Biomedical subjects

M H Gil

Publications and source records attributed to M H Gil.

12 recordsLinked to original sources

PubMed2006

Modification of the biopolymer castor oil with free isocyanate groups to be applied as bioadhesive.

Surgical adhesives have been used for several applications, including haemostasis, sealing air leakages and tissue adhesion. The aim of this work was to develop a biodegradable urethane-based bioadhesive containing free isocyanate groups. This material presents the advantage of being biodegradable, biocompatible and having the capacity of reacting with amino groups present in the biological molecules. A urethane based on castor oil (CO) was synthesized by reaction of the molecule with isophorone diisocyanate (IPD). The characterization of the material was accomplished by different techniques: ATR-FT-IR (attenuated transmittance reflection-Fourier transform infrared), swelling capacity determination, evaluation of the moisture curing kinetics, reaction with aminated substrates and determination of surface energy by contact angle measurement. The study of the urethane thermal properties was performed by DMTA (dynamical mechanical thermal analysis) and TGA (thermogravimetric analysis). The haemocompatibility of the urethane was also evaluated by thrombosis and haemolysis tests.

Biopolymers↗

PubMed2006

Synthesis and characterization of membranes obtained by graft copolymerization of 2-hydroxyethyl methacrylate and acrylic acid onto chitosan.

Chitosan based membranes to be applied on wound healing as topical drug delivery systems were developed by graft copolymerization of acrylic acid (AA) and 2-hydroxyethyl methacrylate (HEMA) onto chitosan using cerium ammonium nitrate as chemical initiator. Evidence for graft copolymerization of the vinyl monomers onto chitosan was obtained by FTIR and DMTA. Swelling degree, cytotoxicity, thrombogenicity and haemolytic activity of these membranes were evaluated. Chitosan-graft-AA-graft-HEMA showed to be the best matrix for drug delivery systems than chitosan-graft-AA because it retains good swelling properties, but the content in HEMA has improved cytocompatibility, hemocompatibility and thrombogenic character.

Acrylates↗

PubMed2006

Structural analysis of dextran-based hydrogels obtained chemoenzymatically.

This work reports the results of structural analysis in novel dextran-acrylate (dexT70-VA) hydrogels generated chemoenzymatically. Porous structure as well as hydrogel surface and interior morphologies were evaluated by mercury intrusion porosimetry (MIP), nitrogen adsorption (NA), and scanning electron microscopy (SEM) analyses, as a function of the degree of substitution (DS), and initial water content used in the preparation of the hydrogel. MIP analysis showed that the overall networks were clearly macroporous with pore sizes ranging from 0.065 to 10 microm. As expected, the average pore size decreased as DS increased and as initial water content decreased. Moreover, the porosity values ranged from 75 up 90%, which shows that these hydrogels present an interconnected pore structure. Nitrogen adsorption analyses showed that the specific surface area of dexT70-VA hydrogels increased either by increasing the DS or by decreasing the initial water content of the hydrogel. SEM results revealed that the surface of hydrogels with lower DS presented either a porous structure or a polymeric "skin" covering the pores, whereas hydrogels with higher DS were totally porous. Furthermore, the interior morphology varied according to the DS and the initial water content of the hydrogels. Finally, the average pore size was also determined from the swelling of hydrogel using a theoretical model developed by Flory-Rehner. The comparison of the SEM and MIP results with the ones obtained by the equilibrium swelling theory of Flory-Rehner shows that this approach highly underestimates the average pore size.

Biocompatible Materials↗

PubMed2006

Surface and hemocompatibility studies of bi-soft segment polyurethane membranes.

Cross-linked urethane/urea membranes with two soft segments were prepared by extending a poly(propylene oxide) based tri-isocyanate-terminated prepolymer (PUR) with polybutadiene diol (PBDO). The ratio of prepolymer and polybutadiene diol was varied to yield cross-linked membranes with different compositions, exhibiting different degrees of phase-separation of the PBDO segments in the bulk and of surface enrichment in PUR. In this work, surface energy and hemocompatibility aspects (hemolysis and thrombosis) of the PUR/PBDO membranes were evaluated. The results showed that the membrane surface energy increased with the PBDO content until 25% of PBDO, and decreased thereafter. The introduction of the second, more hydrophobic, soft segment (PBDO) in the PUR membranes turned hemolytic into non-hemolytic membranes and, for a blood-material contact time of 10 minutes, decreased the thrombogenicity significantly. The 10% PBDO membrane was the least thrombogenic and was also non-hemolytic. The hemolysis degree did not vary significantly with the PBDO content while, for blood-material contact times of 10 minutes, the thrombogenicity increased with an increase in PBDO content above 10%. Membrane thrombogenicity varied with the blood-material contact time. For blood contact times of 10 minutes, all membranes tested were less thrombogenic than glass.

Animals↗

PubMed2002

Enzyme-linked immunofiltration assay used in the screening of solid supports and immunoreagents for the development of an azinphos-methyl flow immunosensor.

Azinphos-methyl (AM), O,O-dimethyl S-[(4-oxo-1,2,3-benzotriazin-3(4H)-yl)methyl] phosphorodithioate, is a dithiophosphorous insecticide extensively used for the control of fruit culture pests. In this work the ELIFA system, initially developed and marketed to substitute conventional ELISA methods, was used for the screening of supports and immunoreagents in the development of a flow immunosensor to AM. The objective was to find the optimal antibody concentration, support quantity and enzymatic tracer concentration to develop a sensitive and reusable immunosensor. The influence of chitosan as protein stabilizing agent was also investigated. We observed that, on the basis of immunosorbent characterization, chitosan-modified silica with immobilized LIB-MFH14 monoclonal antibody (MAb) showed the best sensitivity, with a I(50) value of 6 nM AM. All of the immobilized MAbs either in alkylaminated or chitosan-modified silica showed I(50) values between 10 and 36 nM. Regarding the regeneration capability, the best desorption agent tested was 0.1 M glycine/HCl, pH 2.0, performing in most cases a 100% desorption after just one wash and maintaining the antibody activity even after 20 cycles of regeneration. The chitosan-modified silica seemed to be the best support for this purpose.

Animals↗

PubMed2000

Evaluation of poly(2-hydroxyethyl methacrylate) gels as drug delivery systems at different pH values.

Studies of dynamic and equilibrium swelling, structural characterisation and solute transport in swollen poly(2-hydroxyethyl methacrylate) gels (pHEMA) cross-linked with tripropyleneglycol diacrylate (TPGDA) were done for a wide range of TPGDA concentrations. The influence of the pH on these pHEMA properties was evaluated. In swelling studies it was found that in changing the pH from 6.5 to 12.0, a large increase in swelling occurred, from approximately 48 to 55%, for the lowest concentration of TPGDA (1 mol%), and from 40 to 80% for the highest concentration (10 mol%). Fourier transform infrared (FTIR) and differential scanning calorimetry (DSC) measurements were made after the equilibrium swelling of the gels at different pH values, to explain these results. The advantage of using these gels as controlled drug delivery systems is illustrated using salicylic acid (SA) as a model drug. The loading and the release of the SA were made at different pH values and the results obtained showed that it is possible to modulate the hydrogel performance by controlling an external factor, the pH at which the drug loading and release were performed.

Drug Delivery Systems↗

PubMed1999

Study of the thermal stability and enzymatic activity of an immobilised enzymatic system for the bilirubin oxidation.

In this work, we have studied the immobilisation of the haemoglobin/glucose oxidase coupled enzymatic system in poly(vinyl alcohol) membranes. These are to be used as a reagent phase either in the development of an optical sensor or as an efficient bilirubin (BR) removal reactor. Poly(vinyl alcohol) (PVA) was chosen as the support for this purpose, due to its good biocompatibility, hydrophilicity and non-thrombogenic effects. A hydrogel containing the enzymatic system, consisting of PVA crosslinked with glutaraldehyde, was prepared and characterised by DSC, enzyme activity measurements and release tests. Investigating protein conformational changes as a function of temperature and the enzymatic system activity we have found that, in spite of the destabilizing effect of the glutaraldehyde in the acidic medium, the PVA insolubilisation conditions seem do not perturb either the conformation of the 'native state' nor the enzymatic system activity. Moreover, it was found that PVA/glutaraldehyde membranes offer a simple way to hold enzymatic system, with the possibility of controlling the conditions to obtain either the effective prevention of leaching of the entrapped proteins or the in situ delivery of the haemoglobin.

Bilirubin↗

PubMed1997

Microcapsules prepared from starch derivatives.

In this work, vinyl groups were introduced on soluble starch by reaction with 2-vinyl-4,4-dimethyl-2-oxazolin-5-one. The polysaccharides obtained were characterized by 1H-NMR and DSC. The 1H-NMR spectra showed high degrees of substitution and the DSC thermograms suggest a low crystallinity in the modified starch. The modified starch was used to obtain microcapsules prepared through interfacial crosslinking with DPGDA (dipropyleneglycol diacrylate) by a water-in-oil emulsion polymerization.

Journal Article↗

PubMed1996

Study of an enzyme coupled system for the development of fibre optical bilirubin sensors.

We have established an analytical procedure for bilirubin (BR) determination by following the variation of its yellow absorption band in the presence of the biocatalytic system haemoglobin/glucose oxidase/glucose. The influence of the pH of the medium, the haemoglobin/hydrogen peroxide ratio, and the concentration of the various components in the mixture on the system activity was evaluated. The solubility of the unconjugated BR was analysed. An anionic detergent in Tris buffer, in which it presents a high critical micellar concentration, was selected for the BR/bovine serum albumin cleavage. The Michaelis-Menten constant of the system (after the optimisation of the various parameters) was determined and compared with Km values reported in the literature for similar catalytic systems. Preliminary results obtained with the haemoglobin and glucose oxidase immobilised either in poly(vinyl alcohol) or by crosslinking with glutaraldehyde, show that this system is promising as a future biosensor.

Bilirubin↗

PubMed1992

Covalent binding of urease on ammonium-selective potentiometric membranes.

As part of the development of disposable urea bioselective probes, the covalent binding of urease on ammonium-selective potentiometric membranes has been assessed. Nonactin/bis(1-butylpentyl)adipate/poly(vinylchloride) (PVC) membranes, directly applied to an internal solid contact (conductive epoxy-graphite composite), has been used as a support for covalent immobilization of urease. Two types of all-solid-state construction process have been assayed: thin layers of cellulose acetate (CA) were coated on the PVC ammonium-selective membranes (type 1) and blends of PVC and CA at various ratios were used as ammonium-selective membrane matrices (type 2). Urease was covalently attached to CA via aldehyde groups. These groups were created on the polysaccharide with sodium periodate to which the enzyme was immobilized through a spacer (hexamethylenediamine). The viability of both types of probe for the determination of ammonium ions was assessed after each step of the activation process. Results indicated that type 2 potentiometric probes are altered after the treatment with sodium periodate. Good results were obtained with type 1 probes. Their dynamic concentration range of response to urea was from 2 x 10(-5) to 0.01 M with a sensibility of 50 mV/decade.

Ammonia↗

PubMedDate not supplied

Immobilization of luciferase from a firefly lantern extract on glass strips as an alternative strategy for luminescent detection of ATP.

The bioluminescent reaction catalysed by firefly luciferase has become widely established as an outstanding analytical system for assay of ATP. When used in solution, luciferase is unstable and cannot be re-used, a problem that can be partially circumvented by immobilizing the enzyme on solid substrates. Transparent glass is especially advantageous over alternative immobilizing matrices, since it allows most of the emitted photons to be detected. We report a new method for luciferase immobilization on glass which does not require prior silanization and glutaraldehyde activation, thus saving preparation time and minimizing enzyme inactivation. Our method is based on the co-immobilization by adsorption of luciferase (from a firefly lantern extract) and poly-L-lysine (PL) on non-porous glass strips. Luciferase immobilized in this way exhibits minimal variations in intersample activity, high sensitivity for ATP detection (linear luminescence responses down to 50 nmol/L) and good stability (full activity for at least 60 days when stored at -80 degrees C). PL-mediated immobilization of luciferase on glass strips provides an attractive strategy for the design of specific ATP biosensors, with potential in industry, environmental screening, medicine and biological research.

Adenosine Triphosphate↗

PubMedDate not supplied

Synthesis, characterization, and relaxivity of two linear Gd(DTPA)-polymer conjugates.

Two linear polyamide conjugates of Gd(DTPA)2- were synthesized and characterized by high-resolution nuclear magnetic resonance (NMR) spectroscopy and size exclusion chromatography (SEC). DTPA was copolymerized with two different diamines, 1,6-hexanediamine and trans-1,4-cyclohexanediamine, yielding the polymers DTPA-HMD and DTPA-CHD, with low polydispersity. Their molecular flexibility in solution was studied using 13C spin-lattice relaxation time measurements, indicating that the cyclohexanediamine linking moiety of the DTPA-HMD polymer is more rigid than that of DTPA-CHD. The influence of the flexibility of the linking functionalities on the relaxivity of the Gd3+-DTPA-polymer conjugates was studied by water nuclear magnetic relaxation dispersion (NMRD). The relaxivity of the Gd(DTPA-CHD) polymer was only slightly higher than that of the Gd(DTPA-HMD) polymer, and only two times higher than the usual values for small Gd-DTPA-like chelates. The low relaxivities obtained for both polymers, much lower than expected from the polymer apparent molecular weights, result from their substantial residual flexibility, and also from a too long, nonoptimal, value of the inner-sphere water exchange rate. These polymeric compounds are also cleared very quickly from the blood of rats, indicating that they are of limited value as blood pool contrast agents for MRA.

Animals↗