RESUMO
Activated neutrophils have been implicated in reperfusion injury and the no-reflow phenomenon of intramyocardial arterioles. This study tested the hypothesis that ischemia and activated neutrophils impair coronary endothelial and smooth muscle cell function of epicardial and intramyocardial coronary arteries. Alteration of smooth muscle and endothelial cell function in epicardial coronary arteries (3 mm diameter) and intramyocardial coronary arteries (0.3 mm diameter) was compared by means of a myograph after exposure to ischemia (epicardial, 160 minutes, intramyocardial, 30 minutes), activated neutrophils, and combined ischemia and activated neutrophils. Morphologic studies at the ultrastructural level were done by means of scanning electron microscopy. Epicardial coronary artery function was normal after ischemia, storage with activated neutrophils, and ischemia followed by storage with activated neutrophils. Intramyocardial artery function, however, was altered. Contraction to a 45 mmol/L concentration of potassium chloride after ischemia and storage with activated neutrophils was increased (p = 0.06). Smooth muscle relaxation was significantly decreased after ischemia, but storage with activated neutrophils did not further decrease smooth muscle relaxation. Endothelium-dependent relaxation to bradykinin was significantly decreased after combined ischemia and incubation with activated neutrophils (p < 0.05). Sensitivity to bradykinin was decreased after both ischemia alone (p < 0.05) and activated neutrophils alone (p < 0.05). Similar morphologic alterations were found in epicardial and intramyocardial arteries after ischemia. Activated neutrophils alone minimally damaged endothelial cells of nonischemic intramyocardial and epicardial arteries. Endothelial cells of both arteries exposed to ischemia alone showed evidence of ischemic damage, including endothelial cell blebbing, nuclear bulging, and appearance of large holes in the cell surface. Severe endothelial cell damage was found after combined ischemia and storage with neutrophils: total destruction of cells and exposure of the basal lamina. Endothelial damage, therefore, correlated with artery function in intramyocardial but not in epicardial arteries. These results indicate that ischemia is a prerequisite for severe neutrophil injury of intramyocardial artery endothelium-mediated relaxation. This may explain no-reflow phenomenon in arterioles concurrent with myocardial reperfusion injury.
Assuntos
Vasos Coronários/fisiopatologia , Endotélio Vascular/fisiopatologia , Isquemia/fisiopatologia , Ativação de Neutrófilo , Animais , Vasos Coronários/ultraestrutura , Endotélio Vascular/ultraestrutura , Isquemia/patologia , Microcirculação , Microscopia Eletrônica de Varredura , Contração Miocárdica , Óxido Nítrico/fisiologia , SuínosRESUMO
To assess the impact of in vitro endothelialization on prosthetic graft patency, we performed femorotibial reconstruction in four patients. Polytetrafluoroethylene grafts (6 mm), lined with cultivated autologous endothelial cells, harvested from the veins of the forearm, were used. Autologous endothelial cells were harvested enzymatically and characterized by morphology and factor VII staining. After a cultivation period of 17 to 23 days, the cell count increased from 27 +/- 3 x 10(4) endothelial cells to 5.4 +/- 1.1 x 10(6). Endothelial cell seeding on polytetrafluoroethylene prostheses was then performed. To improve endothelial cell attachment to the graft surface, polytetrafluoroethylene grafts (60 to 70 cm; 6 mm diameter) were precoated with fibrin glue containing fibrin and fibronectin and the fibrinolysis inhibitor aprotinin. Seeding density of 49 +/- 10 x 10(3) endothelial cells per square centimeter yielded a preconfluent monolayer immediately after seeding, as demonstrated by scanning electron microscopy. A second cultivation period of 6 days, after seeding and before implantation, was necessary for establishment of a confluent monolayer and to allow for maturation of the endothelial cell cytoskeleton as well as production and excretion of extracellular matrix. Grafts endothelialized in vitro were implanted in four patients requiring femorotibial reconstruction. Scintigraphic studies with indium 111-labeled platelets demonstrated little or no platelet deposition, indicating persistent endothelialization. All grafts remained patent at 3 months after implantation.
Assuntos
Prótese Vascular , Endotélio/citologia , Politetrafluoretileno , Idoso , Aprotinina/farmacologia , Divisão Celular , Células Cultivadas , Citoesqueleto , Matriz Extracelular , Feminino , Adesivo Tecidual de Fibrina , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de TempoRESUMO
Endothelium-mediated relaxation and smooth muscle function in large coronary arteries are resistant to prolonged global ischemia. We used a small-vessel myograph to test the hypothesis that small intramyocardial artery endothelium and smooth muscle function have greater sensitivity to ischemic injury than large artery endothelium and smooth muscle. Normothermic global ischemia was induced in 15 porcine hearts. Intramyocardial arterial ring segments were assessed at 0, 30, 60, 90, and 120 minutes of ischemia in vitro with a small-vessel myograph. Potassium determined smooth muscle contraction, bradykinin endothelium-mediated relaxation, and sodium nitroprusside direct smooth muscle relaxation. Endothelium-mediated relaxation after 30 minutes of ischemia was similar to control (56% versus 66%) but was impaired at 60, 90, and 120 minutes of ischemia (32%, 11%, and 6%). Smooth muscle contraction was unchanged at 30 and 60 minutes compared with control (56 and 53 versus 63 mm Hg) but was significantly decreased at 90 and 120 minutes (33 and 13 mm Hg). Direct smooth muscle relaxation was significantly decreased at 120 minutes of ischemia compared with control (58% versus 95%). In a previous study, epicardial coronary artery endothelium-mediated smooth muscle vasodilation and direct smooth muscle vasodilation were preserved until 160 minutes of ischemia. After 160 minutes of ischemia, endothelium-mediated relaxation was lost and only direct smooth muscle vasodilation was preserved. In contrast to vasodilation, vasoconstriction was significantly reduced at 140 minutes of ischemia. These data indicate a greater and earlier adverse effect of ischemia on intramyocardial arterial endothelium-mediated relaxation than smooth muscle contraction or relaxation. These data support the hypothesis that there is an early functional endothelial cell injury associated with global ischemia. Relaxation that is endothelium-dependent in intramyocardial arteries is more sensitive to ischemic injury than in epicardial arteries. Unique to this study was the evaluation of small intramyocardial arteries (281 +/- 29 microns) that are the primary sites of coronary vascular resistance. Microvascular endothelial dysfunction after ischemia, therefore, may contribute to the "no-reflow phenomenon" seen during reperfusion injury.
Assuntos
Vasos Coronários/fisiopatologia , Microcirculação/fisiopatologia , Isquemia Miocárdica/fisiopatologia , Animais , Bradicinina/farmacologia , Endotélio Vascular/fisiopatologia , Técnicas In Vitro , Músculo Liso Vascular/fisiopatologia , Nitroprussiato/farmacologia , Suínos , Sistema Vasomotor/fisiopatologiaRESUMO
Bioprosthetic heart valves removed 76 to 150 months after implantation were morphologically investigated to correlate structural alterations with clinical failure modes. Traditional morphologic methods of evaluating valvular heterografts, such as microradiography and electron microscopy, were complemented by undecalcified ground sections, a new technique for analyzing the distribution of mineral deposits. Apart from well-investigated mechanisms that accelerate tissue degeneration, our observations point to additional facts: (1) phagocytosis of collagen fibrils and elastic material by macrophages and foreign body giant cells in areas near tears and perforations and (2) initial calcification indicated by delicate crystals in the intercellular space arranged in close relation to the periodicity of the cross-striation pattern of collagen fibrils. The present report not only calls attention to degenerative changes that are enhanced by mechanical stress but also underlines phagocytosis as an important mechanism in the destruction of bioprosthetic heart valves.
Assuntos
Bioprótese , Células Gigantes de Corpo Estranho/fisiologia , Doenças das Valvas Cardíacas/cirurgia , Próteses Valvulares Cardíacas , Macrófagos/fisiologia , Calcinose/patologia , Colágeno/análise , Feminino , Reação a Corpo Estranho/patologia , Humanos , Masculino , Microscopia Eletrônica , Pessoa de Meia-Idade , Fagocitose , Falha de Prótese , Estresse Mecânico , Fatores de TempoRESUMO
The shear stress resistance of cultured human endothelium was investigated on 6 mm polytetrafluoroethylene vascular grafts. Endothelial cell attachment was promoted by precoating the grafts with fibrin glue, which contained human fibronectin and inhibitors of fibrinolysis (aprotinin and tranexam acid). To evaluate the possible effect of fibrinolysis on cell detachment, seven grafts were lined with adult human saphenous vein endothelial cells (AHSVEC) and 11 with fibrinolytically almost inactive human umbilical vein endothelial cells (HUVEC). Endothelial cell seeding was performed in a microprocessor-controlled rotation device, allowing a low inoculum of 12 X 10(4) endothelial cells/cm2. Grafts were then cultivated for 9 days to enable the maturation of the cytoskeleton, before they were exposed to pulsatile shear stress for 48 hours. A mock circulation simulated the flow patterns and the wall shear forces of the femoral artery. After a 3-hour seeding process, 45% of AHSVEC and 43% of HUVEC were attached to the fibrin matrix, forming a confluent monolayer. After 24 hours of perfusion, a cell loss of 23% in AHSVEC- and of 42% in HUVEC-lined grafts was encountered. In spite of a further cell loss during the following 24 hours of perfusion, the majority of the graft surface was still covered by endothelial cells. Therefore we conclude that fibrin glue is a suitable substrate for the formation of a shear stress-resistant endothelial cell monolayer on polytetrafluoroethylene vascular grafts.
Assuntos
Aprotinina , Prótese Vascular , Endotélio Vascular/fisiologia , Fator XIII , Fibrinogênio , Trombina , Adesivos Teciduais , Ciclo Celular , Combinação de Medicamentos , Endotélio Vascular/citologia , Endotélio Vascular/ultraestrutura , Adesivo Tecidual de Fibrina , Humanos , Técnicas In Vitro , Perfusão , Politetrafluoretileno , Veia Safena/fisiologia , Estresse Mecânico , Veias Umbilicais/fisiologiaRESUMO
BACKGROUND: Pulmonary hypertension after cardiopulmonary bypass is a common problem in pediatric cardiac operations. This study tested the hypothesis that there is a difference between adult and immature pulmonary artery constrictor and dilator responses. METHODS: Reactivity of pulmonary artery ring segments from 22 mature (15 to 19 weeks) and 15 immature pigs (4 to 5 weeks) was tested in a vessel myograph. Potassium as a receptor-independent vasoconstrictor and phenylephrine as an alpha-receptor-mediated vasoconstrictor were used to assess smooth-muscle vasoconstriction. To assess endothelial cell function (nitric oxide production and secretion), we used increasing concentrations of bradykinin or acetylcholine. Sodium nitroprusside was used to produce maximum smooth-muscle relaxation at the end of each experiment. RESULTS: The data demonstrated maturation-independent endothelium and smooth-muscle-mediated vasodilation. Pulmonary artery ring segments from immature pigs had significantly less KCl constriction compared with mature pigs (p < 0.05). In contrast, pulmonary ring segments from immature pigs demonstrated enhanced alpha-receptor-mediated constriction compared with mature pigs. CONCLUSIONS: These data may explain perioperative pulmonary vasoconstriction in pediatric patients.
Assuntos
Agonistas alfa-Adrenérgicos/farmacologia , Fenilefrina/farmacologia , Artéria Pulmonar/fisiologia , Vasoconstrição/efeitos dos fármacos , Envelhecimento/fisiologia , Animais , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/fisiologia , Humanos , Hipertensão Pulmonar/fisiopatologia , Músculo Liso/efeitos dos fármacos , Músculo Liso/fisiologia , Artéria Pulmonar/efeitos dos fármacos , SuínosRESUMO
BACKGROUND: Cardiac rhabdomyoma is the most common primary heart tumor in infants. Spontaneous regression of such tumors is common, particularly with smaller lesions, followed by resolution of symptoms. Based on our data on spontaneous involution, our institutional philosophy has been one of expectant management in the absence of life-threatening symptoms. However, surgical intervention sometimes is required for the extirpation of a rhabdomyoma from the left ventricular outflow tract. METHODS: A retrospective review was conducted of 30 children in whom a rhabdomyoma was diagnosed over a 27-year period. RESULTS: Twenty-three percent (7/30) of the children required surgical extirpation of the tumor from within their left ventricular outflow tract, although a total of 94% had left ventricular involvement. There were no deaths. To date, no child has required reexcision of tumor. CONCLUSIONS: The natural history of rhabdomyoma is one of spontaneous regression (the 23 children who did not undergo surgical intervention are alive and continue to be followed up medically). We recommend surgical excision to alleviate acute outflow tract obstruction with reliance on the tumor's natural history of regression to achieve long-term freedom from reoperation. Although operation has been recognized as lifesaving, we were somewhat surprised to find that greater than 20% of our pediatric population required operative intervention.
Assuntos
Neoplasias Cardíacas/complicações , Rabdomioma/complicações , Obstrução do Fluxo Ventricular Externo/etiologia , Feminino , Seguimentos , Neoplasias Cardíacas/embriologia , Neoplasias Cardíacas/fisiopatologia , Neoplasias Cardíacas/cirurgia , Septos Cardíacos/patologia , Ventrículos do Coração/patologia , Humanos , Lactente , Recém-Nascido , Masculino , Regressão Neoplásica Espontânea , Reoperação , Estudos Retrospectivos , Rabdomioma/embriologia , Rabdomioma/fisiopatologia , Rabdomioma/cirurgia , Taxa de Sobrevida , Obstrução do Fluxo Ventricular Externo/embriologia , Obstrução do Fluxo Ventricular Externo/cirurgiaRESUMO
Thyroid dysfunction has been shown to have a significant impact on hemodynamic status and cardiac function. The purpose of this study was to determine the influence of triiodothyronine (T3) on cardiac functional recovery after ischemia in a dose-dependent manner. Postischemic functional recovery was assessed in isolated rabbit hearts mounted in a modified Langendorff preparation. Left ventricular systolic, diastolic, and peak developed pressures were measured before and after ischemia, and calculated as a percentage of preischemic function. Two cohorts of hearts were studied: the first was exposed to warm ischemia until a myocardial contracture of 4 mmHg was produced; the second cohort was exposed to warm ischemia until a contracture of 15 mm Hg was observed. In each cohort, T3 was added to the perfusion solution after ischemia in a physiologic concentration (2.5 x 10(-9) g/mL; 1 x T3), as well as ten times (2.5 x 10(-8) g/mL; 10 x T3) and a hundred times (2.5 x 10(-7) g/mL; 100 x T3) the physiologic concentration. One group, given the carrier only but without T3, served as the control. Rabbit hearts exposed to a short period of ischemia (4-mmHg diastolic contracture) showed increased recovery with 1 x T3 and 10 x T3. 100 x T3 did not bring about improved left ventricular recovery versus that in the control group. Rabbit hearts in the 15 mm Hg-diastolic contracture cohort showed increased recovery with 10 x T3 but not with 1 x T3. 100 x T3 led to decreased recovery in this cohort versus that in the control group.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Coração/efeitos dos fármacos , Isquemia Miocárdica/fisiopatologia , Tri-Iodotironina/farmacologia , Animais , Circulação Coronária/efeitos dos fármacos , Relação Dose-Resposta a Droga , Edema Cardíaco/etiologia , Coração/fisiopatologia , Técnicas In Vitro , Contração Miocárdica/efeitos dos fármacos , Isquemia Miocárdica/complicações , Coelhos , Tri-Iodotironina/administração & dosagem , Função Ventricular Esquerda/efeitos dos fármacosRESUMO
BACKGROUND: Surgical repair of congenital lesions associated with right ventricular outflow tract obstruction frequently requires the destruction of pulmonary valve (PV) components including the valve annulus. The resultant pulmonary insufficiency may lead to late functional deterioration of right ventricular performance. Acute right ventricular dysfunction has been associated with poor pulmonary runoff, tricuspid valve regurgitation, and pulmonary hypertension. Preservation of PV competence may prevent both early and late right ventricular failure. However, the recent trend towards earlier repair of tetralogy of Fallot (TOF) may preclude preservation of the PV in favor of a transannular patch. We reviewed our experience with surgical repair of TOF to determine if age and/or body size affected the ability to repair the PV. METHODS: We reviewed the clinical records of 50 consecutive children who underwent surgical repair of TOF by one surgeon. The latter 27 patients underwent repair with an intention to preserve their pulmonary valve. In total, 28 patients underwent repair with preservation of their pulmonary valve, and form the basis of this study. Serial echocardiographic assessments were performed early (3 to 6 months) and late (12 months) after surgery. RESULTS: Pulmonary valve preservation was possible in the majority of children (89%) in whom it was intended. Pulmonary valve competence was observed in 68% of children, with only 5 (16%) children demonstrating severe insufficiency at follow-up. Residual right ventricular outflow tract obstruction was present in only 1 child who underwent repair with pulmonary valve preservation at greater than 2 years of age. CONCLUSIONS: Our data suggest that earlier repair of TOF does not preclude preservation of the pulmonary valve and may indeed facilitate repair. The pulmonary valve remains competent at 12 months, with acceptable gradients, and should participate in somatic growth. Pulmonary valve preservation during repair of TOF may prevent free pulmonary insufficiency, progressive right ventricular dilation, and the need for future prosthetic pulmonary valve replacement.
Assuntos
Valva Pulmonar/cirurgia , Tetralogia de Fallot/cirurgia , Fatores Etários , Análise de Variância , Constituição Corporal , Superfície Corporal , Peso Corporal , Pré-Escolar , Ecocardiografia , Ecocardiografia Transesofagiana , Seguimentos , Crescimento , Humanos , Hipertensão Pulmonar/etiologia , Lactente , Complicações Pós-Operatórias , Implantação de Prótese , Valva Pulmonar/patologia , Insuficiência da Valva Pulmonar/etiologia , Estudos Retrospectivos , Tetralogia de Fallot/patologia , Insuficiência da Valva Tricúspide/etiologia , Disfunção Ventricular Direita/etiologia , Obstrução do Fluxo Ventricular Externo/cirurgiaRESUMO
BACKGROUND: Progressive stenosis of the pulmonary veins after repair of total anomalous pulmonary venous drainage is frequently refractory to surgical therapy. METHODS: Retrospective review of 170 consecutive patients treated for total anomalous pulmonary venous drainage identified 13 patients with postrepair pulmonary vein stenosis. Preoperative and operative data were analyzed to define the patterns of progression and efficacy of surgical techniques. RESULTS: Seventeen reoperations were performed in 13 patients. Postrepair pulmonary vein stenosis was most common in the infracardiac and mixed subtypes (p < 0.02). All 4 patients with unilateral stenosis, 2 of whom had progression of stenosis resulting in nearly complete unilateral pulmonary vein occlusion, survived. Six of 9 patients with bilateral disease died (p < 0.05 versus unilateral); 2 of the 3 survivors were repaired with a novel technique creating a sutureless neoatrium without evidence of restenosis at 1.8 years' follow-up. Stenting was uniformly unsuccessful. CONCLUSIONS: In unilateral stenosis, progression of disease may be survivable with loss of single-lung perfusion. Although bilateral disease is lethal in most cases, creation of a sutureless neoatrium has demonstrated short-term freedom from disease progression.
Assuntos
Veias Pulmonares/anormalidades , Veias Pulmonares/patologia , Constrição Patológica , Feminino , Cardiopatias Congênitas/mortalidade , Cardiopatias Congênitas/cirurgia , Humanos , Lactente , Recém-Nascido , Complicações Pós-Operatórias , Veias Pulmonares/cirurgia , Reoperação , Estudos Retrospectivos , StentsRESUMO
BACKGROUND: Recent reports have cited improving results for surgical management of isolated total anomalous pulmonary venous drainage. Complex cases (with other cardiac anomalies) are less frequently reported and are associated with higher mortality. METHODS: Retrospective review identified 170 consecutive patients treated for total anomalous pulmonary venous drainage from 1982 to 1996: 44 cases were "complex" (with significant associated cardiac lesions) and 126 cases were "simple." RESULTS: Operative mortality for simple cases decreased from 26% to 8%, and mortality for complex cases remained constant at 52%. Age, size, and the presence of atrial isomerism were univariate predictors of mortality. Multivariable analysis identified only univentricular hearts and associated cardiac lesions as predictors of operative mortality. Pulmonary artery (n = 16) and arteriopulmonary (n = 7) shunting strategies for complex cases resulted in less than 30% long-term survival. CONCLUSIONS: Despite improvement in survival for simple cases, management of total anomalous pulmonary venous drainage with single-ventricle hearts or other associated cardiac lesions remains problematic.
Assuntos
Cardiopatias Congênitas/complicações , Veias Pulmonares/anormalidades , Adolescente , Criança , Pré-Escolar , Cardiopatias Congênitas/mortalidade , Cardiopatias Congênitas/cirurgia , Humanos , Lactente , Recém-Nascido , Análise Multivariada , Estudos Retrospectivos , Taxa de SobrevidaRESUMO
In this report we have investigated the effects of BAX in enhancing apoptosis in two primary non-small cell lung cancer cell lines. A count of the apoptotic cells by TUNEL staining revealed that almost 70% of BAX over-expressing cells died, while very few apoptotic cells were detectable in the wildtype cells or in the cells transfected with an empty vector. These findings suggest that de-regulated expression of BAX may provide a novel mechanism for initiating cell death in non-small cell lung cancer cells. Further studies are needed to better define the involvement of this protein in the complex mechanism of lung carcinogenesis and to definitely demonstrate the therapeutic utility of targeting this pathway.
Assuntos
Apoptose/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Neoplasias Pulmonares/genética , Proteínas Proto-Oncogênicas c-bcl-2 , Proteínas Proto-Oncogênicas/genética , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Terapia Genética , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Proteínas Proto-Oncogênicas/biossíntese , Transfecção , Células Tumorais Cultivadas , Proteína X Associada a bcl-2RESUMO
Surgical treatment of post-traumatic or neoplastic bone defects often represents a problem in orthopaedic routine. Autologous tissue always stands for the first choice material. The recent therapeutic approaches for tissue repair of bone defects attempt to mimic the natural process of bone repair by delivering a source of cells capable of differentiating into osteoblasts. In this study two different types of human osteoblast cell harvesting were compared in primary cell culture to evaluate the best way to obtain cells for clinical use. Numerous articles describe the characteristics of each one of these systems, but there is no report comparing the influence of these different isolation techniques on cell growth. Cultures from 22 bone specimens obtained from donors were established in two different ways and their proliferation was compared. An enzymatic procedure to extract human osteoblasts (hOBcol) was used in one group; spontaneous cells outgrowth, human osteoblasts (hOBsog) was expected in the other group. Cells of both groups were characterised as osteoblasts by immunohistochemical staining with Bone Morphogenetic Protein-2,4 (BMP-2,4), expression of collagen type I as well as the amount of alkaline phosphatase activity (ALP) detected in the cell cultures. We found that the time needed in primary cultures till confluence was dependent on the age of the donors as well as on the method of cell harvesting. Cells from under 65-year old donors were growing significantly faster by the hOBcol method as compared to hOBsog 20.57 +/- 2.99 days vs. 30.00 +/- 4.36. Cells harvested from donors older than 65 years of age needed 23.88 +/- 2.95 in the hOBcol compared to 34.25 +/- 4.27 days in the other group. In the experimental cultures, after one passage with trypsin/EDTA, there was a significant difference between the two groups. There was an improved cell growth in the hOBsog group found on days 8, 9 and 10 of cultivation. Immunohistochemical staining as well as ALP activity were similar in both groups. In conclusion this study evaluated an important step for a tissue engineering approach to the repair of bone defects, which may have clinical applications in post-traumatic orthopaedic surgery.
Assuntos
Técnicas de Cultura de Células/métodos , Osteoblastos/citologia , Fator de Crescimento Transformador beta , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Fosfatase Alcalina/metabolismo , Antígenos/análise , Células da Medula Óssea/citologia , Proteína Morfogenética Óssea 2 , Proteína Morfogenética Óssea 4 , Proteínas Morfogenéticas Ósseas/imunologia , Calcificação Fisiológica/fisiologia , Cálcio/metabolismo , Divisão Celular , Separação Celular/métodos , Colágeno/biossíntese , Colagenases/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Osteoblastos/metabolismoRESUMO
In the last few years many attempts were made to line artificial vascular grafts with in vitro grown endothelial cell layers and thereby to minimize the risk of thromboembolism. However, adherence and resistance against shear stress forces were not tested under physiological pulsatile shear stress forces. In this paper, a mock-circulation apparatus is described, which simulates various forms of pulsatile shear stress, and which at the same time meets the requirements of cell cultivation. It can be sterilized and needs less than 700 ml of culture medium for priming. The generated flow profile can be adapted to a wide range of shear stress and also to different viscosities of used media. To take account of the different viscosities of culture medium and blood, a computerized calculation of the shear stress pattern was performed. Using the results of this computer model, the flow pattern was modified to obtain normal physiological shear stress when using culture medium. Results of pulse generation and simulation for the superficial femoral artery are presented.
Assuntos
Artérias/fisiologia , Modelos Cardiovasculares , Fluxo Pulsátil/fisiologia , Reologia , Fenômenos Biomecânicos , Artéria Femoral/fisiologia , Estresse Mecânico , Ultrassonografia , ViscosidadeRESUMO
To evaluate the possibility of providing, in vitro, an endothelial lining for artificial hearts, we cultivated adult autologous endothelial cells on two polyurethane and two silicone rubber surfaces. Over the ensuing 11-day period, we investigated the resulting cell proliferation and morphology by means of scanning electron and light microscopy. On the silicone rubber surfaces, seeding of 200,000 human saphenous vein endothelial cells per cm(2) produced an ideal cobblestone monolayer within a single day. In contrast, the polyurethane surfaces displayed an uneven, patchy distribution of endothelial cells. Scanning electron microscopy revealed microvilli and marginal overlapping in both groups. After the first day, the cell count on the polyurethane surfaces increased, whereas the count on the silicone rubber surfaces decreased. Morphologic investigations revealed that the ideally shaped cells initially on the silicone rubber had begun to overspread and subsequently to become detached, leaving denuded spheroid areas. Moreover, cultivation for 11 days on the polyurethane surfaces resulted in an unevenness of cell distribution that far exceeded the unevenness seen on the first day. Thus, despite the fact that materials with a high surface tension (such as silicone rubbers) seem to be ideal for initial cell spreading, subsequent cultivation results in cell detachment and death. On materials with a lower surface tension (such as polyurethanes), the less differentiated monolayers do at least proliferate, although their morphology remains unsatisfactory. Even if adult human endothelial cells should prove shear-stress-resistant, a minimum of 6 to 8 weeks would be required to establish autologous endothelial cell monolayers on the inner surface. Therefore, the endothelialization of artificial hearts is not possible when such hearts are used for urgent "bridging" before cardiac transplantation.
RESUMO
Seventeen male patients undergoing cardiopulmonary bypass (CPBP) surgery for aorto-coronary bypass grafting were investigated by scanning electron microscopy (SEM) for alterations of the surface morphology of circulating platelets. An initial decline in the percentage of unactivated smooth discocytes (SD) to 87 +/- 12% was found after thoracotomy. Three minutes after the onset of CPBP, the percentage of SD had dropped drastically to 59 +/- 13%, and by the 8th minute of CPBP it had dropped to its lowest point (49 +/- 19%). On the other hand, the percentage of shape-changed platelets (SC) increased to 17 +/- 9% after 3 minutes, and the percentage of pseudopod discocytes (PD) to 25 +/- 13% after 8 minutes. Surprisingly, a remarkable recovery of platelet morphology could be observed after even 15 minutes of CPBP, and by the end of bypass 78 +/- 15% of the circulating platelets had regained the smooth discoid (SD) appearance of unactivated platelets. We conclude that this recovery of platelet morphology is due to an increasing insensitivity of the platelets to activating stimuli during the course of CPBP. Our study provides evidence that the only major platelet activation occurs during the first minutes of CPBP, and that CPBP-caused platelet activation is much less pronounced than generally believed.
RESUMO
The beginning of angioplasty dates back to the turn of this century. Two lines of research were pursued over the past 40 years, organic and synthetic replacements of vessels. Autologous replacement, using an endogenous extremity vein, proved to be the most favourable variant. It was found to be not applicable to about 20 per cent of all patients, so that allogenic, semixenogenic or synthetic substitutes were required. Synthetic replacement was found to be characterised by a high rate of longtime closure, attributable to good biocompatibility of synthetic prostheses and low thrombogenic property of their inner surfaces. Endothelialisation of the prosthesis would be a possibility to improve the rate of success in bypass vascular surgery.
Assuntos
Arteriopatias Oclusivas/cirurgia , Prótese Vascular , Veias/transplante , Seguimentos , Humanos , Desenho de PróteseRESUMO
Primary adherence and attachment area of seeded human endothelial cells (EC) were determined on differently coated polytetrafluoroethylene (PTFE) grafts. Cell counts and morphometric analyses were done immediately after 60 minutes of electronically controlled seeding of 3 x 10(4) EC/cm2, as well as after 3 hours of subsequent incubation. Cell adherence and cell spreading were distinctly superior on two surface-covering substrates: fibronectin-treated type I/III collagen and fibrinolytically inhibited fibrin glue. Uncovered, purely fibronectin- or laminin-coated PTFE or type IV collagen treated with the specifically binding glycoprotein laminin showed a far lower EC attachment rate and less pronounced cell spreading. It appears that not only a high surface content of fibronectin but also a smooth PTFE-covering matrix are prerequisites for optimal primary adherence and cell spreading. Because fibrin glue might be fibrinolytically degraded despite its plasmin-inhibiting epsilon-amino-caproic acid compound, type I/III collagen plus fibronectin could provide an optimal precoating substrate for EC lining of PTFE grafts.