Atherosclerosis is an inflammatory disease which develops focally in regions of the vasculature where there is dysfunction of endothelial cells modulated in part by shear stress from flowing blood. To address the clinical crisis of atherosclerosis, tissue engineering has focused on development of a living blood vessel substitute for use as a vascular graft in bypass surgery. Despite substantial progress in understanding the biological basis and developing clinical treatments for cardiovascular disease, critical challenges remain. As a novel strategy to improve understanding of basic human vascular biology and develop superior tissue engineered grafts, this dissertation combines the scientific and clinical approaches by using a tissue engineered blood vessel as a more physiologic in vitro model to study endothelial cell biology. Through the use of transcriptional profiling, results demonstrate significant changes in endothelial cell gene expression using the tissue engineered blood vessel model. Furthermore, the presence of a more physiologic substrate alters the cellular response to shear stress which is a critical mediator of vascular pathology. A case study of endothelial cell function in this system focuses on cell-cell communication through gap junctions. Endothelial cell connexins which form gap junctions are shown to be differentially regulated by substrate and shear stress. Moreover, gap junction communication between endothelial cells is modulated by the mechanical environment. Studies using RNA interference to knockdown expression of individual connexin isotypes demonstrate integrated regulation of connexins yet unique roles in endothelial cell function. Collectively, results exemplify the sensitivity of endothelial cell phenotype to substrate and shear stress and underline the importance of using more physiologic models in the study of basic cell biology.

Abdominal aortic aneurysms are a leading cause of death in the U.S. where 14,000 people die from aneurysm rupture and 178,000 are diagnosed each year. A novel alternative treatment for abdominal aortic aneurysms has been proposed, where a biodegradable polymer scaffold is photopolymerized in situ around the exterior of the aneurysm. This scaffold will mechanically constrain the aneurysm from further expansion, and will deliver a drug, doxycycline, to treat the underlying biological cause of the disease. In order for device development, a suitable polymer must be designed with appropriate mechanical properties, degradation rate, polymerization, and elution rate. Poly(β-amino ester) networks have been proposed as the material of choice; however, many of their structure-property relationships have yet to be determined. Therefore, the overall goal of this work is to determine the structure-property relationships of the poly(β-amino ester) networks in order to advance the design of the treatment, and has been divided into three objectives: (1) understand the structure-property relationships of poly(β-amino ester) networks, specifically the polymerization, degradation rate, and thermo-mechanical properties, (2) determine the impact of doxycycline incorporation on degradation rate and mechanical properties, (3) evaluate the effect of simulated physiological conditions on degradation rate and mechanical properties. In the initial chapters, the fundamental structure-property relationships are established between reactant chemical structure, step-growth polymerization, photopolymerization, thermo-mechanical properties, and degradation rate using a systematic approach of two homologous series of reactants. Further tailoring of degradation rate, water content, and modulus in vitro was performed by using a copolymer network. Doxycycline inhibited photopolymerization due to overlapping absorbance spectra with the photoinitiator, but full network formation occurred by increasing the photoinitiator concentration. Networks displayed varying controlled release rates, and the underlying release mechanism was determined for each network using established methods. In order to increase mechanical properties, a co-monomer, methyl methacrylate, was added to the network to increase the glass transition temperature, toughness, and deformation capacity. These co-networks displayed temporal-control of mechanical properties in simulated physiological conditions, since degradation caused a shift in the glass transition temperature, which changed the mechanical behavior of the network. The temporal-control of mechanical properties was further investigated under degradation conditions in vitro and in vivo. Due to the mechanically active loading environment in vivo, networks displayed a decrease in toughness, yet maintained mechanical properties similar to native biological tissues. These networks establish a multifunctional biomaterials platform with materials that can be easily synthesized, photopolymerized into various geometries, and sustain mechanical properties while undergoing degradation and therapeutic agent release.

Made available in DSpace on 2016-06-02T20:19:05Z (GMT). No. of bitstreams: 1 1361.pdf: 7031010 bytes, checksum: e59579f600a182ade6d5f0cd26523978 (MD5) Previous issue date: 2007-02-27 / Universidade Federal de Sao Carlos / There is a rapid increase in the size of the elderly population due to a complex interaction of medical and epidemiological factors. Also, there is a parallel increase in morbidity associated with age-related delayed tendinopathy healing, and treatment of such acute and chronic condition costs the Health Services over 1 Billion dollars per year. Despite the obvious clinical impact, the basic cellular and molecular mechanisms underlying impaired human tendon healing are largely unknown. However, recent reports have showed the action mechanism of low level laser therapy (LLLT) in producing accelerated tendon healing, but the real effects of this therapy associated with aging effects are not yet known. This study evaluated the effects of the LLLT on repair process of rat calcaneal tendon in two different ages. Eighty male Wistar rats were divided in 2 experimental groups with 40 rats at each one: young group (4 weeks old, body mass = 111.3±34,3g) and mature adult group (27 weeks old, body mass = 385.4±34,3g). The young group was subdivided in subgroup Y3J (12 session, total energy = 101.7mJ, laser 685nm, 5.4W/cm2, 3J/cm2), subgroup Y10J (12 session, total energy = 339.6mJ, laser 685nm, 5.4W/cm2, 10J/cm2); subgroup YPL (placebo treatment) and subgroup YCL (no treatment). The mature adult group was subdivided in subgroup A3J (12 session, total energy = 101.7mJ, laser 685nm, 5.4W/cm2, 3J/cm2), subgroup A10J (12 session, total energy = 339.6mJ, laser 685nm, 5.4W/cm2, 10J/cm2); subgroup APL (placebo treatment) and subgroup ACL (no treatment). All animals had the medial region of right calcaneal tendon totally tenotomized. They were sacrified on the 13th post-operate day and their tendons were surgically removed for a quantitative and qualitative analysis The Atomic Force Microscopy analysis showed better organization quality (alignment, thickness and aggregation) of collagen bounds in mature irradiated groups. However, it was found differences (p < 0.05) in thickness of collagen bundles (TCBs) for several intersections among subgroups. The subgroup A10J presented greatest TCBs (5.69±0.69), and the ACL subgroup presented smallest TCBs (4.51±0.6) in the mature group. The dimensional data showed higher differences in the irradiated subgroup in mature group when compared to the same subgroup in young group. Histological analysis showed increase (p < 0.05) in the number of capillar vessels and fibroblast cell proliferation after LLLT in both young and mature groups, however the higher modulation was observed in the mature group. Minor influence of laser radiation in the young group may be justified by divergence on chemistry, physics and metabolic age-related. Maybe the accumulated oxidative stress resulting from a gradual shift in the redox status of tissues should be a key mechanism underlying the aging process and the higher laser effect noted in the mature group. Age should be considerated an important parameter to LLLT. / O envelhecimento biológico é uma das principais causas para a predisposição do tendão aos processos degenerativos, patológicos e retardo da reparação tecidual.Várias modalidades terapêuticas são utilizadas para promover a aceleração e melhora do reparo tendíneo, dentre elas a terapia laser de baixa intensidade (LLLT). Porém são pouco conhecidos os reais efeitos desta terapia quando se associa um fator potencialmente decisivo na resposta celular como o envelhecimento. Este estudo avaliou através de técnicas de microscopia de luz comum e de força atômica os efeitos da LLLT, aplicada na reparação do tendão calcanear em duas fases maturacionais. Um grupo de ratos jovens (4 semanas de idade, n = 40, massa = 111,3±8,3g), subdividido aleatoriamente em 4 subgrupos contendo cada um 10 animais: J3J (dose 3J/cm2, energia total = 101,7mJ), J10J (10J/cm2, energia total = 339,6mJ), placebo - JPL (dose 0J/cm2), controle - JCL (nenhum tratamento); e um grupo adulto (27 semanas de idade, n = 40, massa = 385,4±34,3g), subdividido aleatoriamente em 4 subgrupos: A3J (dose 3J/cm2, energia total = 101,7mJ), A10J (dose 10J/cm2, energia total = 339,6mJ), placebo - APL (dose 0J/cm2), controle - ACL (sem tratamento). Todos os animais foram submetidos ao procedimento de tenotomia radical do tendão direito, entre a inserção calcanear e a transição miotendínea, sem posterior tenorrafia. O laser utilizado (685nm, 5,4W/cm2) foi aplicado em 12 sessões, uma vez ao dia, em um único ponto sobre a região da lesão. No 13º dia pós-operatório os animais foram sacrificados e seus tendões removidos, processados e analisados qualitativa e quantitativamente, por meio de microscopia de força atômica (MFA) e análise histológica por morfometria. A análise dos dados mostrou significativa alteração (p< 0,05) da resposta do tecido à irradiação laser, tanto de forma idade-dependente como dose-dependente, sendo que, os resultados demonstram maior relevância para o grupo adulto que para o jovem. Comparativamente, o grupo adulto apresentou qualitativamente maior resposta biológica na agregação, organização, alinhamento do colágeno. Quantitativamente, o grupo adulto apresentou melhor resposta para a dose de 10J/cm2 principalmente em relação à angiogênese, contagem de fibroblastos e espessura dos feixes de colágeno. Esses resultados possivelmente podem ter uma relação com o maior estresse oxidativo característico ao incremento da idade.

Objetivo: Medir a pulsatilidade da parede do aneurisma de aorta humano antes e depois de sua correção endoluminal. Método: Cinco aneurismas foram submetidos à perfusão pulsátil antes e depois do implante de uma endoprótese. Resultado: o nível da coluna de água oscilou durante a pulsação com variações de 17, 16, 13, 7 e 25 cm antes da colocação da endoprótese. Depois da prótese, a oscilação diminuiu em todos os casos para 13, 12, 9, 3,5 e 23 cm, respectivamente. Conclusão: A pulsação da endoprótese é transmitida à parede do aneurisma / Objective: To measure the pulsatility of human aortic aneurysms before and after exclusion with endograft. Method: Five aneurysms were submitted to pulsatile perfusion before and after implantation of a bifurcated endograft. Result: The level of the water column oscillated during pulsation, in each case, with an amplitude of 17, 16, 13, 7 and 25 cm before the endograft insertion. After that, the amplitudes dropped to, respectively 13, 12, 9, 3.5 and 23 cm.Conclusion: Pulsation of an endograft is transmitted to the aneurysm wall even in the absence of endoleak

Abstract Endovascular aneurysm repair (EVAR) is a minimally invasive alternative to open surgery to exclude an abdominal aortic aneurysm from the circulation to avert a rupture. The aim of this thesis was to evaluate the early and mid-term results of EVAR using the Zenith® stent-graft (Cook Inc, Bloomington, IN, USA) in asymptomatic and symptomatic abdominal aortic aneurysm (AAA) patients in three Finnish university hospitals. Furthermore, the aim was to study whether preoperative embolization of the inferior mesenteric artery (IMA) before EVAR decreases the incidence of a type II endoleak or has an effect on the aneurysm sac shrinkage. Finally, the results after secondary interventions for a type II endoleak were evaluated. Two hundred six patients underwent elective endovascular repair of an intact AAA. The use of the Zenith® stent-graft was associated with good early and mid-term results. The thirty-day mortality rate (2.9%) was in accordance with other EVAR studies. Only one late aneurysm-related death occurred in this series, whilst no patients died of a late aneurysm rupture. No stent-graft migrations or fractures were observed. Endoleak, defined as persistent blood flow outside the graft and within the aneurysm sac, remains a long-term problem with EVAR. The overall endoleak incidence was 34.6%. A type II endoleak (retrograde perfusion via aortic side branches) occurred in 52 patients (25.4%). EVAR was performed for 14 patients with a symptomatic, unruptured AAA. The median delay from admission to intervention was 4 days. EVAR of a symptomatic, unruptured AAA was associated with a favourable outcome even in patients with a very high operative risk. There were no perioperative deaths. Altogether forty patients treated at Kuopio University Hospital had a patent IMA on preoperative computed tomography (CT) and were treated successfully with coil embolization before EVAR. Thirty-nine patients who underwent EVAR at Oulu University Hospital without preoperative embolization of a patent IMA served as a control group. Preoperative coil embolization of the IMA significantly reduced the incidence of type II endoleaks after EVAR, but the present study failed to show any influence on late postoperative aneurysm sac shrinkage. Overall, 14 patients underwent a secondary intervention to repair the type II endoleak. Ten patients had transarterial embolization and four patients had translumbar embolization. The results were unsatisfactory; clinical success after the first secondary intervention was achieved in only two patients in the transarterial embolization group and three patients in the translumbar embolization group. These results seem to favour direct translumbar embolization rather than transarterial embolization. In conclusion, EVAR with the Zenith® stent-graft is effective in excluding AAAs from the circulation and is associated with good mid-term results.

This thesis deals with method of blood vessels segmentation from retinal images acquired by fundus camera. There is explored possibility of using wavelet transform as fast outline segmentation. The thesis includes study problems of preprocessing input image and decomposition of image using 2D DWT. Furthermore there is explored possibility of parametrical images thresholding that ensue from application of 2D DWT. There are designed algorithms for cleaning off artifacts from rough vessel map of blood vessel structures. The realization of algorithm was solved in programming environment MATLAB. There was created a user control interface in graphic application GUIDE, for easy control of whole segmentation process. In conclusion of thesis is proceeded the discussion of segmentation results for images from DBME database and quantitative evaluation of results for DRIVE database images.

The purpose of this study was to evaluate the effectiveness of aspirin (ASA) and porcine endothelial cell seeding in improving the patency rate of vena cava grafts. Thirty-nine dogs underwent infrarenal vena cava replacement by 10 cm lengths of 8 mm I.D. ringed polytetrafluoroethylene grafts. Thirty-one grafts were seeded with 1-1.5 x 10(6) porcine aortic endothelial cells while eight were not (GIII). Of the seeded group, 16 animals received no ASA (GI), while 15 others (GII) were given ASA (325 mg) daily starting two days preoperatively and continuing until sacrifice. Venograms were performed on the fourth postoperative day. Grafts were harvested 32 days after insertion and evaluated for patency rate and endothelialized surfaces. The 32-day patency rate was significantly higher for GII than for GI and III animals (67% vs. 13 and 25% respectively). Endothelialized surface was higher in GII than Gi and III (67% vs. 16% and 18% respectively). We conclude that endothelial cell seeding alone does not prevent graft closure and that a combination of ASA and cell seeding significantly increases the patency rate of vena cava grafts.

No description available.

A revascularização da oclusão arterial ilíaca crônica com implante de endoprótese vascular é uma nova modalidade terapêutica para os pacientes com aterosclerose obliterante das extremidades. Os objetivos deste trabalho são verificar: os resultados clínico e radiológico do tratamento percutâneo com implante de endoprótese vascular nas oclusões arteriais crônicas do território ilíaco; a influência da aterosclerose e seus principais fatores de risco; os principais sintomas clínicos segundo os estágios de Fontaine e o comportamento das endopróteses vasculares, avaliando as permeabilidades primária e secundária. Foram estudados 67 pacientes, com 69 oclusões arteriais ilíacas crônicas, submetidos a intervenção de janeiro de 1992 a dezembro de 1998, por meio de avaliação clínica, Doppler com medida do índice tornozelo/braço e arteriografia dos membros inferiores. As revascularizações arteriais ilíacas foram realizadas sob anestesia local, utilizando-se as endopróteses tipos Wallstent® e Cragg®. O índice de sucesso técnico foi de 97,10%. A mediana do período de internação foi de dois dias e as complicações mais importantes foram tromboses arteriais (2,99%), roturas arteriais (2,99%) e embolia poplítea (1,49%). O índice tornozelo/braço pré e pós-procedimento demonstrou um incremento, estatisticamente significante, após a intervenção (P = 0,0001), e os eventos durante o seguimento foram as estenoses (5,97%) e tromboses (8,96%) das endopróteses. Houve algum grau de melhora clínica em 92,5% dos pacientes. Os casos que não apresentaram melhora tiveram associação, estatisticamente significante, com cardiopatia associada (P = 0,003) e estágios III e IV de Fontaine (P = 0,022). Não houve associação, estatisticamente significante, entre os fatores analisados e permeabilidade das endopróteses vasculares. A revascularização percutânea com implante de endopróteses vasculares nas oclusões arteriais ilíacas crônicas, demonstrou apresentar bons índices de permeabilidades primária (75%) e secundária (95%) durante o período médio de 30 meses de acompanhamento. / Arterial recanalization with percutaneous transluminal angioplasty and implantation of a vascular endoprosthesis in an occluded iliac artery is a ew therapeutic modality for those patients with obliterative atherosclerosis of the extremities. The objectives of this work are to demonstrate the clinical and radiological results of treatment of chronic occlusions of the iliac arteries; to demonstrate the effects of atherosclerosis as well as primary risk factors; to evaluate the primary clinical symptoms using the Fontaine classification and to study the effectiveness of the vascular endoprostheses with primary and secondary patency rates. Sixty-seven patients with 69 chronically occluded iliac arteries underwent percutaneous intervention from January 1992 throught December 1998. Evaluations included clinical assessment, Doppler examinations with ankle/brachial indexes and bilateral lower extremity arteriograms. The iliac artery recanalizations were performed under local anesthesia and using Wallstent® and Cragg® vascular endoprostheses. Technical success rate was 97.10%. The mean hospitalization time was two days and major complications included arterial thrombosis (2.99%), arterial rupture (2.99%) and popliteal embolization (1.49%). The prior and post procedure ankle/brachial indexes demonstrated a statistically significant increase following intervention (P = 0.0001). During follow-up there was 5.97% of stenosis and 8.96% thrombosis of the endoprostheses. There was clinical improvement in 92.42% of patients. Patients that did not show clinical improvement had association with coronary disease (P = 0.003), Fontaine stages III and IV (P = 0.022) and arterial hypertension (P = 0.087). There was no statistically significant factor associated with lower patency of the endoprostheses. Percutaneous revascularization of chronically occluded iliac arteries with utilization of vascular endoprostheses has shown good primary (75%) and secondary (95%) patency rates during 30 months of mean follow-up period.

A revascularização da oclusão arterial ilíaca crônica com implante de endoprótese vascular é uma nova modalidade terapêutica para os pacientes com aterosclerose obliterante das extremidades. Os objetivos deste trabalho são verificar: os resultados clínico e radiológico do tratamento percutâneo com implante de endoprótese vascular nas oclusões arteriais crônicas do território ilíaco; a influência da aterosclerose e seus principais fatores de risco; os principais sintomas clínicos segundo os estágios de Fontaine e o comportamento das endopróteses vasculares, avaliando as permeabilidades primária e secundária. Foram estudados 67 pacientes, com 69 oclusões arteriais ilíacas crônicas, submetidos a intervenção de janeiro de 1992 a dezembro de 1998, por meio de avaliação clínica, Doppler com medida do índice tornozelo/braço e arteriografia dos membros inferiores. As revascularizações arteriais ilíacas foram realizadas sob anestesia local, utilizando-se as endopróteses tipos Wallstent® e Cragg®. O índice de sucesso técnico foi de 97,10%. A mediana do período de internação foi de dois dias e as complicações mais importantes foram tromboses arteriais (2,99%), roturas arteriais (2,99%) e embolia poplítea (1,49%). O índice tornozelo/braço pré e pós-procedimento demonstrou um incremento, estatisticamente significante, após a intervenção (P = 0,0001), e os eventos durante o seguimento foram as estenoses (5,97%) e tromboses (8,96%) das endopróteses. Houve algum grau de melhora clínica em 92,5% dos pacientes. Os casos que não apresentaram melhora tiveram associação, estatisticamente significante, com cardiopatia associada (P = 0,003) e estágios III e IV de Fontaine (P = 0,022). Não houve associação, estatisticamente significante, entre os fatores analisados e permeabilidade das endopróteses vasculares. A revascularização percutânea com implante de endopróteses vasculares nas oclusões arteriais ilíacas crônicas, demonstrou apresentar bons índices de permeabilidades primária (75%) e secundária (95%) durante o período médio de 30 meses de acompanhamento. / Arterial recanalization with percutaneous transluminal angioplasty and implantation of a vascular endoprosthesis in an occluded iliac artery is a ew therapeutic modality for those patients with obliterative atherosclerosis of the extremities. The objectives of this work are to demonstrate the clinical and radiological results of treatment of chronic occlusions of the iliac arteries; to demonstrate the effects of atherosclerosis as well as primary risk factors; to evaluate the primary clinical symptoms using the Fontaine classification and to study the effectiveness of the vascular endoprostheses with primary and secondary patency rates. Sixty-seven patients with 69 chronically occluded iliac arteries underwent percutaneous intervention from January 1992 throught December 1998. Evaluations included clinical assessment, Doppler examinations with ankle/brachial indexes and bilateral lower extremity arteriograms. The iliac artery recanalizations were performed under local anesthesia and using Wallstent® and Cragg® vascular endoprostheses. Technical success rate was 97.10%. The mean hospitalization time was two days and major complications included arterial thrombosis (2.99%), arterial rupture (2.99%) and popliteal embolization (1.49%). The prior and post procedure ankle/brachial indexes demonstrated a statistically significant increase following intervention (P = 0.0001). During follow-up there was 5.97% of stenosis and 8.96% thrombosis of the endoprostheses. There was clinical improvement in 92.42% of patients. Patients that did not show clinical improvement had association with coronary disease (P = 0.003), Fontaine stages III and IV (P = 0.022) and arterial hypertension (P = 0.087). There was no statistically significant factor associated with lower patency of the endoprostheses. Percutaneous revascularization of chronically occluded iliac arteries with utilization of vascular endoprostheses has shown good primary (75%) and secondary (95%) patency rates during 30 months of mean follow-up period.