Healing process in anastomosis of homologous venous grafts preserved in glutaraldehyde implanted aorta of rabbits

Aim. Autologous vein (AV) is sometimes not suitable or present for a vascular restoration. Homologous vein preserved in glutaraldehyde may be an alternative to AV, but little is yet known about this graft and its healing process after implantation in arteries. The purpose of this study was to compare the initial healing process of glutaraldehyde-tanned homologous venous grafts (group 1) with fresh autologous venous grafts (group 2), at 4 or 15 days.Methods. Forty Norfolk rabbits were allocated in 2 groups of 20 animals each. The grafts was interposed in the infra-renal aorta of the rabbit. Anastomotic tensile strength (TS), hydroxyproline (HP) determination, and histology (HA) were performed.Results. TS increased in both groups, from the 4th to 15th day, (p < 0.01) in both proximal (G1: from 364.5 &PLUSMN; 98.3 g to 491.8 &PLUSMN; 107.3 g; G2: from 366.26 &PLUSMN; 85.15 g to 518.46 &PLUSMN; 82.79 g) and distal anastomosis (GI: from 363.53 &PLUSMN; 96.26 g to 507.32 &PLUSMN; 91.01 g; G2: from 352.30 &PLUSMN; 102.41 g to 528.67 &PLUSMN; 48.58 g), with no difference between the groups. HP did not change (p > 0.10) in this same period and was similar in both groups, in the proximal (GI: from 677.99 +/- 153.98 mug/100 mg to 914.92 +/- 459.83 mug/100 mg; G2: from 668.65 +/- 170.28 mug/100 mg to 669.46 +/- 319.80 mug/100 mg) as well as in the distal anastomosis (G1:from 740.07 +/- 213.53 mug/100 mg to 923.52 +/- 270.57 mug/100 mg; G2: from 737.66 +/- 266.76 mug/100 mg to 707.68 +/- 171.25 mug/100 mg). Initial inflammatory and reparative features of the anastomosis were similar in both groups.Conclusion. We can conclude that the healing process of the glutaraldehyde-tanned homologous vein graft was similar to that of the fresh autologous venous graft.

Glutaraldehyde-treated homologous vein graft as a vein substitute: experimental study in rabbits

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Voltammetric sensor for amoxicillin determination in human urine using polyglutamic acid/glutaraldehyde film

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Biocompatibility studies of anionic collagen membranes with different degree of glutaraldehyde cross-linking

The work describes the biocompatibility and biodegradation studies of anionic collagen membranes casted form collagen gels collagen, that were selective hydrolyzed at the carboxyamide groups, as a function of the degree of cross-links induced by glutaraldehyde. Independently from the degree of cross-links, all membranes studied were characterized by a similar inflammatory response, inversely dependent on glutaraldehyde reaction time, that decreased from the time of the implant. Cell alterations, mineralization or contact necrosis were not observed in any of the membranes studied. Rates for membrane tissue biodegradation were directly related to glutaraldehyde reaction time, and ranged from 30 to periods longer than 60 days, associated with good biocompatibility. Although other properties must be considered, their use in the treatment of periodontal diseases, the biological behavior observed with the 8 h GA cross-linked membrane suggests that, anionic collagen membrane described in this work may be of potential use, not only in association with guided tissue regeneration technique for periodontal tissue reconstruction, but also in other collagen biomaterial applications where controlled biodegradability is required. (C) 1998 Published by Elsevier B.V. Ltd. All rights reserved.

Modification of glassy carbon electrodes with a poly-L-lysine/glutaraldehyde film

A film of poly-L-lysine (PLL) adheres better to the surface of a glassy carbon electrode when the PLL is partially cross-linked by means of glutaraldehyde. A film composition of 97.5% PLL/2.5% glutaraldehyde gives good adhesion and retains the anionic exchange capability of the PLL. The performance of the film was tested with hexacyanoferrate(III) using electrochemical and nonelectrochemical accumulation.

Determination of iodide and idoxuridine at a glutaraldehyde-cross-linked poly-L-lysine modified glassy carbon electrode

The detection limit (about 0.017 mu g mL(-1)) for voltammetric determination of iodide (peak at +0.87 V vs. Ag/AgCl at pH 2) at a glutaraldehyde-cross-linked poly-L-lysine modified glassy carbon electrode involving oxidation to iodine was found to be several orders of magnitude lower than that for the voltammetric determination on a bare glassy carbon electrode. This method was applied successfully to the determination of iodide in two medicinal formulations. Idoxuridine was determined indirectly at the same electrode by accumulating it first at -0.8 V vs. Ag/AgCl. At this potential the C-I bond in the adsorbed idoxuridine is reduced giving iodide, which is then determined at the modified electrode. The method was successfully applied to the determination of idoxuridine in a urine sample.

Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme

A simplified procedure for the preparation of immobilized beta-amylase using non-purified extract from fresh sweet potato tubers is established in this paper, using differently activated agarose supports. Beta-amylase glutaraldehyde derivative was the preparation with best features, presenting improved temperature and pH stability and activity. The possibility of reusing the amylase was also shown, when this immobilized enzyme was fully active for five cycles of use. However, immobilization decreased enzyme activity to around 15%. This seems to be mainly due to diffusion limitations of the starch inside the pores of the biocatalyst particles. A fifteen-fold increase in the Km was noticed, while the decrease of Vmax was only 30% (10.1 U mg-1 protein and 7.03 U mg-1 protein for free and immobilized preparations, respectively). © 2013 Elsevier Ltd.

Clinical evaluation of postoperative sensitivity using self-etching adhesives containing glutaraldehyde

The present clinical study aimed to assess the postoperative sensitivity (POS) after 48 hours and seven days in occlusal restorations bonded with three different adhesive systems, two of them containing glutaraldehyde. The restorative procedures were performed using the three-step etch-and-rinse Adper SBMP-Plus adhesive (SBMP), the two-step etch-and-rinse Gluma Comfort One Bond + Desensitizer adhesive (GC+D) and the all-in-one self-etching/priming I Bond (IB) adhesive, which also has glutaraldehyde in its formula. All cavities were restored with Filtek Supreme nanoparticle composite resin. After 48 hours and seven days the patients were recalled and the postoperative sensitivity evaluated. The data analyzed by non-parametric Friedman test showed no significant differences in POS among the three tested groups after 48 hours and seven days.

Transdentinal cytotoxicity of carbodiimide (EDC) and glutaraldehyde on odontoblast-like cells

Objective: To evaluate the transdentinal cytotoxicity of three different concentrations of carbodiimide (EDC) or 5% glutaraldehyde (GA) on MDPC-23 cells. Methods: Seventy 0.4-mm-thick dentin disks obtained from human molars were adapted to artificial pulp chambers. MDPC-23 cells were seeded on the pulpal surface of the disks. After 48 hours, the occlusal dentin was acid-etched and treated for 60 seconds with one of the following solutions (n=10): no treatment (negative control); 0.1 M, 0.3 M, or 0.5 M EDC; 5% GA; Sorensen buffer; or 29% hydrogen peroxide (positive control). Cell viability and morphology were assessed by methyltetrazolium assay and scanning electron microscopy (SEM), respectively. The eluates were collected after the treatments and applied on MDPC-23 seeded in a 24-well plate to analyze cell death, total protein (TP), and collagen production. The last two tests were performed 24 hours and seven days after the challenge. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (p<0.05). Results: EDC at all test concentrations did not reduce cell viability, while 5% GA did increase cell metabolism. Cell death by necrosis was not elicited by EDC or 5% GA. At the 24-hour period, 0.3 M and 0.5 M EDC reduced TP production by 18% and 36.8%, respectively. At seven days, increased TP production was observed in all groups. Collagen production at the 24-hour period was reduced when 0.5 M EDC was used. After seven days, no difference was observed among the groups. SEM showed no alteration in cell morphology or number, except in the hydrogen peroxide group. Conclusions: Treatment of acid-etched dentin with EDC or GA did not cause transdentinal cytotoxic effects on odontoblast-like cells.

Determination of salicylate in blood serum using an amperometric biosensor based on salicylate hydroxylase immobilized in a polypyrrole-glutaraldehyde matrix

The use of an amperometric biosensor for the salicylate determination in blood serum is described. The biosensor is based on salicylate hydroxylase (EC 1.14.13.1) electropolymerized onto a glassy carbon-working electrode with polypyrrole and glutaraldehyde, to improve the biosensor lifetime. The hexacyanoferrate (II) was also incorporated to work as a redox mediator to minimize possible interferences. The salicylate is enzymatically converted to catechol, which is monitored amperometrically by its electrooxidation at +0.170 V versus SCE (saturated calomel electrode). Salicylate determination was carried out maintaining the ratio between β-NADH and salicylate at 4:1 (30°C). The amperometric response of the biosensor was linearly proportional to the salicylate concentration between 2.3 x 10-6 and 1.4 x 10-5 mol l- 1, in 0.1 mol l-1 phosphate buffer (pH 7.8), containing 0.1 mol l-1 KCl and 5.0 x 10-4 mol l-1 Na2H2EDTA, as supporting electrolyte. The recovery studies, in the presence of several interfering compounds, showed recoveries between 96.4 and 104.8%. The useful lifetime of the biosensor in the concentration range evaluated was at least 40 days, in continuous use. Blood serum samples analyzed by this biosensor showed a good correlation compared to the spectrophotometric method (Trinder) used as reference, presenting relative deviations lower than 7.0%. (C) 2000 Elsevier Science B.V.

Transdentinal cytotoxicity of glutaraldehyde on odontoblast-like cells

This study investigated the transdentinal cytotoxicity of glutahaldehyde-containing solutions/materials on odontoblast-like cells. Dentin discs were adapted to artificial pulp chambers. MDPC-23 cells were seeded on the pulpal side of the discs and the occlusal surface was treated with the following solutions: water, 2% glutaraldehyde (GA), 5% GA, 10% GA, Gluma Comfort Bond+Desensitizer (GCB+De) or Gluma Desensitizer (GDe). Cell viability and morphology were assessed by the Alamar Blue assay and SEM. The eluates were collected and applied on cells seeded in 24-well plates. After 7 or 14 days the total protein (TP) production, alkaline phosphatase activity (ALP) and deposition of mineralized nodules (MN) were evaluated. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (p<0.05). GA solutions were not cytotoxic against MDPC-23. GCB+De (85.1%) and GDe (77.2%) reduced cell viability as well as TP production and ALP activity at both periods. After 14 days, GCB+De and GDe groups produced less MN. Affected MDPC-23 presented deformation of the cytoskeleton and reduction of cellular projections. The treatment with 2.5%, 5% and 10% GA was not harmful to odontoblast-like cells. Conversely, when GA was combined with other components like HEMA, the final material became cytotoxic. Glutaraldehyde has been used to decrease dentin hypersensitivity. This substance is also capable of preventing resin-dentin bond degradation by cross-linking collagen and MMPs. This study showed that GA might be safe when applied on acid etched dentin. However, when combined with HEMA the product becomes cytotoxic.

Glutaraldehyde in bio-catalysts design: a useful crosslinker and a versatile tool in enzyme immobilization

Glutaraldehyde is one of the most widely used reagents in the design of biocatalysts. It is a powerful crosslinker, able to react with itself, with the advantages that this may bring forth. In this review, we intend to give a general vision of its potential and the precautions that must be taken when using this effective reagent. First, the chemistry of the glutaraldehyde/amino reaction will be commented upon. This reaction is still not fully clarified, but it seems to be based on the formation of 6-membered heterocycles formed by 5 C and one O. Then, we will discuss the production of intra- and inter-molecular enzyme crosslinks (increasing enzyme rigidity or preventing subunit dissociation in multimeric enzymes). Special emphasis will be placed on the preparation of cross-linked enzyme aggregates (CLEAs), mainly in enzymes that have low density of surface reactive groups and, therefore, may be problematic to obtain a final solid catalyst. Next, we will comment on the uses of glutaraldehyde in enzymes previously immobilized on supports. First, the treatment of enzymes immobilized on supports that cannot react with glutaraldehyde (only inter and intramolecular cross-linkings will be possible) to prevent enzyme leakage and obtain some enzyme stabilization via cross-linking. Second, the cross-linking of enzymes adsorbed on aminated supports, where together with other reactions enzyme/support crosslinking is also possible; the enzyme is incorporated into the support. Finally, we will present the use of aminated supports preactivated with glutaraldehyde. Optimal glutaraldehyde modifications will be discussed in each specific case (one or two glutaraldehyde molecules for amino group in the support and/or the protein). Using preactivated supports, the heterofunctional nature of the supports will be highlighted, with the drawbacks and advantages that the heterofunctionality may have. Particular attention will be paid to the control of the first event that causes the immobilization depending on the experimental conditions to alter the enzyme orientation regarding the support surface. Thus, glutaraldehyde, an apparently old fashioned reactive, remains the most widely used and with broadest application possibilities among the compounds used for the design of biocatalyst.

Simultaneous fixation using glutaraldehyde and osmium tetroxide or potassium ferricyanide-reduced osmium for the preservation of monogenean flatworms: An assessment for Merizocotyle icopae

Simultaneous fixation was investigated for a marine organism: the monogenean flatworm ectoparasite Merizocotyle icopae. Four protocols for primary fixation were compared: 3% glutaraldehyde alone in OAM cacodylate buffer for a minimum of 2 hours; 1% glutaraldehyde in combination with 1% osmium tetroxide, both in 0.1M cacodylate buffer, until tissues darkened (5-20 minutes); 1% glutaraldehyde in OAM cacodylate buffer in combination with 0.5% potassium ferricyanide-reduced osmium until tissues darkened (5-20 minutes); 1% glutaraldehyde in combination with 1% osmium tetroxide, both in 0.1M cacodylate buffer, for 30 minutes. The study confirms that the standard method for transmission electron microscopic fixation (first listed protocol) routinely applied to platyhelminths is optimal for ultrastructural preservation, but some simultaneous fixation methods (second and third listed protocols) are acceptable when rapid immobilization is needed. Scanning electron microscopic preparations may be improved using simultaneous primary fixation. (C) 2004 Wilcy-Liss, Inc.