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Biomedical subjects

G Embery

Publications and source records attributed to G Embery.

At least 55 records · Page 3Linked to original sources
PubMed1997

Detection and possible biological role of chondroitinase and heparitinase enzymes produced by Porphyromonas gingivalis W50.

Gingival crevicular fluid levels of the glycosaminoglycan (GAG) chondroitin-4-sulphate (C-4-S) have received increased attention as potential indicators of periodontal tissue turnover. However, little is known about the relationship between crevicular fluid connective tissue metabolites and microbial factors. In this study Porphyromonas gingivalis, a periodontopathogen, was investigated for its ability to degrade the GAGs C-4-S, dermatan sulphate (DS) and heparan sulphate (HS) in vitro. The effect of P. gingivalis extracts on the proteoglycans (PG) derived from human gingiva were also investigated. The presence of chondroitinase and heparitinase eliminase enzymes were identified from the vesicle fraction of P. gingivalis W50. These enzymes were extracted from the vesicle fraction by a differential centrifugation technique and partially purified by non-denaturing gel filtration chromatography which revealed heparitinase enzyme peaks at 200 and 150 kDa and chondroitinase at 70 kDa. Gingival proteoglycans for use as substrates were purified using 4 M guanidinium chloride extraction and anion exchange chromatography; these proteoglycans contained 48% DS, 27% C-4-S and 13% HS P. gingivalis chondroitinase and heparitinase enzymes were capable of the degradation of C-4-S and HS but not DS GAGs. The presence of chondroitinase enzymes produced by P. gingivalis may influence levels of connective tissue metabolites in crevicular fluid. Furthermore these enzymes, particularly the heparitinase, may be involved in the initial permeation of the gingival epithelium, permitting the ingress of further microbial virulence factors.

Chondroitin Lyases↗
PubMed1997

Interaction of zinc with a synthetic calcium phosphate mineral.

As zinc has been included in several oral health products as an anticalculus and antiplaque agent, the interaction of zinc with a synthetic phosphate was investigated. The synthetic calcium phosphate used in this study was beta-tricalcium phosphate, or whitlockite, which is a major constituent of mature calculus. The aim of this work was to study the mechanism of uptake of zinc to this mineral. Zinc was readily taken up by the calcium phosphate to a maximum level of 13.9 mumol/m2. The interaction was reversible and followed a Langmuir adsorption isotherm. There was no concomitant release of calcium with zinc uptake. Inclusion of calcium in the exposure solution did however marginally depress the acquisition of zinc (12% max), but fluoride had no significant effect on uptake.

Adsorption↗
PubMed1997

The use of immunohistochemistry in understanding the structure and function of the extracellular matrix of dental tissues.

The availability of monoclonal and polyclonal antibodies directed toward the recognition of epitopes in a variety of extracellular matrix components of the dentition represents a powerful tool in the investigation of the structure and biology of dental tissues in health and disease. The immunolocalization of both whole molecule structures and specific regions of molecules has the potential to yield information on tooth development, the effects of aging, changes in tooth structure during the initiation and progression of the caries process, together with the response of the tooth to restorative treatment. This review reports on current research to elucidate the role of extracellular matrices of enamel, dentin, cementum, and bone. Attention is directed at the use of antibodies toward the small leucine-rich proteoglycans such as decorin and biglycan, in addition to their glycosaminoglycan chains. Antibodies are also being developed toward dental tissue-specific macromolecules such as phosphophoryn and amelogenin; the use of these antibodies will increase our understanding of the role of these macromolecules in mineralized tissues.

Animals↗
PubMed1996

Adsorption of bovine serum albumin on to titanium powder.

The adsorption of bovine serum albumin (BSA) on to titanium powder has been studied as a function of protein concentration and pH, and in the presence of calcium and phosphate ions. Isotherm data have shown that the adsorption process does not follow the Langmuir model (inflection points). The maximum adsorption (Admax) at pH 6.8 is 1.13 +/- 0.21 mg m-2. For the pH dependence of adsorption the amount adsorbed increases with decreasing pH (at pH 5.15 Admax = 1.31 +/- 0.2 mg m-2), indicating that hydration effects are important. Adsorption increases and decreases in the presence of calcium (at pH 6.8 for 0.002 M Admax = 1.73 +/- 0.23 mg m-2) and phosphate (at pH 6.8 for 0.002 M Admax = 0.64 +/- 0.14 mgm-2)ions, respectively, indicating that electrostatic effects are important. The time dependence, isotherm and desorption data provide indirect evidence of possible conformational changes in the BSA molecule.

Adsorption↗
PubMed1996

Modification of the proteoglycans of rat incisor dentin-predentin during in vivo fluorosis.

Proteoglycans (PGs) were isolated from the dentin-predentin of fluorotic and control rat incisor teeth using demineralization in EDTA, followed by extraction with 4 M guanidinium chloride in the presence of protease inhibitors. Differences in the behaviour of fluorotic and control PG were evident during purification by anion exchange chromatography on Q-Sepharose and MONO-Q interfaced with fast protein liquid chromatography. The PG from fluorotic teeth exhibited a more anionic profile, due to changes in glycosaminoglycan characteristics, since no apparent differences were evident between the respective core proteins, both of which were 45 kDa. The constituent glycosaminoglycan chains of fluorotic dentin were of lower molecular size and showed the additional presence of dermatan sulfate and heparan sulfate by comparison to nonfluorotic controls, where only chondroitin-4-sulfate was detected.

Animals↗
PubMed1996

High-performance liquid chromatography analysis of chondroitin sulphate isomers in human whole saliva in a variety of clinical conditions.

OBJECTIVES: Tests have been carried out to assess the level of unsaturated disaccharide isomers obtained from chondroitin sulphate in whole saliva, which contains chondroitin sulphate derived from gingival crevicular fluid (GCF). MATERIALS AND METHODS: Whole saliva was collected from periodontally diseased subjects (PDS), clinically healthy subjects (CHS) and edentulous subjects (ES). Glycosaminoglycans (GAG) were liberated by digestion with Pronase E, and precipitated with cetylpyridinium chloride and ethanol. The unsaturated disaccharides obtained by chondroitinase ACII digestion of the liberated GAG were analysed by high-performance liquid chromatography. The unsaturated disaccharides included delta Di-0S, delta Di-6S and delta Di-4S. RESULTS AND CONCLUSIONS: Analysis of data indicated that delta Di-0S, delta Di-6S and delta Di-4S were found in all PDS samples. The amount (ng ml-1 collected whole saliva) of delta Di-0S, delta Di-6S and delta Di-4S (P < 0.01) indicated significant differences between CHS and PDS whole saliva samples. The quantities of delta Di-0S and delta Di-4S (P < 0.01) indicated significant differences between PDS and ES whole saliva. The amount of delta Di-0S (P < .05) and delta Di-6S (P < 0.01) also indicated significant differences between CHS and ES whole saliva. These results indicate that chondroitin sulphate in PDS and CHS whole saliva is representative of that previously reported in gingival crevicular fluid and so provides a useful and alternative means of assessing the role of GAG as indicators of periodontal disease.

Adult↗
PubMed1996

Hydrolytic and depolymerising enzyme activity of Prevotella intermedia and Prevotella nigrescens.

OBJECTIVE: Prevotella intermedia has been reported to be associated with periodontal disease whilst P. nigrescens has predominantly been isolated from more specific conditions and healthy sites. The aim of the present study was to compare the enzyme activity of these species. MATERIALS AND METHODS: Nine strains of P. intermedia and 12 strains of P. nigrescens were studied. Lipolytic, saccharolytic, nucleolytic and proteolytic activity was determined by traditional microbiological and chromogenic substrate methods. RESULTS: All strains hydrolysed gelatine, casein, DNA and RNA. Lipase activity was produced by all strains except P. nigrescens ATCC 33563T. Lipolytic activity of P. nigrescens strains decreased as the environmental glucose concentration was increased. Only two strains, both P. intermedia, hydrolysed benzyl-arg-rho-nitroanilide. All strains hydrolysed alkaline rho-nitrophenolphosphate (except P. intermedia DAL100), produced glycylprolyl dipeptidase activity and demonstrated elastase-like activity. All but three strains (2 P. intermedia and I P. nigrescens) hydrolysed suc-ala-ala-pro-phe-rho-nitroanilide. Overall, no qualitatively analysed enzyme activity was exclusive to all strains of either species. Quantitatively analysed activity exhibited a high degree of variability both within and between species. CONCLUSIONS: P. intermedia and P. nigrescens degrade natural and synthetic substrates, but intra- and interspecies activity is variable.

Alkaline Phosphatase↗
PubMed1996

Influence of titanium oxide and titanium peroxy gel on the breakdown of hyaluronan by reactive oxygen species.

The molecular events occurring at the interface between titanium and connective tissue were investigated in order to help explain the unique biocompatible properties of titanium implants and their successful osseointegration into bone tissue. In this study the influence of commercially pure titanium and titanium peroxy gels on the breakdown of the connective tissue component and serum derived factor, hyaluronan, by reactive oxygen species (ROS), produced during the insertion of an implant in vivo, was examined. Hyaluronan breakdown was monitored in vitro in the presence of a hydroxyl radical flux, generated in the presence and absence of titanium powder and discs. Parallel studies examined the breakdown of hyaluronan by hydroxyl radicals in the presence of a titanium peroxy gel, prepared by incubation of the titanium powder or discs in concentrated hydrogen peroxide. The hyaluronan degradation products were separated according to their hydrodynamic size by gel exclusion chromatography. Similarly, experiments were also performed examining the degradation of 2-deoxy-D-ribose by a hydroxyl radical flux in order to demonstrate the detrimental potential of the hydroxyl radicals and to provide a measure of the effectiveness of titanium and titanium peroxy gels as scavengers of ROS. Titanium reduced the harmful effects of the hydroxyl radicals on the breakdown of hyaluronan, presumably acting as a scavenger for the reactive species, possibly by absorbing them into its surface oxide layer, which spontaneously forms on the surface. In contrast, the formation of a titanium peroxy gel from the titanium powder or on the surface of titanium discs enhanced breakdown of both the hyaluronan chains and 2-deoxy-D-ribose. The implications of these findings with regards to the biocompatible nature of the titanium and the ability of these implants to successfully osseointegrate are discussed.

Biocompatible Materials↗
PubMed1995

The chemical modification of glycosaminoglycan structure by oxygen-derived species in vitro.

The effect of reactive oxygen species (ROS) on the chemical structure of glycosaminoglycans (GAG) was studied in order to consider their role in connective tissue damage during an inflammatory disease state such as periodontal disease. GAG were exposed to a radical generating system for 1 h and analysed by gel filtration for fragmentation and chemically with respect to uronic acid, hexosamine and sulfate content. Non-sulfated GAG, hyaluronan and chondroitin, were most susceptible to depolymerisation and chemical modification of uronic acid and hexosamine residues by ROS. Depolymerisation and chemical modification of sulfated GAG, chondroitin 4-sulfate, dermatan sulfate and heparan sulfate was significantly less than for non-sulfated GAG. The highly sulfated GAG heparin showed minimal depolymerisation by ROS, but uronic acid residues were readily modified. Analysis of the ROS-exposed residues suggests that uronic acid is capable of degrading to a 3-carbon aldehyde, malondialdehyde. Chondroitin sulfate exposed to ROS resulted in marginal desulfation. The results suggest that the presence of sulfate on the GAG chain may protect the molecule against ROS attack. However, chemical modification of GAG may affect proteoglycan function and be of importance in considering connective tissue destruction in a variety of pathological situations, including periodontal disease.

Chondroitin Sulfates↗
PubMed1995

The influence of fluoride on the adsorption of proteoglycans and glycosaminoglycans to hydroxyapatite.

Proteoglycans and their spatial arms, the glycosaminoglycans, are known to interact with hydroxyapatite, and are considered to have a role in the regulation of mineralization. This study investigates the interactive mechanisms, with particular attention directed at the influence of fluoride on the adsorption process. Proteoglycans and glycosaminoglycans were adsorbed to hydroxyapatite in the presence of fluoride (1-20 ppm range). The adsorbates included a chondroitin 4-sulfate-rich proteoglycan extracted from rat incisor dentine, hyaluronan, chondroitin 4-sulfate, and dermatan sulfate. The order of glycosaminoglycan in decreasing affinity for hydroxyapatite was chondroitin 4-sulfate, dermatan sulfate, and hyaluronan, and the individual glycosaminoglycans showed different responses to the presence of fluoride. Graded increases in fluoride (1-4 ppm) led to 5-40% reduction of glycosaminoglycan adsorption to hydroxyapatite. The proteoglycan showed less affinity for hydroxyapatite, and demonstrated a reduction in adsorption of up to 22% with 20 ppm fluoride. The inhibitory effect of fluoride indicated an electrostatic mechanism, presumably via the calcium sites in the hydroxyapatite lattice.

Adsorption↗
PubMed1995

Adsorption of bovine serum albumin onto hydroxyapatite.

The adsorption of bovine serum albumin (BSA) onto hydroxyapatite (HA) has been studied as a function of protein concentration, pH and ionic strength. Isotherm data (adsorption being a reversible process) have been analysed using the Langmuir model, the adsorption parameters AT (maximum amount of protein adsorbed, mg m-2) and K (affinity constant, L g-1) being calculated for each solution condition (except NaF). For the pH dependence of adsorption, both AT and K increase with decreasing pH, indicating that both electrostatic and hydration effects are important. For the ionic strength dependence, increasing NaCI concentrations result in a slight increase in AT, but K decreases. With increasing CaCI2 concentrations the AT and K values increase, the opposite being true for increasing concentrations of Na2HPO4. NaF both enhances and inhibits adsorption depending on the concentration. Possible reasons for these results are discussed.

Adsorption↗
PubMed1995

Gingival crevicular fluid glycosaminoglycan levels in patients with chronic adult periodontitis.

This study investigated levels of hyaluronan and chondroitin-4-sulphate in the crevicular fluid of patients with chronic adult periodontitis at diseased and healthy sites before and after treatment. The relationship between clinical diagnostic parameters and levels of glycosaminoglycans in gingival crevicular fluid were also analysed. Within each patient, 4 sites either mesial or distal and on single rooted teeth were classified as diseased or healthy using a modified gingival index, pocket depth and attachment loss. Crevicular fluid was collected from each site using glass micropipettes and analyzed for glycosaminoglycan content by cellulose acetate electrophoresis. Significantly higher levels of chondroitin-4-sulphate were detected at diseased sites prior to treatment correlating with increased pocket depth or attachment levels. Following a period of treatment consisting of oral hygiene instruction and root planing, the patients were reassessed for their response to treatment by measuring the modified gingival index, pocket depth, attachment loss and levels of glycosaminoglycans. Analysis of glycosaminoglycan levels at diseased sites that demonstrated a poor response to treatment also demonstrated significantly higher levels of chondroitin-4-sulphate than those sites that responded well to treatment. Hyaluronan levels were less significantly associated with clinically succesful treatment. This study confirmed the use of the sulphated glycosaminoglycan chondroitin-4-sulphate as a potential diagnostic aid of periodontal tissue destruction; however, further longitudinal studies are required to assess their performance.

Adult↗
PubMed1994

A comparative study of translucent apical dentine in vital and non-vital human teeth.

Previous studies have shown that dentinal apical translucency increases with age. Although the mechanism by which apical translucency is produced is still uncertain, it has been suggested that it develops at a slower rate in non-vital teeth. In this investigation the amount and distribution of apical translucent dentine in a group of age-matched vital and non-vital teeth were compared. Fifty-five non-vital (root-filled) teeth and 49 vital teeth were used. Freshly extracted teeth were fixed in 2% glutaraldehyde solution, sectioned longitudinally in the buccolingual plane and ground to a thickness of 150 microns. Each section was photographed in polarized light at a standard magnification against a ruled scale background. Black-and-white prints of standard magnification were produced and translucent apical dentine was plotted using an Apple II digitizer. Variables measured were length, length as a percentage of root length, area and area as a percentage of root area. Data were related to age and tooth type and analysed by Mann-Whitney and regression analysis. Significant differences (p < 0.05) were present when tooth types were pooled. There was more apical translucency present in all types of non-vital teeth when compared with vital teeth at any given age.

Adolescent↗
PubMed1994

Characterization of proteoglycan metabolites in human gingival crevicular fluid during orthodontic tooth movement.

Previous studies have identified glycosaminoglycans in gingival crevicular fluid (GCF) associated with a variety of clinical conditions, notably those involving bone resorptive activity. GCF was here collected from around teeth undergoing active orthodontic movement. Proteoglycan metabolites were purified from GCF by anion-exchange chromatography using fast performance liquid chromatography. Sulphated glycosaminoglycan was associated with the most highly anionic protein fractions IV, V and VI, and biochemical analysis was restricted to these fractions. Analysis included glycosaminoglycan content by cellulose acetate electrophoresis, molecular size by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting and amino acid analyses. Fraction IV contained hyaluronan (18.7%) and chondroitin sulphate (10.9%), fraction V heparan sulphate (29.5%) and chondroitin sulphate (19.6%) and fraction VI chondroitin sulphate only (21.3%). SDS-PAGE revealed two Coomassie blue bands in fraction V of 72 and 60 kDa and two further bands in fraction VI of 71 and 56 kDa. These proteoglycans appeared resistant to digestion by chondroitinase ABC or heparinase III, although the glycosaminoglycan chains underwent degradation after protein-core removal. The molecular mass and amino acid composition of the chondroitin sulphate proteoglycan fractions showed a close similarity to those of human alveolar bone proteoglycan. The presence of heparan sulphate proteoglycan in GCF in association with orthodontic movement is in accord with previous reports. The findings support the view that proteoglycans in GCF are 'biomarkers', notably those associated with active resorption of alveolar bone.

Adolescent↗
PubMed1994

Changes in the composition of glycosaminoglycans during normal palatogenesis in the rat.

The synthesis and hydration of glycosaminoglycans (GAG) has been implicated in the generation of a palatal shelf-elevating force in mammals. This study quantifies the GAG composition in the palatal shelves of Wistar rat fetuses at various stages of palatogenesis. Hyaluronan, heparin sulphate and chondroitin-4-sulphate were detected but not dermatan sulphate or chondroitin-6-sulphate. The distribution of the GAG differed in the anterior and posterior regions (i.e. the presumptive hard and soft palates) and with the stage of development. At the time of palatal-shelf reorientation, there were no significant differences for either the total amount of GAG or for the percentages of specific GAG types between anterior and posterior regions. Indeed, the most marked differences were detected at the stages of histogenesis after shelf elevation. Nevertheless, the results support the view that hyaluronan is involved in palatal-shelf reorientation, its percentage being initially high and decreasing after shelf elevation. No changes were detected for the other (sulphated) GAG during the time of elevation. The findings point to the need to correlate the events during histogenesis with changes in the ground substance of palatal-shelf mesenchyme and indicate that there are different developmental mechanisms within the presumptive hard and soft palates.

Animals↗
PubMed1994

Gingival crevicular fluid: biomarkers of periodontal tissue activity.

The lack of precise clinical criteria for assessment of periodontal disease has led to a search for alternative means of determining active disease sites, prognosis of future sites of breakdown, and response to therapy. This review highlights the potential array of biomarkers present in gingival crevicular fluid and which may relate to existing or predicted tissue regions undergoing metabolic change and derived from bacterial or host-cell-derived products. Among the former may be listed endotoxin, amines, butyrate, and a variety of enzymes and their inhibitors, such as trypsin-like proteases and bacterial collagenase. Arising from host cells is a variety of leucocytic hydrolase enzymes, lactoferrin, and lysozyme. These appear to be useful inflammatory markers and may be distinguished from products of connective tissue breakdown which include collagenous and non-collagenous products, including collagen peptides, osteonectin, and fibronectin. The proteoglycans have found particular favor as biomarkers of possible bone-resorptive activity. Attention has also been directed at the immune response, including comment on immunoglobulins, complement, eicosanoids, and cytokines. This review lists available information on the presence of these in gingival sulcus fluid and wherever possible relates their presence to disease activity.

Biomarkers↗
PubMed1993

The influence of fluoride on proteoglycan structure using a rat odontoblast in vitro system.

Using an in vitro rat incisor odontoblast system, the effect of fluoride on proteoglycans was investigated at both the metabolic and structural level. Incisors were removed from 4-week-old rats, split longitudinally, and the pulps removed. Teeth were incubated at 37 degrees C, 5% CO2 in Eagle's Minimum Essential Medium containing 35S-sulfate for 7 hours in the presence of 0 mM, 3 mM, or 6 mM sodium fluoride. Teeth were demineralized in EDTA, proteoglycan was extracted from the residue with 4 M guanidinium chloride, and further purified by anion exchange chromatography. Uptake of radiolabel was monitored by liquid scintillation counting. The resultant products were examined by cellulose acetate electrophoresis, SDS-PAGE, chondroitinase digestion, and amino acid analysis. Differential effects of fluoride were observed in both metabolism and biochemical characterization of proteoglycans following incubation at the two concentrations. Fluoride decreased uptake of the radiolabel but led to an accumulation of glycosaminoglycan within the proteoglycan of the matrix. Chondroitin sulfate was the predominant glycosaminoglycan identified, with the additional presence of dermatan sulfate and heparan sulfate identified. Dermatan sulfate levels increased in 3 mM-treated teeth. Fluoride-treated proteoglycans had a reduced molecular weight (200-90K to 180-79K); this reduction is primarily a result of smaller glycosaminoglycan chains, with limited reduction in the size of the core protein of 6 mM-treated teeth occurring. Such alterations in the biochemical metabolism and hence structure and function of proteoglycan may be implicated in the hypomineralization seen in fluorosis.

Amino Acids↗