Дисертації з теми "Pressure on tissue"

The overall aim of this thesis was to combine optical methods into a system with the ability to simultaneously measure blood flow changes at different tissue depths. The goal of such a system was to reveal vascular mechanisms relevant to pressure ulcer etiology under clinically relevant conditions and in relation to the evaluation of pressure-redistribution support surfaces. This thesis consists of four quantitative, cross-sectional studies measuring blood flow responses before, during, and after pressure exposure of the sacral tissue. Two optical methods – photoplethysmography and laser Doppler flowmetry – were combined in a newly developed system that has the ability to discriminate blood flows at different tissue depths. Studies I and II explored blood flow responses at different depths in 17 individuals. In Study I the blood flow was related to tissue thickness and tissue compression during pressure exposure of ≥ 220 mmHg. In Study II, the sacral tissue was loaded with 37.5 mmHg and 50.0 mmHg, and the variation in blood flow was measured. Studies III and IV included 42 healthy individuals < 65 years, 38 healthy individuals ≥ 65 years, and 35 patients ≥ 65 years. Study III included between-subject comparisons of blood flow and pressure between individuals in the three study groups lying in supine positions on a standard hospital mattress. Study IV added within-subject comparisons while the individual was lying on four different types of mattress. The studies explored the vascular phenomena pressure-induced vasodilation (PIV) and reactive hyperemia (RH). The most common blood flow response to tissue exposure in this thesis was PIV, although a decrease in blood flow (a lack of PIV) was observed in some individuals. The patients tended to have higher interface pressure during pressure exposure than the healthy groups but no differences in blood flow responses were seen. Our results showed that pressure levels that are normally considered to be harmless could have a significant effect on the microcirculation in different tissue structures. Differences in individual blood flow responses in terms of PIV and RH were seen, and a larger proportion of individuals lacked these responses in the deeper tissue structures compared to more superficial tissue structures. This thesis identified PIV and RH that are important vascular mechanisms for pressure ulcer development and revealed for the first time that PIV and RH are present at different depths under clinically relevant conditions. The thesis also identified a population of individuals not previously identified who lack both PIV and RH and seem to be particularly vulnerable to pressure exposure. Further, this thesis has added a new perspective to the microcirculation in pressure ulcer etiology in terms of blood flow regulation and endothelial function that are anchored in clinically relevant studies. Finally, the evaluation of pressureredistribution support surfaces in terms of mean blood flow during and after tissue exposure was shown to be unfeasible, but the assessment of PIV and RH could provide a new possibility for measuring individual physiological responses that are known to be related to pressure ulcer development.

This study was motivated by a theoretical formulation on mechanobiology of soft and hard skeletal tissue differentiation. To prove this formulation experimentally, I hypothesized that cartilaginous phenotype can be induced in vitro in a seemingly non-cartilaginous cell source from fibrous tissue. In testing this hypothesis, I have focused on cartilage as a target and fibrous tissue as an origin or the source of cell. Four different trials were pursued with one supposition in common, i.e. hydrostatic pressure is one of the main driving forces for chondroinduction in vitro. The first and second trials pertained to the influence of a relatively short and long duration cyclic hydrostatic compression on rat Achilles tendon fibroblasts. The third trial was to examine the effect of two different drugs on cytoskeletal elements of mesenchymal stem cells or mouse embryonic fibroblast lines in pellet cultures combined with the similar duration and/or frequency of cyclic hydrostatic pressure adopted in the aforesaid trials with no pharmacological agents added. Last, attempts were made to implement an advanced technique in molecular biology called 'PCR array' to further quantify expression levels of eighty four pathway-specific genes in mouse TGFbeta/BMP signaling traffic under the same physiological regimen of hydrostatic compression. Results demonstrated that transdifferentation in phenotype from tendon to fibrocartilage may have occurred in vitro in tendon fibroblasts in pellet cultures exposed to hydrostatic pressure. Experiments on the role of the cytoskeleton in mechanotransduction of the applied level of hydrostatic pressure demonstrated that disruption of microfilaments in the presence of cytochalasin-D did not significantly interfere with the anabolic effect of cyclic pressure. However, disruption of microtubule assembly by nocodazole abolished the pressure-induced stimulation in cartilage marker genes. These findings suggest that microtubules, but not microfilaments, are involved in mechanotransduction of hydrostatic pressure by mesenchymal stem cells.

Soft tissue breakdown occurs in association with biochemical changes that can be attributed to a reduction in blood and lymph flow to a localised tissue area in response to applied pressure. The resulting ischaemia can lead to a reduction in available oxygen and accumulation of waste products. Tissue breakdown leading to the development of pressure sores afflicts patients who are already debilitated, although not all patients appear to be equally susceptible. Measurement of sweat biochemistry and blood gas tensions may reflect the biochemical process in the underlying tissues and provide a simple and non-invasive method of investigating the status of soft tissues. The potential of specific sweat metabolites to act as markers of soft tissue status during and following loading has been investigated at a clinically relevant site in healthy volunteers, and in two clinically relevant patient groups. A range of validation procedures were undertaken and a series of parameters derived to investigate the temporal profile of sweat biochemistry, and identify various modes of gas tension response. Investigations at the loaded sacrum of healthy individuals showed a statistically significant increase in sweat lactate, urea, urate and chloride concentrations which were dependent upon the level of externally applied pressure. Mean increases of between 10%-60% were demonstrated for sweat metabolite concentrations at the loaded site compared to the control site for applied pressures in the range 40-120 mmHg. Similar increases were demonstrated in sweat collected from highly loaded tissue areas within the stump socket of lower limb amputees. A threshold value for P02 tension was identified, amounting to a 60% reduction from the unloaded value, which was associated with elevated tissue carbon dioxide levels as well as increased sweat metabolite concentrations in the loaded phase. This finding may provide a useful predictor of soft tissue status during prolonged loading. No pessimist ever discovered the secrets of the stars, or sailed to an uncharted land, or opened a new heaven to the human spirit. Helen Adams

Thesis: S.B., Massachusetts Institute of Technology, Department of Mechanical Engineering, 2015.
Cataloged from PDF version of thesis.
Includes bibliographical references (pages 53-55).
Removing necrotic tissue and foreign materials from wounds is a critical step in the management and treatment of chronic wounds. MIT's BioInstrumentation Laboratory developed a novel debridement technology that uses two high-speed impinging water jets to excise necrotic tissue. However, this device potentially causes accidental injection of water into healthy tissue beneath the wound bed, which can cause injury and necrosis in the healthy tissue. The purpose of this thesis is to explore tissue tension as a solution to reduce the required cutting power and consequently reduce water injection to acceptable levels. After validating the positive effect of tissue tension on the cutting efficiency of the water jet debridement device, we developed a technology that uses angled rolling wheels to tension tissue prior to debridement. This novel tensioner was qualitatively tested and successfully applied local tension at the site of cutting. Suggestions for further testing to improve this device are given. This tissue tensioner shows promise as a complementary appendage to the water jet debridement device.
by Colette P. Abah.
S.B.

This report is a Bachelor thesis in the field of product development and design. It includes a literature review in the field of pressure ulcers and diabetes as well as a design process. The writer of this report, Hanna Länne Rosenlund, is a Mechanical Engineering student at the School of Engineering at Jönköping University. The focus of the education lies within product development and design. Pressure ulcers are a growing health care problem due to an increase in the mean life expectancy as well as an increase in diabetes in the world population. Patients with artificial limbs are often victims of pressure ulcers due to prolonged pressures from the prosthetic sockets on already sensitive areas of the body. Research in the field of pressure-induced injuries is currently taking place at Jönköping University. Their knowledge in finite element modelling and orthopaedic engineering made the research project, PEOPLE, possible. PEOPLE is a collaboration project between the School of Engineering and the School of Health and Welfare at Jönköping University as well as three company partners. In the project they aim to develop a device that will apply pressure to a lower limb while a MR camera takes scans of the limb. The images are later analysed closely by use of the finite element model, which means that all the different tissue properties will be collected for a computer simulation. In that way the tissues reactions to more extreme forms of pressure can be evaluated. This will contribute to the research in hope of eventually being able to predict whether or not a person might be at risk of developing pressure ulcers. A design of the prototype’s chassis was needed to optimize ease of use for both patient and staff, user options to expand research abilities, and sustainability. The design process includes product decomposition, concept generation, conceptual design, brainstorming, design for assembly, and design for component manufacturing, which generated several concepts. The final concept was decided by use of Pugh’s-matrices. The different concepts and the final concept were created in the computer aided design programme, Solid Works. The work resulted in a highly adjustable two-piece concept with optimized ease of use and sustainability due to the use of a Velcro strap. The prototype will come in two different sizes and will be mountable by a developed screwing system and therefore easy to pack, store and replace. It will also contain a new pressure relief system for a more comfortable patient experience. For further development of chassis of this kind, a replaceable pressure relief system would enhance the comfort when usage of larger limbs. When the device will be available for testing, a patient’s point of view can be taken in to consideration for a more reliable thesis and for further optimization of the comfort.
Detta är ett examensarbete på kandidatnivå inom ämnet produktutveckling och design. I arbetet ingår en litterär överblick och sammanfattning av forskning i ämnet angående trycksår och diabetes, samt en designprocess. Författaren, Hanna Länne Rosenlund studerar Maskinteknik med inriktning Produktutveckling och Design på Jönköpings Tekniska Högskola. Trycksår är ett växande problem inom vården på grund av en ökning i medellivslängden samt en ökning av diabetesdiagnoser hos världens befolkning. Patienter med proteser faller ofta offer för trycksår på grund av extrema och långvariga tryckförhållanden där proteserna är lokaliserade. Ett område som redan är känsligare för tryck. Forskning inom tryckframkallande skador pågår just nu på Jönköping University. Deras kunskap inom finita elements modellering samt ortopedingenjörsteknik har gjort detta forskningsprojekt möjligt. Forskningsprojektet heter PEOPLE och är ett samverkningsprojekt mellan Tekniska Högskolan, Hälsohögskolan samt tre företagspartners. Tillsammans siktar de mot att utveckla en prototyp som ska utsätta en lem för ett konstant tryck medan en MR kamera scannar vävnaden. En finit elements modell av lemmen skapas sedan för närmre granskning av vävnaden hos individen. Vävnadens egenskaper samlas sedan för en simulering då man kan utvärdera hur vävnaden skulle reagera på mer extrema former av tryck. På så sätt kan prototypen bidra till forskningen inom ämnet för att förhoppningsvis kunna förutspå ifall en person är vid risk för att utveckla trycksår eller inte. En omkonstruktion av prototypens chassi har utvecklats för att optimera användarvänligheten för både patient och personal, användarmöjligheten för forskningssyfte, samt för att bättre bidra till en mer hållbar lösning. Designprocessen har inkluderat teorier såsom produktnedbrytning, konceptgenerering, konceptutveckling, brainstorming, design for assembly och design for manufacturing som alla har hjälpt till att generera koncept. Det slutgiltiga konceptet valdes med hjälp av Pugh matriser. Koncepten samt det slutliga konceptet skapades i ett CAD (computer aided design) program, Solid Works. Arbetet resulterade i ett justerbart tvådelat koncept med optimerad användarvänlighet och hållbarhet genom att använda sig av ett kardborreband. Prototypen kommer att finnas i två olika storlekar och vara monterbar genom att det går att skruva bort chassit och på så sätt optimera packning, hantering och förvaring. Det kommer också att innehålla ett nyutvecklat system för att underlätta fördelningen av tryck på motsatt sida från indenteringen. För fortsatt utveckling av chassit hade ett utbytbart system för tryckavledning optimerat produkten ytterligare då komforten hade ökat vid användning på större lemmar. När produkten finns tillgänglig för testning i framtiden kommer en patients syn vara möjlig att ta med och på så sätt förstärka trovärdigheten av arbetet samt bidra till fortsatt strävande för komfort.

Development of a Tool for Pressure Ulcer Risk Assessment and Preventive Interventions in Ancillary Services Patients Monica S. Messer Abstract The incidence of nosocomial pressure ulcers has increased 70 percent in U.S. hospitals over the past 15 years despite implementation of preventive guidelines and the wide-spread use of validated risk assessment tools. Most preventive efforts have been focused primarily on patients who are bed-ridden or immobile for extended periods. What has not been well studied or identified is the risk for pressure injury to patients undergoing diagnostic procedures in hospital ancillary units where extrinsic risk factors such as high interface pressures on procedure tables and friction and shear from positioning and transport can greatly magnify the effect of patient-specific intrinsic risk factors which might not otherwise put these patients at high risk on an inpatient unit. The purpose of this study was to develop a risk assessment tool designed explicitly to quantify the combination of these intrinsic and extrinsic risk factors in individual patients undergoing ancillary services procedures, and to identify targeted preventive interventions based on the individual level of risk. Empirically and theoretically-derived risk factors for the tool were tested in a nation-wide hospital database of over 6 million patient discharge records using bivariate and multivariate analysis to identify significant predictors of pressure ulcer outcomes. The statistically significant factors emerging were then used to develop the risk assessment scale. These predictors included; advanced age, diabetes, human immunodeficiency virus infection, sepsis, and fever. The scale was tested for internal validity using the split-sample cross-validation method, and for accuracy using the area under the Receiver Operating Characteristics curve. The optimum score cut point was identified to provide a predictive accuracy of 71 percent. Interventions for the tool were identified from national clinical practice guidelines and aligned in sets based on patient levels of risk identified by the scoring portion of the tool. The entire tool was evaluated for content validity by a panel of five international nurse experts in pressure ulcer prevention and tool development. The content validity index calculated from their ratings was .91 indicating excellent agreement on content validity.

ABSTRACT Implementation of Physiologic Pressure Conditions in a Blood Vessel Mimic Bioreactor System Kevin Mark Okarski Tissue engineering has traditionally been pursued as a therapeutic science intended for restoring or replacing diseased or damaged biologic tissues or organs. Cal Poly’s Blood Vessel Mimic Laboratory is developing a novel application of tissue engineering as a tool for the preclinical evaluation of intravascular devices. The blood vessel mimic (BVM) system has been previously used to assess the tissue response to deployed stents, but under non-physiologic conditions. Since then, efforts have been made to improve the vessel and bioreactor’s ability to emulate in vivo conditions. The ability to tissue engineer constructs similar to their native tissue counterparts is heavily reliant upon controlling the environment and mechanical stimuli the construct is exposed to. Mimicking physiologic conditions influences cellular growth, proliferation, and differentiation. Two important mechanical stimuli are cyclic strain and wall shear stress. Previous work sought to improve these factors within the BVM bioreactor and resulted in the implementation of pulsatile perfusion and increased fluid viscosity. These previous bioreactor design modifications generated pulsatile pressures of approximately 80 mmHg and a wall shear stress of 6.4 dynes/cm2. However, physiologic pressure waveforms were not achieved. Studies in this thesis were carried out to implement an effective means of establishing a more physiologic pressure wave within the bioreactor that is accurate, consistent, and easily adjustable. As a result of conducting the present studies, modifications to the bioreactor system were made that uphold the overall goals of efficacy and efficiency. The desired pressure wave was created by setting the degree of pump tubing occlusion on the 3-roller peristaltic pump head and using a water column to backpressure the bioreactor chamber. Maintaining a desired backpressure within the system necessitated the development of a new bioreactor chamber with increased extraluminal leak pressure resistance. The opportunity was also used to further improve upon the bioreactor chamber design to allow for 360° rotation to reduce cell sedimentation. Modifications to the bioreactor system required quantitative evaluation to assess their impact upon local flow dynamics to the tissue construct. A system model was created and evaluated using computational modeling. Through the work performed in this thesis, pulsatile pressure waves of approximately 120/80 mmHg were successfully established within the bioreactor. The ability to accurately model physiologic pressures will ultimately help yield tissue constructs more similar to native tissues – both healthy and pathological. The newly designed bioreactor chamber and computational model for the system will be helpful tools for implementing or evaluating future bioreactor developments or improvements. While the main objective of the thesis has been completed by creating a system capable of emulating physiologic pressure fluctuations, there still remains room for further improvements in back-pressuring and scaling the system, refining the rotational bioreactor chamber design, and building upon the complexity and accuracy of the computational model.

Polymer scaffold use has become commonplace in tissue engineering strategies. Scaffolds provide sturdy interfaces that securely anchor tissue engineered constructs to their designated locations. Researchers have used scaffolds to provide support to developing tissues as well as a growth template to aid the development of the desired phenotypic structure. In addition to using scaffolds for their mechanical support, scaffolds can be used as a diagnostic tool by attaching sensors. Strain gauge sensors have been attached to scaffolds to monitor compression and elongation. These polybutylterphalate (PBT) scaffolds were used in a cartilage tissue-engineering project for femoral cartilage repair. The aim of this project was to measure native cartilage pressure in normal canine stifle joints using strain gauge scaffolds. By using pressure sensitive films to confirm joint surface pressures determined with strain gauge measurements, "sensate" scaffolds were created to be able to provide in vivo joint loading measurements. An understanding of the in vivo pressures in the menisco-femoral joint space will facilitate the development of tissue engineered cartilage by determining chondrocyte mechanical triggers as well as helping define reasonable expectations for engineered articular cartilage tissue that is required for successful cartilage repair.

Current orthopaedics treatments of bone defects often involve the use of implanted fixatives and/or autograft procedures to restore function to the afflicted area following injury. Fixatives and implants are usually temporary solutions, since they are intrinsically prone to failure. In addition to this, replacing implants involve expensive and invasive procedures that cause great hardship to patients. Whilst autografts can provide an excellent outcome in healing of the initial injury site, donor site morbidity from the autologous bone graft can lead to complications such as infection, chronic pain and an abnormal walking gait. Bone tissue engineering is a field of science aiming to address these limitations by providing in vitro manufactured bone to replace autografts, and also limit the use of temporary fixatives. Hydrostatic force bioreactors are currently being developed within this field to attempt improve the outcome of the tissue engineered bone by mimicking the forces typically experienced by cells in the native bone niche. Based on this principle, it is hoped that such systems will aid the translation of research in bone tissue engineering from the lab to the clinic. This research aims to investigate and validate the use of a hydrostatic force bioreactor for improving the outcome of in vitro manufactured bone using a clinically relative strategy employing human mesenchymal stem cells seeded in 3D scaffolds. The research first describes a validation process to determine the initial response of cells to hydrostatic pressure in monolayer cultures. The outcome of this study indicated that mechanical responsiveness in cells can vary according to cell phenotype and the integrity of the f-actin cytoskeleton. Next it was demonstrated that hydrostatic pressure can improve the outcome of in vitro bone formation by MG-63 human osteoblast like cells, validating the bioreactor as a potential preconditioning platform. Following this, a model of bone formation in hMSCs/collagen scaffolds was described, whereby a predictable rate of bone formation was determined by adjusting cellular distribution and protein concentration in collagen type-1 scaffolds. Finally, an organotypic fracture repair model was established using explanted embryonic chick femurs to test the hypothesis that hydrostatic preconditioning of hMSC/collagen hydrogels can improve the outcome of fracture repair. The results of this study showed that bioreactor stimulation could enhance the outcome of repair using a combination of undifferentiated hMSC/collagen type-1 scaffolds, and global mechanical signalling (stimulation of entire femur constructs). It was then shown that hydrostatic preconditioning of hMSC seeded hydrogels prior to implantation did not increase the rate of in vitro bone formation. Following implantation of the hydrogels into the fracture repair model, it was demonstrated that highly mineralised preconditioned implants actually inhibited the fracture repair process. In addition to this, it was shown that preconditioned implants with a lower level of mineralisation allowed invasion and bone formation by native cells from the host tissue. Collectively, the results implied that the outcome of repair using this model relied on three main factors: the presence of global hydrostatic stimulation; the lineage commitment of hMSCs in collagen scaffolds at the time of implantation; and the permeability and cell invasion capacity of the implant.

Lung diseases, cancers, and trauma can result in injury to the connective tissue lining the lung, i.e., the pleura. Pleural injuries lead to pneumothoraxes or pleural effusions, i.e., air or fluid leaking out of the lung respectively, and potential lung collapse - an immediately life threatening condition. While several bioengineered soft tissue sealants exist on the market, there is only one sealant FDA-approved for use in pulmonary surgery. In addition, very limited techniques are presented in the literature for characterizing the burst properties of hydrogel tissue sealants. For my thesis, I proposed to develop a protocol for characterizing the burst properties of hydrogel sealants using a novel burst pressure test chamber. I further proposed a novel combination of oxidation and methacrylation reactions of alginate for tissue sealant applications, with a particular focus on developing a pulmonary sealant. The proposed research objectives are: 1) To develop protocol for testing hydrogel sealants for soft tissue applications; 2) To verify alginate as a potential for tissue sealant applications; and 3) To optimize an alginate hydrogel sealant and perform ex vivo analysis for a pleural sealant application. Alginate materials with varying degrees of oxidation and methacrylation were synthesized and characterized. Oscillatory rheometry was used to characterize material properties such as viscosity, hydrogel gelation kinetics, and complex moduli. Burst pressure measurements properties and failure mechanisms, i.e. delamination or material failure, were collected for a liquid and dry-state application. Preliminary ex vivo mouse lung model testing demonstrated that methacrylated alginate hydrogels are able to withstand physiological pressures associated with breathing, and failure occurs within the hydrogel for adhesive alginate-based tissue sealants.

All blood vessels in the microvasculature are embedded in loose connective tissue, which regulates the transport of fluid to and from tissues. The intersti-tial fluid pressure (IFP) is one of the forces that control this transport. A lowering of IFP in vivo results in an increased transport of fluid from the circulation into the underhydrated connective tissues, resulting in edema formation. During homeostasis, contractile connective tissue cells exert a tension on the connective tissue fibrous network by binding with β1 in-tegrins, thereby actively controlling IFP. During inflammation, the IFP is lowered but platelet-derived growth factor (PDGF)-BB induces an IFP nor-malization dependent on integrin αVβ3. We demonstrate that extracellular proteins from Streptococcus equi subspecies equi modulated cell-mediated and integrin αVβ3-directed collagen gel contraction in vitro. One of these proteins, the collagen- and fibronectin binding FNE, stimulated contraction by a process dependent on fibronectin synthesis. This study identified a pos-sible novel virulence mechanism for bacteria based on the ability of bacteria to modulate the edema response. Another protein, the collagen-binding pro-tein CNE, inhibited contraction and this led to the identification of sites in collagen monomers that potentially are involved in connecting αVβ3 to the collagen network. PDGF-BB and prostaglandin E1 (PGE1) stimulate and inhibit collagen gel contraction in vitro and normalize and lower IFP, respec-tively. We showed that these agents affected both similar and different sets of actin-binding proteins. PDGF-BB stimulated actin cytoskeleton dynamics whereas PGE1 inhibited processes dependent on cytoskeletal motor and adhesive functions, suggesting that these different activities may partly ex-plain the contrasting effects of PGE1 and PDGF-BB on contraction and IFP. Mutation of the phosphatidylinositol 3’-kinase (PI3K), but not phospholipase C (PLC)γ activation site, rendered cells unable to respond to PDGF-BB in contraction and in activation of the actin binding and severing protein cofilin. Ability to activate cofilin after PDGF-BB stimulation correlated with ability to respond to PDGF-BB in contraction, suggesting a role for cofilin in this process downstream of PDGF receptor-activated PI3K. Many proteins can modulate contraction either by affecting the extracellular matrix and cell adhesions or by altering cytoskeletal dynamics. Knowledge on how these proteins might influence IFP is likely to be of clinical importance for treat-ment of inflammatory conditions including anaphylaxis, septic shock and also carcinoma growth.

Despite lymphatic function regularly being proposed as one of the causative mechanisms in the development of pressure ulcers, there has been a paucity of research regarding the effect of compression on lymphatic function in human tissues. This has motivated the present work, which has developed and validated techniques to safely assess lymphatic function and concomitantly measure the physiochemical response in the skin, directly under and following the application of a uniaxial load in human volunteers. Lymphatic function was assessed through optical imaging of an intradermally injected near-infrared fluorophore, namely indocyanine green (ICG), which is rapidly cleared in lymph. Pro-inflammatory biomarkers were recovered by two minimally invasive collection techniques involving microdialysis and SebutapeTM. Through the application of established image processing concepts to NIR lymphangiography for the first time, a method for objectively quantifying parameters of active lymphatic clearance is also described. The results of the study suggest that applied pressure of 60 mmHg for a period of 40 minutes can lead to changes in both novel pro-inflammatory and lymphatic parameters, not previously described in the literature. During the loading period, induced expression or accumulation of IL-6 and IL-8 in the interstitium was observed (p < 0.01). For IL-6, this upregulation was sustained for over 40 minutes post loading (p < 0.01). The same loading protocol was associated, through categorical analysis, with impaired lymph formation in lymphatic capillaries and disrupted valve function in collecting vessels directly under the load. The velocity of active lymphatic drainage along the loaded pathway was also significantly reduced (p < 0.05).The findings from this thesis support the proposal that impaired lymphatic function, with a corresponding accumulation of harmful biomolecules, are feasible mechanisms which can contribute to the development of pressure ulcers. The present study strongly supports further exploration of these hypotheses using the techniques described.

Doutoramento em Engenharia Química - Especialização em Engenharia de Materiais e Produtos Macromoleculares
The ultra-high hydrostatic pressure processing (UHP) is an expanding technology used mostly in food industry for the pasteurization of foods while preserving their organoleptic properties. This technique may be also applied to introduce structural and functional changes in biomolecules including cellulose. Recent studies have demonstrated the potential of UHP towards cellulosic pulps, namely the ability to promote forced fibre hydration thus improving cellulose accessibility and fibrils reorganization. The main purpose of this thesis was the evaluation of UHP potential for the modification of recycled fibres aiming to improve its performance in the production of tissue paper in collaboration with the biggest national producer Renova FPA, S.A. The present study has identified the structural changes that occurred in cellulose of recycled pulp induced by UHP. Thus the crystalline regions demonstrated to some extent the co-crystallization of appropriately oriented crystallites and the recrystallization of paracrystalline regions as demonstrated by XRD and 13C NMR. In addition, UHP have induced fibrils disaggregation upon forced hydration thus enhancing their accessibility towards water and chemical reagents. A substantial reduction of recycled fibres hornification upon UHP has been suggested. UHP-processed recycled fibres also demonstrated the increment of strongly bound water content, as revealed by thermal analysis and by FTIR of deuterated samples. Changes in physical structure of cellulose caused the enhancement of such properties of recycled pulp as accessibility, hydrophilicity, moisture sorption capacity, surface contact angle, capillarity, among others. The effect of beating (B) before (B-HP) or after the UHP (HP-B) or between two beating stages (B1-HP-B2) on the drainability and mechanical properties were evaluated. In these studies, basic papermaking properties and, especially, the capillarity (up to 112%) were improved. The most advantageous from the point of view of strength properties of tissue paper production was suggested to be the B1-HP-B2 sequence suitable both for recycled fibres and softwood fibres. A synergetic effect of UHP on the beating of recycled cellulosic fibres has been detected allowing up to 50% energy savings in refining. At the same time, the UHP effect on the virgin fibres refinability was much more moderated. The enhanced accessibility of recycled pulp upon UHP was also used to improve its ability towards targeted chemicals, enzymes and nano-sized structures. These experiments were carried out also with dried virgin fibres used as the models of recycled pulp. As concerns enzymes, it was suggested that enzymatic modification improves significantly the papermaking properties of recycled pulp. These improvements were related with selective removal of xylan bound to impurities and to aggregated cellulose fibrils on the fibre surface thus favouring the ensuing swelling and inter-fibre bonding in paper. UHP pretreatment and posterior enzymatic treatment revealed a synergetic effect on the mechanical properties of recycled pulp. This fact was assigned to enhanced accessibility of fibres towards xylanase by forced hydration and favourable rearrangement of cellulosic fibrils in fibres after UHP pre-treatment. The increase of basic strength properties after UHP and promoted by xylanase treatment was up to 30%, being the most pronounced for the tensile strength and the burst resistance. The impregnation of dyes in combination with UHP also has demonstrated an enhancement in impregnation/fixation of dye molecules. However, a high dependence was found on the equilibrium between dye, fixative and cellulosic fibres, depending on the dye used, both with and without UHP. This behaviour was assigned to specificity of dyes molecular structure, affinity, substantivity, among others. The impregnation of humectant compounds applying UHP was also evaluated and the improved hygroscopicity of treated pulp was confirmed. This fact was evidenced by capillarity, water absorption capacity and moisture sorption tests. The effect of UHP on the impregnation of antimicrobial agent in fibres has been carried out using polyhexamethylene biguanide (PHMB). Being a cationic polymer, PHMB was readily absorbed in fibres with and without UHP treatment. However, UHP allowed higher PHMB uptake and better retention after leaching when compared to conventional impregnation without UHP. This was attributed to stronger interactions between cationic PHMB and cellulose due to the deeper penetration of the antimicrobial agent under UHP treatment. However, the amount of PHMB to impregnate in fibres is limited because of its negative effect on paper strength properties due to disruptive action of PHMB as debonding agent. Likewise, silica encapsulated PHMB demonstrated fairly high retention upon UHP and high release during the leaching tests. Being encapsulated, PHMB was not directly bound to cellulose (electrostatic interaction) and the final paper showed at the same time fairly high PHMB release rates. The impregnation with encapsulated perfumes (Dovena New) was also carried out and revealed higher impregnation of these nano-sized structures upon UHP than under conventional conditions. This was reflected by a higher release of volatiles from pulp samples impregnated with stuffed nanoparticles under UHP treatment when compared to pulps samples with free adsorbed nanoparticles
O processamento de alta pressão hidrostática (UHP) é uma tecnologia em expansão, bastante utilizada na indústria alimentar para a pasteurização de alimentos, ao mesmo tempo que preserva as suas propriedades organoléticas. Esta técnica pode ainda ser aplicada na modificação estrutural e funcional de biomoléculas, tais como a celulose. Em estudos recentes demonstrou-se o potencial da UHP para com pastas celulósicas, nomeadamente no que diz respeito à sua capacidade de promover a hidratação forçada das fibras e, consequentemente, melhorar a acessibilidade das mesmas e ainda promover a reorganização das suas fibrilas. Posto isto, o principal objetivo desta dissertação consistiu na avaliação do potencial da UHP na modificação de fibras recicladas e virgens, para alcançar a melhoria da sua performance na produção de papéis tissue, em colaboração com uma das maiores empresas nacionais do ramo, a Renova FPA, SA. No presente estudo foram identificadas alterações estruturais na celulose, tanto em fibras recicladas como em virgens, produzidas pelo UHP. Neste sentido, as regiões cristalinas demonstraram, até certa extensão, a cocristalização de cristalitos convenientemente orientados e a recristalização de regiões paracristalinas, tal como evidenciado por XRD e 13C RMN. Para além disso, a UHP induziu ainda a desagregação das fibrilas, aquando o fenómeno de hidratação forçada, originando maior acessibilidade à água e a reagentes químicos. Sugere-se ainda a ocorrência de uma redução significativa da hornificação das fibras. Fibras processadas por UHP também demonstraram um incremento na presença de água fortemente ligada, tal como sugerido por análises térmicas e FTIR de amostras deuteradas. As alterações produzidas na celulose originaram uma melhoria nas propriedades das fibras, nomeadamente da sua hidrofilicidade, sorção de vapor de água, ângulo de contacto de superfície, capilaridade, etc. O efeito da refinação (B) antes (B-HP), ou depois da UHP (HP-B), ou entre refinações (B1-HP-B2), também foi analisado no que diz respeito à drenabilidade e propriedades mecânicas das fibras. Nestes estudos, as propriedades papeleiras e em especial a capilaridade (112%) revelaram melhorias. Com base nos resultados sugere-se que a sequência B1-HP-B2 seria a mais vantajosa para fibras recicladas e fibras virgens longas. Para além disso, o efeito sinergético da UHP com a refinação sugere ainda a possibilidade de uma poupança de 50% de energia na refinação. As melhorias induzidas pela UHP na acessibilidade de fibras celulósicas foram também utilizadas para otimizar a impregnação de químicos, enzimas e nanoestruturas. No que diz respeito às enzimas, verificou-se que a modificação enzimática de fibras celulósicas promoveu melhorias significativas nas propriedades papeleiras destas, em especial das fibras recicladas. Estes resultados foram relacionados com a remoção seletiva de xilana ligada a impurezas e a fibrilas na superfície das fibras, que consequentemente favoreceu o intumescimento e a ligação entre fibras. A utilização de um pré-tratamento de UHP e um posterior tratamento enzimático revelaram um efeito sinergético nas propriedades mecânicas das fibras celulósicas, em especial nas fibras recicladas. Este facto foi relacionado com o aumento de acessibilidade nas fibras para com a xilanase, como consequência da hidratação forçada e rearranjo favorável das fibrilas originado pela UHP. A melhoria registada nas propriedades mecânicas das fibras, como resultado do pré-tratamento UHP e posterior tratamento enzimático, foi até 30%, tendo demonstrado maior impacto nas propriedades de índices de tração e rebentamento. A impregnação de corantes com UHP também foi realizada e demonstrou uma melhor impregnação/fixação das moléculas de corante. No entanto, verificouse uma grande dependência no equilíbrio entre moléculas de corante, fixador e fibras celulósicas, dependendo no corante utilizado, com e sem alta pressão. Estas variações foram atribuídas à especificidade da estrutura molecular da molécula de corante, à sua afinidade, substantividade, etc. A impregnação de agentes humectantes com UHP também foi avaliada e a melhoria nas propriedades hidrofílicas de fibras celulósicas foi confirmada. Este facto foi evidenciado por testes de capilaridade, sorção de vapor de água e testes de capacidade de sorção de água. O efeito da UHP na impregnação de um agente antimicrobiano também foi estudado utilizando polihexametileno biguanide (PHMB). Sendo um polímero catiónico, o PHMB foi facilmente absorvido pelas fibras celulósicas, com e sem alta pressão. A UHP permitiu ainda uma melhor impregnação/retenção do PHMB, após lixiviação, quando comparado com a amostra impregnada sem UHP. Estes resultados foram relacionados com as fortes interações estabelecidas entre o PHMB catiónico e a celulose, devido a uma impregnação mais profunda do agente antimicrobiano durante o tratamento de UHP. No entanto, o teor de PHMB a impregnar nas fibras é limitado pelos efeitos negativos que este originou nas fibras como consequência da sua ação de interrupção das ligações entre fibras como “agente desligante”. Do mesmo modo, cápsulas de sílica com PHMB também demonstraram uma maior retenção nas fibras celulósicas com a utilização da UHP. Encapsulado, o PHMB não está diretamente em contacto com a celulose (interações electroestáticas), exibindo deste modo uma maior libertação de PHMB (limitação por difusão). A impregnação com perfumes (Dovena New) encapsulados também foi realizada e revelou uma maior impregnação destas nanoestrutras, com a utilização da UHP, relativamente à metodologia convencional. Isto foi refletido pela maior libertação de voláteis por parte das amostras impregnadas com cápsulas com perfumes.

The material properties of skin are of great importance to a variety of fields such as dermatology and reconstructive surgery. Relatively little infrastructure and expertise exists locally in South Africa for testing biological tissue. The difficulties of testing the material properties of skin are the non-uniformity and anisotropy across specimen location and subjects. This anisotropy is most commonly measured by tensile testing of samples cut in different orientations. However, the individual samples at different orientations would be extracted from slightly different locations on the same subject, which will naturally vary in response. Bulge testing is a method of determining response to tension in different directions at the same location, by applying biaxial tension. It uses a positive pressure applied to a peripherally clamped specimen to deform the specimen in a balloon type manner. In this project, bulge testing apparatus was designed and built for the purpose of testing skin and membrane tissue, under biaxial tension. The testing apparatus consists of a syringe pump to control inflation of a specimen, which is clamped in an inflation chamber. Digital Image Correlation (DIC) was used to capture the 3D deformation fields of the specimen, and hence infer the strain fields. To simplify commissioning testing, a commercial silicone elastomer suited for skin prosthetics, was used to manufacture specimens for uniaxial and bulge experimental testing. Two types of bulge specimens were manufactured, standard round specimens and elliptical specimens. The round specimens were used to compare their material response to uniaxial tests and the elliptical bulge specimens were used to simulate the anisotropic response of skin. The method of analysis used in this project is based on using DIC and curvature calculations at multiple points to calculate membrane stresses in principal directions. The method of calculating principal curvatures from DIC is adapted from the work by Machado et al. [1] that calculated Gaussian curvature using the first and second fundamental forms of a surface. In total 18 round, 6 elliptical and 10 uniaxial specimens were tested and the material properties were found to vary slightly between each specimen. The spread in data between the uniaxial and bulge tests was found to be very similar with the bulge data showing 10 % spread at 1.2 stretch and constant 8 % spread above 1.2 stretch and the uniaxial data showing increasing spread from 7 % to 15 %. The curvature results showed very clear principal directions of curvature for the elliptical specimens. This demonstrated that the method used in this project is capable of clearly extracting the orientations of stiffer fibre directions of skin and other collagenous tissue.

L'obésité et le diabète sont associés à des complications cutanées, en particulier des fonctions dermiques impliquées dans le maintien de la résistance mécanique de la peau. Cependant, les mécanismes par lesquels l'obésité provoque la fragilité du tissu cutané sont peu étudiés. Notre travail consistait à évaluer les effets d'un régime hypercalorique sur la microcirculation cutanée et les conséquences sur le développement de lésions cutanées. Les études ont été réalisées chez des souris C57Bl6/J développant une obésité induite par une alimentation enrichie en graisse et en sucre pendant 2, 4, 12 et 20 semaines. Les propriétés de la microcirculation cutanée sont évaluées par les variations du flux sanguin en réponse : 1) à l'acétylcholine afin de déterminer la vasodilatation endothélium-dépendante, 2) au nitroprussiate de sodium afin de déterminer la vasodilatation endothélium-indépendante, 3) à l'application de la pression locale afin de déterminer la vasodilatation induite par la pression (PIV). Ces études sont complétées par des explorations métaboliques (IPGTT, IPITT), morphologiques, immuno-histologiques et biochimiques pour caractériser les quatre modèles. Enfin, chaque modèle a été testé pour l'incidence d'ulcère de pression par l'application d'une pression unique de 85 mmHg. Dans ce travail de thèse, nous avons mis en évidence de nombreux mécanismes de compensation accompagnant l'évolution de l'obésité et qui permettent de maintenir les fonctions vasculaires intactes et permettent de s'adapter à l'environnement inflammatoire induit par l'alimentation hypercalorique. La PIV est un outil de diagnostic nous permettant d'évaluer l'intégrité de la fonction neurovasculaire et prédire l'incidence de lésions cutanées
Obesity and diabetes are associated to skin pathophysiology, particularly the dermal functions involved in maintaining the skin’s mechanical strength. The underlying mechanisms in pressure ulcer during obesity remain unclear. In this study we evaluated the effects of a hypercaloric diet on the cutaneous microcirculation and its consequences on pressure sores. C59Bl6/J mice were fed a high fat and high sugar diet during 2, 4, 12 and 20 weeks. Microvascular properties were assessed by measuring the skin blood flow variations in response to 1) acetylcholine in order to determine the endothelium-dependent vasodilation, 2) sodium nitroprusside in order to determine the endothelium-independent vasodilation, 3) local pressure application in order to determine the pressure-induced vasodilation (PIV). Each model was characterized for metabolic assessment (IPGTT, IPITT), hisological, immune-histological and biochemical measurements. Finally, each model was tested for pressure ulcer incidence with a 85 mmHg pressure application on skin layers. In this study, we found that obesity is a pathology in constant evolution that is associated with many compensatory mechanisms for maintaining vascular functions and for the adaptation of the skin tissue to an inflammatory environment induced by a hypercaloric diet. PIV has become a useful tool for pressure ulcer prediction

Hypercholesterolemia is a risk factor for osteoporosis but the underlying mechanism is unknown. Previous evidence suggests that osteoporosis results from an impaired regulation of osteoblasts by fluid pressure fluctuations in the bone matrix. Recently, our laboratory showed that enhanced cholesterol in the cell membrane, due to hypercholesterolemia, alters leukocyte mechanosensitivity. We predict a similar link between osteoblasts and hypercholesterolemia leading to osteoporosis. Specifically, we hypothesize that extracellular cholesterol modifies the osteoblast sensitivity to pressure. MC3T3-E1 cells were exposed to hydrodynamic pressures regimes (mean=40mmHg, amplitude=0-20mmHg, frequency=1Hz) for 1-12 hours. To assess the impact of membrane cholesterol enrichment, cells were pre-treated with 0-50 µg/mL cyclodextran:cholesterol conjugates. We assessed the pressure effects on mitosis and F-actin stress fiber formation (SFF) of cells. Exposure of cells to 50/30 mmHg pressure transiently increased the number of cells in the S- and G2M-phases of mitosis after 6 and 12 hours, respectively. Relative to controls, osteoblast-like cells exposed to all pressures exhibited significantly (p

Background Right ventricular (RV) diastolic function has been associated with outcomes for patients with pulmonary hypertension; however, the relationship between biomechanics and hemodynamics in the right ventricle has not been studied. Methods and Results Rat models of RV pressure overload were obtained via pulmonary artery banding (PAB; control, n=7; PAB, n=5). At 3 weeks after banding, RV hemodynamics were measured using a conductance catheter. Biaxial mechanical properties of the RV free wall myocardium were obtained to extrapolate longitudinal and circumferential elastic modulus in low and high strain regions (E-1 and E-2, respectively). Hemodynamic analysis revealed significantly increased end-diastolic elastance (E-ed) in PAB (control: 55.1 mm Hg/mL [interquartile range: 44.785.4 mm Hg/mL]; PAB: 146.6 mm Hg/mL [interquartile range: 105.8155.0 mm Hg/mL]; P=0.010). Longitudinal E1 was increased in PAB (control: 7.2 kPa [interquartile range: 6.718.1 kPa]; PAB: 34.2 kPa [interquartile range: 18.144.6 kPa]; P=0.018), whereas there were no significant changes in longitudinal E-2 or circumferential E-1 and E-2. Last, wall stress was calculated from hemodynamic data by modeling the right ventricle as a sphere: (stress = Pressure x radius/2 x thickness Conclusions RV pressure overload in PAB rats resulted in an increase in diastolic myocardial stiffness reflected both hemodynamically, by an increase in E-ed, and biomechanically, by an increase in longitudinal E-1. Modest increases in tissue biomechanical stiffness are associated with large increases in E-ed. Hemodynamic measurements of RV diastolic function can be used to predict biomechanical changes in the myocardium.

This study explored hydrostatic pressure as a mechanobiological parameter to control in vitro endothelial cell tubulogenesis in 3-D hydrogels as a model microvascular tissue engineering approach. For this purpose, the present investigation used an endothelial spheroid model, which we believe is an adaptable microvascularization strategy for many tissue engineering construct designs. We also aimed to identify the operating magnitudes and exposure times for hydrostatic pressure-sensitive sprout formation as well as verify the involvement of VEGFR-3 signaling. For this purpose, we used a custom-designed pressure system and a 3-D endothelial cell spheroid model of sprouting tubulogenesis. We report that an exposure time of 3 days is the minimum duration required to increase endothelial sprout formation in response to 20 mmHg. Notably, exposure to 5 mmHg for 3 days was inhibitory for endothelial spheroid lengths without affecting sprout numbers. Moreover, endothelial spheroids exposed to 40 mmHg also inhibited sprouting activity by reducing sprout numbers without affecting sprout lengths. Finally, blockade of VEGFR-3 signaling abolished the effects of the 20-mmHg stimuli on sprout formation. Based on these results, VEGFR-3 dependent endothelial sprouting appears to exhibit a complex pressure dependence that one may exploit to control microvessel formation.

The symptoms in Idiopathic Adult Hydrocephalus Syndrome (IAHS) – gait disturbance, incontinence, and cognitive deficit – correlate anatomically to neuronal dysfunction in periventricular white matter. The pathophysiology is considered to include a cerebrospinal fluid (CSF) hydrodynamic disturbance, including pressure oscillations (“B waves”), in combination with cerebrovascular disease. IAHS and Subcortical Arteriosclerotic Encephalopathy (SAE) show clinical similarities, which constitutes a diagnostic problem. The aim of this thesis was to investigate biochemical markers in CSF, possibly related to the pathophysiology, and their usefulness in diagnosis, to investigate the effect of ICP changes on glucose supply and metabolism in periventricular deep white matter, and to present criteria for objective, computerised methods for evaluating the content of B waves in an intracranial pressure (ICP) registration. CSF samples from 62 IAHS patients, 26 SAE patients, and 23 controls were analysed for sulfatide, total-tau (T-tau) hyperphosphorylated tau (P-tau), neurofilament protein light (NFL), and beta-amyloid-42 (Aß42). In ten IAHS patients, recordings of ICP, brain tissue oxygen tension (PtiO2), and samplings of brain extracellular fluid from periventricular white matter by way of microdialysis were performed, at rest and during a CSF infusion and tap test. Microdialysis samples were analysed for glucose, lactate, pyruvate, glutamate, glycerol, and urea. Patterns before and after spinal tap were analysed and changes from increasing ICP during the infusion test were described. The long term ICP registration was used to evaluate two computerised methods according to optimal amplitude threshold, monitoring time, and correlation to the manual visual method. In CSF, NFL was elevated in both IAHS and SAE patients, reflecting the axonal damage. In a multinominal logistic regression model, the combined pattern of high NFL, low P-tau and low Aß42 in CSF was shown to be highly predictive in distinguishing between IAHS, SAE and controls. Analysis of microdialysis samples for glucose, lactate, and pyruvate showed, in combination with PtiO2, a pattern of low-grade ischemia. After the spinal tap of CSF, the pattern changed, indicating increased glucose metabolic rate. During the infusion test, there were prompt decreases in the microdialysis values of glucose, lactate and pyruvate during ICP increase, but no sign of hypoxia. The values normalised immediately when ICP was lowered, indicating that the infusion test is not causing damage. One of the computerised methods, with an amplitude threshold set to 1 mm Hg, was shown robust in evaluating B wave content in an ICP registration. At least 5 hours registration time was needed. The highly predictive pattern of biochemical markers in CSF indicates a possibility of identifying simple tests in diagnosing and selecting patients for surgical treatment. The results of microdialysis and PtiO2 indicate low-grade ischemia in the periventricular white matter, which is ameliorated from CSF removal, and that glucose supply and metabolism are sensitive to short-term ICP elevations, thus proposing a link between ICP oscillations and symptoms from neuronal disturbance. A computerised method for evaluation of B waves is a prerequisite for evaluating the impact of pressure oscillations in the pathophysiology of IAHS.

This thesis aimed at studying mechanisms involved in control of tissue fluid homeostasis during inflammation.

The interstitial fluid pressure (P_IF) is of importance for control of tissue fluid balance. A lowering of P_IF in vivo will result in a transport of fluid from the circulation into the tissue, leading to edema. Loose connective tissues that surround blood vessels have an intrinsic ability to take up fluid and swell. The connective tissue cells exert a tension on the fibrous network of the tissues, thereby preventing the tissues from swelling. Under normal homeostasis, the interactions between the cells and the fibrous network are mediated by β1 integrins. Connective tissue cells are in this way actively controlling P_IF.

Here we show a previously unrecognized function for the integrin αVβ3, namely in the control of P_IF. During inflammation the β1 integrin function is disturbed and the connective tissue cells release their tension on the fibrous network resulting in a lowering of P_IF. Such a lowering can be restored by platelet-derived growth factor (PDGF) -BB. We demonstrated that PDGF-BB restored P_IF through a mechanism that was dependent on integrin αVβ3. This was shown by the inability of PDGF-BB to restore a lowered P_IF in the presence of anti-integrin β3 IgG or a peptide inhibitor of integrin αVβ3. PDGF-BB was in addition unable to normalize a lowered P_IF in β3 null mice. Furthermore, we demonstrated that extracellular proteins from Streptococcus equi modulated αVβ3-mediated collagen gel contraction. Because of the established concordance between collagen gel contraction in vitro and control of P_IF in vivo, a potential role for these proteins in control of tissue fluid homeostasis during inflammation could be assumed. Sepsis and septic shock are severe, and sometimes lethal, conditions. Knowledge of how bacterial components influence P_IF and the mechanisms for tissue fluid control during inflammatory reactions is likely to be of clinical importance in treating sepsis and septic shock.

L’obésité et le diabète sont associés à des complications, notamment une fragilité cutanée. Celle-ci pourrait être associée à une altération du métabolisme du glucose et à une augmentation du tissu adipeux sous-cutané. Il a été montré que l’hypoderme pourrait avoir une fonction spécifique pour la peau. Notre travail a consisté d’une part à caractériser l’adiposité hypodermique et à étudier son influence dans la réactivité du tissu cutané en réponse à des pressions faibles ou fortes chez des souris obèses et d’autre part à étudier le processus de cicatrisation. Les fortes pressions augmentent la fragilité cutanée ce qui peut aboutir à des lésions, notamment des ulcères de pression. Les études ont été réalisées chez des souris C57BL/6J développant une obésité induite par une alimentation riche en lipides et en sucres pendant 4 et 12 semaines. La taille des adipocytes de l’hypoderme a été mesurée ainsi que leur réponse lipolytique en présence ou non d’insuline. La réactivité tissulaire a été évaluée en mesurant les variations du flux sanguin en réponse : 1) à l’application d’une pression locale faible afin de déterminer la vasodilatation induite par la pression (PIV), 2) par iontophorèse d’acétylcholine ou de nitroprussiate de sodium. Afin d’explorer le processus spécifique d’ulcère de pression chez les souris obèses diabétiques, une compression par cycles d’ischémie-reperfusion a été réalisée. Ces études ont été complétées par des explorations métaboliques, histologiques et biochimiques. Par ailleurs nous avons déterminé l’impact de l’augmentation de l’adiposité sur des fibroblastes dermiques in vitro afin de mieux comprendre le processus de cicatrisation.Dans ce travail de thèse, nous avons mis en évidence une augmentation de l’adiposité hypodermique associée à une insulinorésistance tissulaire et systémique. Nous avons également montré un retard de cicatrisation en fonction de l’évolution de l’obésité et des réponses micro vasculaires diminuées post cicatrisation par rapport à une peau non lésée
Obesity and diabetes led to complications, including skin fragility. Skin fragility could be associated to a glucose metabolism alteration and a subcutaneous adipose tissue increase. It has been shown that hypodermis could have a specific function for the skin. Our work consisted on the one hand in characterizing the dermal adiposity and studying its involvement in the skin tissue reactivity in response to low or high pressures in obese mice and, on the other hand, in studying the healing process. High pressures increase cutaneous fragility which can lead to skin wounds, in particular pressure ulcers. This study was realized using C57BL/6J male mice with a diet-induced obesity. C57BL/6J mice were fed a high fat and high sugar diet during 4 or 12 weeks. Hypodermis adipocytes size was measured as well as their lipolytic response in presence or absence of insulin. The skin tissue reactivity was assessed measuring the skin blood flow variations in response to 1) a local pressure application in order to determine the pressure-induced vasodilation (PIV), 2) an acetylcholine or sodium nitroprusside iontophoresis. To examine the specific mechanism of the pressure ulcer in obese diabetic mice, a compression with ischemia-reperfusion cycles was realized. Metabolic assessment, histological and molecular biological studies were carried out to characterize each stage of healing through obesity. Furthermore, we determined the adiposity increase on dermal fibroblasts in vitro to better understand the healing process. In this thesis work, we have highlighted a hypodermis adiposity linked to a tissue and a systemic insulin resistance. We have also showed a delayed healing depending on the evolution of the obesity. The microvascular responses were decreased post healing compared to a non-wounded skin

The neurological wake-up test, NWT, is a clinical monitoring tool that can be used to evaluate the level of consciousness in patients with traumatic brain injury (TBI) and subarachnoid haemorrhage (SAH) during neurocritical care (NCC). Since patients with severe TBI or SAH are often treated with mechanical ventilation and sedation, the NWT requires that the continuous sedation is interrupted. However, interruption of continuous sedation may induce a stress response and the use of the NWT in NCC is controversial. The effects of the NWT on intracranial pressure (ICP) and cerebral perfusion pressure (CPP) were evaluated in 21 patients with TBI or SAH. Compared to baseline when the patients were sedated with continuous propofol sedation, the NWT resulted in increased ICP and CPP (p<0.05). Next, the effects of the NWT on the stress hormones adrenocorticotrophic hormone (ACTH), cortisol, epinephrine and norepinephrine were evaluated in 24 patients. Compared to baseline, the NWT caused a mild stress response resulting in increased levels of all evaluated stress hormones (p<0.05). To compare the use of routine NCC monitoring tools, the choice of sedation and analgesia and the frequency of NWT in Scandinavian NCC units, a questionnaire was used. The results showed that all 16 Scandinavian NCC units routinely use ICP and CPP monitoring and propofol and midazolam were primary choices for patient sedation in an equal number of NCC units. In 2009, the NWT was not routinely used in eight NCC units whereas others used the test up to six times daily. Finally, intracerebral microdialysis (MD), brain tissue oxygenation (PbtiO2) and jugular bulb oxygenation (SjvO2) were used in 17 TBI patients to evaluate the effect of the NWT procedure on focal neurochemistry and cerebral oxygenation. The NWT did not negatively alter interstitial markers of energy metabolism or cerebral oxygenation. In conclusion, the NWT induced a mild stress response in patients with TBI or SAH that did not result in a detectable, significant secondary insult to the injured brain. These results suggest that the NWT may safely be used as a clinical monitoring tool in the NCC of severe TBI and SAH in a majority of patients.

This thesis explores physiological changes occurring after acute brain injury. The first two chapters focus on traumatic brain injury (TBI), a significant cause of disability and death worldwide. I discuss the evidence behind current management of secondary brain injury with emphasis on partial brain oxygen tension (PbtO2) and intracranial pressure (ICP). The second chapter describes a subgroup analysis of the effect of hypothermia on ICP and PbtO2 in 17 patients enrolled to the Eurotherm3235 trial. There was a mean decrease in ICP of 4.1 mmHg (n=9, p < 0.02) and a mean decrease in PbtO2 (7.8 ± 3.1 mmHg (p < 0.05)) in the hypothermia group that was not present in controls. The findings support previous studies in demonstrating a decrease in ICP with hypothermia. Decreased PbtO2 could partially explain worse outcomes seen in the hypothermia group in the Eurotherm3235 trial. Further analysis of PbtO2 and ICP guided treatment is needed. The third chapter focuses on delayed cerebral ischaemia (DCI) after aneurysmal subarachnoid haemorrhage (aSAH), another form of acute brain injury that causes significant morbidity and mortality. I include a background of alpha-calcitonin gene-related peptide (αCGRP), a potential treatment of DCI, along with results from a systematic review and meta-analysis of nine experimental models investigating αCGRP. The meta-analysis demonstrates a 40.8 ± 8.2% increase in cerebral vessel diameter in those animals treated with αCGRP compared with controls (p < 0.0005, 95% CI 23.7 to 57.9). Neurobehavioural scores were reported in four publications and showed a Physiological responses to brain tissue hypoxia and blood flow after acute brain injury standardised mean difference of 1.31 in favour of αCGRP (CI -0.49 to 3.12). I conclude that αCGRP reduces cerebral vessel narrowing seen after SAH in animal studies but note that there is insufficient evidence to determine its effect on functional outcomes. A review of previous trials of αCGRP administration in humans is included, in addition to an original retrospective analysis of CSF concentrations of αCGRP in humans. Enzyme-linked immunosorbent assay of CSF (n = 22) was unable to detect αCGRP in any sample, which contrasts with previous studies and was likely secondary to study methodology. Finally, I summarise by discussing a protocol I designed for a dose-toxicity study involving the intraventricular administration of αCGRP to patients with aSAH and provide some recommendations for future research. This protocol was based upon the systematic review and was submitted to the Medical Research Council's DPFS funding stream during the PhD.

The effects of mechanical forces on endothelial cell function and behavior are well documented, but have not been fully characterized. Specifically, fluid pressure has been shown to elicit physical and chemical responses known to be involved in the initiation and progression of endothelial cell-mediated vascularization. Central to the process of vascularization is the formation of tube-like structures. This process—tubulogenesis—is essential to both the physiological and pathological growth of tissues. Given the known effects of pressure on endothelial cells and its ubiquitous presence in the vasculature, we investigated pressure as a magnitude-dependent parameter for the regulation of endothelial tubulogenic activity. To accomplish this, we exposed two- and three-dimensional bovine aortic endothelial cell (BAEC) cultures to static pressures of 0, 20, and 40 mmHg for 3 and 4 days. The most significant findings were: (1) cells in two-dimensional culture exposed to 20, but not 40, mmHg exhibited significantly (p < 0.05) increased expression of both VEGF-C and VEGFR-3, and (2) cells in three-dimensional culture exposed to 20, but not 40, mmHg exhibited significant (p > 0.05) increases in endothelial sprouting. These findings evidence the utility of pressure as a selective modulator of tissue microvascularization in vitro and implicates pressure as factor in pathological tubulogenesis in vivo.

In vitro tissue engineering is a method for developing living and functional tissues external to the body, often within a device called a bioreactor to control the chemical and mechanical environment. However, the quality of bone tissue engineered products is currently inadequate for clinical use as the implant cannot bear weight. In an effort to improve the quality of the construct, hydrostatic pressure, the pressure in a fluid at equilibrium that is required to balance the force exerted by the weight of the fluid above, has been investigated as a mechanical stimulus for promoting extracellular matrix deposition and mineralisation within bone tissue. Thus far, little research has been performed into understanding the response of bone tissue cells to mechanical stimulation. In this thesis we investigate an in vitro bone tissue engineering experimental setup, whereby human mesenchymal stem cells are seeded within a collagen gel and cultured in a hydrostatic pressure bioreactor. In collaboration with experimentalists a suite of mathematical models of increasing complexity is developed and appropriate numerical methods are used to simulate these models. Each of the models investigates different aspects of the experimental setup, from focusing on global quantities of interest through to investigating their detailed local spatial distribution. The aim of this work is to increase understanding of the underlying physical processes which drive the growth and development of the construct, and identify which factors contribute to the highly heterogeneous spatial distribution of the mineralised extracellular matrix seen experimentally. The first model considered is a purely temporal model, where the evolution of cells, solid substrate, which accounts for the initial collagen scaffold and deposited extracellular matrix along with attendant mineralisation, and fluid in response to the applied pressure are examined. We demonstrate that including the history of the mechanical loading of cells is important in determining the quantity of deposited substrate. The second and third models extend this non-spatial model, and examine biochemically and biomechanically-induced spatial patterning separately. The first of these spatial models demonstrates that nutrient diffusion along with nutrient-dependent mass transfer terms qualitatively reproduces the heterogeneous spatial effects seen experimentally. The second multiphase model is used to investigate whether the magnitude of the shear stresses generated by fluid flow, can qualitatively explain the heterogeneous mineralisation seen in the experiments. Numerical simulations reveal that the spatial distribution of the fluid shear stress magnitude is highly heterogeneous, which could be related to the spatial heterogeneity in the mineralisation seen experimentally.

Fluid pressures regulate endothelial cell (EC) tubulogenic activity involving fibroblast growth factor 2 (FGF-2) and its receptor, FGF receptor 2 (FGFR2). Our lab has recently shown that sustained 20 mmHg hydrostatic pressure (HP) upregulates EC sprout formation in a FGF2-dependent fashion. This upregulation of sprout formation may be due to enhanced FGF-2 / FGFR2 interactions in the presence of 20 mmHg HP. We hypothesize that exposure of ECs to 20 mmHg sustained HP enhances FGF-2 binding kinetics. We used a custom hydrostatic pressure system, immunofluorescence, and FACS to quantify FGF-2 binding by ECs in the absence or presence of a range of HPs for 30 minutes. Relative to cells maintained under control pressure, ECs exposed to 20, but neither 5 nor 40 mmHg, displayed a significant increase in binding affinity to FGF-2. EC binding of VEGF-A, another angiogenic growth factor, was unaffected by similar pressure stimuli. Additional studies showed that pressure-selective FGF-2 binding was independent of FGFR2 surface expression. These results implicate the FGF-2 axis in the pressure-sensitive, magnitude-dependent angiogenic processes which we have previously described. The present study provides novel insight regarding the involvement of FGF-2 signaling and interstitial pressure changes in various microvascular physiological and pathobiological processes.

The purpose of this thesis is to examine the effects of inhaled corticosteroid drugs (ICs) on the voice due to their frequent use in treating an increasing prevalence of asthma disorders. As part of a larger five-year study, the focus of this thesis is specifically on whether 8 weeks of in vivo exposure to ICs will cause changes in the sustained subglottal pressure, sustained airflow, and visual-perceptual ratings of edema and erythema in excised rabbit larynges. Researchers administered either ICs or a control nebulized isotonic saline solution to 22 rabbits in vivo, sacrificed them, and harvested their larynges for benchtop research. While ensuring proper tissue preservation, researchers then finely dissected the larynges to expose the true vocal folds and run phonation trials. Dependent variables included continuous acoustic signals (Hz), subglottal pressure (cm H2O), and airflow (L/min) data for 15 phonation trials per rabbit larynx. Researchers also collected still image photographs at this time and subsequently normalized them for use in the visual-perceptual portion of this thesis. For visual-perceptual ratings, raters used a 0-3 equal appearing interval scale to rate aspects of edema and erythema on left and right vocal fold and arytenoid tissues. Results indicate that, when compared to control larynges exposed to nebulized isotonic saline, experimental larynges treated with ICs require significantly higher subglottal pressure to maintain phonation, p < .05. Mean sustained phonation for experimental larynges is 11.24 cm H2O compared to 8.92 cm H2O for that of control larynges. Phonation trials for experimental larynges have significantly higher sustained airflow with a mean of 0.09 L/min compared to 0.07 L/min for that of control larynges, p < .05. Surprisingly, experimental larynges have higher average fundamental frequencies with less variability (mean: 519 Hz, standard deviation: 66 Hz) than that of control larynges (mean: 446 Hz, standard deviation: 130 Hz). On visual-perceptual ratings, experimental larynges have significantly higher severity ratings on all eight items rated, p < .0001 - p = .0305. Based on these results, it is concluded that ICs cause significant damage to rabbit vocal folds, as evidenced by higher sustained pressure, higher airflow, and higher severity ratings for experimental versus control larynges. The dependent variables in this thesis are novel in benchtop model research and demonstrate a unique perspective on this research question. Thus, this thesis informs future phonation, benchtop, and visual-perceptual research.

Outcome after traumatic brain injury (TBI) depends on the extent of primary cell death and on the development of secondary brain injury. The general aim of this thesis was to find strategies and quality systems to minimize the extent of secondary insults in neurointensive care (NIC). An established standardized management protocol system, multimodality monitoring and computerized data collection, and analysis systems were used. The Uppsala TBI register was established for regular monitoring of NIC quality indexes. For 2008-2010 the proportion of patients improving during NIC was 60-80%, whereas 10% deteriorated. The percentage of ‘talk and die’ cases was < 1%. The occurrences of secondary insults were less than 5% of good monitoring time (GMT) for intracranial pressure (ICP) > 25 mmHg, cerebral perfusion pressure (CPP) < 50 mmHg and systolic blood pressure < 100 mmHg. Favorable outcome was achieved by 64% of adults. Nurse checklists of secondary insult occurrence were introduced. Evaluation of the use of nursing checklists showed that the nurses documented their assessments in 84-85% of the shifts and duration of monitoring time at insult level was significantly longer when secondary insults were reported regarding ICP, CPP and temperature. The use of nurse checklist was found to be feasible and accurate.  A clinical tool to avoid secondary insults related to nursing interventions was developed. Secondary brain insults occurred in about 10% of nursing interventions. There were substantial variations between patients. The risk ratios of developing an ICP insult were 4.7 when baseline ICP ≥ 15 mmHg, 2.9 when ICP amplitude ≥ 6 mmHg and 1.7 when pressure autoregulation ≥ 0.3. Hyperthermia, which is a known frequent secondary insult, was studied. Hyperthermia was most common on Day 7 after admission and 90% of the TBI patients had hyperthermia during the first 10 days at the NIC unit. The effects of hyperthermia on intracranial dynamics (ICP, brain energy metabolism and BtipO2) were small but individual differences were observed. Hyperthermia increased ICP slightly more when temperature increased in the groups with low compliance and impaired pressure autoregulation. Ischemic pattern was never observed in the microdialysis samples. The treatment of hyperthermia may be individualized and guided by multimodality monitoring.

Abstract The principal aim of a population-based cross-sectional survey was to generate information on the lifetime occurrence of prostatitis in Finnish men and their exposure to the disease, and also on the influence of prostatitis-related fears and disturbances on their sexual life. A second aim was to develop and clinically validate a new diagnostic tool for differential diagnosis between the forms of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), especially between patients belonging to categories IIIA and IIIB in the new NIH (National Institutes of Health) clinical classification. Altogether 1832 men out of 2500 aged 20–59 years chosen randomly from the two most northerly provinces of Finland (Oulu and Lapland) participated in the epidemiological study, a response rate of 75%. The overall lifetime prevalence of prostatitis was 14.2%. The risk of having had the disease increased with age, being 1.7 times greater in the men aged 40–49 years than in those aged 20–39 years, and 3.1 times greater in those aged 50–59 years. More than a quarter of the 261 men who had or had had prostatitis symptoms (27%) suffered from them at least once a year, while 16% suffered from chronic prostatitis symptoms throughout the year. 63% of the men with prostatitis had their worst symptoms during the wintertime (November–march). 17% of the men with chronic prostatitis reported a constant fear of undetected prostate cancer. Erectile dysfunction was reported by 43% of the symptomatic men and decreased libido by 24%. Self-assessment of personality showed that the men with prostatitis were more often busy and nervous and had a meticulous attitude to life and problems than were the non-symptomatic men. 197 patients with chronic prostatitis/chronic pelvic pain syndrome participated in three clinical case-control studies during the years 1995–2000, at Oulu University Hospital, the District Hospital of Oulainen and Seinäjoki Central Hospital. The first prostatic tissue pressure measurement (PTPM) study included 34 patients and 9 controls. A novel method was developed to measure intraprostatic tissue pressure with a Stryker® intracompartmental pressure monitor. The PTPM showed a clear increase (p < 0.001) in the patients with symptoms of prostatitis and benign prostatic enlargement (BPE) relative to the controls and the patients with BPE but without pain symptoms. The second PTPM study included 42 patients with chronic prostatitis symptoms without significant BPE and 12 new controls. Significantly higher pressure readings (p < 0.001) were recorded at all three measurement points in the patients than in the controls. 48 new patients and 12 new controls were enrolled for the third PTPM study, the purpose of which was to confirm the results of the previous ones and to compare the prostatic tissue pressures of two clinical groups (IIIA and IIIB). The prostatic tissue pressure was again significantly higher in the patients with chronic prostatitis symptoms than in the controls (p < 0.001). An interesting finding was that prostatitis patients belonging to clinical category IIIA had significantly higher tissue pressures (p < 0.01) than those in category IIIB, probably reflecting more severe inflammation in the prostatic tissue. This new PTPM method provides a more precise and/or exact tool for differential diagnosis between the forms of pelvic pain and CP/CPPS.

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro
O consumo excessivo de gorduras tem sido implicado na gênese da obesidade e de diversas comorbidades relacionadas a esta doença. O objetivo do presente estudo foi avaliar o efeito do treinamento resistido em ratos alimentados com dieta hiperlipídica nas seguintes variáveis: composição corporal, adiposidade, área de adipócitos, expressão do fator de crescimento do endotélio vascular (VEGF) e dos receptores ativados por proliferadores de peroxissomo tipo gama (PPAR-γ) no tecido adiposo retroperitoneal, níveis pressóricos e atividade da metaloproteinase-2 (MMP-2) no ventrículo esquerdo. Foram avaliados 32 ratos machos Wistar divididos em quatro grupos experimentais (n=8/cada) de acordo com o tipo de dieta e o treinamento: sedentário (SED; dieta padrão), sedentário obeso [SED-OB; dieta hiperlipídica (30% de gordura)], treinamento resistido (TR; dieta padrão) e treinamento resistido obeso (TR-OB; dieta hiperlipídica). Após o desmame (dia 21), os animais foram submetidos à dieta experimental durante 24 semanas. Doze semanas após início da dieta, os grupos TR e TR-OB iniciaram o período de 12 semanas de treinamento resistido. Este era composto por escaladas em uma escada vertical de 1.1 metros com pesos atados a cauda num total de três sessões de treinamento por semana (Segundas, Quartas e Sextas-feiras) com 4 a 9 escaladas/sessão e 8 a 12 movimentos dinâmicos a cada subida. O treinamento resistido induziu reduções significativas na massa corporal, na massa gorda, no percentual de gordura, na área de adipócitos e nos níveis pressóricos e ainda promoveu aumento de massa magra e na expressão de VEGF e PPAR-γ em ambos os grupos treinados. Além disso, foi observada uma maior atividade da MMP-2 no ventrículo esquerdo nos grupos treinados (TR e TR-OB). Nossos achados demonstram que o treinamento resistido foi capaz de promover mudanças positivas na composição corporal, na atividade da MMP-2 no ventrículo esquerdo, nos níveis pressóricos e em mediadores enzimáticos do tecido adiposo retroperitoneal sugerindo que esta modalidade de exercício pode ser uma ferramenta útil para prevenir as alterações induzidos pelo consumo de uma dieta hiperlipídica.
The purpose of the present study was to evaluate the effect of resistance training on body composition, adiposity, adipocyte area, vascular endothelium growth factor (VEGF) and peroxisome proliferator-activated receptor-gamma (PPAR-γ) in adipose tissue, systolic and diastolic blood pressures (BP), and activity of muscle MMP-2 in left ventricle. We evaluated 32 Wistar rats divided into four experimental groups (n=8/each) according to diet and exercise status: sedentary (SED; standard diet), sedentary obese [SED-OB; hiperlipidic diet (30% of fat)], resistance training (RT; standard diet) and resistance training RT obese (RT-OB; hiperlipidic diet). After weaning (day 21), animals were subjected to the experimental diet according to their groups during 24 weeks. A 12-week resistance-training period was used, during which the animals climbed a 1.1-m vertical ladder with weights attached to their tails. The sessions were performed three times/week (Mondays, Wednesdays and Fridays), with 49 climbs/session and 812 dynamic movements/climb with two rest minutes. The RT promoted significantly reductions of body and fat masses, fat percentage, adipocytes area and systolic and diastolic BPs, associated with higher expression of VEGF and PPAR-γ in retroperitoneal adipose tissue and higher fat free mass in both trained groups. Furthermore, it was observed higher muscle MMP-2 activity of the left ventricle in trained animals. This study demonstrated that, resistance training induced positive changes in body composition, MMP-2 activity in left ventricle, systolic and diastolic BPs and adipose tissue metabolism, suggesting that it can be a useful tool to prevent the physiopathological changes induced by high-fat diet consumption.

mestrado em Engenharia Química
Este trabalho consiste no estudo das alterações das propriedades papeleiras através da modificação estrutural de pasta kraft de Eucalyptus globulus pela implementação de alta pressão hidrostática (TH) de modo a que as propriedades da pasta sejam as adequadas para a sua aplicação, papel tissue de baixa gramagem. Este trabalho consiste no estudo da aplicação da alta pressão hidrostática ao nível das fibras celulósicas de modo a verificar qual a influência que este tipo de tratamento apresenta nas propriedades papeleiras, tais como as mecânicas e estruturais. Assim, começou-se por implementar um TH em pasta kraft não refinada, tendo em estudo quatro gamas de pressão diferentes, 4000 – 7000 bar, numa pasta com uma consistência de 1,5%. Com esta aplicação verificaram-se melhorias nas propriedades mecânicas da pasta, sendo que os melhores resultados foram obtidos aquando aplicada uma pressão de 6000 bar, onde se obteve uma taxa de crescimento de 2% para o índice de rotura à tração, 1% no alongamento percentual na rotura, 1% no índice de rigidez à tração e 6% no índice de rebentamento, sendo que para estas propriedades esta gama de pressão foi a única que apresentou valores superiores ao da pasta referência. No índice de rasgamento, a única gama de pressão que apresentou uma taxa de crescimento positiva foi a 4000 bar, com cerca de 1%. De um modo geral, a pressão que apresentou melhores resultados foi a de 6000 bar, sendo esta a gama utilizada para a continuação dos estudos. De modo a verificar qual a influência da alta pressão na pasta kraft, foi refinada num moinho PFI em quatro gamas de refinação 0,500,1000 e 2000 revoluções, pasta sem tratamento de alta pressão e pasta sujeita prévio tratamento de alta pressão a 6000 bar. Assim, verificou-se que as propriedades mecânicas da pasta kraft aumentaram com o número de revoluções quer em pasta sujeita a TH quer em pasta somente refinada, embora estas sejam superiores para pasta somente refinada, à exceção do índice de rasgamento que apresenta uma taxa de crescimento de 41% para uma refinação de 500 revoluções em pasta sujeita a TH a 6000 bar e essa taxa de crescimento em pasta somente refinada só é atingida para uma refinação de 1000 revoluções. De um modo geral o tratamento hiperbárico não é vantajoso nas propriedades mecânicas quando aplicado em conjunto com a refinação. De modo a compreender melhor o efeito da alta pressão, foi feito um estudo ao grau de cristalinidade em pasta somente sujeita a alta pressão e em pasta refinada com prévio TH a 6000 bar. Neste estudo verificou-se que quando aplicada o TH a 4000 bar o grau de cristalinidade diminui, aumentando para as pressões seguintes progressivamente, facto este que se deve a ocorrência de rearranjos ao nível das cadeias de celulose, uma vez que as zonas paracristalinas sofrem recristalização. O mesmo não acontece na pasta refinada com prévio TH a 6000 bar, pois o grau de cristalinidade diminui, o que se pode dever ao facto da refinação degradar a parede celular. Como o futuro da indústria de pasta de papel está cada vez mais direcionado para papéis de baixa gramagem, papel tissue, foi inserido neste trabalho uma componente de estudo das alterações das propriedades papeleiras deste tipo com a aplicação da alta pressão hidrostática. Ao realizar este estudo verificou-se que a velocidade e a capacidade de absorção é superior no caso da pasta refinada sujeita a prévio TH a 6000 bar em relação à pasta somente refinada. A maciez apresenta o seu máximo em pasta não sujeita a qualquer tratamento, 86.2, diminuindo com a aplicação da alta pressão, tendo o seu mínimo para uma pressão de 5000 bar, 81,8, voltando a aumentar até atingir a pressão de 7000 bar.
This work consists in the study of the changes of the paper proprieties through the structural modification of the kraft pulp coming from Eucalyptus globulus by the implementation of high hydrostatic pressure (HT) so that the proprieties of the pulp be the suitable ones to its application, tissue paper of low grammage. This work aims at studying the application of the HT concerning the level of the cellulosic fibres in order to check which influence this kind of treatment presents in the paper proprieties, such as the mechanic and structural ones. Thus, a HT was first implemented in the kraft pulp which was not refined, having studied from ranges of different pressures, from 4000 to 7000 bar, in a pulp with a consistency of 1,5%. With this application some improvements were noticed concerning the mechanic proprieties of the pulp and the best results were obtained when a pressure of 6000 bar was applied, during which is obtained a raise of 2% for the tensile breaking length, 1% for the percentage on the elongation at break, 1% for the tensile strength and 6% for the burst index, having in mind that for these proprieties this range of pressure was the only one which presented values that were superior to the ones of the pulp reference. In the tear index, the only pressure range that presented a positive growth rate was the 4000 bar with about 1%. In a general way, the pressure that presented the best results was the one of 6000 bar which was the one used to the keeping on of studies. In order to verify which was the influence of the high pressure in the kraft pulp, this one was refined in a PFI mill according to four ranges of refining: 0,500,1000 and 2000 revolution, pulp without HT and pulp subjected to previous HT according to 6000 bar. Thus, it was noticed that the mechanical proprieties of the pulp were increased according to the number of revolutions in both kinds of pulps although these mechanical proprieties are superior in a pulp which is only refined, except the tear index which presents a growth rate of 41% concerning a refining of 500 revolutions in the pulp subjected to a HT according to 6000 bar. This growth rate concerning the pulp that is only refined is only obtained according to a refining of 1000 revolutions. In a general way the HT is not advantageous in the mechanical proprieties when it is applied together with the refining. In order to better understand the effect of the HT, a study was done to the level of crystallinity in the pulps only subjected to the HT and in the refined pulp with a previous HT according to 6000 bar. In this study it was applied a HT according to 4000 bar and that it increased when the following pressures where progressively increased, which was due when some rearrangements were done at the level of the cellulose chains, since the paracrystallines areas go through recrystallization. The same does not happen in the refined pulp submitted to a previous HT according 6000 bar, because the level of crystallinity decreases, which is due to the fact that the cell wall. As the future of the pulp industry is directing itself to low grammage paper, tissue paper, it was inserted in this work a study component that changes the papermaking properties of this type of paper using the application of high hydrostatic pressure . In the process of this study we found that the speed and absorption capacity is higher in the case of refined pulp with a previous HT according to 6000 bar relative to the refined pulp alone. The softness presents its maximum in the pulp not subjected to any kind of treatment 86,2, decreasing according to an application of HT, having its minimum according to a pressure of 5000 bar, 81,8, starting increasing again till it reaches a pressure of 7000 bar.

BACKGROUND Cardiac surgery patients have some of the highest reported incidence and prevalence of pressure injuries (PI). A growing subset of cardiac surgery include patients with end-stage heart failure who undergo ventricular assist device (VAD) or total artificial heart (TAH) surgery. The risk of PI and their natural history of development in this population are unknown and the specific risk factors for PI development remain unexplored. OBJECTIVES To perform a systematic review of the literature to identify the incidence and risk factors of PI development in patients undergoing VAD-TAH surgery and thereby inform study design and variables in an eight-year retrospective study of all patients undergoing VAD-TAH surgery at a large academic university medical center. METHODS The preferred reporting items for systematic reviews and meta-analyses or PRISMA statement guided this systematic review. Quality of evidence was determined using the Johns Hopkins Nursing Evidence-Based Practice Rating Scale. Two reviewers independently appraised manuscripts matching the eligibility criteria for study inclusion. Four databases including PubMed, CINAHL, Web of Science, Google Scholar, and hand searches of journals based on reference lists from included studies were utilized. Initial results of this primary search revealed zero studies that met inclusion and this search methodology was confirmed by medical librarian consultation. Therefore, a follow up retrospective study was necessary to identify incidence of PI in the VAD-TAH population. However, a secondary search, dropping keywords of VAD-TAH and instead focusing on studies of on-pump cardiac surgery and mixed surgical studies where cardiac surgery patients were included, was conducted to establish variables to guide a retrospective study of all VAD-TAH surgeries between 2010-2018. The retrospective study evaluated the incidence of pressure ulcers by case, patient and incidence density for each of the respective 1000 patient days during the study period. Univariate statistics are reported by four different VAD-TAH devices. Variables significant in bivariate analysis were entered in a stepwise logistic regression model. RESULTS In the systematic review, 312 articles were identified from the databases with eight additional articles from hand searches. Following abstract review, 208 were excluded for not meeting inclusion criteria or study quality metrics. 77 articles were read in full, with 61 excluded, leaving 16 articles for inclusion. 31 risk factors were identified for PI development in on-pump cardiac surgery patients with 11 risk factors which were identified as significant in multivariate analysis for inclusion in the retrospective study.

L’escarre est une pathologie liée à l’immobilité des patients, accidentelle ou associée à des comorbidités. Les premières lésions apparaissent dans le muscle avant de se développer en plaie cutanée sans que les mécanismes physiopathologiques de cette atteinte ne soient encore connus. L’objectif principal de cette thèse était d’identifier des voies de signalisation intervenant de manière précoce dans le développement des escarres au travers d’une étude transversale. Nous formulons l’hypothèse qu’une compression musculaire induit une altération de l’homéostasie calcique musculaire par atteinte des canaux calciques du réticulum sarcoplasmique (les récepteur de la ryanodine de type 1, RyR1) conduisant à la lésion du tissu musculaire et une inflammation du tissu sous-cutané.Sur un modèle animal de compression de 2 heures, à 100 mmHg, nous avons identifié une initiation des voies apoptotiques et une augmentation du stress oxydant des muscles de la paroi abdominale. Le RyR1 y est hyper-nitrosylé et hyper-oxidé et sa protéine régulatrice, calstabin1 se dissocie sous l’action de ce remodelage, ce qui entraîne une fuite calcique du réticulum sarcoplasmique vers le cytosol. Cette dysfonction n’est pas réversible à 3 jours post-compression mais il est possible de la prévenir en traitant les souris avec un rycal qui bloque la déplétion de la calstabin1. En clinique, chez une cohorte de patients paraplégiques, porteurs d’escarres, nous avons identifiés un remodelage du RyR1 dans les muscles paralysés (comparaison intra patient avec une biopsie saine) et une hypoxie des tissus sous la lésion médullaire. La dissociation de la calstabin1 au RyR1 a pu être corrélée à la pression moyenne et maximale exercée sur la peau de la zone sacrée du patient allongé en regard du muscle biopsié.Ce travail de thèse a permis de préciser les voies de signalisation intervenant de manière précoce dans le développement des escarres dans le muscle squelettique. Une compression mécanique induit une augmentation du stress oxydant, un remodelage du RyR1 et une dysfonction du canal à cause de la perte de l’interaction RyR1/calstabin1. Ces résultats ouvrent des perspectives intéressantes sur des traitements préventifs pharmacologiques et de suivi non-invasif qui permettront de retarder l’apparition des premières lésions musculaires
Pressure ulcer is a pathology related to patient immobility, which can be either accidental or incidental to comorbidities. The first damage are located in muscle tissue before developing in cutaneous breakdown per an unclear pathophysiology. The core objective of my PhD was to identify the early signaling pathways involved in pressure ulcer development, through a transversal study. We hypothesized that muscle compression will induce a calcium imbalance in muscles by a dysfunction of calcium channels from the sarcoplasmic reticulum (Ryanodine receptor isoform 1, RyR1) which will lead to muscle damage and sub-cutaneous inflammation.Mice model of a 100 mmHg, 2 hours compression of abdominal muscles was used to identify the apoptotic pathway initiation and a rise of oxidative stress. RyR1 is hyper-nitrosylated and hyper-oxydated thus this remodeling induces depletion of RyR1 stabilizing protein, calstabin1, and the resulting leaky phenotype increases intracellular calcium concentration. This channel functional impairment was not reversible up to 3 days post-compression but it was possible to prevent it through rycal treatment, protecting the binding calstabin1/RyR1. In a clinical trial, we identified from a paraplegic population with existing pressure ulcers, a RyR1 remodeling in paralyzed muscles (intra patient comparison with a healthy muscle biopsy) and a hypoxia of tissues below the spinal cord injury. Calstabin1 dissociation was correlated to the mean and peak pressure intensity of interface pressure applied over the sacrum skin of the bedridden patient directly above the biopsy location.This thesis project focused on early signaling pathways participating in pressure ulcer in skeletal muscle. A mechanical strain induces an increase of the intracellular redox state, post translational RyR1 modifications and a channel dysfunction because of calstabin1 depletion. The significance of my work is to propose both pharmacology and non-invasive monitoring solutions to prevent first muscle damage in pressure ulcer development

Thesis (M. S.)--Chemical and Biomolecular Engineering, Georgia Institute of Technology, 2005.
Dr. Athanassios Sambanis, Committee Member ; Dr. Timothy M. Wick, Committee Member ; Dr. Ajit P.Yoganathan, Committee Chair. Includes bibliographical references.

Vid flertalet hjärtsjukdomar kan det förekomma störningar i den diastoliska funktionen, detta tillstånd benämns diastolisk dysfunktion. Detta innebär att fyllnadstrycken i vänsterkammare ökar på grund av nedsatt eftergivlighet i kammaren. Bedömning av diastolisk funktion, hos patienter som utvecklat förmaksflimmer, är en utmaning inom ekokardiografi. Detta beror på att förmaksflimmer innebär utebliven förmakskontraktion, oregelbunden längd av hjärtcykeln och förmaksdilatation, vilket försvårar bedömningen. Syftet med studien var att med ekokardiografi studera diastoliska parametrar hos patienter med förmaksflimmer för att studera om dessa kan användas vid bedömning av den diastoliska vänsterkammarfunktionen hos denna patientgrupp. I studien inkluderades 37 deltagare med förmaksflimmer som var remitterade för en ekokardiografisk undersökning med olika frågeställningar. Pulsad doppler teknik och vävnads doppler teknik användes för att registrera följande diastoliska parametrar: förmaksvolym, mitralisinflöde (E-vågshastigheten) och myokardiets diastoliska hastigheter (e´). Utöver dessa uppskattades även ejektionsfraktion, hjärtfrekvens, hypertrofi och trycket i lilla kretsloppet, som togs med vid bedömningen. Mann-Whitneys test visade att det förelåg ett starkt statistiskt samband mellan fyllnadstrycket (E/e´) och E-vågen, e ´samt förmaksvolym (p = <0,05). Signifikant resultat erhölls även för sambandet mellan PA-tryck och fyllnadstryck (p = 0,014) genom ett chitvå-test. Vidare gav multipel linjär regression utslag på E-vågen och e´. Analysen visade att det förelåg en hög förklaringsgrad för E-vågen (p = <0,001) och e´ (p = 0,008). Sammanfattningsvis visade resultaten att ekokardiografi kan användas för diagnostik av förmaksflimmerpatienter avseende fyllnadstryck där förmaksvolym, E-vågshastigheten och e´ anses vara bästa parametrarna.
In the majority of heart diseases disturbances in the diastolic function may occur, this condition is called diastolic dysfunction. This means that the left ventricular filling pressure increases due to reduced compliance in the chamber. The assessment of diastolic function in patients who have developed atrial fibrillation (AF), is a challenge in echocardiography. This is a result of AF which involves absence of atrial contraction, irregular length of the cardiac cycle and left atrium dilatation that complicates the assessment. The aim of this study was to observe the diastolic echocardiographic parameters in patients with AF to examine if these can be used in the assessment of diastolic left ventricular function in this population. The study included 37 participants with AF who were remitted for an echocardiographic examination due to various concerns. Pulsed Doppler technique and tissue Doppler technique was used to record the following diastolic parameters: atrial volume, mitral inflow velocity (E) and the myocardial diastolic velocity (e'). Ejection fraction, heart-rate, hypertrophy and pulmonary artery pressure were also estimated and included in the assessment. Mann-Whitneys test showed that there was a strong statistical correlation between the filling pressure (E/e') and E, e' and atrial volume (p = <0.05). Significant results were also obtained for the relation between pulmonary artery pressure and the filling pressure (p = 0.014) by a chi-square test. A multiple linear regression showed association between E and e'. The analysis showed that there was a significant value of coefficient of determination for E (p = <0.001) and e' (p = 0.008). In conclusion, the results showed that echocardiography can be used for diagnosis of AF patients regarding filling pressures, where atrial volume, E velocity and e' are considered to be the best parameters.

Обсяг дипломної роботи складає 72 сторінки, містить 64 ілюстрації, 17 таблиць. Загалом було опрацьовано 23 літературних джерела. Актуальність: на сьогоднішній день не існує повноцінного інструменту для зварювання легень, який зміг би гарантовано впоратись з особливо твердими її частинами, такими як бронхи, замість цього використовують загальні інструменти для зварювання м’яких тканин. Структура легень неоднорідна, окрім паренхіми, бронхів, альвеол, трахеї, кровоносних судин легені, під час зварювання, легені заповнені повітрям, що привносить складнощі в процес зварювання. Це вимагає від інструменту підвищених параметрів надійності та міцності. Мета: модель та дослідження на статичне навантаження інструменту для зварювання легень. Завдання: – огляд літератури по темі зварювання легень; – оглянути інструменти для зварювання легень та результати проведених операцій по зварюванню легень та інших живих тканин; – обрати середовище моделювання інструменту для зварювання легень та визначити умови проведення даної процедури; – створити модель інструменту для зварювання легень та провести дослідження інструменту на статичне навантаження на стиск; – проаналізувати отримані результати.
Scope of the diploma is 72 pages, contains 63 illustrations, 17 tables. 23 sources were totally processed. Relevance: nowadays there is no full-fledged tool for welding the lungs, which could be guaranteed to cope with particularly hard parts of lungs, such as the bronchi, common tools for welding soft tissues are used instead. The structure of the lungs is heterogeneous. Except the parenchyma, bronchi, alveoli, trachea, blood vessels, the lungs are filled with air, which causes difficulties during welding process. Everything mentioned above requires from the tool improved parameters of reliability and durability. Objective: to model and study the static load of a lung welding tool. Task: – review of literature on lung welding; – inspect lung welding tools and the results of operations performed to weld lungs and other living tissues; – select the modeling environment of the lung welding tool and determine the conditions for this procedure; – create a model of a lung welding tool and conduct a study of the tool for static compressive load; – analyze the results obtained.

Diese Arbeit stellt das Konzept der kompressionssensitiven Magnetresonanzelastographie vor. Kompressionssensitive MRE analysiert die Ausbreitung von Kompressionswellen und liefert dadurch Erkenntnisse über die Kompressionseigenschaften eines Mediums auf Grundlage eines poroelastischen Modells. Anomalien bei der Regulation des Gewebedrucks stehen in Zusammenhang mit verschiedenen Krankheitsbildern, wie Normaldruck-Hydrozephalus und Pfortader-Hypertonie. Statischer Druck spielt als Porendruck eine zentrale Rolle in den poroelastischen Wellengleichungen; die kompressionssensitive MRE könnte daher ein nichtinvasives Diagnoseinstrument darstellen, das die durch konventionelle Scherwellen-Elastographie gewonnenen Informationen um weitere Aspekte ergänzt. Diese Arbeit beschreibt die Entwicklung einer schnellen Singleshot-EPI-Bildgebungssequenz, mit deren Hilfe die durch propagierende Druckwellen hervorgerufene volumetrische Verzerrung quantifiziert werden kann. Die Validierung der kompressionssensitiven MRE erfolgte an verschiedenen Systemen: an porösen Gelphantomen, an der menschlichen Lunge in zwei Atemzuständen, in einer ex-vivo Schafsleber bei unterschiedlichen hydrostatischen Drücken und schließlich am menschlichen Gehirn. Die Ergebnisse belegen, dass die Stärke der induzierten volumetrischen Verzerrung sensitiv gegenüber Druckänderungen ist, wohingegen die Scherverzerrung keine derartige Abhängigkeit aufweist. In einer weiteren Studie wurde intrinsische Pulsation des menschlichen Hirns anstelle einer externen Vibrationsquelle ausgenutzt. Dabei erzeugte die arterielle Pulswelle eine kurze lokale Expansion des Hirnparenchyms; in der sich anschließenden diastolischen Phase erfolgte eine langsame Rückkehr zum Ausgangszustand. Aus den gemessenen volumetrischen Verzerrungen wurden durch Inversion der Druckwellengleichung numerische Werte für den Druckwellenmodul M berechnet; Rauschen wurde als primäre Ursache für die systematische Unterschätzung von M identifiziert.
This thesis introduces the concept of compression-sensitive Magnetic Resonance Elastography. Compression-sensitive MRE detects the propagation of pressure waves, providing insight into the compressibility of a material based on a poroelastic tissue model. Poroelastic models incorporate compressibility through interaction of compartments, even as each individual compartment remains incompressible. Hydrostatic tissue pressure abnormalities are associated with a number of diseases, such as normal pressure hydrocephalus or hepatic portal hypertension. Since pore pressure plays a central role in the poroelastic wave equations, compression-sensitive MRE could potentially serve as a diagnostic tool, providing information complimentary to shear-wave MRE data. This thesis describes the development of a fast single-shot EPI MR sequence capable of quantifying volumetric strain induced by external vibrations. Compression-sensitive MRE was validated in porous gel phantoms, in the human lung at two different respiratory states, in an ex vivo sheep liver at varying levels of hydrostatic pressure, and finally in human liver and brain. Results illustrate that compression-sensitive MRE is capable of quantifying volumetric strain in phantoms and in human organs. It was found that volumetric strain was sensitive toward pressure changes associated with different physiological states, whereas shear strain remained constant. In an additional study, pulsation of the human brain, driven by the heart cycle, was used as the actuation source instead of the external vibration generator. Results indicate local expansion of brain parenchyma upon the arrival of the arterial pulse wave, followed by a slow return to the initial state during the diastolic phase. Numerical values for the pressure wave modulus M were calculated from measured volumetric strain through inversion of the pressure wave equation. Measurement noise was identified as the primary effect causing a severe underestimation of M.

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The aim of this study was to evaluate the effects of 30 days exposure to CdCl2 100 mg L-1 on the tissue distribution of this metal and the effect on myocardial contractility. Male Wistar rats were randomly divided in two groups: control and treated. Blood pressure was measured weekly during the exposure. At the end of treatment the animals were anesthetized for hemodynamic assessment and sacrificed for in vitro measurements using the technique of isolated muscles. Tissue samples were sent for determination of cadmium using atomic absorption spectrometry technique. The blood concentration of cadmium in the treated group was attained 40 μg L-1, higher than the biological index allowed by existing laws in the world. The main sites of deposition of the metal were the kidneys and liver. Since first week of exposure, the blood pressure of the treated group was high and remained increased until the end of the treatment. Hemodynamic evaluation showed an increase in systolic blood pressure (Control: 114 ± 5 vs Treated: 127 ± 3 mmHg), diastolic blood pressure (Control: 63 ± 2 vs Treated: 81 ± 4 mmHg), left ventricular (Control: 127 ± 2 vs Treated: 140 ± 4 mmHg) and heart rate (control: 333 ± 8 vs. Treaty: 377 ± 7 mmHg) and a reduction in end-diastolic pressure of the right ventricle (Control: 6.4 ± 0.8 vs Treated : 4.1 ± 0.3 mmHg). In vitro, treatment with cadmium did not change the inotropic state (contractile force). The direct exposure of the metal on isolated muscles showed a reduction in contractile force development at concentrations above 5 μmol L-1 of CdCl2. These results demonstrate that cadmium is a metal with great potential for developing of hypertension
O objetivo deste trabalho foi avaliar os efeitos da exposição por 30 dias a CdCl2 100 mg L-1 sobre a distribuição tecidual deste metal e a consequência sobre a contratilidade miocárdica. Foram utilizados ratos Wistar separados aleatoriamente em dois grupos: controle e tratado. A pressão arterial foi mensurada semanalmente no decorrer da exposição. Ao final do tratamento os animais foram anestesiados para avaliação hemodinâmica e sacrificados para avaliação, in vitro, através da técnica de músculos isolados. As amostras teciduais e cardíacas foram encaminhadas para determinação do teor de cádmio através da técnica de Espectrometria de Absorção Atômica. A concentração sanguínea de cádmio no grupo tratado foi de, aproximadamente, 40 μg L-1, valor acima do índice biológico permitido por leis mundiais vigentes. Os principais sítios de deposição do metal foram os rins e o fígado. Desde a primeira semana de exposição, a pressão arterial do grupo tratado mostrou-se elevada e assim permaneceu ao longo das semanas seguintes. A avaliação hemodinâmica evidenciou o aumento da pressão arterial sistólica (Controle: 114 ± 5 vs Tratado: 127 ± 3 mmHg), da diastólica (Controle: 63 ± 2 vs Tratado: 81 ± 4 mmHg), da ventricular esquerda (Controle: 127 ± 2 vs Tradado: 140 ± 4 mmHg) e da frequência cardíaca (Controle: 333 ± 8 vs Tratado: 377 ± 7 mmHg) e, uma redução da pressão diastólica final do ventrículo direito (Controle: 6,4 ± 0,8 vs Tratado: 4,1 ± 0,3 mmHg). In vitro, o tratamento com cádmio não alterou o estado inotrópico (força contrátil). A exposição direta de músculos isolados ao metal demonstrou uma redução no desenvolvimento de força contrátil em concentrações superiores a 5 μmol L-1 de CdCl2. Esses resultados demonstram que o cádmio é um metal com potencial para desenvolvimento de hipertensão arterial