Biomedical subjects

L Bordenave

Publications and source records attributed to L Bordenave.

At least 55 records · Page 3Linked to original sources

PubMed1997

[Alteration of myocardial viability and systemic ventricular dysfunction after Senning procedure].

After a Senning procedure for transposition of the great arteries (TGA), systolic dysfunction of the right ventricle (RV) is common. Pre and peroperative chronicale hypoxia may be the cause of this ventricular myocardial alteration. In order to detect abnormalities of myocardial viability and to study their relationship to RV function, the authors studied 41 patients (pts), 11.3 +/- 3 years after a Senning procedure. All patients underwent myocardial scintigraphy of the RV under basal conditions. 1 hour after injection of 1.5 mCi of Thallium 201. The RV ejection fraction (n = 41) and at peak effort (n = 25). Exercise ability and aerobic capacity were assessed by exercise testing (Bruce) with gas exchange measurement and compared with 41 normal matched subjects. The cardiorespiratory response to exercise was altered in the Senning group : duration of effort (10.5 +/- 2 vs 13.2 +/- 2 min; p < 0.0001), peak VO2 (33 +/- 5 vs 44.4 +/- 6 ml/min/kg; p < 0.0001), anaerobic threshold (6 +/- 1 vs 8.4 +/- 1.9 min; p < 0.05) were lower compared to controls. Abnormalities of RV viability were observed in 18/41 pts (44%), moderate in 12 cases and severe in 6 cases. Resting and exercise RVEF were significantly lower in patients with myocardial defects (45 +/- 5 vs 51 +/- 7%; p < 0.0005, and 49 +/- 9 vs 58 +/- 9%; p < 0.05 respectively). In conclusion, after the Senning procedure for TGA, scintigraphic abnormalities of the systemic ventricule are common and associated with an alteration of systolic function at rest and on exercise.

Adolescent↗

PubMed1996

Optimization of use of UEA-1 magnetic beads for endothelial cell isolation.

Most endothelial cells (EC) in the body belong to the microvasculature. Isolation and subsequent culture of these microvessel EC contributes greatly to our understanding of the heterogeneity and vascular specificity that exist between one organ site and another. However, a major obstacle is the overgrowth of contaminating cells (fibroblasts, pericytes, smooth-muscle cells) in cultures. Since 1990 the use of magnetic beads in combination with either a lectin, Ulex europaeus agglutinin-1 (UEA-1), or a monoclonal antibody has represented a powerful tool for the isolation/purification of microvessel EC. In the former case, operative conditions remain to be optimized to obtain pure cultures of EC. We have performed studies to optimize conditions of use for magnetic beads coated with UEA-1. Incubating beads with cells, the influences are studied of time, temperature, cell concentration, and number of beads per target cell for two cell types, human umbilical vein EC (HUVEC) and skin fibroblasts (HSF), either isolated or mixed. The effect of the last parameter was also checked on the behavior of cells undergoing proliferation after isolation. Results, expressed as isolation efficiency (from 40% to 90%) allowed us to select a 15-min incubation time at 4 degrees C with rotary agitation, an optimal concentration of 4 x 10(5) cells/ml, and an optimal cell:bead ratio of 1:3. From a mixed cell population and in these conditions, even very low HUVEC:HSF proportions of 2.5:97.5 allowed us to obtain a pure HUVEC population in subsequent culture.

Culture Techniques↗

PubMed1996

Ability of various inserts to promote endothelium cell culture for the establishment of coculture models.

To select an insert suitable for human umbilical vein endothelial cell (HUVEC) culture, we compared several available inserts of 0.2 to 0.45 micron porosity: Cellagen (ICN), Transwell-COL (Costar), Millicell-HA and CM (Millipore), Anopore (Nunc), Cyclopore (Falcon) in comparison with a control surface (Thermanox). The requirements were: (i) to promote attachment, adhesion and proliferation of HUVEC (judged by [3H]thymidine incorporation into DNA at days 1, 3, 7); (ii) to allow HUVEC visualization by inverted, fluorescence microscopy for uptake of DiI-Ac-LDL and scanning electron microscopy, performed at day 9 after seeding. Because Transwell and Cellagen are collagen precoated and CM has to be coated for cell culture, we performed collagen coating (types I + III or IV) for non-pretreated inserts for the purpose of comparison. Our preferences comprise Transwell-COL, Cyclopore not coated or coated (whatever the collagen type), and Cellagen. However, on a quality/price ratio criterion, Cyclopore, even uncoated, is the insert of choice. The HA, CM and Anopore inserts, even coated, do not allow HUVEC growth but do not alter positive uptake of acetylated LDL.

Cells, Cultured↗

PubMed1996

Functional upregulation of granulocytes labeled with technetium-99m-HMPAO and indium-111-oxinate.

UNLABELLED: Indium-111-oxinate-labeled granulocytes have been used in vivo for several years for the detection of abscesses. Technetium-99-m-hexamethylpropyleneamine oxime (99mTc-HMPAO) labeling has more recently been described. METHODS: The influence of radiolabeling by both radiotracers on adhesion glycoprotein CD11b quantification was studied in quiescent and formyl-methionylleucylphenylalanine (fMLP)-activated neutrophils (PMN). Adhesion was assessed on human umbilical endothelial cells (HUVEC) as well as the repercussion of the granulocyte labeling on HUVEC viability (neutral red) and metabolic activity (MTT). Chemotaxis of PMN was evaluated by measuring migration under agarose with fMLP as chemoattractant. We also measured phagocytosis and the production of hydrogen peroxide induced by staphylococcus aureus. RESULTS: Whereas whole functional integrity is maintained after labeling, most of the functions (CD11b expression, adhesion, HUVEC metabolic activity) are up-regulated while chemotaxis is decreased in the presence of both radiotracers. Indium-111-oxinate induces larger alterations than 99mTc-HMPAO. CONCLUSION: These data were obtained in normal volunteers. In patients, alterations due to the in vitro labeling procedure, in addition to potential functional alterations caused by the underlying pathology, should be taken into account during image interpretation.

Cells, Cultured↗

PubMed1995

Beware of commercial human thrombins used to stimulate cultured endothelial cells.

In the field of in vitro biocompatibility testing, the investigation of cell response at the interface with a biomaterial is of great importance; there is a need for standard conditions and thus of well-defined and reliable sources of materials for an objective evaluation of cellular function. Thrombin is often used in vitro as a stimulating agent to check the specific functions of cultured endothelial cells. In the present work, and in order to select a thrombin of commercial origin, two criteria were borne in mind: purity towards the presence of the von Willebrand factor (vWF) and effectiveness towards vWF release by human umbilical venous endothelial cells (HUVEC) that have been submitted to four human commercial thrombins. We detected the presence of vWF in some thrombin solutions that have not yet been in contact with HUVEC. The different thrombins contained vWF antigen ranging from less than 0.1 mUnit per NIH unit of thrombin (from Diagnostica Stago and Sigma Chemical Co.) to 10-20 mUnit per NIH unit of Fibrindex thrombin (from Ortho Diagnostic Systems). Thus, if vWF is present in commercial thrombins, it contributes to and overestimates the vWF appearance in the media resulting from cell stimulation. Consequently, we fixed on thrombin from Diagnostica Stago for further studies involving HUVEC on biomaterials.

Biocompatible Materials↗

PubMed1994

Evaluation of cell colonization on biomaterials: preventing cell attachment to plastic containers.

In biocompatibility evaluation involving cell culture models, we use samples of biomaterials of different forms and sizes. During cell seeding onto biomaterials of an inadequate size to cover the bottom of the culture wells completely, cells have the opportunity to attach to the plastic. As described in this report with two culture models and two biomaterials, we use an agarose gel sublayer to prevent this phenomenon.

Biocompatible Materials↗

PubMed1994

In vitro study of the hemocompatibility of superparamagnetic contrast agent for magnetic resonance imaging.

Five different nanoparticles, potentially useful in magnetic resonance imaging (MRI) after venous administration, were studied for their hemocompatibility. The in vitro methodology evaluated these materials by several parameters: cytotoxicity towards cells cultured in vitro, aggregation ability of platelets, hemolysis inducibility, intrinsic and extrinsic coagulation pathway activation, and complement activation. With the proposed clinical dose, regardless of the cell type used (murine cell line or human endothelial cells) no toxicity was observed. The presence of the particles in blood did not produce any considerable damage: either hemolysis or platelet aggregation or blood coagulation were recorded. However, a slight decrease in aggregation ability of platelets was noticed as well as an increase in partial thromboplastin time. Because of the quick removal of the particles from the bloodstream, these phenomena must be short-lived, thus avoiding significant adverse clinical effects.

Animals↗

PubMed1994

An ex vivo original test using radiotracers for evaluating haemocompatibility of tubular biomaterials.

The haemocompatibility of a vascular prosthesis can be estimated as the result of its interaction with blood components. The authors describe an ex vivo canine shunt for evaluating isotopic haemocompatibility in blood-wall interactions. Methods employing radioisotopic tracers can be used to dynamically monitor the adsorption of labelled blood cells and proteins on different biomaterial surfaces. This ex vivo test should enable materials to be assessed for quality according to two thrombogenic criteria: (i) number of adhered platelets mm-2 s-1; (ii) quantity of adsorbed fibrinogen expressed as microgram mm-2 s-1, which would provide the basis for a scale of haemocompatibility.

Animals↗

PubMed1994

A canine ex vivo shunt for isotopic hemocompatibility evaluation of a NHLBI DTB primary reference material and of a IUPAC reference material.

Factors determining the thrombogenic response to particular artificial surfaces were investigated ex vivo in a canine shunt model. Methods using radioisotopic tracers made it possible to dynamically monitor the deposition of labelled blood cells and proteins on a NHLBI.DTB primary reference material polydimethylsiloxane (PRM.PDMS) and on a IUPAC reference material polyvinyl chloride (IUPAC.PVC). On the one hand, leukocyte affinity tau s(leu) (number of deposited leukocytes mm-2s-1) was not significantly different between IUPAC.PVC (tau s(leu) = 1.2-2.5) and PRM.PDMS (tau s(leu) = 1.5-3.4) and the fibrinogen adsorption rate varied from 33 to 48.10(-5) micrograms mm-2s-1 for both these materials. On the other hand, platelet affinity tau s(plat) (number of deposited platelets mm-2s-1) was significantly different (p < 0.05) for IUPAC.PVC and PRM.PDMS (tau s(plat)PVC = 683 +/- 200 > tau s(plat)PDMS = 327 +/- 80). Scanning electron micrographs of adherent platelets, red cells and leukocytes after blood contact ex vivo were performed after each experiment. This preliminary work contributes not only to quantify the adsorption of different radiotracers, but also to evaluate the superficial distribution of the labelled biological species on the inner surface of the tested biomaterials.

Animals↗

PubMed1993

Endothelial cell compatibility testing of three different Pellethanes.

There is a need for viable small diameter vascular grafts, the luminal surface of which could be seeded by endothelial cells (ECs) to prevent thrombosis. In order to select candidates for EC seeding before implantation, the in vitro cytocompatibility of three different Pellethanes (polyetherurethanes) using human ECs was investigated. The methodology included two stages depending on either direct contact between cells and materials or contact between cells and material extracts, obtained under standardized conditions. By the latter method, we observed a cytotoxic effect on cell growth with 2363-55 D Pellethane extract at a 50% (v/v) concentration in the nutrient medium, likely provoked by leachables and correlated with the lowest levels of tPA, PAI1, and vWF antigens in the supernatants. By the former method, we studied EC attachment and growth. Morphology was studied by classical means and completed by scintigraphy and microautoradiography after 111Indium-labeling of the EC monolayer. Differentiation was determined by the release of vWF antigen and measurement of vWF activity (multimeric organization) after human thrombin stimulation. Despite an inhibition of proliferation for both 55 D and 75 D types (compared to the control), a functional monolayer of ECs was obtained on 75 D. Pellethane 75 D could be the best support for in vitro endothelization.

Blood Vessel Prosthesis↗

PubMed1993

Radioimmunodetection of rat and rabbit cartilage using a monoclonal antibody specific to link proteins.

Biodistribution analysis using [125I]Fab-6F3 specific to link proteins from human articular cartilage performed in rats by autoradiography showed a high concentration of radioactivity in all cartilaginous tissues. Preliminary immunoscintigraphic assays were performed in rabbits. Front and side view images of whole animals exhibited high uptake in cartilage tissue of the knee articulation, in the invertebral disk and the humeral head. This fixation was still detected 24 h post-injection, although high washout of radioactivity was observed.

Animals↗

PubMed1993

Mathematical analysis of the upper respiratory tract from an anthropometric study.

In order to design a new, performed tracheal tube adapted to the shape of the upper respiratory tract, we have undertaken an anthropometric study from lateral x-rays of the neck in 130 patients with the head in a fixed position. In order to assess different clinical situations, we studied three groups of patients: group 1 = sitting, no tracheal tube; group 2 = supine, no tracheal tube; group 3 = supine, trachea intubated. We defined a standard coordinate system and determined mathematical curves for segments of the upper respiratory tract by a polynomial regression method. With these data it was possible to study the effects of physiological variations on the shape of the curves. It was then possible to determine at which region changes occur and the factors influencing these changes. We found that the relative position of the larynx was constant, whereas the hypopharynx exhibited the greatest change with the position of the head. These observations should allow us to construct a new, performed tracheal tube with elastic compliance properties to fit the changes occurring in the region of the hypopharynx.

Adolescent↗

PubMed1992

New artificial connective matrix-like structure made of elastin solubilized peptides and collagens: elaboration, biochemical and structural properties.

Several improvements of the basic reaction between elastin peptides and type III collagen, specially the addition of heparan sulphate proteoglycans, are presented. The consequent elaboration of either cell culture support or membranes are described and illustrated by scanning electron microscopy. The material produced possesses composition, very well organized lamellar structure and some properties quite close to natural membranes such as subendothelial tissue.

Biocompatible Materials↗

PubMed1992

New artificial connective matrix-like structure: thrombogenicity and use as endothelial cell culture support.

The recently described artificial connective matrix made of elastin solubilized peptides, type I+III collagens and connective proteins is shown to have structural and biological properties very close to the natural arterial subendothelium: the capacity to promote endothelial cell cultures maintaining their phenotype expression and its non-thrombogenicity. This new bioactive composite material could be used to replace arteries.

Biocompatible Materials↗

PubMed1992

A new apheresis system using a copolymer (polyvinyl alcohol triethylene glycol diacrylate) for removal of LDL from dog whole blood.

Studies were undertaken to determine the effectiveness of a copolymer composed of PVA-TEGDA (Poly Vinyl Alcohol TriEthylene Glycol) as a plasma-cholesterol lowering procedure. For a comparative study, five dogs underwent plasmapheresis including the transfusion bag containing gel in the plasma line, while three control dogs underwent the same plasmapheresis without gel. Numerous biological determinations were performed firstly in whole blood circulation before and after treatment over 10 days, and secondly in plasma before and after LDL binding on the gel. In the whole blood circulation, the average significant depletion of cholesterol levels was 31-51% for treated, 0-16% for control dogs and the average significant depletion of LDL cholesterol was 26-75% for treated and 0-3% for control dogs. Gel was therefore able to bind 121-217 mg of total cholesterol and 34-70 mg of LDL cholesterol per gram of gel. Lipid and lipoprotein levels rebounded 3-4 days after treatment. Adverse effects were not observed during all plasmapheresis. No significant differences between control plasmapheresis and gel-plasmapheresis were obtained for blood cell counts while lengthened coagulation times were observed during 24 h. Complement was not significantly activated by the copolymer as shown by a same decreased activity in the blood stream of all dogs: in fact, CH 50 depletion in the gel incubated plasma was due to a protein adsorption on the hydrogel. This new approach for LDL apheresis appears to be a promising new technique.

Acrylic Resins↗

PubMed1992

Cytocompatibility study of NHLBI primary reference materials using human endothelial cells.

Four different materials, low density polyethylene (LDPE), polydimethylsiloxane (PDMS), polyvinylcholoride (PVC) and cellulose, were selected by the Devices and Technology Branch of the National Heart Lung and Blood Institute (NHLBI) as primary reference materials for blood contacting. Among the wide variety of tests proposed to assess hemocompatibility of short-term blood contacting catheters, it was desirable to rule out whether these materials could release toxics for vascular cells of the physiological environment. Thus, the cytocompatibility of these materials have been checked towards human umbilical vein endothelial cells: the method used avoids direct contact between cells and materials but evaluates the effect of possible toxic substances leached from materials. These substances were obtained under defined conditions according to a standard. The results show that the extracts of cellulose and LDPE provoke an important cytotoxic effect on the endothelial cell cultures, while the extracts of PDMS and PVC allow the obtention of endothelial cell lining of the reference surface, with a correct global metabolic activity and the intracellular presence of von Willebrand factor.

Biocompatible Materials↗

PubMed1992

Physical and biological characteristics of the main biomaterials used in pelvic surgery.

Our study compared mechanical and biological properties of four materials classically used in surgery: polyethylene terephtalate (Mersilene), polypropylene (Marlex), polytetrafluoroethylene (Teflon) and expanded one (Gore-Tex) and polyaramide (Kevlar). No deterioration for polytetrafluoroethylene and polypropylene under irradiation was observed when materials were treated by physical means. Mechanical tests showed that all these materials could bear more than 50 N. Such a high tensile strength is never reached in visceral physiology. Results of graft elongation during tensile strength test shows two classes: a first one that includes high elongation grafts (Gore-Tex and Marlex) and a second one that includes low elongation grafts (Mersilene and Kevlar). As these materials have many potential uses in surgery, we have performed cytotoxicity tests. Material extracts were obtained under standardized conditions, and we have looked at a potentially toxic effect of substances eventually leached from the materials towards cells cultured in vitro. None of the material extracts listed above were cytotoxic except for untreated Kevlar. Toxicity disappeared when Kevlar was treated with methanol. As suspected, untreated Kevlar contains toxic additives introduced during the manufacture of this textile. Thus, in spite of good mechanical properties, Kevlar should not be used in pelvic surgery on account of its lower bicompatibility. These results shows that the choice of the grafts by surgeons must be relevant in a given application.

Drug Screening Assays, Antitumor↗

PubMed1991

In vitro evaluation of an epoxy resin's cytocompatibility using cell lines and human differentiated cells.

The cytocompatibility of a polyepoxy resin (Elf Aquitaine) has been studied using both cell lines and human differentiated cell cultures. The human models were gingival fibroblasts and bone osteoblasts, while the cell lines were Hela cells and 3T3 Balb/c cells. Basal cytocompatibility was assessed by estimation of the cell proliferation, total cell protein content, cell membrane sub-lysis, and cell attachment and spreading. Specific cytocompatibility concerning human osteoblasts, from both alveolar and trabecular bone, was determined by measuring the intracellular alkaline phosphatase activity. Resin colonization by the cells was studied by both TEM and SEM. The behaviour of the two cell lines reveals a significant level of discrepancy, whereas the behaviour of human cells, whatever the model, is comparable; however, osteoblasts look more sensitive. Moreover, the results show that this epoxy resin exhibits a moderate cytocompatibility which could be the result of the cytotoxicity of early released products, associated with the considerable surface roughness.

Alkaline Phosphatase↗