Syftet med denna litteraturstudie var att via litteraturen beskriva nutritionens betydelse för sårläkning. Studien genomfördes som en deskriptiv litteraturstudie där metoden bestod av att söka vetenskapliga artiklar i databaserna Medline, Cinahl och Academic Search Elite. Sjutton artiklar valdes ut. Resultatet visade att patienter med bensår eller trycksår i många fall inte når upp till de näringsrekommendationer som finns. Det visade sig vara vanligt att dessa patienter ligger vid gränsen för att vara undernärda. Det upptäcktes att det är sällan som kostintaget täcker energibehovet. Framför allt var zinknivåerna genomgående väldigt låga och det visar även att det är brist på en hel del andra ämnen. Ämnen som visat sig speciellt fördelaktiga för sårläkning är protein, vitamin C, zink samt arginin. För patienter med brännskador däremot har det visat sig att saltet ornithine α-ketoglutarate minskar proteinkatabolismen samt förbättrar proteinsyntesen, vilket är en förutsättning för sårläkning. Slutsatsen med föreliggande studie var att nutritionen är viktig och kan ha en avgörande betydelse för sårläkning. / The purpose with this literature study was that through literature describe the nutrition’s significance for healing wounds. The study was made as a descriptive literature study where the method was conducted by searching scientific articles in the databases Medline, Cinahl and Academic Search Elite. Seventeen articles were chosen. The result showed that patients with leg ulcers or pressure sores in many cases don’t attain the recommendations for nutrition that are available. It showed that it is normal that these patients are on the border of being undernourished. It was discovered that the nutrient intake rarely covers the energy needs. Above all the zinc levels were throughout very low and that shows that there is also lack of some other substances. The substances that have been shown especially good for healing wounds are protein, vitamin C, zinc and arginine. However, for patients with burn injuries the salt ornithine α-ketoglutarate has been shown to reduce protein catabolism and also improve protein synthesis, which are essential to healing wounds. The conclusion from this study was that nutrition may be an important but also a determining factor to healing wounds.

The plasminogen activator (PA) system has been shown to be intimately involved in wound healing. However, the role of this system in the initiation and resolution of inflammation during healing process remained to be determined. The aims of this thesis were to investigate the molecular mechanism underlying the interaction between the PA system and the inflammatory system during wound healing and to explore the therapeutic potential of plasminogen in various wound-healing models. The role of plasminogen in the inflammatory phase of the healing process of acute and diabetic wounds was studied first. Our data showed that administration of additional plasminogen to wild-type mice accelerates the healing of acute wounds. After injury, both endogenous and exogenous plasminogen are bound to inflammatory cells and are transported to the wound site, which leads to activation of inflammatory cells. In diabetic db/db mice, wound-specific accumulation of plasminogen does not take place and the inflammatory response is impaired. However, when additional plasminogen is injected, plasminogen accumulates in the wound, the inflammatory response is enhanced, the signal transduction cascade is activated and the healing rate is significantly increased. These results indicate that administration of plasminogen may be a novel therapeutic strategy to treat different types of wounds, especially chronic wounds in diabetes. The role of plasminogen at the later stage of wound healing was also studied in plasminogen-deficient mice. Our data showed that even if re-epithelialization is achieved in these mice, a prolonged inflammatory phase with abundant neutrophil accumulation and persistent fibrin deposition is observed at the wound site. These results indicate that plasminogen is also essential for the later phases of wound healing by clearing fibrin and resolving inflammation. The functional role of two physiological PAs during wound healing was further studied in a tympanic membrane (TM) wound-healing model. Our data showed that the healing process was clearly delayed in urokinase-type PA (uPA)-deficient mice but not in tissue-type PA (tPA)-deficient mice. Less pronounced keratinocyte migration, abundant neutrophil accumulation and persistent fibrin deposition were observed in uPA-deficient mice. These results indicate that uPA plays a central role in the generation of plasmin during the healing of TM perforations. Finally the therapeutic potential of plasminogen in the TM wound-healing model was studied. Our data showed that local injection of plasminogen restores the ability to heal TM perforations in plasminogen-deficient mice in a dose-dependent manner. Plasminogen supplementation also potentiates healing of acute TM perforations in wild-type mice, independent of the administration method used. A single local injection of plasminogen in plasminogen-deficient mice can initiate healing of chronic TM perforations resulting in a closed TM with a continuous but rather thick outer keratinocyte layer. Three plasminogen injections lead to a completely healed TM with a thin keratinizing squamous epithelium covering a connective tissue layer that can start to reorganize and further mature to its normal appearance. In conclusion, our results suggest that plasminogen is a promising drug candidate for the treatment of chronic TM perforations in humans.  Taken together, our data indicate that plasminogen is a novel inflammatory regulator that promotes wound healing.

Rapid wound healing is important to regain the skins protective function after injury. Studies have shown that enamel matrix proteins (EMP) have many desirable effects which may accelerate wound healing [Bosshardt et al. 2008]. Polymers (Polymer A, B and C) were formed into a mat form, with or without incorporated enamel matrix derivative (EMD) (Collaboration partner). The materials may be suitable for wound care and drug delivery systems. Protein release tests were performed on samples incubated in physiological-like solution using pyrogallol red staining, ultraviolet (UV) spectrophotometer and high-performance liquid chromatography (HPLC). Protein was detected in Polymer A material samples, compared to a reference material sample, using pyrogallol red staining. An in vitro experiment showed that normal human dermal fibroblasts (NHDF) cultivated with Polymer A material (with EMD) had significantly higher viability than NHDF cultivated with reference material (Polymer A without EMD) and comparable viability to fibroblasts grown with either 0.1 mg EMD in solution or with 10% fetal calf serum. Images taken of Polymer A material, with incorporated Fluorescein isothiocyanate- (FITC) labeled EMD, indicate a homogenous distribution of EMD peptides and/or EMD aggregates throughout the material. A dressing which contains an active substance may have clinical promise for wound care applications.

In this study, it was aimed to produce sericine/collagen composite membranes and to investigate their properties as a wound dressing. Different membrane compositions were prepared by casting and solvent evaporation method. After initial studies for optimization of ratios, membrane groups at two different thicknesses were prepared for each selected ratio and cross-linked with 3 % (w/v) glutaraldehyde (GTA). Considering the wound dressing requirements, equilibrium degree of swelling (EDS), water vapor transmission rate (WVTR), oxygen permeability, mechanical properties, in situ degradation, microbial penetration and cytotoxicity of membranes were examined. The EDS of membranes had a range of 14.91 to 4.37 (g/g) and increased significantly with the presence of sericin. There was no obvious relationship between the sericin ratio of membranes and WVTR, but the increase in membrane thickness decreased WVTR significantly. Thin and sericin containing membranes had statistically better oxygen permeabilities. Sericin deteriorated the tensile strength and elongation of membranes statistically. Cross-linked groups were resistant to hydrolytic degradation through 4 weeks of incubations. None of the membranes were penetrable to bacteria owing to their dense structure. For cytotoxicity studies, 3T3 fibroblasts and keratinocytes were seeded on membranes separately, and analyzed with MTT assays, and light microscopy and scanning electron microscopy (SEM). As regards to MTT assay, keratinocytes proliferated significantly on membranes and reached to high confluence within 7 days. Similarly, fibroblasts also showed high proliferation on membranes. Light microscopy and SEM analysis showed that both cells could attach, grow and spread on membranes. Also, cells gained their characteristic morphology after 1 day and formed flattened structure within 7 days.

High-dose morphine has been extensively used in the control of postoperative and cancer pain. Patients receiving prolonged administration of high-dose morphine are known to be associated with certain cardiovascular complications and tissue regeneration defects. This research thesis aims to investigate the biological effects and molecular mechanisms of high-dose morphine on the vascular endothelial function, angiogenesis and wound regeneration using murine models of morphine-dependence and cultured endothelial cell assays. Mice were subjected to placebo or morphine (20 mg/kg, i.p.) injection for consecutive 14 days. Aortas were harvested for assessment of vasomotor function by isometric force recordings. Protein expression p47phox (a major subunit of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase) was determined by Western blotting. Generation of superoxide anions was detected under confocal microscope. Endothelium-dependent relaxations to acetylcholine were significantly reduced in morphine-treated animals, but were normalized by superoxide scavenging. Fluorescent densities of dihydroethidium and expression of p47phox were increased in the aorta of morphine-treated mice. In the second part of this thesis, the candidate determined the effects of high-dose morphine on angiogenesis and mobilization of endothelial progenitor cells (EPCs) in a mouse model of excisional wound injury. Excisional wound was created on control and morphine-dependent mice. Wound healing was compared by measuring the final-to-initial wound area ratio. Generation of superoxide anions in the wound was determined by luminol-enhanced chemiluminescence. Circulating mononuclear cells were isolated and measured for EPC (defined as CD34+/CD133+ cell) counts. In vivo and in vitro measurements of angiogenesis following morphine treatment were performed using the Matrigel assay. The results showed that wound closure was significantly reduced in mice treated with morphine when compared with controls, and higher levels of superoxide anions were generated in these wounds. High-dose morphine reduced numbers of circulating EPCs following creation of excisional wound. Matrigel assay showed impaired angiogenesis in animals and reduced capillary tube formation in cultured endothelial cells treated with high-concentration of morphine. Collectively, this research thesis demonstrated a number of novel findings. First, high-dose of morphine impairs vascular endothelial function by increased production of vascular superoxide anions. Activation of NADPH oxidase may be the molecular mechanisms responsible for reduced bioavailability of endothelium-derived NO. Second, systemic administration of high-dose morphine delays healing of excisional wounds and impairs angiogenesis. This antiangiogenic effect is associated with increased superoxide anions production and impaired mobilization of EPCs. In line with direct endothelial dysfunction, impaired angiogenesis and EPC mobilization resulted from high-dose morphine treatment may cause increased cardiovascular morbidity in human subjects receiving higher therapeutic dose of morphine.

Thesis (Ph. D.)--Case Western Reserve University, 2004. / [School of Medicine] Department of Neurosciences. Includes bibliographical references. Available online via OhioLINK's ETD Center.

Thesis (Ph. D.)--University of Washington, 2003. / Vita. Includes bibliographical references (leaves 198-210).

Zugl.: Berlin, Freie Univ., Diss., 2008 / Includes bibliographic references.

Hyperbaric oxygen (HBO) therapy is the intermittent inhalation of 100% oxygen at a pressure greater than one atmosphere absolute. It is an effective treatment for various inflammatory conditions, including chronic wounds which are characterized by an excessive influx of neutrophils and their prolonged persistence at the wound site. Neutrophil apoptosis and clearance have been shown to be required for resolution of inflammation. The mechanisms by which HBO aids wound healing are well documented, but its effects on cellular inflammatory response are not well understood particularly with respect to neutrophils. The hypothesis presented in this thesis is that increased oxygenation via HBO assists chronic wound healing by enhancing non-inflammatory neutrophil defences and cell death through apoptosis. An investigation was carried out into the effects of HBO on neutrophil antimicrobial function and apoptosis using differentiated HL-60 cells as an in vitro neutrophil model. The data clearly showed that a single HBO treatment for 90 min caused an increase in the oxidative burst activity of neutrophil-like cells as shown by increased NBT staining, superoxide (cytochrome c reduction) and H2O2 production (Kruskal-Wallis, P < 0.05), and phagocytosis of Staphylococcus aureus. HBO treatment displayed a pro-apoptotic effect, enhancing caspase 3/7 activity both in the presence and absence of a TNF-α stimulus (Kruskal-Wallis, P < 0.05) and causing morphological changes (observed using Giemsa and SYBR® Safe staining) associated with apoptosis. Although no consistent pattern was observed, both hyperoxia and pressure alone seemed to contribute to both the increase in antimicrobial activity and the increase in apoptosis induced by HBO in these neutrophil-like cells (Chapters 4 and 5). HBO-enhanced neutrophil clearance by macrophages was investigated using bovine neutrophils and monocyte-derived macrophages (MDMФ). A single 90 min HBO exposure significantly increased the clearance of fresh and 22 h-aged neutrophils by MDMФ (two-way ANOVA, P < 0.05), suggesting an increase in phosphatidylserine (PS) exposure in apoptotic neutrophils after HBO treatment (Chapter 6). Importantly, a long-term repetitive exposure to HBO in patients with chronic wounds caused a significant decrease in the antioxidant enzyme defence system (one-way repeated measures ANOVA, P < 0.05), plasma TNF-α and IL-1β after 30 HBO sessions, with down regulation of expression of the anti-apoptotic factors, NF-B and Bcl-2 (Chapter 7). These findings may go some way towards explaining the effectiveness of HBO treatment not only for chronic wounds but also for other inflammatory conditions that may be affected by this treatment.

published_or_final_version / Pharmacology / Master / Master of Philosophy