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Biomedical subjects

Y F Missirlis

Publications and source records attributed to Y F Missirlis.

At least 19 recordsLinked to original sources

Adhesion of slime producing Staphylococcus epidermidis strains to PVC and diamond-like carbon/silver/fluorinated coatings.

Staphylococcus epidermidis has emerged as a pathogen associated with infections of implanted medical devices. Bacterial adhesion is a crucial step in infection on biomaterial surfaces. To quantitatively determine the relationship between poly (vinyl chloride) (PVC) surface properties and bacterial adhesion, we have compared attachment of slime-producing S. epidermidis strains on PVC and various coatings under flow conditions. Bacterial adhesion and colonization was quantified by counting the viable organisms on the adherent surface as well as by scanning electron microscopy, epifluorescence microscopy and atomic force microscopy. Fluorination of the PVC surface encourages S. epidermidis adhesion whereas; diamond-like carbon (DLC) and especially silver (Ag) coatings seem to inhibit its adhesion. In most materials, the number of adherent bacteria decreased with the increase of shear rate. These results indicate that bacterial adhesion is influenced by the chemical properties of the polymeric surfaces, the surface roughness and the associated flow conditions.

Bacterial Adhesion↗

Liposome coated stents: a method to deliver drugs to the site of action and improve stent blood-compatibility.

A method to correct stent related complications non-invasively, is the local delivery of therapeutic agents. Different drugs have been delivered on stents, after being either dispersed or encapsulated in polymeric materials, and placed on stents to form drug-eluting-stents (DE-stents). Investigation of possibility to cover polymer - coated metallic stents, with liposomal drugs, for preparation of novel DE-liposome-coated-stents, has been initiated few years ago. In this context our research has been focused on answering the following questions: (i) Can liposomes be applied as coatings on polymer covered stents? (ii) Can drug release from liposome coated-stents be controlled? And: (iii) how is haemo-compatibility of stents affected? The results of the experiments carried out demonstrate that liposomal formulations of drugs can be used as coating systems of polymer covered stents for achieving sustained release of drugs at the site of interest. By modifying liposome characteristics, different amounts of drugs may be placed on the stents and their release rates can be adjusted for maximum therapeutic benefit. Finally, haemocompatibility of stents is highly improved (mainly in terms of cell adhesion and activation of coagulation system), when stents are coated with heparin-encapsulating -DRV liposomes.

Blood↗

Concise review of mechanisms of bacterial adhesion to biomaterials and of techniques used in estimating bacteria-material interactions.

This article reviews the mechanisms of bacterial adhesion to biomaterial surfaces, the factors affecting the adhesion, the techniques used in estimating bacteria-material interactions and the models that have been developed in order to predict adhesion. The process of bacterial adhesion includes an initial physicochemical interaction phase and a late molecular and cellular one. It is a complicated process influenced by many factors, including the bacterial properties, the material surface characteristics, the environmental factors, such as the presence of serum proteins and the associated flow conditions. Two categories of techniques used in estimating bacteria-material interactions are described: those that utilize fluid flowing against the adhered bacteria and counting the percentage of bacteria that detach, and those that manipulate single bacteria in various configurations which lend themselves to more specific force application and provide the basis for theoretical analysis of the receptor-ligand interactions. The theories that are reviewed are the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory, the thermodynamic approach and the extended DLVO theory. Over the years, significant work has been done to investigate the process of bacterial adhesion to biomaterial surfaces, however a lot of questions still remain unanswered.

Bacteria↗

Comparison of haemocompatibility improvement of four polymeric biomaterials by two heparinization techniques.

Two surface heparinization procedures, introduced by Bamford and Al-Lamee (Polymer 22 (1996) 4885; 13 (1994) 2844) and Seifert et al. (J. Mater. Sci.: Mater. Med. 7 (1996) 465), respectively, were applied to four commercially available biomaterials (silicone rubber, polyethylene, polypropylene and polyvinylchloride) in order to compare their efficiency in improving haemocompatibility. The indirect method (Bamford and Al-Lamee) produced a much better heparinization yield-10.5% maximum, compared to the direct one (Seifert et al.), of only 0.20% maximum. Both methods provided a better response of the heparinized biomaterials compared to the uncoated ones in terms of platelet retention and a significantly better response in terms of activation of the coagulation system, suggesting that heparin molecules remained biologically functional in both cases. The results were particularly interesting in the case of polyvinylchloride where the maximum immobilization yield was obtained by the indirect method resulting also to a pronounced haemocompatibility improvement. Scanning electron microscopy studies confirmed adhered platelet morphology whereas atomic force microscopy was used to examine surface morphology of heparinized and reference materials surface.

Blood Coagulation↗

Assessment of techniques used in calculating cell-material interactions.

A review of the techniques used in measuring the forces of deadhesion of cells that have been adhering on substrate surfaces is presented. Two categories of techniques are described, those that utilize fluid flowing against the adhered cells and counting the percentage of cells that detach (global tests) and the manipulation of single cells in various configurations which lend themselves to more specific force application and provide the basis for theoretical analysis of the receptor-ligand mechanics.

Biocompatible Materials↗

Influence of test protocol in determining the blood response to model polymers.

A multi-parametric, multi-center evaluation of three polymers was performed measuring their response to blood contact. The purpose of this study was to pinpoint differences in tests performed for assessing "basic" hemocompatibility on identical materials at different centers and attempt to rationalize. Assays for platelet adhesion, activation, aggregability and activation of the coagulation system in addition to an ex vivo patency assay were performed at four centers across Europe, using protocols favored by each center for determining the blood-contacting performance of a biomaterial. Three polymers were chosen for their expected blood response spanning the range of undesirable to desirable: ethylenevinylacetate (EVA), polyvinylchloride (PVC) and PVC modified with polyethylene oxide (PEO). The assays were ranked in terms of their efficacy compared to cost and simplicity. A correlation between assays was calculated, indicating the ability of one test to correctly determine the blood response compared to another. Some assays were unable to distinguish between materials, but of the assays which could, the materials were ranked in the following order: EVA; PVC; PVC-PEO, EVA producing the most undesirable response. It is concluded that many commonly used assays for determining hemocompatibility are inappropriate, but there are simple and reliable test methods available which correlate well with the more sophisticated protocols.

Journal Article↗

Effect of surface roughness of hydroxyapatite on human bone marrow cell adhesion, proliferation, differentiation and detachment strength.

Initial attachment of osteoblast cells and mineralization phenomena are generally enhanced on rough, sandblasted substrata. In the present work the effect of surface roughness of hydroxyapatite (HA) on human bone marrow cell response was investigated. Human bone marrow cells were plated onto HA disc-shaped pellets, prepared from synthetic HA powder. The pellets were sintered and polished with SiC paper 180-, 600- and 1200-grit, resulting in three surface roughness grades. Cell adhesion, proliferation and differentiation (evaluated with the expression of ALP activity) were determined following various incubation periods. Cell detachment strength was determined as the shear stress required to detach a given quantity of the adherent cells from the different substrata, using a rotating disc device that applied a linear range of shear stresses to the cells. The cells attached and grew faster on culture plastic in comparison with HA. No statistically significant differences were observed in the expression of ALP activity on all three HA surfaces and culture plastic. Cell adhesion, proliferation and detachment strength were surface roughness sensitive and increased as the roughness of HA increased. The percentage of the adherent cells decreased in a sigmoidal mode as a function of the applied shear stress. In conclusion, surface roughness of HA generally improved the short- and longer-term response of bone marrow cells in vitro. This behavior could be explained by the selective adsorption of serum proteins.

Aged↗

Effect of surface roughness of the titanium alloy Ti-6Al-4V on human bone marrow cell response and on protein adsorption.

The effect of surface roughness of the titanium alloy Ti-6Al-4V (Ti alloy) on the short- and long-term response of human bone marrow cells in vitro and on protein adsorption was investigated. Three different values in a narrow range of surface roughness were used for the substrata (R(alpha): 0.320, 0.490 and 0.874 microm). Cell attachment, cell proliferation and differentiation (alkaline phosphatase specific activity) were determined past various incubation periods. The protein adsorption of bovine serum albumin and fibronectin, from single protein solutions, on rough and smooth Ti alloy surfaces was examined with two methods, X-ray photoelectron spectroscopy (XPS) and radiolabeling. Cell attachment and proliferation were surface roughness sensitive and increased as the roughness of Ti alloy increased. No statistically significant difference was observed in the expression of ALP activity on all three Ti alloy surfaces and culture plastic. Both methods, XPS and protein radiolabeling, showed that human serum albumin was adsorbed preferentially onto the smooth substratum. XPS technique showed that the rough substratum bound a higher amount of total protein (from culture medium supplied with 10% serum) and fibronectin (10-fold) than did the smooth one. The cell attachment may be explained by the differential adsorption of the two proteins onto smooth and rough Ti alloy surfaces.

Adsorption↗

Effect of cytokines and colony-stimulating factors on passive polymorphonuclear leukocyte deformability in vitro.

Cytokines are potent polymorphonuclear leukocyte (PMN) activators and can decrease their deformability. We evaluated passive PMN deformability using the micropipette method after incubation with different concentrations of lipopolysaccharide (LPS), interleukins (IL-) 1, 6, 8 and 10, tumour necrosis factor (TNF), granulocyte (G) and granulocyte-macrophage (GM) colony-stimulating factors (CSF). TNF, IL-1, G-CSF, GM-CSF and, to a lesser degree, IL-6 significantly and in a dose-dependent fashion decrease PMN deformability. LPS had no direct effect on PMN deformability. When cytokines at concentrations with no effect on deformability were combined they increased PMN rigidity. The findings suggest that several cytokines and CSF impair directly, and not by activation alone, PMN deformability.

Cell Size↗

Polymorphonuclear leukocyte rigidity is defective in patients with chronic renal failure.

BACKGROUND: The purpose of the study was to investigate the rigidity of polymorphonuclear leukocytes (PMNs) in non-dialysed chronic renal failure (CRF) and haemodialysis (HD) patients. METHODS: PMN rigidity as well as tumour necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL-1beta) plasma levels were assessed in 10 early-stage CRF, 10 late-stage non-HD, and 10 HD patients, before and during dialysis. In HD patients both cellulose acetate and polysulphone membranes were used. Ten healthy subjects served as controls. Rigidity was tested by counting the deformability in morphologically passive PMNs by the micropipette method. Cytokine levels were measured by enzyme-linked immunosorbent assay. RESULTS: PMN rigidity was significantly increased in end-stage CRF patients regardless of HD but not in early-stage CRF. In HD patients PMN rigidity increased significantly 60 min after initiation of HD. There was an increase of TNF-alpha and IL-1beta levels in end-stage non-HD and HD patients and a further increase at 60 min after initiation of HD. The percentage of morphologically activated PMNs was increased only during dialysis. The nature of the HD membrane had no influence on rigidity, PMN activation, or cytokine production. CONCLUSIONS: The results indicate that PMN rigidity is defective in end-stage chronic CRF patients and is further increased 60 min after initiation of HD, regardless of the nature of the HD membrane used. PMN activation, increased TNF-alpha and IL-1beta levels, or a direct PMN impairment may cause the observed cell rigidity.

Cytokines↗

Neutrophil deformability in patients with sepsis, septic shock, and adult respiratory distress syndrome.

OBJECTIVE: To investigate the deformability of morphologically active and passive neutrophils in patients with sepsis (SP), septic shock (SS), and adult respiratory distress syndrome (ARDS). DESIGN: Prospective, observational study. SETTING: A university hospital intensive care unit and research laboratory. PATIENTS: Six patients with sepsis, six patients with septic shock, and six patients with ARDS. Eight healthy volunteers and eight ventilated but noninfected patients served as controls. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Morphologically passive and active neutrophil deformability as defined by the micropipette method was significantly decreased in patients with SP, SS, and ARDS associated with sepsis as compared with both control groups. Neutrophils from SS and ARDS patients were significantly more rigid as compared with neutrophils from SP patients but they did not differ from each other. The percentage of activated neutrophils was significantly higher in SP, SS, and ARDS patients. Increased passive neutrophil rigidity was significantly attenuated after coincubation with cytochalasin D. Tumor necrosis factor-alpha and interleukin-1beta serum levels were significantly higher in SP, SS, and ARDS patients. CONCLUSIONS: The entire neutrophil population is less deformable in SP, SS, and ARDS patients. The decreased deformability of passive neutrophils suggests that a direct mechanism involving actin polymerization, distinct from cell activation, is involved. These observations may be important in the mechanism of impaired vascular flow in patients with sepsis.

Adult↗

Test methodology for characterizing in vitro biodegradation.

In this paper mechanical tests for the characterization of the time-dependent behaviour of absorbable osteosynthetic materials are described. The tensile test is performed according to International Standard Organization (ISO) 3268/1978 and provides Young's modulus, tensile strength, elongation at tensile strength, and rupture force. The bending test is performed according to ISO 178/1975 and gives the flexural stress at various deflections, the force and deflection at break and the initial bending modulus. The torsional test is carried out according to ISO 458-1 and gives torsional load. The bending and torsional stress relaxation are measured. Relaxation modulus and the creep compliance are then determined. The parameters to be recorded in a cyclic bending test include the number of cycles or, in the case of sample survival, the remaining force at break at a bending test. The cyclic torsion test is carried out according to ISO 537/1989. The parameters to be recorded are the number of cycles or, in case of sample survival, the remaining force at a torsional test at 45 deg. Simulation tests provide a comparative assessment of the degradation phenomena under cyclic mechanical loading in bending or in torsion at elevated temperatures.

Biodegradation, Environmental↗

Stress relaxation behaviour of trabecular bone specimens.

The present study defines several conditions under which stress relaxation tests can be performed and investigates the viscoelastic behaviour of trabecular bone in compression through a series of stress relaxation tests at three strain levels and in three loading directions of each cubic specimen. A viscoelastic model is proposed to characterize the behaviour of trabecular bone and a spectrum of relaxation times is determined. Trabecular bone from the femoral head is non-linearly viscoelastic and displays anisotropic behaviour, which cannot be more symmetric elastically than orthotropic.

Aged↗

Deformability of the erythrocyte membrane in patients with myelodysplastic syndromes.

One of the major determinants of erythrocyte survival is membrane deformability, and an important intrinsic parameter of membrane deformability is the shear elastic modulus (mu) with higher mu values corresponding to increased membrane rigidity. Using a micropipette technique, we determined the shear elastic modulus of erythrocytes from 21 patients with myelodysplastic syndromes (MDS). Ten thalassemic patients and 15 healthy subjects served as controls. The shear elastic modulus of MDS erythrocytes was very significantly increased in all the patients studied, reflecting the rigidity of the membrane; the value of mu was also significantly higher in comparison with thalassemic cells. These data point to a fundamental change in the mechanical properties of the erythrocyte membrane in MDS. Biochemical studies of the membrane composition are clearly needed.

Aged↗

Aortic valve mechanics. Part II: a stress analysis of the porcine aortic valve leaflets in diastole.

The stress analysis of the porcine aortic valve leaflets in diastole at 80 mm Hg pressure in-vitro is presented. Incorporation of local geometrical asymmetry, material inhomogeneity, anistropy and non-linearity are applied. The stress theory used is a modified form of the thin membrane stress theory for a homogeneous linearly elastic and orthotropic lamina. Modifications are made so that the Hooke's law constitutive equations of stress may be applied to the inhomogeneous, non-lineary elastic and orthotropic thin (membrane) aortic valve leaflets. Stress calculations are made on the premise that the valve is in pre-transition (i.e. low elastic modulus) in the circumferential direction and post-transition (i.e. high elastic modulus) in the radial direction. It is shown that sigmaCIR less than 1 gm/mm2, and for most of the noncoronary leaflet, 0 less than sigmaRAD less than 30 gm/mm2. The areas of highest stress concentrations are in the areas of mutual leaflet coaptation near the Node of Arantii. A progressive increase of radial stresses from the sinus-annulus edge toward the node is observed.

Animals↗