Healthy Gingival Crevice Research Articles

Characterization of the soft-tissue wall lining residual periodontal pockets and implications in periodontal wound healing

AimTo characterize the soft-tissue wall of remaining periodontal pockets for wound healing-related parameters versus healthy gingival crevices in the same individuals.Materials and methodsGingival tissues collected from the diseased interface of pockets (GT biopsies) and from healthy gingival crevices (G biopsies) were subjected to RT2-profiler PCR Array for wound healing-related markers and network analysis of differentially expressed genes. Lymphangiogenesis-related gene expression was determined by qRT-PCR. The migration potential of mesenchymal stem cells isolated from GT biopsies (GT-MSCs) and G biopsies (G-MSCs) was evaluated by the scratch- and the transwell migration assays. The total collagen protein content was determined in GT-MSCs and G-MSCs homogenates.ResultsGene-ontology analysis on significantly upregulated genes expressed in GT biopsies revealed enrichment of several genes involved in processes related to matrix remodeling, collagen deposition, and integrin signaling. No significantly expressed genes were seen in G biopsies. Regarding lymphangiogenesis-related genes, GT biopsies demonstrated greater expression for PROX1 than G biopsies (p = 0.05). Lower migration potential (p < 0.001), yet greater production of collagen protein (p = 0.05), was found for GT-MSCs over G-MSCs.ConclusionDifferential expression patterns of various molecular pathways in biopsies and cell cultures of diseased versus healthy gingival tissues indicate a potential of the former for tissue remodeling and repair.Clinical relevanceIn the course of periodontitis, granulation tissue is formed within a periodontal defect in an attempt to reconstruct the site. Following treatment procedures periodontal granulation tissue remains inflamed but appears to retain healing potential.

Microbiota in Human Periodontal Abscess Revealed by 16S rDNA Sequencing.

Periodontal abscess is an oral infective disease caused by various kinds of bacteria. We aimed to characterize the microbiota composition of periodontal abscesses by metagenomic methods and compare it to that of the corresponding pocket and healthy gingival crevice to investigate the specific bacteria associated with this disease. Samples from abscess pus (AB), periodontal pocket coronally above the abscess (PO), and the gingival crevice of the periodontal healthy tooth were obtained from 20 periodontal abscess patients. Furthermore, healthy gingival crevice samples were obtained from 25 healthy individuals. Bacterial DNA was extracted and 16S rRNA gene fragments were sequenced to characterize the microbiota and determine taxonomic classification. The beta-diversity analysis results showed that the AB and PO groups had similar compositions. Porphyromonas gingivalis, Prevotella intermedia, and other Prevotella spp. were the predominant bacteria of human periodontal abscesses. The abundances of Filifactor alocis and Atopobium rimae were significantly higher in periodontal abscesses than in the periodontal pocket, suggesting their association with periodontal abscess formation. In conclusion, we characterized the microbiota in periodontal abscess and identified some species that are positively associated with this disease. This provides a better understanding of the components of periodontal abscesses, which will help facilitate the development of antibiotic therapy strategies.

Vascular Fragility and the Endothelial Glycocalyx in the Tissues Lining the Healthy Gingival Crevice.

The aim of this study is to investigate vascular fragility and endothelial glycocalyx (ECG) in images of the healthy gingival crevice (GC). The ECG is a surface layer lining the luminal walls of blood vessels composed of proteoglycans and glycoproteins with other components. Degradation of the layer has been shown to drastically affect microvascular flow, permeability, and immune function. Videomicroscopic images from tissues lining the healthy GC in 20 healthy volunteers were analyzed to determine the presence of microhemorrhage related to branched dilated microvessels. Imaging of capillary ECG in human labial mucosa was achieved by the use of a 1-mm-diameter 0.5 numerical aperture gradient index objective lens and one-sided oblique illumination using light at 450 nm. This allows resolution of red blood cells within the capillary and the endothelial cell wall of the lumen. The cell-free plasma gap between the red cell and endothelial wall represents the ECG. In the healthy GC there was no evidence of microhemorrhage in participants that had only simple capillary loops (seven of 20). Microhemorrhage was present in the majority of participants with branched dilated microvessels (11 of 13) (McNemar Test for equal distribution, P = 0.05). A cell-free plasma gap representing the ECG could be imaged in the superficial tissues lining the labial sulcus. No such cell-free plasma gap could be detected in the branched dilated microvessels in tissues lining the GC. There is an association between the presence of vascular microhemorrhage and branched dilated microvessels in the tissues lining the clinically healthy GC. Loss of ECG might be part of the inflammatory process in these tissues.

Analysis of proteins in human gingival crevicular fluid by mass spectrometry

Gingival crevicular fluid is a bodily fluid transuded from periodontal tissues into the gingival crevice and periodontal pocket, and contains many species of components. Proteins in gingival crevicular fluid have been studied as markers for periodontal diseases. Mass spectrometric analysis is used for the analyses of proteins, lipids, saccharides and metals, and expected as an approach for disease diagnosis. For better analysis of the protein components in gingival crevicular fluid, we investigated proteins in gingival crevicular fluid samples from the healthy gingival crevice and periodontal pocket using mass spectrometry. Gingival crevicular fluid samples were collected from subjects who gave their informed consent and were periodontally healthy or had diseased pockets. These samples were electrophoretically separated, and each fraction on the gels was analysed by nano liquid chromatography coupled with tandem mass spectrometry. Antimicrobial peptides detected in gingival crevicular fluid were confirmed by western blotting. One hundred and four proteins were detected in gingival crevicular fluid samples from both healthy sites and sites of periodontitis; 64 proteins were contained only in gingival crevicular fluid from healthy sites and 63 proteins were observed only in gingival crevicular fluid from periodontitis sites. These proteins were blood-, cytoskeleton-, immunity-, inflammation- and lipid-related proteins and enzymes. Some proteins, including ceruloplasmin, glycogen phosphorylase, glutathione S-transferase, phosphoglycerate mutase, psoriasin, S100A11 and resistin, were identified for the first time in gingival crevicular fluid. Antimicrobial peptides, such as lactoferrin, α1-antitrypsin, lipocalin, S100A7, S100A8, S100A9 and cathelicidin, were observed by mass spectrometry and western blotting. Multiple protein components in gingival crevicular fluid were analysed at the same time using mass spectrometry, and this approach may be useful for the diagnosis of periodontal diseases.

Profiling of bacterial flora in crevices around titanium orthodontic anchor plates

The aims of this study were to characterize the microflora in crevices around titanium orthodontic anchor plates using anaerobic culture and molecular biological techniques for bacterial identification, and to compare the microbial composition between crevices around anchor plates and gingival crevices. Samples from crevices around titanium anchor plates and healthy gingival crevices of 17 subjects (aged 20-29) were cultured anaerobically, and isolated bacteria were identified by 16S rRNA sequencing. The average logarithm colony-forming units/ml were 6.84, 7.51 and 8.88 in healthy anchor plate crevices, inflamed anchor plate crevices and healthy gingival crevices, respectively, indicating that the bacterial density of anchor plate crevices was lower than that of healthy gingival crevices. Of 184 strains isolated from healthy anchor plate crevices of seven subjects, 108 (59%) were anaerobic bacteria, while 73 (40%) were facultative bacteria. Predominant isolates were Gram-negative rods, such as Campylobacter (12%), Fusobacterium (10%) and Selenomonas (10%), and Gram-positive facultative bacteria, such as Actinomyces (17%) and Streptococcus (8.2%). Of 133 strains isolated from inflamed anchor plate crevices of three subjects, 110 (83%) were anaerobic bacteria, while predominant isolates were Gram-negative rods, such as Prevotella (47%), Fusobacterium (33%) and Campylobacter (16%). On the other hand, of 146 strains isolated from healthy gingival crevices of seven subjects, 98 (67%) were facultative bacteria, while 45 (31%) were anaerobic bacteria. Predominant isolates were Gram-positive facultative bacteria, such as Actinomyces (37%) and Streptococcus (20%). These results suggest that the environment in crevices around titanium orthodontic anchor plates is anaerobic and supportive of anaerobic growth of bacteria, which may trigger inflammation in the tissue around the plates.

Neutrophils die in the gingival crevice, periodontal pocket, and oral cavity by necrosis and not apoptosis.

Neutrophils play important roles in the homeostasis of periodontal tissues. However, remarkably little is known about how these cells live and die in the gingival crevice and periodontal pocket. We have examined whether subgingival neutrophils die by necrosis or apoptosis and have begun to study the mechanisms controlling the functional lifespan of these cells. Neutrophils collected from clinically healthy gingival crevices, periodontal pockets, and the oral cavity (saliva) were examined with Hoechst dye 33342, propidium iodide/eithidium bromide, and FITC-annexin V to identify live, dead, and apoptotic cells. Blood neutrophils were cultured for 17 to 20 hours in the presence or absence of gingival crevice washings (GC-w) to study the effect of GC-w on neutrophil apoptosis. In addition, endotoxin was removed from GC-w by affinity resin to investigate the contribution of LPS to the inhibitory effect of GC-w on blood neutrophils. The percentage of dead neutrophils in all subgingival sites and in all oral samples far exceeded the percentage of apoptotic neutrophils. In all 3 locations, approximately 30% of neutrophils were dead, whereas less than 1% of neutrophils were apoptotic. We conclude that the majority of neutrophils which lose their viability within gingival crevices, periodontal pockets and the oral cavity die by necrosis and not by apoptosis. Washings obtained from clinically healthy gingival crevices (GC-w) variably, but significantly, delayed apoptosis of peripheral blood neutrophils (mean suppression 45.7% +/- SD 22.3). Removal of endotoxin from GC-w significantly reduced this inhibitory effect. Our findings provide insights into the mechanisms of neutrophil death and the control of the functional lifespan of neutrophils in gingival crevices and periodontal pockets and therefore into the pathogenesis of periodontal diseases.

Calprotectin in gingival crevicular fluid correlates with clinical and biochemical markers of periodontal disease.

Clinical and biochemical markers of periodontal disease have been used for precise objective diagnosis of periodontal inflammation. Interleukin 1beta (IL-1beta) and prostaglandin E2 (PGE2), inflammatory factors, levels in gingival crevicular fluid (GCF) of patients with periodontal disease are elevated and have been studied as biochemical markers. The levels of calprotectin, a leukocyte protein, in body fluids of patients with some inflammatory diseases are raised. Recently, we detected calprotectin in GCF and its concentrations in periodontal pockets were higher than those in healthy gingival crevices. In this study, we investigated the correlations between GCF calprotectin levels and clinical indicators (probing depth and bleeding on probing, BOP), and the IL-1beta or PGE2 levels in GCE Probing depth and BOP at 130 sites of 110 subjects with periodontal or other oral diseases were examined, then GCF samples were collected and their calprotectin, IL-1beta and PGE2 were determined by ELISA. The calprotectin level correlated positively with the probing depth and was significantly higher at BOP-positive than BOP-negative sites. There were significant, positive correlations between the calprotectin and IL-1beta or PGE2 concentrations. These results indicate that the calprotectin level in GCF correlates well with clinical and biochemical markers of periodontal disease and suggest that calprotectin may be useful for evaluating the extent of periodontal inflammation.

Preliminary Evaluation of the Use of Methylene Blue, a Redox Dye, in the Treatment of Chronic Periodontitis

It is well established that chronic periodontitis is associated with the accumulation of subgingival plaque [1]. The microflora of this plaque is extremely complex and varies from niche to niche. Thus, the sparse subgingival flora of the healthy crevice consists largely of aerobic and facultative bacteria (mainly gram-positive cocci and rods). In gingivitis, the relative numbers of gram-negative cocci and rods increase, and in periodontitis, there is a further increase in the number of anaerobic gram-negative rods. [2]. Some of these organisms have been strongly implicated in the pathogenesis of periodontal disease; Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Wolinella recta, and spirochetes are examples. Current methods of treating periodontitis involve mechanical removal of plaque, with occasional adjunctive chemical therapy with agents such as chlorhexidine or the tetracyclines [3, 4]. However, antimicrobial agents are not the only means of controlling the growth of anaerobic bacteria. Survival and growth of such organisms in an ecosystem is, in part, dependent on the existence of a low redox potential [5-7]. Studies have shown that redox potentials as low as -300 mV exist in periodontal pockets, thereby enabling the survival of oral anaerobic bacteria in this ecosystem [8, 9]. The redox potential of the healthy gingival crevice has been measured by Kenney and Ash to be +70 mV [8], with a small population of anaerobic bacteria. Thus, by raising the redox potential of the periodontal pocket, it should be possible to create an environment incompatible with the growth of anaerobic periodontal pathogens. Various substances are available that might achieve this, such as redox dyes (like methylene blue) and transition metal ions (like ferric ions) [10, 11]. The aim of the present study was to determine whether

Association of oral Bacteroides with gingivitis and adult periodontitis.

The microbial flora of healthy gingival crevices and of periodontal pockets was studied to determine if a relation exists between the presence of Bacteroides asaccharolyticus and/or saccharolytic subspecies of B. melaninogenicus and the severity of the disease. Seventy‐eight samples were collected with sterile absorbent points from 40 individuals. The samples were immediately placed in reduced transport fluid. After dispersal by sonic treatment, the samples were put into an anaerobic chamber, serially diluted and plated on modified MM10 medium. After incubation, plates were examined and major bacterial types were counted. Black‐pigmented colonies were selected for further characterization. Samples from patients with severe inflammation (gingival index, GI = 3) showed a greater proportion of Gram‐negative anaerobic rods and 31.8 % of these organisms were identified as B. asaccharolyticus. This organism was absent in healthy sulci. It was recovered from mildly inflamed sites (GI = 1 or 2) but less often than B. melaninogenicus ss intermedius. B. melaninogenicus ss melaninogenicus was found only occasionally and in low numbers.

Rifampin as a selective agent for isolation of oral spirochetes.

Spirochetes indigenous to the healthy gingival crevice of the human mouth were isolated directly from colonies in agar medium containing rifampin as a selective agent.

The concentration of proteins in human gingival crevicular fluid

The amount and concentration of protein present in fluid obtained from individual healthy and inflamed gingival crevices have been examined. It was found that an artefactual hyperbolic relationship existed between protein concentration and fluid volume in fluid originating from healthy crevices, resulting in over‐estimation of protein content. In fluid originating from inflamed crevices, no statistically significant correlation existed between protein concentration and fluid volume, which is to be expected for an inflammatory exudate. The fluid dynamics of the crevicular region is therefore probably similar to that of the rest of the body.

THE BACTERIAL STATE OF DIFFERENT REGIONS WITHIN THE CLINICALLY HEALTHY GINGIVAL CREVICE.

THE BACTERIAL STATE OF DIFFERENT REGIONS WITHIN THE CLINICALLY HEALTHY GINGIVAL CREVICE.

The Occurrence of Bacteria within the Clinically Healthy Gingival Crevice

The Journal of PeriodontologyVolume 32, Issue 3 p. 198-202 The Occurrence of Bacteria within the Clinically Healthy Gingival Crevice J. B. Gavin, J. B. Gavin Department of Basic Dental Sciences, University of Otago Dental School, Dunedin, New Zealand.Search for more papers by this authorA. A. Collins, A. A. Collins Department of Basic Dental Sciences, University of Otago Dental School, Dunedin, New Zealand.Search for more papers by this author J. B. Gavin, J. B. Gavin Department of Basic Dental Sciences, University of Otago Dental School, Dunedin, New Zealand.Search for more papers by this authorA. A. Collins, A. A. Collins Department of Basic Dental Sciences, University of Otago Dental School, Dunedin, New Zealand.Search for more papers by this author First published: 01 July 1961 https://doi.org/10.1902/jop.1961.32.3.198Citations: 17AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume32, Issue3July 1961Pages 198-202 RelatedInformation

An Evaluation of the Antimicrobial Effect of the Fluid Exudate from the Clinically Healthy Gingival Crevice

An Evaluation of the Antimicrobial Effect of the Fluid Exudate from the Clinically Healthy Gingival Crevice