Biomedical subjects

G Pascual

Publications and source records attributed to G Pascual.

26 records · Page 2Linked to original sources

PubMed1999

Effect of relaparotomy through previously integrated polypropylene and polytetrafluoroethylene experimental implants in the abdominal wall.

BACKGROUND: The appearance of new pathologies affecting abdominal organs after implant of a prosthesis to repair an abdominal wall defect may necessitate reintervention. The aim of this study was to compare the behavior of two types of biomaterial widely used in clinical practice, polypropylene (PL) and polytetrafluoroethylene (ePTFE), after a second laparotomy involving the implant. The behavior, in terms of tensile resistance and integration with tissues, of intact prostheses was compared to that of prostheses subjected to opening and repair. METHODS: A defect (7x5 cm) involving all tissue layers was created in the anterior abdominal wall of 24 male New Zealand rabbits. These defects were repaired with a reticular, macroporous PL mesh (Marlex, Bard Card., Madrid, Spain) or a laminar, micro/macroporous ePTFE prosthesis (Mycro Mesh, W.L. Gore, Flagstaff, AZ) of similar size to the defect. Four study groups were established: Intact PL/Intact ePTFE (n = 6 each): animals implanted with a PL or ePTFE prosthesis and sacrificed 90 days after implant; Repaired PL/Repaired ePTFE (n = 6 each): animals implanted with a PL or ePTFE prosthesis subjected to midlongitudinal relaparotomy through the center of the prosthesis 90 days postimplant, followed by repair with continuous polypropylene 4/0 suture. Animals in repaired groups were sacrificed 90 days after the second intervention. Specimens comprised of prosthesis and neoformed tissue were subjected to light and scanning electron microscopy. In addition, 2 cm-wide strips, consisting of the prosthesis and anchorage tissue, were subjected to biomechanical analysis using an Instron tensiometer (Instron, Canton, MA). The results obtained were statistically compared using the Mann-Whitney U-test. RESULTS: The intact PL implants were fully infiltrated by dense, disorganized, well-vascularized scar tissue with fibers concentric to the mesh monofilaments. The appearance of the repaired PL prostheses was similar, with establishment of neoformed tissue in repaired areas of the prosthesis such that both cut edges of the prosthesis were joined together. In contrast, intact ePTFE prostheses were encapsulated by organized tissue with fibers running parallel to the surface of the biomaterial. Repaired ePTFE prostheses including sutured areas were similarly encapsulated. But the edges of the sutured middle area did not fuse. Tensile resistance values of intact and repaired PL prostheses were similar (intact, mean, 34.78 Newtons; repaired, mean, 34.74N, p>0.001). Tensile resistance values of intact ePTFE implants were significantly different to those of the repaired ePTFE prostheses (intact, mean, 22.64N; repaired, mean, 17.21N, p<0.001). Breakage of both types of PL specimen strips was restricted to recipient tissue while breakage of intact ePTFE specimens occurred in the areas of anchorage to the abdominal wall. Rupture of repaired ePTFE specimens took place in the sutured central areas of the prostheses. CONCLUSIONS: We conclude that relaparotomy through an existing PL prosthesis previously integrated with the abdominal wall does not affect the tissue integration process or the tensile resistance of the implant. When the relaparotomy involves an ePTFE prosthesis, however, although the repair process itself is unaffected, significant loss in tensile strength is incurred. In addition, relaparotomy through both types of biomaterial is likely to result in the neoformation of adhesions in the areas of the prosthesis subjected to opening and repair but, in general, the number of adhesions formed in the presence of intact or repaired polypropylene implants was larger than that observed with the use of ePTFE.

Abdominal Muscles↗

PubMed1999

Arterial damage induced by cryopreservation is irreversible following organ culture.

OBJECTIVES: The aim of the present study was to investigate the changes which occur to the arterial wall following cryopreservation and thawing and to determine whether these changes are reversible after a week of culture in an organ bath. MATERIALS AND METHODS: Rat iliac arterial segments were cryopreserved. Once thawed, the arterial segments were cultured for a period of 0, 1, 2, 4 or 7 days. Freshly isolated rat iliac vessels cultured for 7 days served as the control group. Evaluation was made of ultrastructural changes, the expression of metalloproteinase activity (MMP-1, MMP-3 and MMP-9) and the apoptotic state of cells. RESULTS: The freezing-thawing process induced damage to the arterial segments compared to fresh control vessels. After 1 week of culture, arteries showed a high degree of tissue degeneration. Only a few individual endothelial cells remained on the luminal surface. There was a gradual increase in the proportion of apoptotic cells. The sequential expression of MMP-1 during the first 2 days and subsequent expression of MMP-3 and MMP-9 were of most significance. CONCLUSIONS: Cryopreservation induced damage to the vessels which could not be reversed by organ culture. The changes observed in the expression of metalloproteinases may be indicative of the degenerative process which occurs in the extracellular matrix.

Animals↗

PubMed1999

Neoperitoneal formation after implantation of various biomaterials for the repair of abdominal wall defects in rabbits.

OBJECTIVE: To study the interfaces between the visceral peritoneum and some of the biomaterials used to repair defects in the abdominal wall. DESIGN: Animal study. SETTING: School of medicine, Spain. MATERIAL: 48 New Zealand white rabbits divided into 4 groups of 12 each. INTERVENTIONS: Full thickness defects 50 x 70 mm were created in the abdominal wall and repaired with polytetrafluoroethylene (PTFE, Soft Tissue Patch), one of two polypropylene patches (Marlex and Prolene), or lyophylised dura mater (Lyo-Dura). 3 animals from each group were killed at 14, 30, 60 and 90 days and specimens examined by light microscopy, scanning electron microscopy, and immuno histochemistry by labelling of macrophages with RAM-11, a specific monoclonal antibody (MoAb). MAIN OUTCOME MEASURES: Infection, healing, development of adhesions, and histological appearance of the interface. RESULTS: Tissues responded similarly to materials of similar structures. Layered prostheses (PTFE and Lyo-Dura) caused formation of a well organised neoperitoneum with few adhesions to the abdominal viscera (loose adhesions in 2 animals in each group), whereas the mesh prostheses generated a disorganised neoperitoneum with many adhesions (Marlex loose adhesions 3, firm 8, and integrated 1; Prolene loose adhesions 2, firm 8, and integrated 2). Lyo-Dura was associated with the formation of areas of calcification. Labelling of macrophages with the MoAb showed that they were in direct contact with all materials studied. CONCLUSIONS: Layered biomaterials with little or no porosity (PTFE and Lyo-Dura) are the most suitable of the four for implantation in sites where the prosthesis is in contact with the visceral peritoneum, because they induce minimal adhesions.

Abdominal Muscles↗

PubMed1999

IkappaB kinase (IKK)-associated protein 1, a common component of the heterogeneous IKK complex.

Activation of the transcription factor NF-kappaB is controlled by the sequential phosphorylation, ubiquitination, and degradation of its inhibitory subunit, IkappaB. We recently purified a large multiprotein complex, the IkappaB kinase (IKK) signalsome, which contains two regulated IkappaB kinases, IKK1 and IKK2, that can each phosphorylate IkappaBalpha and IkappaBbeta. The IKK signalsome contains several additional proteins presumably required for the regulation of the NFkappaB signal transduction cascade in vivo. In this report, we demonstrate reconstitution of IkappaB kinase activity in vitro by using purified recombinant IKK1 and IKK2. Recombinant IKK1 or IKK2 forms homo- or heterodimers, suggesting the possibility that similar IKK complexes exist in vivo. Indeed, in HeLa cells we identified two distinct IKK complexes, one containing IKK1-IKK2 heterodimers and the other containing IKK2 homodimers, which display differing levels of activation following tumor necrosis factor alpha stimulation. To better elucidate the nature of the IKK signalsome, we set out to identify IKK-associated proteins. To this end, we purified and cloned a novel component common to both complexes, named IKK-associated protein 1 (IKKAP1). In vitro, IKKAP1 associated specifically with IKK2 but not IKK1. Functional analyses revealed that binding to IKK2 requires sequences contained within the N-terminal domain of IKKAP1. Mutant versions of IKKAP1, which either lack the N-terminal IKK2-binding domain or contain only the IKK2-binding domain, disrupt the NF-kappaB signal transduction pathway. IKKAP1 therefore appears to mediate an essential step of the NF-kappaB signal transduction cascade. Heterogeneity of IKK complexes in vivo may provide a mechanism for differential regulation of NF-kappaB activation.

Amino Acid Sequence↗

PubMed1999

Role of macrophages in myocardial apoptosis following cardiac transplant. Influence of immunosuppressive treatment.

Cytotoxic T cells may induce myocardial apoptosis by histiocyte activation during rejection following allogenic heart transplant. The aim of the present investigation was to evaluate the macrophage response and its relationship to the programmed death of cardiomyocytes in rejection and during cyclosporin-A (CsA) treatment. An abdominal, heterotopic heart transplant rat model was used establishing two groups: singenic (ST) and allogenic (AL) transplant. 5 mg/kg/day (s.c.) CsA (Sandimun) was administered to half of the animals in each group. Morphological and structural analysis was performed 7, 14, 21, 30, 50 and 100 days post-transplant. Macrophages were detected using the monoclonal antibody (ED1). The TUNEL method was used to visualise apoptotic cells. Two weeks after ST in animals without immuno-suppressive treatment, the transplanted myocardium had been extensively infiltrated by inflammatory cells, many of which were ED1-positive. At 21 days follow-up, the number of labelled cells had fallen. In animals treated with CsA the amount of ED1-positive cells was lower than that seen in the anterior group. Only a few isolated cells of the infiltrate were TUNEL-positive. In the AT group, rejection took place between 9-15 days in the untreated animals. The myocardium was highly infiltrated by mononuclear cells. Some were ED1-positive. Small groups of apoptotic cells were visible in the infiltrate and in some vessel lumens. Rejection was resolved in animals treated with CsA. The macrophage response diminished during follow-up in a similar way to that occurring in the ST. Few cells showed TUNEL positivity. It may be concluded that: a) CsA treatment diminishes the amount of infiltrated macrophages; b) animals receiving ST or AT, show a low level of apoptosis; c) in the present model, the apoptosis of cardiomyocytes does not appear to be induced by macrophages; and d) in this model it is not possible to relate apoptosis and rejection.

Animals↗

PubMed1998

Effects of ischaemia-reperfusion and cyclosporin-A on cardiac muscle ultrastructure.

The present study investigates the effects on the cardiac muscle cell of two of the determining factors for the success of organ transplant; ischaemia-perfusion and immunosuppressive treatment with cyclosporin-A (CsA). To this end an abdominal, heterotopic heart transplant model in singenic Sprague-Dawley rats was employed. Three study groups were established: Group I (control, n = 15) animals undergoing heart transplant without treatment; Group II (n = 15) animals undergoing heart transplant and subjected to a daily dose of CsA in a cremophor vehicle (Sandimun) (5 mg/kg/sc); Group III (n = 15): animals undergoing heart transplant and administered a daily dose of pure CsA (5 mg/kg/sc). Recipient animals were sacrificed 7, 14, 21, 30 and 50 days after transplant. During the post-operative period, heart function was assessed by daily abdominal palpation. Graft specimens obtained at each follow-up period were subjected to light and transmission electron microscopy. Immunohistochemical analysis of specimens was performed using the rat macrophage-specific monoclonal antibody MCA-341. The ischaemia/reperfusion process induced considerable alteration to cardiac muscle cells of control animals. Effects, apparent after the first week of transplant, included mitochondrial swelling and loss of cristae, hypertrophy of the sarcoplasmic reticulum and structural changes to sarcomeres. Two weeks after transplant, the myocardium was infiltrated by inflammatory cells. These effects diminished 30 days post-transplant. Cardiac tissues of treated animals (groups II and III) showed similar behaviour although, in the latter group, mitochondrial damage was greater and intense myocardial fibrosis took place. Infiltration of cardiac muscle by white blood cells did not take place until 3 weeks post-implant. These results indicate: a) The ultrastructural changes detected in cardiac fibres of animals of the three study groups were attributable to the ischaemia/reperfusion process rather than to treatment with CsA; b) CsA appears to augment mitochondrial damage and myocardial fibrosis; c) the inflammatory response was delayed and reduced by the immunosupressant; and d) the cremophor administration vehicle did not seem to exert an independent toxic effect on the myocardium.

Animals↗

PubMedDate not supplied

Modulation of PECAM-1 (CD31) expression in human endothelial cells: effect of IFNgamma and IL-10.

Intercellular contacts formed between endothelial cells (EC) permit the formation of a confluent monolayer and play a major role in the recruitment and the migration of leukocytes in the inflammatory response. It is currently accepted that cytokines are responsible for the signals involved in the induction of such endothelial alterations. The platelet EC adhesion molecule (PECAM-1), a specific component of EC junctions, is one of many molecules which participate in the regulation of EC interaction with blood components. Given that the regulatory mechanisms which affect the expression of this adhesion molecule are only partially understood, the aim of the present study was to compare the effects of two antagonistic inflammatory cytokines, interferon (IFN)-gamma and interleukin (IL)-10, on the expression of PECAM-1. Human umbilical vein EC grown to subconfluence were stimulated with IL-10 (10 ng/ml) and/or IFNgamma (100 U/ml) for 24 h. PECAM-1 expression was determined by FACScan and immunofluorescence. Morphological analysis of the cell cultures was performed by optical and scanning electron microscopy. In the presence of IL-10, no changes in cell growth and morphology or in the intensity of PECAM-1 expression were observed. However, when the cultures were treated with IFNgamma, the EC acquired a fibroblast-like appearance, growth was disorganized and PECAM-1 disappeared from cell junctions. The mean intensity expression and the percentage of EC expression of this antigen were analyzed by flow cytometry and significantly decreased after culture in the presence of IFNgamma. The simultaneous addition of IFNgamma and IL-10 to the EC cultures induced modifications similar to those observed in the presence of IFNgamma alone. Regulation of the expression of PECAM-1 with the subsequent functional implications seems to be dependent upon the IFNgamma signal and it is unaffected by IL-10. The different effects shown by IL-10 and IFNgamma on the expression of PECAM-1 in EC could reflect opposite regulatory actions of the inflammatory response induced by these cytokines.

Blotting, Western↗

PubMedDate not supplied

A prospective and immunologic study of factory workers exposed to TDI (toluene diisocyanate).

Thirty factory workers whose annual exposure to TDI amounted to 280 hours were examined. Seven of them had been removed from their jobs for presenting respiratory symptomatology and a further five were removed for presenting bronchial asthma. Their medical histories were consulted and further measures were taken such as a radiological thorax study, total IgE, TDI, MDI and HDI RAST, a basal spirometric study and finally a provocation test. The Rast proved negative in every case. In the spirometric study carried out on the provocation test, four cases showed a significant decrease in the FEV1 over 20% and over 40% in the FMEF and PEFR. There was no connection between the four patients who presented an elevated total IgE and the four who presented a positive provocation test. The provocation test proved negative in five of the seven patients removed from their places of work. Two of the four workers who responded positively to the provocation test remained in their places of work during the TDI foam test without showing any symptoms. The patients who presented symptoms did not appear to present bronchial obstruction during the provocation test. Nonetheless, patients who seemingly had not shown any symptoms presented what appeared to be bronchial obstruction.

Adult↗