The Effect of Cross-linking Efficiency of Drug-Loaded Novel Freeze Gelated Chitosan Templates for Periodontal Tissue Regeneration.

Innovative strategies for periodontal regeneration have been the focus of research clusters across the globe for decades. In order to overcome the drawbacks of currently available options, investigators have suggested a novel concept of functionally graded membrane (FGM) templates with different structural and morphological gradients. Chitosan (CH) has been used in the past for similar purpose. However, the composite formulation of composite and tetracycline when cross-linked with glutaraldehyde have received little attention. Therefore, the purpose of the study was to investigate the drug loading and release characteristics of novel freeze gelated chitosan templates at different percentages of glutaraldehyde. These were cross-linked with 0.1 and 1% glutaraldehyde and loaded with doxycycline hyclate. The electron micrographs depicted porous morphology of neat templates. After cross-linking, these templates showed compressed ultrastructures. Computerized tomography analysis showed that the templates had 88 to 92% porosity with average pore diameter decreased from 78 to 44.9 µm with increasing concentration. Fourier transform infrared spectroscopy showed alterations in the glycosidic segment of chitosan fingerprint region which after drug loading showed a dominant doxycycline spectral composite profile. Interestingly, swelling profile was not affected by cross-linking either at 0.1 and 1% glutaraldehyde and template showed a swelling ratio of 80%, which gained equilibrium after 15 min. The drug release pattern also showed a 40 µg/mL of release after 24 h. These doxycycline-loaded templates show their tendency to be used in a functionally graded membrane facing the defect site.

Hemoglobin-based oxygen carrier microparticles: synthesis, properties, and in vitro and in vivo investigations.

Bovine hemoglobin microparticles (Hb-MPs) as suitable oxygen carriers are fabricated easily by three key steps: coprecipitation of Hb and CaCO(3) to make Hb-CaCO(3)-microparticles (Hb-CaCO(3)-MPs), cross-linking by glutaraldehyde (GA) to polymerize the Hb and dissolution of CaCO(3) template to obtain pure Hb-MPs. The Hb entrapment efficiency ranged from 8 to 50% corresponding to a hemoglobin quantity per Hb-MP of at least one-third of that in one erythrocyte. The Hb-MPs are spherical, with an average diameter of 3.2 µm and high oxygen affinity. The methemoglobin level was increased after preparation, but can be reduced to less than 7% with ascorbic acid. Phagocytosis assays showed low immunogenicity of Hb-MPs if the particles were cross-linked with low concentration of GA and treated with sodium borohydride. Magnetite-loaded Hb-MPs circulated up to 4 days after intravenous application.

Detección de glutaraldehído en tubos anestésicos dentales mediante espectrofotometría UV-VIS / Detection of gluteraldehyde in dental anesthetics tubes using UV-Vis spectrophotometry

El proceso de esterilización de tubos anestésicos se realiza mediante una solución de glutaraldehído activado al 2 por ciento, pero el émbolo o la membrana de goma del tubo anestésico puede permitir una difusión del compuesto esterilizante. El objetivo del estudio es detectar la presencia de glutaraldehído dentro de tubos anestésicos después de aplicar protocolo de esterilización en frío (Normas de Desinfección MINSAL, 2008) mediante espectroscopía de absorción molecular. Al someter los tubos de anestésico al protocolo de esterilización podemos observar que existe una interacción entre el anestésico y la solución esterilizadora de glutaraldehído activado al 2 por ciento, entre los 220 y 250 nm, además se observa una laxitud en la membrana semipermeable después de la exposición por 10 horas al agente esterilizante. El glutaraldehído activado al 2 por ciento toma contacto con el anestésico mediante su filtración por el émbolo o diafragma.

The sterilization process is performed anesthetic tube with a solution of 2 percent activated gluteraldehyde, but the piston or diaphragm anesthetic tube allows a diffusion of sterilizing compound. The objective of this study is to detect the presence of gluteraldehyde into tubes after applying anesthetic cold sterilization protocol (Normas de Desinfección MINSAL) by molecular absorption spectroscopy. By making the pipes of anesthetic to the sterilization protocol we see that there is an interaction between the anesthetic and sterilizing solution of gluteraldehyde to 2 percent , between 220 and 250 nm, in addition there is a laxity in the semipermeable membrane after exposure for 10 hours a sterilizing agent. The activated 2 percent gluteraldehyde made contact with the anesthetic through its filtration by the piston or diaphragm.

Kinetic characterization and comparison of various protein crosslinking reagents for matrix modification.

We have characterized the relative efficacies of a number of protein crosslinking agents that have the potential for use in the crosslinking of proteinaceous matrices both in vitro and in vivo. The crosslinkers tested were; L: -threose (LT), Genipin (GP), Methylglyoxal (MG), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), proanthrocyanidin (PA) and glutaraldehyde (GA). The relative effectiveness of the crosslinkers with regard to their saturating concentrations was: GA > PA > EDC > MG = GP >> LT. Most of the crosslinkers displayed a pH dependence and were more effective at more alkaline pH. At optimal pH and saturating conditions, the relative reaction rates of the crosslinkers were: PA = GA > EDC > GP > MG >> LT.

Application of the glutaraldehyde test in cattle.

In a cohort study involving 62 cows from an experimental farm, the kinetics of the glutaraldehyde test (GAT) according to Sandholm (1974) was examined by testing samples of EDTA blood, lithium heparinate blood, serum, and plasma taken at various intervals. Total protein was measured in serum, and fibrinogen was determined in plasma. Gamma globulin was measured by electrophoresis. All glutaraldehyde tests were performed in duplicates, and the relation of the two results was used as measurement of precision. Optimal cut-off of the GAT time was determined as the zenith of the sum of sensitivity and specificity of various intervals for detecting combinations of gamma globulin and fibrinogen levels above 32 g/l. Precision was the best in EDTA blood. The relation between coagulation time and gamma globulin plus fibrinogen is best described by an exponential curve. The maximum value for the sum of sensitivity and specificity was found at 7 and 8 min. Seven days was the shortest interval observed between a negative test result (>15 min) and a strongly positive test result (<3 min). Twenty-one days was the shortest interval observed between a strongly positive test result and a negative test result. EDTA blood should be used for the GAT. A cut-off of 8 min yields the highest efficiency. Test results must be viewed in light of clinical findings.

Effects of glutaraldehyde exposure on human health.

Glutaraldehyde (GA) is widely used in the industrial, scientific and biomedical fields. Many adverse health effects on humans have been reported in association with biomedical uses of GA, with 2-3.5% aqueous GA solution generally used for cold sterilization and GA exposure ranges of 0.001 to 2.6 ppm for this type of use. GA is metabolized extensively to CO(2), but urinary excretion of it is low. Sensory irritant effects, sensitization of skin and respiratory organs and other symptoms have been reported among endoscopy nurses and medical radiation technologists. The prevalence of chronic bronchitis and nasal symptoms in humans is significantly correlated with peak concentrations of GA exposure. The extent of primary skin irritation depends on the duration and site of contact, and the severity of symptoms is dose-related. Chronic inhalation affects the nose and respiratory tract, and lesions become severe with prolonged duration of exposure. Increases in neither mortality nor tumor incidence have been found in workers with less than 0.2 ppm GA exposure, no evidence of carcinogenic activity has been obtained in experimental animal studies. There has been no clear evidence of genetic toxicity of GA in either in vitro or in vivo studies, and neither developmental nor reproductive toxicity has been found in humans or animals. To prevent hazards from GA exposure, use of closed-system, fully automated washing machines is recommended, since numerous symptoms have been found in individuals with less than 0.05 ppm GA exposure, the recommended peak exposure limit in many countries.

Ionic diffusion and space charge polarization in structural characterization of biological tissues.

In this study, a new approach to the analysis of the low-frequency (1-10(7) Hz) dielectric spectra of biological tissue, has been described. The experimental results are interpreted in terms of ionic diffusion and space charge polarization according to Sawada's theory. The new presentation of dielectric spectra, i.e. ([Formula: see text]) [Formula: see text] has been used. This method results in peaks which are narrower and better resolved than both the measured loss peaks and an alternative loss quantity [Formula: see text]. The presented method and Sawada's expression have been applied to the analysis of changes in the spatial molecular structure of a collagen fibril network in pericardium tissue exposed to glutaraldehyde (GA), with respect to the native tissue. The diffusion coefficient of ions was estimated on the basis of a dielectric dispersion measurement for an aqueous NaCl solution with a well-calibrated distance between the electrodes. The fitting procedure of a theoretical function to the experimental data allowed us to determine three diffusive relaxation regions with three structural distance parameters d(s), describing the spatial arrangement of collagen fibrils in pericardium tissue. It has been found that a significant decrease in the structural distance d(s) from 87 nm to 45 nm may correspond to a reduction in the interfibrillar distance within GA cross-linked tissue.

In vivo biocompatibility of gelatin-based hydrogels and interpenetrating networks.

The in vivo host response to two gelatin-based hydrogel systems of varying crosslinking modalities and loaded with the anti-inflammatory agent dexamethasone sodium phosphate was investigated. Either gelatin was chemically crosslinked with glutaraldehyde, or polyethyleneglycol diacrylate was photopolymerized around gelatin to form interpenetrating networks. The subcutaneous cage implant system was utilized to determine differential leukocyte concentrations in the inflammatory exudate surrounding the materials as indices for biocompatibility and drug efficacy in vivo. Most of the crosslinked gelatin-based materials, either via glutaraldehyde fixation or interpenetrating network formation, elicited stronger inflammatory responses than either of the starting materials, gelatin and polyethyleneglycol diacrylate. In general, dexamethasone delayed and intensified the inflammatory response. The loss of material mass did not correlate directly with the degree of cellular inflammatory response, but increased with longer implantation time and decreased with more extensive fixation.

Detection of nitric oxide in the guinea pig inner ear, using a combination of aldehyde fixative and 4,5-diaminofluorescein diacetate.

Localization of nitric oxide (NO) production sites in the inner ear of the guinea pig was investigated using a combination of glutaraldehyde fixative and a new fluorescence NO indicator. 4,5-diaminofluorescein diacetate (DAF-2DA). The cochlea and vestibular end organs were examined to locate NO production sites. The fluorescence persisted after glutaraldehyde fixation and embedding with water-soluble resin. NO production in the cochlea was observed in the outer and inner hair cells, nerve endings, nerve fibers and supporting cells of the organ of Corti, stria vascularis, spiral ligament, ganglion cells, etc. In the vestibular end organs, both type I and type II sensory cells, nerve fibers, blood vessels and dark cells displayed fluorescence. This localization was exactly identical to that of NO synthase. Thus, detection of intracellular NO production by using a combination of glutaraldehyde fixation and DAF-2DA is useful for examining the function of NO in cells, both in situ and in vivo.

Ecotoxicology of glutaraldehyde: review of environmental fate and effects studies.

Glutaraldehyde is a biocide used in many industrial applications with potential releases to the environment. This review discusses the environmental fate and effects data on this important biocide. Information drawn from this review indicates that glutaraldehyde is acutely toxic to aquatic organisms. Glutaraldehyde is equally toxic to warm water and cold water fish, but is slightly more toxic to freshwater fish than salt water fish. The acute toxicity of glutaraldehyde for avian species is comparable to that for mammalian species. The toxicity of glutaraldehyde is not appreciably increased with repeated long-term exposures. Results from environmental partitioning studies indicate that glutaraldehyde tends to remain in the aquatic compartment and has little tendency to bioaccumulate. Aqueous solutions of glutaraldehyde are stable at room temperature under acidic to neutral conditions, and to sunlight, but unstable at elevated temperatures, and under alkaline conditions. Glutaraldehyde is readily biodegradable in the freshwater environment and has the potential to biodegrade in the marine environment. Aquatic metabolism studies suggest that glutaraldehyde, under aerobic conditions, is metabolized to CO(2) via glutaric acid as an intermediate. Under anaerobic conditions, glutaraldehyde is metabolized to 1,5-pentanediol. Pretreatment with sodium bisulfite is the best method for inactivating glutaraldehyde prior to disposal to treatment systems.

The anticalcific effect of glutaraldehyde detoxification on bioprosthetic aortic wall tissue in the sheep model.

BACKGROUND: Increasing concentrations of glutaraldehyde (GA) lead to a decreased rather than increased calcification of bioprosthetic aortic wall tissue. This study determined to what extent the benefit of better cross-linking is masked by the intrinsic propensity of GA towards calcification. MATERIALS AND METHODS: Porcine aortic roots were immediately fixed at the abattoir at three different concentrations of GA (0.2%, 1.0%, and 3.0% for 1 week at 4 degrees C). Subsequently, roots underwent a GA extraction process using high volumes of Urazole solution (acetic acid buffer, pH 4.5, 37 degrees C, 1 week) followed by NaBH4 reduction (2 days, 37 degrees C). Roots were implanted in the distal aortic arch of young sheep for 6 weeks and 6 months. Calcium analysis was quantitatively done by atomic absorption spectrophotometry and qualitatively assessed by light microscopy on Von Kossa stains. RESULTS: There was a distinct anticalcification effect of GA detoxification after 6 weeks (56.8% to 97.9%; 95% confidence interval [CI]), which stabilized on a more moderate level after 6 months of implantation (19.1% to 31.6%; 95% CI). The most pronounced effect of GA extraction was seen in 0.2% fixed tissue, where aortic wall calcification was mitigated by 97% and 32% after 6 weeks and 6 months, respectively. Mitigation of aortic wall calcification was 71% (6 weeks) and 21% (6 months) in the 3.0% GA group. The combined effect of higher cross-link density and detoxification achieved an 82% (6 weeks) and 48% (6 months) reduction of calcium levels in the 3.0% GA group. In long-term implants (6 months), detoxification alone on top of standard 0.2% GA fixation was as effective (from 174.1 +/- 11.9 microg/mg without detoxification to 119.3 +/- 19.3 microg/mg with detoxification) as 3.0% fixation (114.8 +/- 10.0 microg/mg without detoxification to 91.3 +/- 11.5 microg/mg with detoxification). CONCLUSION: We were able to determine in the circulatory sheep model to what degree the intrinsic procalcific effect of GA counteracts the protective effect of higher cross-link density. Our study also established that the effect of detoxification is particularly pronounced in commercial low-grade fixation.

Meio CE e glutaraldeído na preservaçäo de plaquetas para o controle de qualidade em hematimetria / CE medium and gluutaraldehyde in platelets preservation for control quality in hematology

Controles comerciais para todas as medidas hematológicas só se tornaram disponíveis no início da década de 60, sendo que ainda há falta de padronizaçäo apropriada para diversas medidas envolvendo componentes celulares. As amostras comerciais de células consistem de células de sangue humano ou de outros animais, alteradas para retardar a deterioraçäo. De um modo geral, os fabricantes utilizam para essas células os termos: fixadas, tamponadas, estabilizadas ou preservadas, para indicar o seu processo de preparo, sem contudo descrevê-lo. Em trabalhos anteriores, desenvolveu-se amostras adequadas ao controle de qualidade em hematimetria, estáveis para os valores do eritrograma durante 100 dias, pela preservaçäo de eritrócitos em meio CE, composto por glicose, NaCl, citrato de sódio, fosfato monohidrógeno de sódio, KCl, EDTANa2, albumina bovina, clormicetina, neomicina e cortisona e pela fixaçäo parcial com glutaraldeído [ Leonart, Rev. Bras. Anál. Clín. 21: 111, 1989 ]. O uso de soluçöes aditivas para o armazenamento de concentrados de plaquetas tem aumentado durante os últimos anos e se tornado frequente na prática clínica. Em testes preliminares, realizados com o meio CE para a preservaçäo de plaquetas humanas, observamos estabilidade para a sua contagem durante até 35 dias [ Emendörfer et al, Rev. Bras. Anál. Clín. 31:18, 1999]. A partir de 20 amostras de sangue venoso em EDTAK2 e da obtençäo do plasma rico em plaquetas, testou-se a fixaçäo parcial em glutaraldeído, com posterior ressuspensäo das plaquetas em meio CE, obtendo-se valores estáveis para contagem em Coulter Counster T890 durante até 50 dias. Foram estudadas modificaçöes na composiçäo do meio CE nas concentraçöes de 1 a 6 g/gL de albumina bovina, obtendo-se maior estabilidade para as amostras de plaquetas preservadoras na concentraçäo de 6 g/dL, como empregada no meio CE original. No entanto, em amostras fixadas com glutaraldeído näo se observou diferenças na contagem das plaquetas durante 50 dias, independentemente da concentraçäo de albumina bovina. Os resultados obtidos sugerem que a fixaçäo parcial com glutaraldeído pode ser adequada para a estabilizaçäo de plaquetas em amostras de controle de qualidade.

Experimental gluing of lung parenchyma in rats.

Gelatin-resorcinol-dialdehyde adhesive has been developed from a gelatin-resorcinol-formaldehyde adhesive by replacing the formaldehyde with two less histotoxic dialdehydes, ethandial and pentandial. The aim of the present study was to evaluate the usefulness of this modified composition in gluing defects in lung parenchyma. In 40 male Wistar rats a standardized lung incision 1.0 cm in length and 0.8 cm in depth were closed by application of gelatin-resorcinol-dialdehyde adhesive. For macroscopic and microscopic examination 4 animals were sacrificed on each of postoperative days 2, 7, and 14 and 14 animals on each of postoperative days 28 and 120. Macroscopic examination revealed a tight closure of the parenchymal defects in all postoperative stages. Initially by an adhesive layer and later on by granulation tissue and scar tissue respectively. On microscopic examination an inflammatory tissue response with polymorphonuclear neutrophils and macrophages predominating was found 2 days postoperatively. After 7 days multinucleated giant cells appeared. On postoperative day 14 the tissue response presented a distinct granulomatous character with multinucleated giant cells persisting. After 28 days remnants of adhesive surrounded by granulation tissue were detectable. On postoperative day 120 the adhesive had been completely resorbed and the parenchymal defect was replaced by fibrous scar tissue. The gelatin-resorcinol-adhesive proved effective in tight closure of lung parenchyma in rats. The adhesive is resorbed completely and does not interfere with parenchymal healing.

Formaldehyde replaces glutaraldehyde in porcine bioprosthetic heart valves.

BACKGROUND AND AIMS OF THE STUDY: In the production of porcine bioprostheses, the initial glutaraldehyde treatment is often followed by a short incubation in formaldehyde to ensure sterility of the valve. It is assumed that the glutaraldehyde cross links are stable and that the formaldehyde step does not alter the glutaraldehyde incorporated. The objective of this study was to determine whether the formaldehyde interacts with the tissue to cause changes in the glutaraldehyde composition. MATERIALS AND METHODS: Two methods of tissue treatment were investigated: (i) fresh porcine leaflet tissue was treated with glutaraldehyde, followed by storage in formaldehyde, (ii) tissue processed in glutaraldehyde and transferred to formaldehyde for six hours was returned to glutaraldehyde for storage. The content of the two aldehydes was estimated by high performance liquid chromatography (HPLC), using an adaptation of the method developed by Hughes et al, which measures the acid labile Schiff bases formed between the collagen and the aldehyde. RESULTS: The initial content of glutaraldehyde in the tissue declined from 63 +/- 10 nmol/mg dry weight to 21 +/- 4 nmol/mg dry weight when the leaflets were placed in formaldehyde for 24 hours. The initial uptake of formaldehyde was 800 +/- 144 nmol/mg dry weight after 24 hours and this declined to 370 +/- 33 nmol/mg dry weight over a 16 week period of storage in formaldehyde. By this stage, the level of glutaraldehyde had decreased to 2.4 +/- 0.2 nmol/mg dry weight. There was a sharp decline in the glutaraldehyde concentration from 89 +/- 6 nmol/mg dry weight to 14 +/- 1 nmol/mg dry weight when the tissue was placed in 4% formaldehyde solution for six hours. The formaldehyde uptake was 770 +/- 54 nmol/mg dry weight. After return to 0.625% glutaraldehyde solution the formaldehyde concentration declined whilst the glutaraldehyde concentration initially increased. CONCLUSIONS: These results show that the formaldehyde reacts with the epsilon amino groups of lysine which had not reacted with glutaraldehyde, probably for steric reasons; and that formaldehyde replaces some glutaraldehyde in the tissue by a mass action effect. The tissue concentration of both aldehydes subsequently declined over the study period.

Pulmonary capillaries are more resistant to stress failure in dogs than in rabbits.

We previously showed that stress failure of pulmonary capillaries occurs at transmural pressures of approximately 50 cmH2O (40 mmHg) and above in rabbit lung. In this study, we examined whether pulmonary capillaries are more resistant to failure in dogs than in rabbits. This might be expected because of the greater athletic ability of dogs and therefore their presumably greater tolerance to large cardiac outputs and higher pulmonary vascular pressures. The lungs of 12 anesthetized mongrel dogs [22.1 +/- 5.2 (SD) kg] were perfused in situ with autologous blood and then with saline-dextran (5 min) and glutaraldehyde solution (10 min), all three perfusions at the same preset transmural pressure of 32.5, 72.5, 92.5, or 112.5 cmH2O. In dogs, the stress failure curves relating break number per millimeter of epithelium and endothelium were right shifted by approximately 40 cmH2O compared with rabbits. Blood-gas barrier thickness was significantly greater than in rabbits at 32.5 cmH2O, and unlike in rabbits, neither total nor interstitial thickness increased significantly with increasing pressure. These results indicate that pulmonary capillaries are more resistant to stress failure in dogs than rabbits.

Glutaraldehyde in dentistry--a review.

The disinfective and fixative properties of glutaraldehyde are now widely investigated. Glutaraldehyde is effective against micro-organisms and their spores. Recently, studies have shown the effectiveness of glutaraldehyde against the HIV virus. 2% glutaraldehyde is now recommended for the sterilisation of surgical instruments, operating areas, dental impressions and root canals during endodontic therapy. Studies have also shown that glutaraldehyde is an effective fixative with minimum side effects, limited penetration and quick acting. Pulpotomy studies using glutaraldehyde as the fixative agent produce high success rates. The important feature is the vital pulpal tissue at the apical third suggesting its limited penetration. The small amounts that get distributed systemically are quickly metabolised and excreted in the urine or exhaled as carbon dioxide.

The microvascular distribution of cardioplegic solution in the piglet heart. Retrograde versus antegrade delivery.

The uniform distribution of cardioplegic solution to all areas of the microvasculature is considered critical for myocardial protection. Despite this, little information exists regarding the ability of retrogradely infused cardioplegic solution to perfuse the microvasculature of the heart. In this report, the microvascular distribution of retrogradely delivered cardioplegic solution was studied by means of a technique to quantitatively demonstrate capillary perfusion. Duroc piglet hearts were subjected to either antegrade (n = 4) or retrograde (n = 8) perfusion fixation with 2.5% glutaraldehyde and subsequently perfused with NTB-2 (an intracapillary marker). The results indicate that retrogradely delivered NTB-2 consistently perfused the anterior half of the intraventricular septum and the anterior and lateral free walls of the left ventricle but inconsistently perfused the posterior half of the intraventricular septum, the posterior wall of the left ventricle, and a small paraseptal region of the right ventricle. The remainder of the right ventricle was not perfused. In contradistinction, all regions of the heart were consistently perfused by the antegrade technique. In regions of the heart in which retrograde microvascular perfusion occurred, no statistical difference was found in the quantitative degree of capillary perfusion achieved by either the antegrade or retrograde technique. These results have important implications for planning strategies of myocardial protection and suggest that further investigation concerning the microvascular distribution of retrogradely delivered cardioplegic solution in human beings is merited.

Delineation of cytotoxic concentrations of two dentin bonding agents in vitro.

Until adhesiveness of dentin bonding agents and other restorative materials to dental structures can be assured, microleakage into resulting "gaps" and dentin permeability will remain major concerns in cases of pulpal irritation. The objectives of the present study were to (a) delineate the kinds and levels of metabolic cytotoxicity of the GLUMA and Scotchbond 2 systems as well as glutaraldehyde and 2-hydroxyethylmethacrylate, and (b) compare the effects of these same materials after diffusion through dentin discs approximately 0.5-mm thick. In monolayer cultures, glutaraldehyde was much more cytotoxic than 2-hydroxyethylmethacrylate. However, GLUMA sealer and Scotchbond 2 adhesive exhibited similar cytotoxicity in monolayer cultures. After diffusion through dentin, glutaraldehyde and 2-hydroxyethylmethacrylate effects were diluted 14.7 and 26.7 times, respectively. The postdiffusional effects of the GLUMA and Scotchbond 2 systems were not significantly different and less than those effects in monolayer cultures. This study should help in the evaluation of possible causes of pulpal irritation following restorative procedures.

Penetration of glutaric dialdehyde into human dentine as measured by changes in dentine microhardness and dentine dimensions.

Microhardness indentation lengths were measured on human dentine sections unfixed and fixed with glutaric dialdehyde (GDA) at pH 7. Fixed dentine was found to be softer than unfixed dentine. Indentation length increased by approximately 10% after fixation in a buffered 5% GDA solution at pH 7. Using this effect the variation of indentation length in the direction of the penetrating GDA as a function of fixation time was measured. Describing the penetration in the mineralized matrix as diffusion in a plane sheet, a diffusion coefficient of 0.5.10(-12); m2/s was calculated. Dimensional changes of the dentine due to fixation were also measured. Dentine was found to expand in both the direction parallel (approximately 0.4%) and perpendicular (approximately 2%) to the tubules.