Biomedical subjects

R Bareille

Publications and source records attributed to R Bareille.

54 records · Page 3Linked to original sources

PubMed1997

Opposing actions of BMP3 and TGF beta 1 in human bone marrow stromal cell growth and differentiation.

BMP3 and TGF beta 1 were found to induce opposite effects on human bone marrow stromal cells both in cell proliferation and cell differentiation and on calcium deposition onto the extracellular matrix. Moreover, BMP3 may exert, in part, the inhibitory effect of TGF beta 1 through decreasing the affinity of TGF-beta for its receptors which are now identified in these human bone cells.

Activin Receptors, Type I↗

PubMed1996

Colonization of a calcium phosphate/elastin-solubilized peptide-collagen composite material by human osteoblasts.

The evaluation of the suitability of a new biomaterial as a possible substitute for bone tissue is described here. This biomaterial is based on calcium phosphate particles linked to an artificial connective matrix, the elastin-solubilized peptides (ESP) associated with type I and III collagens. This work demonstrates the feasibility of shaping this composite material into discs, describes its microstructural characteristics, and evaluates its capacity as a substrate for the proliferation of human osteoblasts without loss of their phenotypic expression.

Adsorption↗

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

The resorption of bone-implanted corals varies with porosity but also with the host reaction.

Three different exoskeletons of coral species Porites asteroides (P), Montastrea annularis (M), and Dichocoenia stokesi (D) were implanted for 2-20 weeks in rabbits. At 2, 4, 8, or 20 weeks, the exoskeletons presented variations in their resorptions depending on the species. To understand the variations in the decreasing speed of the implants despite their similar chemical composition, a study of the surface and architecture of the coral was carried out using scanning electronic microscopy, porosity was evaluated, and growth and differentiation of osteogenic cells cultured in vitro were observed for more than 1 month. At the cellular level, the surface of the implants was identical. Three-dimensional structures of the implants were variable, but the porosity values [P = 42.7%, M = 40.7%, and D = 17.4%] could not completely account for the differences in the resorbing process of the species. Standard histologic studies performed at 2, 4, 8, and 20 weeks after implantation produced the same pattern with P or M, showing aspects of rapid resorption; however, with D there were images resembling those of a foreign-body reaction. It seems that when resorption is not quick enough, a foreign body reaction develops which further slows down the process. This work focuses on the importance of porosity when using coral as bone substitute.

Animals↗

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

Osteogenin (bone morphogenic protein 3) inhibits proliferation and stimulates differentiation of osteoprogenitors in human bone marrow.

Treatment of human bone marrow osteoprogenitors with osteogenin (BMP-3; at 1, 2.5 and 10 ng/ml) caused dose- and time-dependent inhibition of DNA synthesis and cell proliferation. Simultaneously, osteogenin stimulated type I collagen synthesis and cAMP production. Addition of osteogenin to the cell culture increased intracellular alkaline phosphatase activity and osteocalcin synthesis, with maximal stimulation at 2.5 ng/ml. Simultaneous addition of 2.5 ng/ml osteogenin and 1,25 dihydroxy vitamin D3 (10(-8) M) enhanced the stimulation observed in osteocalcin synthesis. The experiments reported here demonstrate the significant "in vitro" influence of osteogenin in the stimulation of osteogenic phenotype in osteoprogenitor cells which have been isolated from human bone marrow and cloned. These results support a reciprocal relationship between cell growth inhibition and expression of osteoblast differentiation.

Adult↗

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

Human bone marrow stromal cells express an osteoblastic phenotype in culture.

This study reports the selection and characterization of osteogenic precursors from human bone marrow which were isolated by two "clonings" and successive subculturing. These cell lines express alkaline phosphatase activity. Gel electrophoresis of [3H]-proline labeled cultures showed that the cloned cells produce only type I collagen. They synthetize osteocalcin and osteonectin. They respond to 1,25 dihydroxy vitamin D3 by increasing osteocalcin synthesis and secretion, and to parathyroid hormone by increasing cyclic AMP synthesis. After the third subculture in the absence of beta-glycerophosphate, these cell lines formed lots of clusters which exhibit high alkaline phosphatase activity and positive von Kossa staining. X-ray energy spectrum shows that these cells are surrounded by "budding" structures containing calcium and phosphorus with a ratio Ca:P identical to those of pure hydroxyapatite. This process was associated with 45Ca uptake into the cells. All these data support the selection of osteogenic cells which may be of considerable clinical importance.

Adult↗

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↗

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

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

Biodistribution study of a monoclonal antibody specific to link proteins from human cartilage.

We report the use of monoclonal antibody 6F3 prepared against link proteins from human articular cartilage to elucidate the distribution of these glycoproteins within connective and other tissues. By immunohistochemical analysis, we showed that only the Fab fragment could reach the antigenic site in human articular sections. Cross-reactivity of the antibody 6F3 with link proteins purified from rat articular cartilage allowed us to carry out a biodistribution analysis in vivo in the rat. The time course of whole blood and plasma showed maximal activity 6 h after the 20 micrograms i.v. injection of [125I]Fab-6F3. Urinary excretion seems to be a high route of elimination. Moreover, we noticed no radioactive uptake across the blood-brain barrier. A significant fixation of labeled antibody Fab-6F3 was observed in noncartilaginous connective tissues such as aorta, skin and lung. As expected, specific and increased radioactivity was observed in all cartilage tissues, this increase was significantly higher 6 h after the [125I]Fab-6F3 injection than in the other connective tissues.

Animals↗

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↗

PubMed1990

Biocompatibility of polyacrylamide microcapsules implanted in peritoneal cavity or spleen of the rat. Effect on various inflammatory reactions in vitro.

The biocompatibility of polyacrylamide microcapsules was investigated by implanting microcapsules in the peritoneal cavity or the spleen of rats. The capsules were retrieved every four weeks for twenty weeks. They remained isolated and free in the peritoneal cavity, but led to a slight inflammatory reaction in the spleen. These results were confirmed, in vitro, by determinations of chemiluminescence and interleukin I levels, and by evaluation of attachment and growth of human fibroblasts. The microcapsules did not enhance these inflammatory reactions, and they were a poor support for cellular proliferation.

Acrylic Resins↗