , Coleta de Tecidos e Órgãos/métodos , Adulto , Feminino , Seguimentos , Humanos , Articulação do Joelho/cirurgia , Masculino , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/cirurgia , ReoperaçãoRESUMO
BACKGROUND/AIMS: Autologous chondrocyte (CC) transplantation has the disadvantages of requiring two surgical interventions and in vitro expansion of cells, implying the risk of cellular dedifferentiation. Our clinical aim is to develop a one-step procedure for autologous CC transplantation, i.e. harvesting, isolation and reimplantation of CC performed in one single surgical procedure. Platelet-rich plasma (PRP) is a source of autologous growth factors reported to have mitogenic effects. The objective of this study was to test the influence of PRP as an autologous scaffold on freshly isolated CC and mesenchymal stem cells (MSC). METHODS: CC and MSC were subjected to two- or three-dimensional (3D) growth systems, either with or without PRP. Chondrogenic differentiation was determined via quantification of collagen type II mRNA and immunohistochemical staining. RESULTS: We observed a proliferative effect for MSCs exposed to PRP in monolayer culture and an increase in the expression of chondrogenic markers when cells are exposed to a 3D environment. CCs exposed to PRP show a decrease in the chondrogenic phenotype with increasing proliferative activity. CONCLUSION: PRP has a proliferative effect on CCs and MSCs. In a one-step procedure for autologous CC transplantation, this might be an advantage over other scaffold materials, but confirmation in in vivo studies is required.
Assuntos
Diferenciação Celular/fisiologia , Condrócitos/citologia , Células-Tronco Mesenquimais/citologia , Plasma Rico em Plaquetas/fisiologia , Animais , Proliferação de Células , Células Cultivadas , Feminino , Imuno-Histoquímica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , OvinosRESUMO
This article is about the evaluation of possible differences in biomechanical or histomorphological properties of bone healing between saw osteotomy and random fracturing after 6 months. A standardized, 30 degrees oblique monocortical saw osteotomy of sheep tibia was carried out, followed by manual fracture completion of the opposed cortical bone. Fixation was performed by bridge plating (4.5 mm, LCDCP, broad). X-rays were taken immediately after surgery and at the end of the study. Polychrome fluorescent staining was performed according to a standardized protocol in the 2nd, 4th 6th, 10th, 14th, 18th, 22th and 26th week. Ten sheep were comprehensively evaluated. Data for stiffness and histomorphology are reported. The average bending stiffness of the operated bone was higher (1.7 (SD 0.3) with plate (MP) vs. 1.5 without plate) than for the intact bone (1.4 (SD 0.2), though no significant differences in bending stiffness were observed (P>0.05). Fluorescence staining revealed small numbers of blood vessels and less fragment resorption and remodeling in the osteotomy gap. Bone healing after saw osteotomy shows a very close resemblance to 'normal' fracture healing. However, vascular density, fragment resorption, fragment remodeling, and callus remodeling are reduced at the osteotomy.