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Biomedical subjects

L Bordenave

Publications and source records attributed to L Bordenave.

At least 37 records · Page 2Linked to original sources

Development of RGD peptides grafted onto silica surfaces: XPS characterization and human endothelial cell interactions.

The attachment of human umbilical vein endothelial cells (HUVECs) on substrates that had been covalently grafted with the cell adhesion peptides Arg-Gly-Asp (RGD) was investigated. This approach was used to provide substrates that are adhesive to cells even in the absence of serum proteins and to cells that have had no prior treatment of the surface with proteins that promote cell adhesion. We wanted to improve control of cellular interactions with cell-adhesive materials by providing fixedly bound adhesion ligands. Silica was examined as a model surface. The peptides were grafted using three different steps: grafting of aminosilane molecules; reaction with a maleimide molecule; and immobilization of cell-binding peptides containing the RGD sequence. The RGD-grafted surface was characterized by X-ray photoelectron spectroscopy (XPS) and contact-angle measurements.

Amino Acid Sequence↗

Review of 10 years of the clinical use of brain natriuretic peptide in cardiology.

Ten years ago brain natriuretic peptide (BNP), the second compound of a family of polypeptide hormones named natriuretic peptides was identified. This peptide has great pathophysiologic importance as a stress-induced cardiac hormone secreted from ventricles, and it rises in several cardiac diseases. It promotes natriuresis and diuresis, acts as a vasodilator, and antagonizes the vasoconstrictor effects of the renin-angiotensin-aldosterone system. The measurement of this peptide in blood by immunoassay has shown promise over the past decade in clinical diagnosis and prognosis. Because heart failure is a major health problem worldwide, BNP is proposed as a biochemical marker that might provide a useful screening test to select patients for further cardiac investigations. Such a hormone assay is inexpensive and available. The implications of BNP in diagnosis, prognosis, and therapy will be reviewed.

Animals↗

In vitro and in situ intercellular adhesion molecule-1 (ICAM-1) expression by endothelial cells lining a polyester fabric.

In order to improve long-term patency of vascular grafts, the promising concept of endothelial cell seeding is actually under investigation. Our laboratory tested a polyester coated with albumin and chitosan which permits a rapid colonization by human umbilical vein endothelial cells (HUVEC) and it seems relevant to test in vitro the expression of adhesive molecules expressed by cells with regard to the inflammatory process. We studied intercellular adhesion molecule-1 (ICAM-1) expression and focused our work on the determination of ICAM-1 sites expressed per adherent cell lining the biomaterial, thus in situ, in comparison to control HUVEC on plastic wells: the results obtained by binding experiments were correlated to flow cytometry analyses and showed that the polyester does not induce a proinflammatory state and that HUVEC covering the structure are able to respond to a stimulus.

Antibodies, Monoclonal↗

In vitro cytocompatibility of radio-opacifiers used in ureteral endoprosthesis.

Ureteral endoprostheses are urinary catheters made of polymeric biomaterials made radio-opaque through the addition of X-ray absorbing additives such as barium, bismuth, tantale or tungsten. The aim of this work was to study the in vitro toxicity of solutions of these radio-opacifiers using two cell culture models. Primary-cultures of human urothelial cells (HUC) arising from normal adult urinary tract and permanent urothelial cell line were used. Solutions at different dilutions were placed into the wells containing monolayers of confluent cells. After 24 h incubation period, the solutions were removed and cell viability and cell metabolic activity tests were performed (Neutral Red assay and MTT assay). At a concentration lower than 1 mg l(-1) the different radio-opacifiers used showed no toxicity. From 1 to 3 mg l(-1) one can note a significant dose-dependent decrease of cell metabolic activity of solely HUC for barium chloride. At 3 mg l(-1) one can note a significant deleterious effect on HUC metabolic activity, with bismuth and tantale. For tungsten, there is no deleterious effect, but on the contrary a significant increase in HUC metabolic activity at a 0.5 mg l(-1) concentration. None of the solutions did provoke alterations in HUC viability for concentrations less than 3 mg l(-1). Interestingly, for permanent cell line one can note a solely significant decrease of cell viability at 3 mg l(-1) for tantale. All the other tested salts on permanent cell line were not significantly different from controls for cell viability as well as cell metabolic activity. HUC culture model may be of relevance for the screening of radio-opacifiers intended for ureteral endoprostheses.

Adult↗

In vitro assessment of endothelial cell adhesion mechanism on vascular patches.

Endothelial cell (EC) seeding of small caliber vascular grafts prior to their implantation has proved to significantly improve long-term patency in humans. We have previously demonstrated that a monolayer of EC could be obtained on type I collagen-coated knitted ultrathin polyster grafts (InterVascular, La Ciotat, France). Thus, the aim of the present work was to understand the nature of cell adhesion mechanisms involved in the cell /biomaterial interface, using HemaCarotid (InterVascular) patches made of type I collagen-coated knitted ultrathin polyster (type I collagen is used to coat patches to attain low permeability). By means of quantitative attachment tests, adhesion blocking assays, RT-PCR for the expression of beta1 integrin mRNA, indirect immunofluorescence with antivinculin antibody, we were able to show that EC are able to adhere to such surfaces by the means (non-unique) of cell surface receptors of the beta1 integrin group. However, the latter are probably downregulated at the cell/biomaterial interface.

Journal Article↗

Clinical performance of vascular grafts lined with endothelial cells.

The replacement of arteries with purely synthetic vascular prostheses often leads to the failure of such reconstructions when small-diameter or low-flow locations are concerned, due in part to the thrombogenicity of the internal graft surface. In order to improve long-term patency of these grafts, the concept of endothelial cell seeding has been suggested because this metabolically active endothelial surface plays major roles in preventing in vivo blood thrombosis and because vascular grafts placed in humans do not spontaneously form an endothelial monolayer whereas they do in animal models. The composite structure resulting from the combination of biologically active cells to prosthetic materials thus creates more biocompatible vascular substitutes. To achieve endothelialization of synthetic vascular grafts, previous efforts aimed at "one-stage" procedure (adding autologous endothelial cells to the graft at the time of implantation) in the 1980's seemed clinically feasible but results of reported clinical trials were controversial and mostly disappointing. An alternative method is an in vitro complete and preformed endothelial lining at the time of implantation: the "two-stage" procedure which implies harvest and culture of autologous endothelial cells. Up to date, the latter approach demonstrated its superiority in terms of significantly increased patency of the grafts that underwent endothelialization eight years earlier.

Blood Vessel Prosthesis↗

Expansion and differentiation of haemopoietic progenitor cells on endothelialized hydroxyapatite under static conditions.

We have analysed the potential of an endothelialized hydroxyapatite matrix (HAP), a synthetic bone substitute, as a cellularized support for the expansion of haemopoietic progenitor cells. Scanning electron microscopy (SEM) showed that the endothelial cells (EC) tended to form a monolayer fitting closely to the matrix, and that the progenitors adhered to the EC layer. Endothelialized HAP supported the proliferation and differentiation of the progenitors with the addition of the exogenous cytokines IL-1, and IL-3. The expanded cell population essentially belonged to the myeloid and monocytic lineages, with a smaller percentage of the megakaryocyte lineage. In comparative experiments CD34+ progenitors were expanded on endothelialized tissue culture flasks, and a significant higher viability of the expanded cells was found with the endothelialized HAP. A high percentage (approx. 40%) of mature granulocytes was generated in accordance with the presence of differentiating cytokines such as G-CSF and GM-CSF at high concentrations in the coculture medium. Other cytokines that could be detected were IL-6, M-CSF, SCF, flt3-ligand. More than 50% of the expanded cell population was able to phagocytose bacteria and to generate an oxydative burst. These data indicate that cellularized HAP may be a useful matrix for stromal cell-based expansion systems.

Cell Differentiation↗

[Nasal washing with Physiomer ... 10 years later: 1988-1998].

The authors present the story of Physiomer. It began in 1988 in Goemar's 'Laboratory of the Sea', with an original idea of making a product manufactured from sea water and transforming it into an isotonic solution, while preserving all the elements of natural sea water, specially the trace elements and the alkaline PH. Scientific analysis has shown that the chemical composition of the product is true to the original, and that it has a positive action on cellular growth in respiratory tract mucosa in experimental culture, and it is very well tolerated. Clinical tests using nasal washings have confirmed beneficial action on the nasal and sinus mucosa in medical disease and after surgery. Tests have been carried out on 410 cases, 344 of which were treated with Physiomer, and on 199 cases after surgery on the nose or sinuses. Physiomer nasal washings have proved their value. It now remains for all the potential qualities of sea water to be demonstrated, using other preparations or other combinations.

Adult↗

A novel potential application for 99mTc-HMPAO: endothelial cell labeling for in vitro investigation of cell-biomaterial interactions.

UNLABELLED: Good adherence of endothelial cells (ECs) seeded on vascular prostheses and cell retention under flow conditions are important factors to consider in the use of functionalized prostheses in vascular surgery. Because 111In-oxine radiolabeling presents disadvantages, we wondered whether, because of its well-known physical properties, 99mTc-hexamethyl propyleneamine oxime (HMPAO or exametazime) could be used. METHODS: The cytotoxicity of unlabeled HMPAO and 99mTc-HMPAO at increasing concentrations and activities was tested on monolayers of the EC line EA-hy-926. The influence of temperature and time on tracer incorporation into cells was also tested. The optimal labeling conditions were applied to evaluate the retention of ECs seeded on polyester grafts under flow conditions by gamma camera detection. RESULTS: The activity of 10 MBq/10(6) cells corresponding to 4.5 microg/10(6) cells of unlabeled HMPAO, applied for 3 h at 37 degrees C (cellular uptake = 18%), was the best compromise between the maintenance of cell viability and metabolic activity and efficient detection by the gamma camera. Spontaneous leakage was observed and analyzed by high-performance liquid chromatography. A cell loss of 13% after 180-min exposure to shear stress was obtained. CONCLUSION: Our data thus indicate the feasibility of using such a radiolabeling technique to investigate EC-biomaterial interactions.

Blood Vessel Prosthesis↗

First use of cultured human urothelial cells for biocompatibility assessment: application to urinary catheters.

For several years, studies performed to estimate in vitro biocompatibility of urinary catheters have been carried out using permanent cell lines. But for a rational design of the testing procedure, the cell culture model should relate to the material application. This work presents the results of a probe study designed to obtain an in vitro model of normal human urothelial cells (HUC) and to test the relevance of this system in cytocompatibility experiments of urinary catheters currently used.A comparison is made with continuous cell lines, the use of which is recommended by normalization bodies. We exposed monolayers of HUC (well characterized for their proliferation, qualitative evaluation, and quantitative measurement of cytokeratins) and two continuous human cell lines to liquid extracts (either pure or diluted in the culture medium) of nine available catheters, including positive (latex) and negative controls, for a 24 h incubation. Then colorimetric assays (Neutral Red and MTT) were performed. The extracts of two polyurethanes provoked a significant toxic effect on HUC only, suggesting differences in sensitivity between the models used. This effect could be due to the presence of a great amount of barium (used as a radioopacifier) in extracts, as highlighted by results of absorption emission spectroscopy. A culture model of HUC may be of relevance for the screening of materials intended for urological practice.

Biocompatible Materials↗

Endothelial cells lining polyester fabric express pro-coagulant phenotype in vitro.

The paper deals with the in vitro assessment of endothelial cell (EC) phenotype covering an albumin- and chitosan-coated polyester fabric and shows that resting ECs express a pro-coagulant phenotype by releasing a high von Willebrand factor level and expressing low thrombomodulin surface activity, despite maintaining an adequate response to stimulating agents.

Biocompatible Materials↗

An in vitro biocompatibility evaluation of double-J stents.

OBJECTIVES: For several years, studies performed to estimate in vitro biocompatibility of urinary catheters have been carried out using permanent cell lines. However, for a rational design of the testing procedure, the cell culture model should depend on the material application. We assess the biocompatibility of 13 double-J stents using an in vitro model of normal human urothelial cells (HUC). This article aims to mimic in vitro, on HUC monolayers, the close contact existing in vivo between the urothelium and double-J stents and to evaluate the subsequent effect on these cells. METHODS: Fragments of each stent were deposited into the wells containing confluent HUC, with close contact maintained between the material and the cells. The same procedure with either no material or fragments of latex catheter was undertaken to provide the negative and positive controls, respectively. The contact was maintained for 1, 3, and 8 days. At the end of the incubation period, fragments of stent were removed and cell activity tests were performed (neutral red assay, MTT assay, and cell proliferation). RESULTS: One of the silicone stents is significantly deleterious on HUC as determined by three tests after 8 days of contact. For two copolymers, a tendency to increase cell proliferation was noted. Concerning polyurethanes, we observed significant decreases in HUC viability and cell metabolic activity for five stents after 8 days of contact. All seven polyurethane stents significantly inhibited cell proliferation. CONCLUSIONS: The HUC culture model may be of relevance for the screening of materials intended for use as double-J stents.

Biocompatible Materials↗

[The biocompatibility of catheters and stents used in urology].

Biocompatibility can be interpreted as the optimal combination of a series of interactions occurring at the material-tissue interface as soon as these two systems are in contact. It is a multifactorial interface property which integrates all of the phenomena involved in a biological environment i.e. absence of toxicity of the material for the body and absence of degradation of the material by the body. Biocompatibility can be evaluated in a normative context by using in vivo techniques in animals or in vitro techniques using cell cultures allowing the study of cytotoxicity (related to a concept of safety) and cytocompatibility (related to biological acceptability) of a material. Because of their intimate contact with the urothelium throughout implantation, the biocompatibility of catheters and stents constitutes a major requirement. This review presents the current data reported in the literature concerning the evaluation of the biocompatibility of materials used in urology. The main problems encountered are alterations of the urothelium, such as erosions or, on the contrary, mucosal hyperplasia, and the existence of incrustations developing on these materials.

Animals↗

Efficacy and safety of a low-molecular-weight heparin and standard unfractionated heparin for prophylaxis of postoperative venous thromboembolism: European multicenter trial.

A randomized, double-blind multicenter trial was performed to compare the safety and efficacy of a new low-molecular-weight heparin (LMWH) (LU 47311, Clivarine) and standard unfractionated heparin for the prophylaxis of postoperative venous thromboembolism. Altogether 1351 patients scheduled to undergo abdominal surgery were included. Main outcome measures included the incidence of thromboembolic events (deep vein thrombosis, pulmonary embolism, or both) and bleeding complications, including wound hematoma. A total of 655 patients received 1750 anti-Xa IU of LMWH plus a placebo injection daily; 677 patients received 5000 IU of unfractionated heparin (UFH) twice a day. Both drugs were found to be equally effective, as 4.7% of patients in the LMWH group and 4.3% in the UFH group developed postoperative thromboembolic complications. However, the incidence of bleeding complications was significantly reduced in the LMWH group: 55 (8.3%) patients in the LMWH group and 80 (11.8%) in the UFH group developed bleeding complications, a relative risk (RR) of 0.70 (95% CI 0.51-0.97;p = 0.03); wound hematoma occurred in 29 (4.4%) of the LMWH group compared with 55 (7.7%) in those in the UFH group for an RR of 0.57 (95% CI 0.37-0.88;p = 0.01). This study confirmed that a very low dose of 1750 anti-Xa IU daily of this new LMWH is as effective as 10,000 IU of UFH for preventing postoperative deep vein thrombosis. At this dose its administration is associated with a significant reduction in the risk of bleeding including wound hematoma.

Adult↗

Up-regulation of thrombomodulin is inducible on an endothelialized polyester.

The concept of endothelial cell seeding of vascular prostheses is designed as a method for improving long-term patency substitutes because endothelium is considered as the haemocompatible surface of reference. The assessment of the functionality of cells lining a biomaterial is thus of crucial importance. We have reported encouraging results concerning the ability of a polyester coated with albumin and chitosan (M 11) to be lined by a confluent monolayer of cultured human endothelial cells (EC). The aim of the present work was to study the expression of thrombomodulin (membrane glycoprotein responsible for anticoagulant activity) in EC lying on M.11 by anticoagulant activity and mRNA level with and without stimulation. Results obtained in basal conditions showed that EC on M.11 have a comparable expression of TM mRNA when compared to control (EC on tissue culture plates) despite a lower TM surface activity for EC on the biomaterial. In terms of ratio (stimulated cells to unstimulated ones) the response in activity for EC on M.11 is comparable to that of the control. These results indicate that cells lying on M.11 are able to respond to physiological-like stimuli, despite a tendency for these cells to express a procoagulant phenotype when compared with control EC.

Journal Article↗

Ex vivo expansion of haematopoietic progenitors on an endothelialized hydroxyapatite matrix.

Autologous haematopoietic progenitor cell (HPC) transplantation is increasingly used to restore haematopoiesis after high-dose chemotherapy treatments. The present study was designed to analyse the ability of hydroxyapatite (HAP) seeded with endothelial cells (EC) to support the proliferation and differentiation of CD34+ HPC in static culture conditions. HAP is endothelializable as assessed by scanning electron microscopy and time-course DNA synthesis analysis using tritiated thymidine incorporated in EC isolated from human umbilical vein cord. Short-term coculture experiments in which CD34+ cells isolated from human cord blood were seeded on endothelialized HAP, were performed. Results show that endothelialized HAP is permissive to CD34+ cell expansion with a maximum expansion obtained between days 7 and 14 of coculture in the presence of IL-1 and IL-3 when compared with other experiments omitting either EC or interleukins. From morphological analyses, the expanded cell population mainly belonged to the myelocytic lineage with 33% mature cells (polymorphonuclear neutrophils and monocytes) at day 14 of coculture. The immature HPC could remain trapped within HAP while giving rise to a more mature progeny that exit from HAP microenvironment.

Journal Article↗

Initial evaluation of CYFRA 21-1 diagnostic performances as a urinary marker in bladder transitional cell carcinoma.

PURPOSE: CYFRA 21-1, an immunoradiometric assay developed for the detection of a soluble cytokeratin 19 fragment, is evaluated for its diagnostic performance in urine of patients with transitional cell carcinoma. MATERIALS AND METHODS: CYFRA 21-1 was investigated in serum and urine of 128 patients, including 48 with bladder transitional cell carcinoma (group 1), 44 with other urological pathological conditions (group 2) and 36 free of urothelial disease (group 3). Urinary cytopathology was also performed. RESULTS: Mean urinary CYFRA was 123.5 +/- 53, 11.9 +/- 4.8 and 2.3 +/- 0.2 ng./ml. for groups 1 to 3, respectively, and was significantly different. From the receiver operating characteristics curve, the optimal combination of 96% sensitivity and 74% specificity was determined for a threshold value of 4 ng./ml. while overall cytopathology sensitivity was 43%. CONCLUSIONS: Urinary CYFRA 21-1 may be a useful marker for diagnosing transitional cell carcinoma.

Aged↗