<p>Osnovni cilj rekonstrukcije prednjeg ukr&scaron;tenog ligamenta kolena je dobijanje pune stabilnosti kolena u celom obimu pokreta. Bez obzira na razvoj operativne tehnike i rehabilitacije i dalje postoji jedan broj pacijenata koji nije u potpunosti zadovoljan rezultatom rekonstrukcije prednjeg ukr&scaron;tenog ligamenta kolena, te je potrebno izvr&scaron;iti ponovnu operaciju. Smatra se da je najče&scaron;ći uzrok rane ponovne nestabilnosti lo&scaron;a pozicija kalema, tj. pozicija tunela koja nije na anatomskom mestu. Većina hirurga koji se bave ovom problematikom procenjuju poziciju kalema u golenjači na osnovu standardnih radiograma: prednje-zadnje i bočne projekcije, &scaron;to svakako nije dovoljno precizno. U ekonomski&nbsp; razvijenim zemljama poziciju kalema određuju analizirajući snimke kompjuterizovane tomografije (CT) ili magnetne rezonance (MRI). Prvenstveno zbog smanjenja doze zračenja kojoj se izlažu pacijenti prilikom kompjuterizovane tomografije, ali i iz ekonomskih razloga bilo bi korisno razvijanje jednog lako dostupnog, jeftinog ali preciznog i jednostavnog metoda određivanja položaja kalema u golenjači. Osnovni cilj ovog istraživanja bio je da se dokaže značaj pozicije kalema u golenjači i mogućnost svakodnevne kliničke primene novorazvijenog kompjuterskog programa u cilju određivanja prostorne pozicije kalema u golenjači iz samo dva standardna radiograma. Studija je bila eksperimentalno-prospektivnog karaktera. Eksperimentalni deo istraživanja sproveden je na Fakultetu tehničkih nauka (FTN) u Novom Sadu na Departmanu za proizvodno ma&scaron;instvo gde je na osnovu standardnih RTG snimaka (prednje-zadnje i bočne projekcije) razvijen kompjuterski program za određivanje prostornog položaja kalema u golenjači. U cilju verifikacije novorazvijenog programa realizovano je prostorno određivanje položaja kalema u golenjači obradom standardnih RTG snimaka (prednje-zadnje i bočne projekcije) kod 10 pacijenata kod kojih je izvr&scaron;ena primarna rekonstrukcija prednjeg ukr&scaron;tenog ligamenta kolena. Dobijeni rezultati su upoređeni sa položajem kalema na CT snimcima koji su obezbeđeni za sve pacijente. Na taj način je eksperimentalnim putem utvrđena preciznost novorazvijenog softvera u određivanju prostornog položaja kalema u golenjači. Drugi deo istraživanja bilo je prospektivno kliničko ispitivanje koje je sprovedeno na Klinici za ortopedsku hirurgiju i traumatologiju Kliničkog centra Vojvodine u Novom Sadu. Ispitivanu grupu je činilo 120 pacijenata, oba pola, sa prekidom prednjeg ukr&scaron;tenog ligamenta levog ili desnog kolena. Metodom slučajnog izbora pacijenti su podeljeni u četiri grupe od po 30 pacijenata prema veličini sagitalnog ugla (S) bu&scaron;enja kanala u golenjači (S 60&ordm;-69,9&ordm; i S 70&ordm;-80&ordm;) i prema veličini transverzalnog ugla (T) bu&scaron;enja kanala u golenjači (T 60&ordm;-69,9&ordm; i T 70&ordm;-80&ordm;). Grupa I 30 pacijenata (S 60&ordm;-69,9&ordm; i T 60&ordm;-69,9&ordm;), grupa II 30 pacijenata (S 60&ordm;-69,9&ordm; i T 70&ordm;-80&ordm;), grupa III 30 pacijenata (S 70&ordm;-80&ordm; i T 60&ordm;-69,9&ordm;), grupa IV 30 pacijenata (S 70&ordm;-80&ordm; i T 70&ordm;-80&ordm;). Svi pacijenti su godinu dana nakon operacije pro&scaron;li klinička ispitivanja po bodovnim skalama (Tegner bodovna skala, Lysholm bodovna skala i IKDC standard) i artrometrijska merenja. Rezultati dobijeni merenjem položaja kalema u golenjači, kliničkim ispitivanjima i artrometrijskim merenjima poređani su unutar svake grupe posebno, a izvr&scaron;eno je i poređenje dobijenih rezultata između svih grupa. U ispitivanje su uključeni samo oni pacijenti koji su dali potpisani informisani pristanak da učestvuju u ispitivanju nakon detaljnog upoznavanja sa planiranom procedurom. Svaki pacijent je bio informisan o svrsi i načinu sprovođenja istraživanja, kao i o pregledima i merenjima koja će biti vr&scaron;ena. Statističkom analizom rezultata utvrđeno je da veličina gre&scaron;ke prostornog određivanja položaja kalema u golenjači posle rekonstrukcije prednjeg ukr&scaron;tenog ligamenta kolena novorazvijenim kompjuterskim programom nije statistički značajna, a kompjutersko određivanje položaja kalema u golenjači omogućava iste rezultate kao i CT snimci. Sumiranjem zaključaka nakon sveobuhvatne analize dobijenih rezultata istraživanja, može se zaključiti da pozicija kalema u golenjači posle rekonstrukcije prednjeg ukr&scaron;tenog ligamenta kolena utiče na postoperativni funkcionalni rezultat.</p> / <p>The main goal of reconstruction of the anterior cruciate ligament of the knee is to obtain complete knee stability in the full range of movement. Regardless of the development of operational techniques and rehabilitation, there is still a number of patients who are not completely satisfied with the result of reconstruction of the anterior cruciate ligament, and a re-operation is required. It is believed that the most common cause of the instability is the bad position of the graft, i.e. position of the tunnel that is not at the anatomical place insertion. Most surgeons who deal with this problem evaluate the position of the graft in the tibia based on standard radiograms: anterior-posterior and lateral projections, which is not precise enough. In economically developed countries, the position of the graft is determined by analyzing images of computerized tomography (CT) or magnetic resonance (MRI). Primarily due to a decrease in the radiation dose exposed to patients during computerized tomography, but also for economic reasons, it would be useful to develop an easily accessible, inexpensive but precise and simple method for determining the position of the graft in the tibia. The main goal of this examination was to prove the significance of the position of the graft and the possibility of daily clinical use of the newly developed computer program in order to determine in-space position of the graft in only two standard radiograms. The study was experimental-prospective. The experimental part of the research was conducted at the Faculty of Technical Sciences (FTN) in Novi Sad at the Department of Production Engineering, where a computer program for determining in-space position of the graft in the tibia was developed on the basis of standard RTG images (anterior-posterior and lateral projections). In order to verify the newly developed program, in-space determination of the position of the graft in the tibia by processing standard RTG images (anterior-posterior and lateral projections) was performed in 10 patients in which the primary reconstruction of the anterior cruciate ligament was performed. The obtained results were compared with the position of the grafts on CT images provided to all patients. In this way, the accuracy of newly developed software in determining in-space position of the graft in the tibia was determined experimentally. The second part of the study was a prospective clinical trial conducted at the Clinic for Orthopedic Surgery and Traumatology at the Clinical Center of Vojvodina in Novi Sad. The investigated group consisted of 120 patients, both sexes, with a break of the anterior cruciate ligament of the left or right knee. By random selection, patients were divided into four groups of 30 patients according to the size of the sagital angle (S) of the tunnel drilling in the tibia (S 60&ordm;-69,9&ordm; and S 70&ordm;-80&ordm;) and according to the transversal angle (T) of the drill tunnel in the tibia ( T 60&ordm;-69.9&ordm; and T 70&ordm;-80&ordm;). Group I 30 patients (S 60&ordm;-69.9&ordm; and T 60&ordm;-69.9&ordm;), group II 30 patients (S 60&ordm;-69.9&ordm; and T 70&ordm;-80&ordm;), group III 30 patients (S 70&ordm;-80&ordm; and T&ordm;60&ordm; -69.9&ordm;), group IV 30 patients (S 70&ordm;-80&ordm; and T 70&ordm;-80&ordm;). All of the patients underwent clinical trials by scales (Tegner score scale, Lysholm score scale and IKDC standard) and arthrometric measurements one year after surgery. The results obtained by measuring the position of the graft in the tibia, clinical trials and arthrometric measurements were classified separately in each group, and obtained results were compared between all groups. The study included only those patients who gave signed informed consent to participate in the study after being thoroughly informed about planned procedure. Each patient was informed about the purpose and method of conducting the research, as well as the examinations and measurements to be performed. Statistical analysis of the results showed that the size of the error in measuring in-space determination of the position of the graft in tibia after reconstruction of the anterior cruciate ligament of the knee by a newly developed computer program was not statistically significant, and the computer determination of the position of the graft in the tibia provides the same results as the CT images. Summarizing the conclusions after a comprehensive analysis of the obtained results of the study, it can be concluded that the position of the graft in tibia after reconstruction of the anterior cruciate ligament affects the postoperative functional result.</p>

Epidermal wound healing requires the coordinated involvement of complex cellular and biochemical processes. In the case of epidermal wounds associated with burns, the healing process may be less than optimal and may take a significant amount of time, possibly resulting in infection and scarring. An innovative method to assist in the repair of the epidermis (the outer layer of skin) is to use an aerosolised apparatus. This method involves taking skin cells from an area of the patient's undamaged skin, culturing the cells in a laboratory, encouraging them to rapidly proliferate, then harvesting and separating the cells from each other. The cells are then sprayed onto the wound surface. We investigate this novel treatment strategy for the healing of epidermal wounds, such as burns. In particular, we model the application of viable cell colonies to the exposed surface of the wound with the intent of identifying key factors that govern the healing process. Details of the evolution of the colony structure are explored in this two-dimensional model of the wound site, including the effect of varying the initial population cluster size and the initial distribution of cell types with different proliferative capacities. During injury, holoclones (which are thought to be stem cells) have a large proliferative capacity while paraclones (which are thought to be transient amplifying cells) have a more limited proliferative capacity. The model predicts the coverage over time for cells that are initially sprayed onto a wound. A detailed analysis of the underlying mathematical models yields novel mathematical results as well as insight into phenomena of healing processes under investigation. Two one-dimensional systems that are simplifications of the full model are investigated. These models are significant extensions of Fisher's equation and incorporate the mixed clonal population of quiescent and active cells. In the first model, an active cell type migrates and proliferates into the wound and undergoes a transition to a quiescent cell type that neither migrates nor proliferates. The analysis yields the identification of the key parameter constraints on the speed of the healing front of the cells on this model and hence the rate of healing of epidermal wounds. Approximations for the maximum cell densities are also obtained, including conditions for a less than optimal final state. The second model involves two active cell types with different proliferative capacity and a quiescent cell type. This model exhibits two distinct behaviours: either both cell types coexist or one of them dies out as the wound healing progresses leaving the other cell type to fill the wound space. Conditions for coexistence are explored.

Objetivos: O objetivo desse estudo foi avaliar comparativamente o potencial osteogênico in vitro de células obtidas do ramo mandibular (RM, área doadora) e do seio maxilar enxertado com uma mistura de RM e osso bovino inorgânico (OBI), previamente à instalação de implantes de titânio (SM, sítio do seio maxilar enxertado). Material e Métodos: As células foram obtidas de três pacientes submetidos a procedimentos de aumento do soalho do seio maxilar com a proporção de 1:1 de RM e OBI (GenOx Inorg®). No momento da realização dos enxertos no seio maxilar e após 08 meses, antes da inserção dos implantes de titânio, fragmentos ósseos foram colhidos do RM e do SM, respectivamente, e submetidos à digestão enzimática com tripsina e colagenase para obtenção de células primárias. As células foram subcultivadas e crescidas sob condições osteogênicas por até 21 dias, tendo sido avaliados os seguintes parâmetros: proliferação/viabilidade celular, expressão gênica de marcadores osteoblásticos, atividade de fosfatase alcalina (ALP) e conteúdo de cálcio, por extração do vermelho de Alizarina. Culturas primárias derivadas do RM foram expostas ao GenOx Inorg® por 7 dias, quando se avaliou a atividade de ALP. Os resultados foram comparados por ANOVA two-way, seguido do teste de Tukey, ou pelo teste de Mann-Whitney. Resultados: Culturas do SM exibiram uma redução significante do potencial osteogênico se comparado ao de culturas do RM, com um aumento progressivo na proliferação celular associado a uma redução da expressão dos marcadores osteoblásticos, da atividade de ALP e do conteúdo de cálcio. A exposição do GenOx Inorg® às células primárias derivadas do RM inibiram a atividade de ALP. Conclusão: Esses resultados sugerem que o uso do GenOx Inorg® em associação a fragmentos do RM para aumento do soalho do seio maxilar inibe a diferenciação de células osteoblásticas no sítio de inserção de implantes de titânio após 8 meses de enxertia. / Objectives: This study aimed to comparatively evaluate the in vitro osteogenic potential of cells obtained from the mandibular ramus (MR, autogenous bone donor site) and from the maxillary sinus bone grafted with a mixture of anorganic bovine bone (ABB) and MR prior to titanium implant placement (MS, grafted implant site). Material and methods: Cells were obtained from three patients subjected to maxillary sinus floor augmentation with a 1:1 mixture of ABB (GenOx Inorg®) and MR. At the time of the sinus lift procedure and after 8 months, prior to implant placement, bone fragments were taken from MR and MS, respectively, and subjected to trypsin-collagenase digestion for primary cell culturing. Subcultured cells were grown under osteogenic condition for up 21 days and assayed for proliferation/viability, osteoblast marker mRNA levels, alkaline phosphatase (ALP) activity and calcium content/Alizarin red staining. ALP activity was also determined in primary explant cultures exposed to GenOx Inorg® (1:1 with MR) for 7 days. Data were compared using the two-way ANOVA followed by the Tukey test; otherwise, the Mann-Whitney test was used. Results: MS cultures exhibited a significantly lower osteogenic potential compared with MR cultures, with a progressive increase in cell proliferation together with a downregulation of osteoblast markers, reduced ALP activity and calcium content. Exposure of MR-derived primary cultures to GenOx Inorg® inhibited ALP activity. Conclusion: These results suggest that the use of GenOx Inorg® in combination with MR fragments for maxillary sinus floor augmentation inhibits the osteoblast cell differentiation at the implant site in the longterm.

Les endoprothèses sont devenues une solution thérapeutique de choix pour les anévrismes aortiques. Même si leur bénéfice à long terme face à la chirurgie ouverte reste incertain, elles permettent de réduire la mortalité et les complications postopératoires précoces. Elles sont toutefois soumises à un certain nombre de contraintes anatomiques. Il y a encore quinze ans, elles étaient contre-indiquées dans le traitement des anévrismes débutant au niveau ou au-dessus des artères rénales, à savoir les anévrismes juxta et suprarénaux (AJSR) et les anévrismes thoraco-abdominaux (ATA). Le développement d'endoprothèses fenêtrées et/ou branchées (EPFB) a permis de proposer une solution endovasculaire à ces patients. Il s'agit d'endoprothèses munies d'orifices ou de branches latérales destinées à maintenir la perfusion des artères rénales et viscérales. Plusieurs études rétrospectives ont rapporté des résultats encourageants à court et moyen terme. Il s'agit néanmoins de techniques complexes, exposées à la survenue de problèmes techniques au niveau des artères cibles (rénales et viscérales) et des voies d'accès fémorales. Les résultats des EPFB nous semblent améliorables par une optimisation de la sélection des patients, par des innovations techniques apportées aux dispositifs implantables, ainsi que l'amélioration des techniques d'imagerie et de navigation endovasculaire. En raisons des délais de fabrication (> 6 semaines), les patients nécessitant un traitement rapide (anévrismes douloureux ou volumineux) échappent par ailleurs aux EPFB manufacturées.Les objectifs de cette thèse étaient: (1) d'évaluer dans notre expérience la fréquence et la nature des problèmes techniques peropératoires, et d'analyser leur impact sur les suites cliniques immédiates; (2) d'évaluer la faisabilité et les performances sur modèle in vitro de la navigation endovasculaire utilisant un système de navigation électromagnétique (StealthStation®, Medtronic); (3) d'évaluer la faisabilité et la sureté de la navigation robotique au cours de la mise en place d'une EPFB; (4) d'analyser les résultats à court terme des endoprothèses avec fenêtres créées à façon par le chirurgien pour des patients porteurs d'anévrismes complexes nécessitant un traitement rapide.Au cours de ce travail, nous avons tout d'abord montré qu'en début d'expérience (113 premiers patients), les problèmes techniques peropératoires n'étaient pas rares (30 % des patients) et que ceux-ci aboutissaient dans plus de la moitié des cas à des complications postopératoires. Ces problèmes techniques sont certes en partie évitables par l'amélioration de la sélection des patients et l'expertise de l'équipe soignante, mais nous pensons que le développement de nouveaux outils d'imagerie et de navigation pourrait également jouer un rôle clé. C'est sur ce dernier point que nous avons accentué nos recherches. Bien qu'une amélioration des performances techniques de la navigation électromagnétique soit nécessaire, nous avons pu montrer qu'elle permettait de fournir des informations complémentaires à la fluoroscopie, notamment lors des étapes de déploiement de l'endoprothèse et de cathétérisme des artères cibles. Plus que concurrente de la fluoroscopie, elle pourrait être utilisée de façon combinée dans le but de diminuer l'irradiation par les rayons X et d'améliorer les performances de la navigation. Nous avons également pu montrer les bénéfices potentiels de la navigation robotique lors de la mise en place d'endoprothèses branchées et ses limites actuelles pour assister de façon efficace les procédures fenêtrées. Enfin, dans une série limitée (n=11) de patients porteurs d'ATA, nous avons montré que l'utilisation d'endoprothèses avec fenêtres créées à façon par le chirurgien était à même de fournir des résultats précoces comparables (mortalité hospitalière : 9%, ischémie médullaire : 9%) à ceux des EPFB, et constituait une solution à considérer pour les patients nécessitant un traitement rapide. / Aortic stent grafts have gained widespread acceptance for the treatment of aortic aneurysms. When compared with open repair, they have been shown to reduce significantly postoperative mortality and complication rates. However, these benefits remain uncertain in the long-term and aortic stent grafts are still limited by anatomic restrictions. Fifteen years ago, patients with complex aneurysms such as juxtarenal, suprarenal and thoracoabdominal aneurysms were not eligible for endovascular repair. With the development of fenestrated and branched stent grafts (FBSG), it is now possible to offer an endovascular solution to these patients. FBSG contains holes (fenestrations) or lateral branches. Their purpose is to allow the proximal margin of the device to sit higher than standard infrarenal devices and allow uninterrupted blood flow to renal and visceral vessels. Retrospective studies have shown encouraging short-term and mid-term results. However, fenestrated/ branched stent grafting remains a complex procedure, associated with significant risks of technical problems such as target vessel loss and difficulties with iliac or femoral accesses. Results of FBSG could be further improved by optimizing patient selection, technical innovation to optimize currently available devices, and developments of new imaging and navigation tools. In addition, due to manufacturing delays (6-10 weeks), FBSG are not applicable to high-risk patients who need rapid treatment (painful or large aneurysms).The objectives of this thesis were: (1) to evaluate the incidence and define the nature of unexpected intraoperative adverse events and analyse their impact on the postoperative outcome; (2) to evaluate in vitro the feasibility and the performances of electromagnetic navigation using a new device (StealthStation®, Medtronic); (3) to evaluate the feasibility and safety of robotic navigation for target vessel cannulation; (4) to analyse short-term results of physician modified stent grafts for patients with complex aortic aneurysms needing rapid treatment.During this work, we showed that intraoperative adverse events were not rare (30% of the first 113 patients) during our early experience. They were responsible for postoperative complications in more than 50 % of cases. Optimal patient selection and increased experience of the medical staff may reduce the incidence of technical problems. However, we believe the development of new imaging modalities and navigation tools could play a key role to limit their occurrence. Although further developments are mandatory, the use of electromagnetic navigation as a complementary imaging modality, as tested with the StealthStation®, might be beneficial in terms of radiation exposure, cannulation performances, and accuracy in the positioning of FBSG. We also showed that robotic cannulation of renal and visceral vessels during complex endovascular aortic procedures is feasible and safe. In our experience, it was more effective for branches than for fenestrations. Finally, we used physician modified stent grafts in a limited series of eleven patients with thoracoabdominal aortic aneurysms. With a 9% in-hospital mortality rate and a 9% rate of spinal cord ischemia, they provided similar results to those of manufactured FBSG, suggesting they might be an option to consider for high-risk patients needing rapid treatment.

The growing incidences of orthopedic problems globally have created a huge demand for strong bioactive materials for bone tissue engineering. Over the years, studies have shown chemical, physical, and mechanical properties of biomaterials influence the cellular interactions at the material-tissue interface, which subsequently controls biological response to materials. Strong biomaterials with surface properties that actively direct cellular response hold the key for engineering the next generation orthopedic implants. With its unique properties graphene can be used to reinforce poly (ε-caprolactone) (PCL) to prepare strong and bioactive polymer nanocomposites for bone tissue regeneration. The thesis entitled ―Engineering bioactive and multifunctional graphene polymer composites for bone tissue regeneration” systematically studies the effect of different chemically functionalized and metal-graphene hybrid nanoparticles in PCL composites for bone tissue engineering. The thesis comprises of seven chapters. Chapter 1 is an outline review on the impact of graphene and graphene derived particles to prepare supporting substrates for tissue regeneration and the associated cell response to multifunctional graphene substrate. This chapter discusses how cells interact with different graphene based particles and the interplay between cells performance and multifunctional properties of graphene based substrates. Chapter 2 describes the role, if any, of the functionalization of graphene on mechanical properties, stem cell response and bacterial biofilm formation. PCL composites of graphene oxide (GO), reduced GO (RGO) and amine-functionalized GO (AGO) were prepared at different filler contents (1%, 3% and 5%). Although the addition of the nanoparticles to PCL markedly increased the storage modulus, this increase was higher for GO and AGO than with RGO. In vitro cell studies revealed that the AGO and GO particles significantly increased human mesenchymal stem cell (hMSC) proliferation. AGO was most effective in augmenting stem cell osteogenesis leading to mineralization. Bacterial studies revealed that interaction with functionalized GO induced bacterial cell death due to membrane damage which was further accentuated by amine groups in AGO. The synergistic effect of oxygen containing functional groups and amine groups on AGO-reinforced composites renders the optimal combination of improved modulus, favorable stem cell response and biofilm inhibition desired for orthopaedic applications. In Chapter 3, toward preparing strong multi-biofunctional materials, poly(ethylenimine) (PEI) conjugated graphene oxide (GO_PEI) was synthesized using poly(acrylic acid) (PAA) as spacer and incorporated in PCL at different fractions. GO_PEI significantly promoted proliferation and formation of focal adhesions in hMSCs on PCL. GO_PEI was highly potent in inducing stem cell osteogenesis leading to 90% increase in alkaline phosphatase activity and mineralization over neat PCL with 5% filler content and was 50% better than GO. Remarkably, 5% GO_PEI was as potent as soluble osteo-inductive factors. Increased adsorption of osteogenic factors due to the amine and oxygen containing functional groups on GO_PEI augment stem cell differentiation. GO_PEI was also highly efficient in imparting bactericidal activity with 85% reduction in counts of E. coli colonies compared to neat PCL at 5% filler content and was more than twice as efficient as GO. This may be attributed to the synergistic effect of the sharp edges of the particles along with the presence of the different chemical moieties. Thus, in contrast to using labile biomolecules, GO_PEI based polymer composites can be utilized to prepare bioactive resorbable biomaterials for fabricating orthopedic devices for fracture fixation and tissue engineering. Chapter 4 describes the preparation of hybrid nanoparticles of graphene sheets decorated with strontium metallic nanoparticles and its advantages in bone tissue engineering. Strontium-decorated reduced graphene oxide (RGO_Sr) nanoparticles were synthesized by facile reduction of graphene oxide and strontium nitrate. X-ray diffraction, transmission electron microscopy, and atomic force microscopy revealed that the hybrid particles were composed of RGO sheets decorated with 200 – 300 nm metallic strontium particles. Thermal gravimetric analysis further confirmed the composition of the hybrid particles as 22 wt% of strontium. Macroporous tissue scaffolds were prepared incorporating RGO_Sr particles in PCL. The PCL/RGO_Sr scaffolds were found to elute strontium ions in aqueous medium. Osteoblast proliferation and differentiation was significantly higher in the PCL scaffolds containing the RGO_Sr particles in contrast to neat PCL and PCL/RGO scaffolds. The increased biological activity can be attributed to the release of strontium ions from the hybrid nanoparticles. This study demonstrates that composites prepared using hybrid nanoparticles that elute strontium ions can be used to prepare scaffolds with osteoinductive property. These findings have important implications for designing the next generation of biomaterials for use in tissue regeneration. Chapter 5 discusses the use of hybrid graphene-silver particles (RGO_Ag) to reinforce PCL and compared with PCL/RGO and PCL/Ag composites containing RGO and silver nanoparticles (AgNPs), respectively. RGO_Ag hybrid particles were well dispersed in the PCL matrix unlike the RGO and AgNPs due to enhanced exfoliation. RGO_Ag led to 77 % increase in the modulus of PCL and provided a conductive network for electron transfer. Electrical conductivity increased four orders of magnitude from 10-11 S/cm to 10-7 S/cm at 5 wt % filler that greatly exceeded the improvements with the use of RGO and AgNP in PCL. RGO_Ag particles reinforced in PCL showed sustained release of silver ions from the PCL matrix unlike the burst release from PCL/Ag. PCL/RGO_Ag and PCL/RGO composites were non-toxic to hMSCs and supported osteogenic differentiation unlike the PCL/Ag composites which were highly toxic at ≥3% filler content. The PCL/RGO_Ag composites exhibited good antibacterial effect due to a combination of silver ion release from the AgNPs and the mechanical rupture induced by the RGO in the hybrid nanoparticles. Thus, the synergistic effect of Ag and RGO in the PCL matrix uniquely yielded a multifunctional material for use in implantable biomedical devices and tissue engineering. Chapter 6 presents investigation of potential differences in the biological response to graphene in polymer composites in the form of 2D substrates and 3D scaffolds. Results showed that osteoblast response to graphene in polymer nanocomposites is markedly altered between 2D substrates and 3D scaffold due to the roughness induced by the sharp edges of graphene at the surface in 3D but not in 2D. Osteoblast organized into aggregates in 3D scaffolds in contrast to more well spread and randomly distributed cells on 2D discs due to the macro-porous architecture of the scaffolds. Increased cell-cell contact and altered cellular morphology led to significantly higher mineralization in 3D scaffolds compared to 2D. This study demonstrates that the cellular response to nanoparticles in composites can change markedly by varying the processing route. Chapter 7 summarizes the important results and future directions of the work. This chapter provides general conclusions arising from this study, and makes suggestions for future work designed to provide a greater understanding of the in vivo response in terms of bio-distribution of the released functionalized graphene from the scaffold or substrate must be assessed with special attention on their accumulation or excretion.

Objetivos: O objetivo deste estudo é analisar a perviedade da artéria torácica interna direita (ATID) pediculada, anteroaórtica em anastomose para a região anterior do coração na cirurgia de revascularização do miocárdio (RM), em relação à artéria torácica interna esquerda (ATIE). Métodos: No período de dezembro de 2008 a dezembro de 2011, 100 pacientes foram selecionados para serem submetidos a cirurgia de RM sem circulação extracorpórea (CEC), de forma prospectiva. Eles foram agrupados em Grupo 1 (G-1) e Grupo 2 (G-2), cada um com 50 pacientes, com randomização por computador e conhecimento da técnica no início da cirurgia. No G-1, os pacientes receberam ATIE para a região anterior do coração e complementação da RM com a ATID livre para ramos da circunflexa (CX) e outros enxertos arteriais ou venosos para a coronária direita (CD) e/ou ramos. Os pacientes do G-2 receberam ATID pediculada para a região anterior do coração e complementação da RM com ATIE, pediculada, para ramos da CX e outros enxertos arteriais ou venosos para a CD e/ou ramos. A perviedade das artérias torácicas internas direita e esquerda foi avaliada através de angiotomografia coronária multislice, 64 canais, no 6º mês de pós-operatório. Resultados: Os dois grupos eram semelhantes quanto aos dados clínicos de pré-operatório, como exemplo: diabetes mellitus, hipertensão arterial sistêmica, obesidade. Os dois grupos apresentaram predominância do sexo masculino com 75,6% e 88% nos grupos 1 e 2, respectivamente. Cinco pacientes migraram do G-1 para o G-2 em virtude de doença ateromatosa na aorta ascendente e um deles foi excluído por ter que utilizar enxerto composto. A média de anastomoses distais no G-1 foi de 3,48 (DP=0,72), e no G-2 foi de 3,20 (DP=0,76). Não ocorreu mediastinite em nenhum paciente. Uma paciente do G-1 apresentou osteomielite, e necessitou de intervenção cirúrgica. Dois pacientes do G-1 foram submetidos a reoperação por sangramento. Os resultados das angiotomografias coronarianas com 96 pacientes re-estudados mostram que todas as ATIs, fosse a direita ou a esquerda, utilizadas pediculadas para a região anterior do coração encontravam-se sem oclusões ou estenoses, configurando 100% de perviedade. No G-1, um enxerto livre da ATID para ramos da CX apresentava oclusão total, em dois pacientes havia estenose leve, em um deles havia estenose moderada na anastomose proximal na aorta ascendente e outro apresentava diminuição de calibre na sua porção distal. Em três pacientes o enxerto de segmento de veia safena para ramos da CD se encontravam ocluídos. No G-2, dois pacientes apresentavam oclusão total na ATIE pediculada para ramos da CX, e outro apresentava estenose moderada na porção distal da ATIE utilizada sequencial para dois ramos marginais. Em dois pacientes o enxerto de segmento de veia safena para ramos da CD se encontravam ocluídos. Não houve óbitos em nenhum dos grupos. Conclusão: A cirurgia de RM com utilização da ATID pediculada, anterógrada para o RIA, apresenta resultado semelhante ao da ATIE utilizada para essa mesma coronária. / Objective: To analyze the patency of the pedicled, anteroaortic, right internal mammary artery (RIMA) anastomosed to the left anterior descending (LAD) and branches in coronary artery bypass graft surgery (CABG), in comparison with the left internal mammary artery (LIMA). Methods: From December 2008 to December 2011, 100 patients were selected to undergo a prospective off-pump coronary artery bypass graft surgery and were randomly divided by computer into Group 1 (G-1) and Group 2 (G-2), so that the technique was known at the beginning of the surgery. In each group, with 50 patients, the patency of both right and left internal mammary arteries, which were used pedicled to the LAD, was comparatively studied through coronary computed tomography angiography. G-1 had 50 patients who received the LIMA to the LAD or LAD/diagonal (sequential) and had the CABG complemented with the free RIMA to circumflex branches and other arterial or venous grafts to the right coronary artery (RCA) and/or branches. G-2 had 50 patients who received the pedicled RIMA to the LAD or LAD/diagonal (sequential) and had the CABG complemented with the pedicled LIMA to circumflex branches and other arterial or venous grafts to the RCA and/or branches. Results: Both groups were similar in pre-operative clinical data, such as: diabetes mellitus, systemic arterial hypertension, obesity. Also, there was predominance of males in both groups, with 75,6% and 88% in Groups 1 and 2 respectively. Five patients were switched from G-1 to G-2 owing to atheromatous disease in the ascending aorta, and one of them was dropped for having to use composite graft. The average of distal anastomosis in G-1 was 3,48 (standard deviation (SD=0,72) and in G-2 was 3,20 (SD=0,76). Mediastinitis didn\'t occur in any patient. A patient from G-1 had osteomyelitis that required surgical intervention. Two patients from G-1 underwent reoperation because of bleeding. The 64-slice coronary computed tomography angiography was performed in the 6th postoperative month; 96 patients have been re-studied so far and all pedicled IMAs to the LAD were patent. In G-1 a free RIMA graft to the circumflex branches presented total occlusion, another two had a discreet stenosis and in one moderate at the proximal anastomosis and one more had a string signal at the distal portion. In G-2 two patients had total occlusion of the pedicled LIMA to circumflex artery branches, and another one presented moderate stenosis at its distal portion. In two patients the saphenous vein graft to the RCA branches were occluded. There were no deaths in any of the groups. Conclusion: The CABG surgery using the pedicled, anteroaortic RIMA to the LAD has a similar outcome to that of the LIMA used for this same coronary.

Pacientes submetidos ao transplante de medula óssea, apresentando hematúria/cistite hemorrágica, tiveram amostras de urina analisadas pela microscopia eletrônica. Esta foi a técnica escolhida de pesquisa viral pela sua confiabilidade. Noventa em 402 pacientes submetidos ao transplante de medula óssea neste serviço apresentaram hematúria/cistite hemorrágica (incidência de 22%). O estudo por microscopia eletrônica foi realizado em 72 destes pacientes com hematúria/cistite hemorrágica (grupo de estudo), identificando 55,6% (40/72) de positividade viral. Foram também estudadas amostras de urina de 12 pacientes submetidos ao transplante de medula óssea sem hematúria/cistite hemorrágica (grupo controle); houve associação significante entre a presença de vírus e hematúria/cistite hemorrágica (p<0,01 - Teste exato de Fisher). No grupo com hematúria/cistite hemorrágica, 65% (26/40) dos vírus detectados pertenciam à família Poliomaviridae, 30% (12/40) à Adenoviridae, e 5% (2/40) foram positivos para ambas as famílias. Houve associação entre a positividade para vírus e a presença da doença enxerto-contra-hospedeiro (p=0,05-x2) e o de início tardio (>dia+21) da H/CH (P=0,04-x2), bem como entre a doença enxerto-contra-hospedeiro e a severidade da H/CH (p=0,04-x2). O presente estudo mostra que a microscopia eletrônica é uma ferramenta útil para detecção de vírus na urina de pacientes submetidos ao transplante de medula óssea apresentando hematúria/cistite hemorrágica e que o vírus provavelmente tem um papel no desenvolvimento do sangramento urinário, o qual é agravado pelo doença enxerto-contra-hospedeiro. A microscopia eletrônica deveria ser realizada rotineiramente para guiar as outras técnicas de detecção viral, como a imunocitoquímica e biologia molecular. / Patients submitted to bone marrow transplantation presenting hematuria/hemorrhagic cystitis had the urine analyzed by electron microscopy. It was the elected technique for viral search because its reliability. Ninety from 402 patients submitted to bone marrow transplantation at this service showed hematuria/hemorrhagic cystitis (incidence of 22%). Electron microscopy study was performed in 72 of these patients with hematuria/hemorrhagic cystitis (study group), identifying 55.5% (40/72) of viral positivity. It was also study the urine of 12 patients submitted to bone marrow transplantation without hematuria/hemorrhagic cystitis (control group); there was a significant association between the presence of virus and hematuria/hemorrhagic cystitis (p<0.01). In the hematuria/hemorrhagic cystitis group, 65% (26/40) of the virus belonged to Poliomaviridae family, 30% (12/40) to Adenoviridae, and 5% (2/40) to both families. There was association between the status of graft-versus-host disease and positivity for virus (p=0.05), as well as between graft-versus-host disease and the severity of hematuria/hemorrhagic cystitis (p=0.04). The present study shows that electron microscopy is a useful tool for detection of virus in urine of patients submitted to bone marrow transplantation presenting hematuria/hemorrhagic cystitis and that the virus probably play a role in the development of the urinary bleeding, which is aggravated by graft-versus-host disease. Electron microscopy should be performed routinely to guide the other techniques for viral detection, like immunocitochemistry and molecular biology.

Pacientes submetidos ao transplante de medula óssea, apresentando hematúria/cistite hemorrágica, tiveram amostras de urina analisadas pela microscopia eletrônica. Esta foi a técnica escolhida de pesquisa viral pela sua confiabilidade. Noventa em 402 pacientes submetidos ao transplante de medula óssea neste serviço apresentaram hematúria/cistite hemorrágica (incidência de 22%). O estudo por microscopia eletrônica foi realizado em 72 destes pacientes com hematúria/cistite hemorrágica (grupo de estudo), identificando 55,6% (40/72) de positividade viral. Foram também estudadas amostras de urina de 12 pacientes submetidos ao transplante de medula óssea sem hematúria/cistite hemorrágica (grupo controle); houve associação significante entre a presença de vírus e hematúria/cistite hemorrágica (p<0,01 - Teste exato de Fisher). No grupo com hematúria/cistite hemorrágica, 65% (26/40) dos vírus detectados pertenciam à família Poliomaviridae, 30% (12/40) à Adenoviridae, e 5% (2/40) foram positivos para ambas as famílias. Houve associação entre a positividade para vírus e a presença da doença enxerto-contra-hospedeiro (p=0,05-x2) e o de início tardio (>dia+21) da H/CH (P=0,04-x2), bem como entre a doença enxerto-contra-hospedeiro e a severidade da H/CH (p=0,04-x2). O presente estudo mostra que a microscopia eletrônica é uma ferramenta útil para detecção de vírus na urina de pacientes submetidos ao transplante de medula óssea apresentando hematúria/cistite hemorrágica e que o vírus provavelmente tem um papel no desenvolvimento do sangramento urinário, o qual é agravado pelo doença enxerto-contra-hospedeiro. A microscopia eletrônica deveria ser realizada rotineiramente para guiar as outras técnicas de detecção viral, como a imunocitoquímica e biologia molecular. / Patients submitted to bone marrow transplantation presenting hematuria/hemorrhagic cystitis had the urine analyzed by electron microscopy. It was the elected technique for viral search because its reliability. Ninety from 402 patients submitted to bone marrow transplantation at this service showed hematuria/hemorrhagic cystitis (incidence of 22%). Electron microscopy study was performed in 72 of these patients with hematuria/hemorrhagic cystitis (study group), identifying 55.5% (40/72) of viral positivity. It was also study the urine of 12 patients submitted to bone marrow transplantation without hematuria/hemorrhagic cystitis (control group); there was a significant association between the presence of virus and hematuria/hemorrhagic cystitis (p<0.01). In the hematuria/hemorrhagic cystitis group, 65% (26/40) of the virus belonged to Poliomaviridae family, 30% (12/40) to Adenoviridae, and 5% (2/40) to both families. There was association between the status of graft-versus-host disease and positivity for virus (p=0.05), as well as between graft-versus-host disease and the severity of hematuria/hemorrhagic cystitis (p=0.04). The present study shows that electron microscopy is a useful tool for detection of virus in urine of patients submitted to bone marrow transplantation presenting hematuria/hemorrhagic cystitis and that the virus probably play a role in the development of the urinary bleeding, which is aggravated by graft-versus-host disease. Electron microscopy should be performed routinely to guide the other techniques for viral detection, like immunocitochemistry and molecular biology.

Synthetic vascular grafts are useful to bypass diseased arteries. The long-term failure of synthetic grafts is primarily due to intimal hyperplasia at the anastomotic sites. The accelerated intimal hyperplasia may stem from a compliance mismatch between the host artery and the graft since commercially available synthetic conduits are much stiffer than an artery. The objective of this thesis is to design a method for fabricating a vascular graft that mechanically matches the patients native artery over the expected physiologic range of pressures. The creation of an optimized mechanical graft will hopefully lead to an improvement in patency rates. The mechanical equivalency between the graft and the host artery is defined locally by several criteria including the diameter upon inflation, the elasticity at mean pressure, and axial force. A single parameter mathematical for a thin-walled tube is used to describe of the final mechanical behavior of a synthetic graft. For the general problem, the objective would be to fabricate a mechanics-matching vascular graft for each host artery. Typically, fabrication parameters are set initially and the properties of the fabricated graft are measured. However, by modeling the entire fabrication process and final mechanical properties, it is possible to invert the situation and let the typical output mechanical values be used to define the fabrication parameters. The resultant fabricated graft will then be mechanically matching. As a proof-of-concept, several prototype synthetic grafts were manufactured and characterized by a single Invariant to match a canine artery. The resultant graft equaled the diameter upon inflation, the elasticity at mean pressure, and axial force of the native canine artery within 6%. An alternative to making an individual graft for each artery is also presented. A surgeon may choose the best graft from a set of pre-manufactured grafts, using a computer program algorithm for best fit using two parameters in a neighborhood. The design optimization problem was solved for both canine carotid and human coronary arteries. In conclusion, the overall process of design, fabrication and selection of a mechanics matching synthetic vascular graft is shown to be reliable and robust.

Objetivos: O objetivo deste estudo é analisar a perviedade da artéria torácica interna direita (ATID) pediculada, anteroaórtica em anastomose para a região anterior do coração na cirurgia de revascularização do miocárdio (RM), em relação à artéria torácica interna esquerda (ATIE). Métodos: No período de dezembro de 2008 a dezembro de 2011, 100 pacientes foram selecionados para serem submetidos a cirurgia de RM sem circulação extracorpórea (CEC), de forma prospectiva. Eles foram agrupados em Grupo 1 (G-1) e Grupo 2 (G-2), cada um com 50 pacientes, com randomização por computador e conhecimento da técnica no início da cirurgia. No G-1, os pacientes receberam ATIE para a região anterior do coração e complementação da RM com a ATID livre para ramos da circunflexa (CX) e outros enxertos arteriais ou venosos para a coronária direita (CD) e/ou ramos. Os pacientes do G-2 receberam ATID pediculada para a região anterior do coração e complementação da RM com ATIE, pediculada, para ramos da CX e outros enxertos arteriais ou venosos para a CD e/ou ramos. A perviedade das artérias torácicas internas direita e esquerda foi avaliada através de angiotomografia coronária multislice, 64 canais, no 6º mês de pós-operatório. Resultados: Os dois grupos eram semelhantes quanto aos dados clínicos de pré-operatório, como exemplo: diabetes mellitus, hipertensão arterial sistêmica, obesidade. Os dois grupos apresentaram predominância do sexo masculino com 75,6% e 88% nos grupos 1 e 2, respectivamente. Cinco pacientes migraram do G-1 para o G-2 em virtude de doença ateromatosa na aorta ascendente e um deles foi excluído por ter que utilizar enxerto composto. A média de anastomoses distais no G-1 foi de 3,48 (DP=0,72), e no G-2 foi de 3,20 (DP=0,76). Não ocorreu mediastinite em nenhum paciente. Uma paciente do G-1 apresentou osteomielite, e necessitou de intervenção cirúrgica. Dois pacientes do G-1 foram submetidos a reoperação por sangramento. Os resultados das angiotomografias coronarianas com 96 pacientes re-estudados mostram que todas as ATIs, fosse a direita ou a esquerda, utilizadas pediculadas para a região anterior do coração encontravam-se sem oclusões ou estenoses, configurando 100% de perviedade. No G-1, um enxerto livre da ATID para ramos da CX apresentava oclusão total, em dois pacientes havia estenose leve, em um deles havia estenose moderada na anastomose proximal na aorta ascendente e outro apresentava diminuição de calibre na sua porção distal. Em três pacientes o enxerto de segmento de veia safena para ramos da CD se encontravam ocluídos. No G-2, dois pacientes apresentavam oclusão total na ATIE pediculada para ramos da CX, e outro apresentava estenose moderada na porção distal da ATIE utilizada sequencial para dois ramos marginais. Em dois pacientes o enxerto de segmento de veia safena para ramos da CD se encontravam ocluídos. Não houve óbitos em nenhum dos grupos. Conclusão: A cirurgia de RM com utilização da ATID pediculada, anterógrada para o RIA, apresenta resultado semelhante ao da ATIE utilizada para essa mesma coronária. / Objective: To analyze the patency of the pedicled, anteroaortic, right internal mammary artery (RIMA) anastomosed to the left anterior descending (LAD) and branches in coronary artery bypass graft surgery (CABG), in comparison with the left internal mammary artery (LIMA). Methods: From December 2008 to December 2011, 100 patients were selected to undergo a prospective off-pump coronary artery bypass graft surgery and were randomly divided by computer into Group 1 (G-1) and Group 2 (G-2), so that the technique was known at the beginning of the surgery. In each group, with 50 patients, the patency of both right and left internal mammary arteries, which were used pedicled to the LAD, was comparatively studied through coronary computed tomography angiography. G-1 had 50 patients who received the LIMA to the LAD or LAD/diagonal (sequential) and had the CABG complemented with the free RIMA to circumflex branches and other arterial or venous grafts to the right coronary artery (RCA) and/or branches. G-2 had 50 patients who received the pedicled RIMA to the LAD or LAD/diagonal (sequential) and had the CABG complemented with the pedicled LIMA to circumflex branches and other arterial or venous grafts to the RCA and/or branches. Results: Both groups were similar in pre-operative clinical data, such as: diabetes mellitus, systemic arterial hypertension, obesity. Also, there was predominance of males in both groups, with 75,6% and 88% in Groups 1 and 2 respectively. Five patients were switched from G-1 to G-2 owing to atheromatous disease in the ascending aorta, and one of them was dropped for having to use composite graft. The average of distal anastomosis in G-1 was 3,48 (standard deviation (SD=0,72) and in G-2 was 3,20 (SD=0,76). Mediastinitis didn\'t occur in any patient. A patient from G-1 had osteomyelitis that required surgical intervention. Two patients from G-1 underwent reoperation because of bleeding. The 64-slice coronary computed tomography angiography was performed in the 6th postoperative month; 96 patients have been re-studied so far and all pedicled IMAs to the LAD were patent. In G-1 a free RIMA graft to the circumflex branches presented total occlusion, another two had a discreet stenosis and in one moderate at the proximal anastomosis and one more had a string signal at the distal portion. In G-2 two patients had total occlusion of the pedicled LIMA to circumflex artery branches, and another one presented moderate stenosis at its distal portion. In two patients the saphenous vein graft to the RCA branches were occluded. There were no deaths in any of the groups. Conclusion: The CABG surgery using the pedicled, anteroaortic RIMA to the LAD has a similar outcome to that of the LIMA used for this same coronary.