scholarly journals Clinically Healthy Human Gingival Tissues Show Significant Inter-individual Variability in GCF Chemokine Expression and Subgingival Plaque Microbial Composition

2021 ◽  
Vol 2 ◽  
Author(s):  
Shatha Bamashmous ◽  
Georgios A. Kotsakis ◽  
Sumita Jain ◽  
Ana M. Chang ◽  
Jeffrey S. McLean ◽  
...  

Aim: Clinically healthy gingival tissue is maintained through controlled regulation of host defense mechanisms against plaque biofilm overgrowth. One key component is the transit of neutrophils from the vasculature into gingival tissue where the expression of different neutrophil chemokines are tightly regulated. This cross-sectional study examines the inter-individual variability in chemokine profiles within gingival crevicular fluid (GCF) in relation to the subgingival bacterial community in a state of gingival health.Methods: Gingival crevicular fluid and subgingival plaque samples were collected from mesiobuccal surfaces of all six Ramfjord teeth of 20 systemically healthy individuals (14.55 ± 1.67 years). A multiplex immunoassay was carried out to quantify the expression of 40 different chemokines in the healthy gingival tissue. Neutrophils were assessed indirectly by myeloperoxidase (MPO) in GCF using traditional ELISA. Characterization of healthy subgingival plaque was conducted with the Illumina Miseq targeting the 16S rRNA gene.Results: In health, there are distinct variations within individual gingival crevicular fluid chemokine expression profiles, as well as in the concentration of neutrophils, that divided the participants into high or low chemokine expressing groups. Specifically, key differences were identified within MIF (2683.54 ± 985.82 pg per 30-s sample), IL-8/CXCL8 (170.98 ± 176.96 pg per 30-s sample), Gro-α/CXCL1 (160.42 ± 94.21 pg per 30-s sample), ENA-78/CXCL5 (137.76 ± 76.02 pg per 30-s sample), IL-1β (51.39 ± 37.23 pg per 30-s sample), TNF-α (1.76 ± 1.79 pg per 30-s sample), and IFN-γ (0.92 ± 0.54 pg per 30-s sample). Of these identified chemokines, the highest correlation was associated between IL-8/CXCL8 and neutrophils (r = 0.54, p = 0.014). Furthermore, species characterization of healthy subgingival plaque revealed significant inter-individual variability that identified two unique groups unrelated to the previously identified chemokine groups.Conclusion: The lack of concordance between the microbial composition and chemokine profile during health may be a reflection of the unique microbial composition of each individual coupled with variations within their host response, emphasizing the vast complexity of the defense mechanisms in place to maintain gingival health.

Author(s):  
J Hanker ◽  
E.J. Burkes ◽  
G. Greco ◽  
R. Scruggs ◽  
B. Giammara

The mature neutrophil with a segmented nucleus (usually having 3 or 4 lobes) is generally considered to be the end-stage cell of the neutrophil series. It is usually found as such in the bone marrow and peripheral blood where it normally is the most abundant leukocyte. Neutrophils, however, must frequently leave the peripheral blood and migrate into areas of infection to combat microorganisms. It is in such areas that neutrophils were first observed to fragment to form platelet-size particles some of which have a nuclear lobe. These neutrophil pseudoplatelets (NPP) can readily be distinguished from true platelets because they stain for neutrophil myeloperoxidase. True platelets are not positive in this staining reaction because their peroxidase Is inhibited by glutaraldehyde. Neutrophil pseudoplatelets, as well as neutrophils budding to form NPP, could frequently be observed in peripheral blood or bone marrow samples of leukemia patients. They are much more prominent, however, in smears of inflammatory exudates that contain gram-negative bacteria and in gingival crevicular fluid samples from periodontal disease sites. In some of these samples macrophages ingesting, or which contained, pseudoplatelets could be observed. The myeloperoxidase in the ingested pseudoplatelets was frequently active. Despite these earlier observations we did not expect to find many NPP in subgingival plaque smears from diseased sites. They were first seen by light microscopy (Figs. 1, 3-5) in smears on coverslips stained with the PATS reaction, a variation of the PAS reaction which deposits silver for light and electron microscopy. After drying replicate PATS-stained coverslips with hexamethyldisilazane, they were sputter coated with gold and then examined by the SEI and BEI modes of scanning electron microscopy (Fig. 2). Unstained replicate coverslips were fixed, and stained for the demonstration of myeloperoxidase in budding neutrophils and NPP. Neutrophils, activated macrophages and spirochetes as well as other gram-negative bacteria were also prominent in the PATS stained samples. In replicate subgingival plaque smears stained with our procedure for granulocyte peroxidases only neutrophils, budding neutrophils or NPP were readily observed (Fig. 6).


2019 ◽  
Vol 44 (5) ◽  
pp. 452-458 ◽  
Author(s):  
R Arif ◽  
JB Dennison ◽  
D Garcia ◽  
P Yaman

SUMMARY Statement of Problem: The long-term effect of the presence of porcelain laminate veneers (PLVs) on the health of the surrounding gingival issues is not available in the restorative literature. Purpose: To assess the long-term effect of PLVs on the health of the surrounding gingival tissues. A secondary aim was to correlate gingival crevicular fluid (GCF) scores with clinical parameters used for gingival health assessment in teeth treated with PLVs. Methods and Materials: Patients who received PLVs placed at the Graduate Restorative Clinic within a seven- to 14-year period were recalled for clinical evaluations. Periodontal measurements including gingival index (GI), periodontal pocket depth (PPD), gingival recession (GR), and clinical attachment level (CAL) were measured using a standard probe and indices. Gingival Crevicular Fluid (GCF) was measured with a Periotron machine (Periotron 8000, Oraflow Inc), using Periopaper (Periopaper Gingival Fluid Collection Strip, Oraflow Inc.) for fluid collection. Photographs of any observed clinical defect were taken. Data were tabulated using Excel 2010 (Microsoft Corp). Statistical analysis for all descriptive statistics was performed using SPSS 21 (SPSS Software, IBM Corp.) and Stata SE 13 (Stata Software, StataCorp). Repeated-measures analysis of variance (ANOVA) was done to test for statistical significance of the mean pocket depths between the restored and unrestored surfaces of the veneered teeth. The significance level for all tests was p<0.05. Pearson's correlation coefficient was performed for testing statistical significance between GCF and GI and between GCF and PPD. Results: The frequency distribution of the GI included 47 PLVs (43%) with normal gingiva, 16 (15%) with mild inflammation, and 46 (42%) with moderate inflammation and bleeding on probing. The average PPD on the facial surface of the maxillary and mandibular PLVs was 2.17 mm and 2.16 mm, respectively. On the lingual surface, the average PPD was 2.10 mm for maxillary and 2.22 mm for mandibular PLVs. Gingival recession was seen in 27% of the evaluated PLVs. The repeated-measures ANOVA revealed p≥0.136, showing no statistical difference in the mean pocket depths between restored facial and unrestored lingual surfaces of the veneered teeth. A moderate correlation (r=0.407) was found between GCF and GI, which was significant at p<0.001. No correlation (r=0.124) was found between GCF and PPD, which was not significant at p=0.197. Conclusions: Gingival response to the evaluated PLVs was in the satisfactory range, with overall GI scores ranging between normal and moderate inflammation, pocket depths ranging from 1 to 2 mm, and recession present in 27% of the evaluated PLVs. No statistically significant difference was found between the mean pocket depths of the restored and unrestored surfaces of veneered teeth (p≥0.136). A moderate correlation was found between GCF and GI.


2021 ◽  
Vol 11 (6) ◽  
pp. 555
Author(s):  
Hammam Ibrahim Fageeh ◽  
Hytham N. Fageeh ◽  
Shankargouda Patil

Background: Periodontitis is an inflammatory condition of the tooth-supporting structures initiated and perpetuated by pathogenic bacteria present in the dental plaque biofilm. In periodontitis, immune cells infiltrate the periodontium to prevent bacterial insult. Macrophages derived from monocytes play an important role in antigen presentation to lymphocytes. However, they are also implicated in causing periodontal destruction and bystander damage to the host tissues. Objectives: The objective of the present study was to quantify the cytokine profile of gingival crevicular fluid (GCF) samples obtained from patients with periodontitis. The study further aimed to assess if GCF of periodontitis patients could convert CD14+ monocytes into macrophages of destructive phenotype in an in vitro setting. The secondary objectives of the study were to assess if macrophages that resulted from GCF treatment of monocytes could affect the synthetic properties, stemness, expression of extracellular matrix proteins, adhesion molecules expressed by gingival stem cells, gingival mesenchymal stromal cells, and osteoblasts. Methods: GCF, blood, and gingival tissue samples were obtained from periodontitis subjects and healthy individuals based on specific protocols. Cytokine profiles of the GCF samples were analyzed. CD14+ monocytes were isolated from whole blood, cultured, and treated with the GCF of periodontitis patients to observe if they differentiated into macrophages. Further, the macrophages were assessed for a phenotype by surface marker analysis and cytokine assays. These macrophages were co-cultured with gingival stem cells, epithelial, stromal cells, and osteoblasts to assess the effects of the macrophages on the synthetic activity of the cells. Results: The GCF samples of periodontitis patients had significantly higher levels of IFN gamma, M-CSF, and GM-CSF. Administration of the GCF samples to CD14+ monocytes resulted in their conversion to macrophages that tested positive for CD80, CD86, and CD206. These macrophages produced increased levels of IL-1β, TNF-α, and IL-6. Co-culture of the macrophages with gingival stem cells, epithelial cells, and stromal cells resulted in increased cytotoxicity and apoptotic rates to the gingival cells. A reduced expression of markers related to stemness, extracellular matrix, and adhesion namely OCT4, NANOG, KRT5, POSTN, COL3A1, CDH1, and CDH3 were seen. The macrophages profoundly affected the production of mineralized nodules by osteoblasts and significantly reduced the expression of COL1A1, OSX, and OCN genes. Conclusion: In periodontitis patients, blood-derived monocytes transform into macrophages of a destructive phenotype due to the characteristic cytokine environment of their GCF. Further, the macrophages affect the genotype and phenotype of the resident cells of the periodontium, aggravate periodontal destruction, as well as jeopardize periodontal healing and resolution of inflammation.


2021 ◽  
pp. 105125
Author(s):  
Adomas Rovas ◽  
Alina Puriene ◽  
Kristina Snipaitiene ◽  
Egle Punceviciene ◽  
Benita Buragaite-Staponkiene ◽  
...  

2019 ◽  
Vol 8 (5) ◽  
pp. 630 ◽  
Author(s):  
Kaja Eriksson ◽  
Guozhong Fei ◽  
Anna Lundmark ◽  
Daniel Benchimol ◽  
Linkiat Lee ◽  
...  

This study aimed to investigate the periodontal health of patients with established rheumatoid arthritis (RA) in relation to oral microbiota, systemic and oral inflammatory mediators, and RA disease activity. Forty patients underwent full-mouth dental/periodontal and rheumatological examination, including collection of blood, saliva, gingival crevicular fluid (GCF) and subgingival plaque. Composition of plaque and saliva microbiota were analysed using 16S rRNA sequencing and levels of inflammatory mediators by multiplex-immunoassay. The majority of the patients (75%) had moderate or severe periodontitis and the rest had no/mild periodontitis. Anti-citrullinated protein antibody (ACPA) positivity was significantly more frequent in the moderate/severe periodontitis (86%) compared to the no/mild group (50%). No significance between groups was observed for RA disease duration or activity, or type of medication. Levels of sCD30/TNFRSF8, IFN-α2, IL-19, IL-26, MMP-1, gp130/sIL-6Rß, and sTNF-R1 were significantly higher in serum or GCF, and April/TNFSF13 was significantly higher in serum and saliva samples in moderate/severe periodontitis. The microbial composition in plaque also differed significantly between the two groups. In conclusion, the majority of RA patients had moderate/severe periodontitis and that this severe form of the disease was significantly associated with ACPA positivity, an altered subgingival microbial profile, and increased levels of systemic and oral inflammatory mediators.


Sign in / Sign up

Export Citation Format

Share Document