chondroitin-sulfates and Gingivitis

To measure the levels of hCAP18/LL-37 in gingival crevicular fluid from patients with periodontal diseases compared with healthy controls and to determine the correlation between hCAP18/LL-37 and chondroitin sulphate (CS) levels in patients with periodontitis.. Gingival crevicular fluid samples from 51 patients and 25 healthy volunteers were analysed for the hCAP18/LL-37 levels by immunoblotting and were determined for the CS levels by the competitive enzyme-linked immunosorbent assay.. Tris buffer pH 9.85 was selected to recover hCAP18/LL-37 from Periopaper strips, in which the percentages of recovery were around 70%. The median levels of hCAP18/LL-37 in the aggressive and the chronic periodontitis (CP) groups were significantly greater than those in the gingivitis and the healthy groups (p < 0.05). Significant correlations between the unprocessed 18-kDa fragment and CS levels (r = 0.650; p < 0.001) and between the mature 4.6-kDa fragment and CS levels (r = 0.502; p < 0.001) were observed only in the CP group.. The significant correlations between the hCAP18/LL-37 and the CS levels were found in CP, but not in aggressive periodontitis. The presence versus absence of such correlations may be clinically applicable to help clinicians distinguish between two distinct types of periodontitis.

Topics: Adolescent; Adult; Aged; Aggressive Periodontitis; Antimicrobial Cationic Peptides; Biomarkers; Cathelicidins; Chondroitin Sulfates; Chronic Periodontitis; Enzyme-Linked Immunosorbent Assay; Epitopes; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Immunoblotting; Lipopolysaccharides; Male; Middle Aged; Multigene Family; Periodontium; Young Adult

The purpose of this study was to compare two biochemical markers, which have been previously used to determine the degrees of alveolar bone destruction, in evaluating periodontal disease severity.. The WF6 epitope of chondroitin sulfate (CS) and the alkaline phosphatase (ALP) levels were determined in gingival crevicular fluid (GCF) samples collected from patients with various degrees of disease severity, including ten patients with gingivitis (50 gingivitis sites) and 33 patients with chronic periodontitis (including gingivitis, slight, moderate, and severe periodontitis sites; n = 50 each), as well as from ten healthy volunteers (50 healthy sites) by Periopaper strips. The levels of CS and ALP were measured by an ELISA and a fluorometric assay, respectively.. The results demonstrated low levels of CS and ALP in non-destructive and slightly destructive periodontitis sites, whereas significantly high levels of these two biomolecules were shown in moderately and severely destructive sites (p < 0.05). Although a significant difference in CS levels was found between moderate and severe periodontitis sites, no difference in ALP levels was found. Stronger correlations were found between CS levels and periodontal parameters, including probing depth, loss of clinical attachment levels, gingival index and plaque index, than between ALP levels and these parameters.. It is suggested that the CS level is a better diagnostic marker than the ALP level for evaluating distinct severity of chronic periodontitis.

Topics: Adult; Alkaline Phosphatase; Alveolar Bone Loss; Antibodies, Monoclonal; Biomarkers; Chondroitin Sulfates; Chronic Periodontitis; Cross-Sectional Studies; Dental Plaque Index; Epitopes; Female; Gingival Crevicular Fluid; Gingival Recession; Gingivitis; Humans; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Periodontium

To determine the levels of chondroitin sulphate (CS) WF6 epitope, recognized by WF6 monoclonal antibody, in gingival crevicular fluid (GCF) from different stages of periodontal disease and healthy periodontium, and to correlate those levels with clinical parameters.. GCF samples, collected from 389 sites, were analysed for the WF6 epitope levels by the competitive enzyme-linked immunosorbent assay.. The median WF6 epitope level was significantly higher in chronic periodontitis sites (n=185) than in healthy and gingivitis sites (n=204) (p<0.001), whereas the median levels did not significantly differ between healthy (n=65) and gingivitis sites (n=139). The median level in severe periodontitis sites (n=60) was significantly higher than that in moderate periodontitis sites (n=63) (p=0.019). Similarly, the median level in moderate periodontitis sites was significantly higher than that in slight periodontitis sites (n=62) (p=0.001). The WF6 epitope levels significantly correlated with probing depth (r=0.777, p=0.001) and loss of clinical attachment level (r=0.814, p=0.001).. Elevated CS WF6 epitope levels in GCF are associated with severity of periodontitis. The WF6 antibody may therefore be clinically applied to monitor disease severity and progression.

Topics: Adult; Aged; Alveolar Bone Loss; Antibodies, Monoclonal; Biomarkers; Case-Control Studies; Chondroitin Sulfates; Chronic Periodontitis; Epitopes; Gingival Crevicular Fluid; Gingivitis; Humans; Middle Aged; Periodontal Pocket; Reference Values

In inflammatory gingival diseases, cytokines have been demonstrated to play critical roles by coordinating the stimulation of immunological and connective tissue cells. The activities of these cells, degrading and remodeling extracellular matrices, constitute the major pathological and repair processes. Thus, elucidating cellular and molecular events occurring in inflamed connective tissues is crucial for the understanding and treatment of inflammation. In order to test a hypothesis that proinflammatory cytokines affect metabolism of major extracellular matrix molecules, we studied metabolism of proteoglycans (PGs) by human gingival fibroblasts (HGF) under the influence of interleukin-4 (IL-4) as a model of gingivitis. HGF in cell culture were metabolically radiolabeled using [3H]glucosamine and [35S]sulfate in the presence or absence of IL-4, and the labeled PGs were analyzed by chromatographic techniques. The incorporation of 35S into PGs increased with IL-4 both in media and cell layer. At 100 ng/ml of IL-4, the increment of 35S incorporation over control culture was 16-39% (p<0.001) in media and 12-35% (p=0.01) in cell layer. The 35S-labeled macromolecules were PGs containing heparan sulfate (HS) and chondroitin sulfate (CS) chains. From the molecular weight and glycosaminoglycan composition analyses, versican and perlecan-type and biglycan and decorin-type were very likely to be the major PG constituents both in media and cell layer. IL-4 stimulated synthesis of versican and perlecan-type more potently than biglycan and decorin-type. With IL-4 treatment, the ratio of CSPG/HSPG decreased in media and increased in cell layer. This ratio suggested that syndecan family HSPGs were also present in HGF. In conclusion, IL-4 stimulated accumulation of CS/HSPGs in human gingival fibroblasts.

Topics: Adult; Biglycan; Cell Culture Techniques; Chondroitin Sulfate Proteoglycans; Chondroitin Sulfates; Chromatography, Agarose; Chromatography, Ion Exchange; Decorin; Extracellular Matrix Proteins; Female; Fibroblasts; Gingiva; Gingivitis; Glucosamine; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Humans; Interleukin-4; Lectins; Lectins, C-Type; Membrane Glycoproteins; Molecular Weight; Proteoglycans; Radiopharmaceuticals; Sulfur Radioisotopes; Syndecans; Transforming Growth Factor beta; Tritium; Versicans

The aim of this project was to determine the in vivo effects of tooth movement with nickel-titanium archwires on the periodontium during the early stages of orthodontic treatment. The extent of tooth movement, severity of gingival inflammation, pocket probing depth, gingival crevicular fluid (GCF) flow, and the amount of the chondroitin sulphate (CS) glycosaminoglycan (GAG) component of the GCF of one maxillary canine in each of 33 patients treated with a pre-adjusted appliance were measured before and at four stages during the first 22 weeks of treatment. The methods involved the use of a reflex metrograph to determine the type of tooth movement and electrophoresis to quantitate the CS in the GCF. It was found that GCF flow increased after 4 weeks of tooth movement whereas the increase in the amount of CS in the GCF, which is taken to be indicative of periodontal tissue turnover, occurred at the later stage of 10 weeks. Teeth which showed the greatest amount of tooth movement continued to express large amounts of CS in large volumes of GCF until 22 weeks, whilst the CS levels in those teeth moving to a smaller extent declined. These data suggest that nickel-titanium archwires may produce a super-elastic plateau effect in vivo on canine teeth, which are initially displaced from the arch such that large amounts of tooth movement occur in the first 22 weeks of treatment.

Topics: Adolescent; Adult; Analysis of Variance; Child; Chondroitin Sulfates; Cuspid; Dental Alloys; Elasticity; Electrophoresis, Cellulose Acetate; Female; Follow-Up Studies; Gingival Crevicular Fluid; Gingivitis; Humans; Male; Maxilla; Nickel; Orthodontic Wires; Periodontal Index; Periodontal Pocket; Periodontium; Stress, Mechanical; Titanium; Tooth Movement Techniques

Counts of cultivable Porphyromonas gingivalis, assays of microbial proteases and the concentration in gingival crevicular fluid of proteoglycan metabolites were investigated at periodontitis and gingivitis sites in 16 patients with chronic adult periodontitis before and after treatment. Two periodontitis sites and two gingivitis sites were selected from each patient on the basis of a clinical examination. Gingival crevicular fluid from each site was analysed for the concentrations of the glycosaminoglycans chondroitin-4-sulphate and hyaluronan and subgingival plaque samples were analysed for cultivable P. gingivalis and microbial trypsin-like proteases assayed by benzoyl-DL-arginine-naphthylamide (BANA) hydrolysis. Significantly higher concentrations (p = 0.007) of chondroitin-4-sulphate were found at periodontitis than gingivitis sites but there was no significant difference in hyaluronan (p = 0.36) between these sites. Although the majority of periodontal sites were P. gingivalis-negative (23/32), there were significantly higher concentrations of chondroitin-4-sulphate (p = 0.05) and hyaluronan (p = 0.04) at the P. gingivalis-positive, compared to negative, periodontitis sites. At BANA-positive periodontitis sites there were also higher concentrations of chondroitin-4-sulphate (p = 0.0015) and hyaluronan (p = 0.0001) than at BANA-positive gingivitis sites. There was a significant decrease in concentrations of chondroitin-4-sulphate and hyaluronan at periodontitis sites after treatment. This study lends support to the hypothesis that P. gingivalis may be actively involved in the destruction of connective tissue components at culture-positive sites but shows that elevated concentrations of connective tissue breakdown products may occur in gingival crevicular fluid from periodontal sites where this organism is absent.

Topics: Adult; Benzoylarginine-2-Naphthylamide; Chondroitin Sulfates; Chronic Disease; Colony Count, Microbial; Dental Plaque; Endopeptidases; Gingival Crevicular Fluid; Gingivitis; Glycosaminoglycans; Humans; Hyaluronic Acid; Periodontitis; Porphyromonas gingivalis

Gingival crevicular fluid (GCF) was collected from around canine teeth before orthodontic treatment, during retraction, and in retention. The aim was to investigate the changes in the flow of GCF and its glycosaminoglycan (GAG) components at these three stages of orthodontic treatment, and relate them to tooth movement, gingival inflammation, and other clinical parameters recorded at the time of GCF sampling. GAG in GCF samples, collected for a 15-minute period into microcapillary tubes, were separated and identified electrophoretically, stained with Alcian blue and quantitated using a laser densitometer. The increase in GCF volume during orthodontic tooth movement and the decrease during retention are only partly due to changes in the severity of gingival inflammation. Levels of the GAG component chondroitin sulphate found in GCF samples taken during retention appear to be related to the duration of retention, particularly the length of time that the fixed appliance is passive before debonding. GCF analysis may assist the clinician to conduct this phase of orthodontic retention more reliably.

Topics: Adolescent; Adult; Child; Chondroitin Sulfates; Cuspid; Dental Debonding; Female; Gingival Crevicular Fluid; Gingivitis; Glycosaminoglycans; Humans; Hyaluronic Acid; Longitudinal Studies; Male; Orthodontic Appliances; Orthodontic Retainers; Time Factors; Tooth Movement Techniques

In this study, gingival crevicular fluid (GCF) was collected from around a canine tooth, in children, before and during orthodontic tooth movement. The aim was to identify and quantify the glycosaminoglycan (GAG) components of GCF and relate them to tooth movement, gingival inflammation, plaque accumulation, pocket probing depth and GCF volume recorded at the site of sampling. GAG in GCF samples, collected for a 15-min period into microcapillary tubes, were separated electrophoretically, stained with Alcian blue and quantified using a laser densitometer. 2 GAG components of hyaluronic acid (HA) and chondroitin sulphate (CS) were identified. The increase in GCF volume during orthodontic tooth movement was only partly due to increased gingival inflammation. GAG levels varied with different types of orthodontic tooth movement. In GCF, levels of CS, in particular, may reflect the changes in the deeper periodontal tissues which could be monitored during orthodontic tooth movements.

Topics: Adolescent; Adult; Chi-Square Distribution; Child; Chondroitin Sulfates; Cuspid; Dental Plaque Index; Dental Stress Analysis; Female; Gingival Crevicular Fluid; Gingivitis; Glycosaminoglycans; Humans; Hyaluronic Acid; Longitudinal Studies; Male; Orthodontic Appliances; Orthodontics, Corrective; Periodontal Index; Periodontium

Glycosaminoglycans were extracted from normal, inflamed and phenytoin induced overgrowth of human gingival tissue by proteolysis and alcohol precipitation. Extracts were run in a Dowex-1 column and the fractions were treated with mucopolysaccharidases. Cellulose acetate electrophoresis was carried out with or without enzyme digestion for identification of individual glycosaminoglycans. Glycosaminoglycans were found to be decreased in inflammation but were observed to increase in the overgrowth. Hyaluronic acid was found to be increased in both the pathological conditions. Dermatan sulphate, chondroitin sulphate and heparan sulphate were observed to be decreased in inflammation. In overgrowth, dermatan sulphate and chondroitin sulphate were found to increase while the presence of heparan sulphate was not significant. The changes in the pattern of individual glycosaminoglycan in the two varied conditions are discussed.

Topics: Chondroitin Sulfates; Dermatan Sulfate; Gingiva; Gingivitis; Heparitin Sulfate; Humans; Hyaluronic Acid; Phenytoin

The effect of inflammation on the distribution of chondroitin sulfate and dermatan sulfate proteoglycans was assessed after normal and inflamed human gingivae were stained with monoclonal antibodies against these extracellular matrix macromolecules. The tissues were obtained following periodontal surgery and reacted with specific antibodies after pre-treatment with chondroitinase ACII or chondroitinase ABC, and staining was visualized by the immunoperoxidase technique. The results indicated that these two proteoglycans were present in both the 4-sulfated and 6-sulfated isomeric forms. While chondroitin sulfate appeared to be uniformly distributed throughout the connective tissue, dermatan sulfate showed greater intensity of staining in the areas immediately subjacent to the epithelium. Positive staining for chondroitin sulfate was noted in the intercellular spaces of the epithelium. In inflamed tissues, there was significant staining associated with 4-sulfated dermatan sulfate and chondroitin sulfate, but this had lost the structured pattern of staining noted in normal sections. The 6-sulfated isomeric forms were greatly reduced in inflamed tissues and tended to show a predilection to be localized within the perivascular tissues. In the inflamed tissues, there was intense staining for chondroitin sulfate associated with the infiltrating inflammatory cells. These findings corroborate earlier biochemical studies on normal and inflamed gingival tissues. The specific tissue localization of dermatan sulfate and chondroitin sulfate in tissues damaged by inflammation indicates that, as opposed to the large loss of collagenous material noted during inflammation, there is not a corresponding large loss of proteoglycan. Indeed, at specific inflammatory foci, the intensity of staining for these macromolecules may intensify.

Topics: Adult; Antibodies, Monoclonal; Chondroitin Sulfates; Connective Tissue; Dermatan Sulfate; Epithelium; Gingiva; Gingivitis; Humans; Immunoenzyme Techniques; Middle Aged; Proteoglycans

Proteoglycans were extracted from human gingiva with 2 M CaCl2. The extracts were examined by gel filtration on Sephacryl S-400 in 2 M CaCl2 under dissociative conditions. The 280 nm absorbance profiles of clinically uninflamed, inflamed and severely-inflamed tissues showed that material was present with molecular weights of between 2 X 10(6) or greater, and 16,000. Proteoglycans were examined by cellulose-acetate electrophoresis with subsequent identification of the constituent glycosaminoglycans after protease digestion, and finally by chondroitinase AC digestion of the liberated glycosaminoglycans. The relative proportion of each glycosaminoglycan was calculated by scanning each cellulose-acetate sheet on an integrating densitometer. Heparan sulphate was found only in fraction I (mol. wt 2 X 10(6) or greater), together with hyaluronic acid and chondroitin-4-sulphate, these being present in all of the glycosaminoglycan-containing fractions (I-IV). Dermatan sulphate was absent from fraction I, but present in II-IV, apparently existing on the same protein core as chondroitin-4-sulphate. The relative proportions of these two glycosaminoglycans was related to molecular size, and with the degree of inflammation for a given molecular species.

Topics: Chondroitin Sulfates; Chromatography, Gel; Dermatan Sulfate; Electrophoresis, Cellulose Acetate; Gingiva; Gingivitis; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Proteoglycans

Topics: Chondroitin Sulfates; Electrophoresis, Cellulose Acetate; Gingival Crevicular Fluid; Gingivitis; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Periodontal Diseases

Topics: Alveolar Process; Animals; Bone Resorption; Chondroitin Sulfates; Dermatan Sulfate; Dogs; Gingivitis; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid; Periodontitis