Sites In Periodontitis Patients Research Articles

An Original Research on Healthy and Diseased Sites in Periodontitis Patients with Dental Fluorosis

Abstract Introduction: This study evaluated the levels of clinical and biochemical parameters in patients with periodontitis and dental fluorosis. Additionally, using cone-beam computed tomography (CBCT), radiographic alveolar bone alterations in the jaws of periodontal patients were explored in terms of changes to the cortical bone thickness, trabecular bone, and bone density. Materials and Methods: The gingival crevicular fluid (GCF) was taken from a total of 40 participants who had periodontitis and dental fluorosed teeth in order to estimate the levels of sialic acid (SA) and chondroitin sulfate (CS) in glycosaminoglycans using colorimetric analysis. Results and Discussion: The total evaluation for bone density was considerably greater in the FD group (657.60 ± 270.91) than the fluorosed healthy (FH) group (526.34 ± 260.03) (P < 0.00) when various alveolar CBCT data were taken into account. The thick dense trabecular pattern was found to occur in 80% of the fluorosed diseased (FD) group as compared to 63% in the FH group (P < 0.017). The grade-wise and overall assessment was done. The FD group shows an increase in plaque index (PI), gingival bleeding index (GBI), community periodontal index-probing pocket depth, and SA than the FH group, whereas CS has no significant difference in both these groups. Moreover, GCF SA levels are highest (686.33) (P < 0.005 S) in FD and GCF CS levels in the FH group (47.05) and FD group (46.52) almost similar, and hence P = 0.686 which is statistically nonsignificant. Conclusion: Nevertheless, the need of the hour is to conduct studies on various endemic fluoride belts globally and appreciate the hidden role of fluorosis. However, fluorosis proved to be an environmental risk factor and further studies are required to decipher its destructive role systemically as well.

Origin of MMP-8 and Lactoferrin levels from gingival crevicular fluid, salivary glands and whole saliva

BackgroundPathologically elevated levels of matrix metalloproteinase-8 (MMP-8) and Lactoferrin in oral fluids have been associated with the presence of gingivitis/periodontitis. This study aimed to assess the origin of MMP-8 and Lactoferrin in periodontitis patients and to identify the degree to which conventional clinical parameters correlate with their presence.MethodsA total of ten periodontitis and ten healthy patients were included in this study. Whole saliva (stimulated and unstimulated), parotid/sublingual glandular fluid and gingival crevicular fluid from pockets and sulci were tested for MMP-8 and Lactoferrin and protein concentrations were quantified using an ELISA assay. Clinical parameters were checked for potential associations with MMP-8 and Lactoferrin levels.ResultsPeriodontal patients presented higher concentrations of MMP-8 and Lactoferrin in pockets than other sources (P = 0.03). Lactoferrin measurement was higher in the parotid compared to sublingual glandular fluid in periodontitis patients (P = 0.03). Increased probing pocket depth was positively correlated with high MMP-8 and Lactoferrin levels.ConclusionsPeriodontal pockets appear to be the major source of active matrix metalloproteinase and Lactoferrin, which also may also enter the oral cavity through the salivary glands. Since clinically healthy sites in periodontitis patients also had elevated biomarker levels, gingival crevicular fluid biomarker testing may be more predictive of future tissue breakdown than conventional clinical parameters.

Sclerostin and WNT-5a gingival protein levels in chronic periodontitis and health.

Wnt signaling pathways regulate osteoblast differentiation and bone formation and are associated with inflammatory responses driven by innate and adaptive immunity via the NF-κB pathway. The aim of this study was to compare the levels of sclerostin (SOST), WNT-5a, and TNF-α between chronic periodontitis and periodontally healthy sites and determine their value as diagnostic markers of chronic periodontitis. In a cross-sectional assessment 25 chronic periodontitis cases and 25 periodontally healthy controls were selected upon clinical and radiographic periodontal evaluation. Gingival crevicular fluid (GCF) was collected cross-sectionally from diseased and healthy sites in periodontitis patients and from healthy sites in each control subject. In a subgroup analysis, ten patients with generalized moderate and severe chronic periodontitis and ten generalized periodontally healthy individuals were included. The protein levels of SOST, WNT-5a, and TNF-α in GCF were measured by sandwich ELISA. The Shapiro-Wilk test was utilized to assess the normality of the distribution and non-parametric comparisons were performed. The protein levels of SOST were significantly higher in the generalized moderate and severe chronic periodontitis subgroup when compared to the generalized healthy (P=0.002), while the WNT-5a and TNF-α GCF total amounts were similar (P>0.05). Diseased sites in the periodontitis patients exhibited significantly higher total protein levels of WNT-5a than in healthy sites (P=0.017), whereas no differences were detected for SOST and TNF-α (P>0.05). The total protein levels of SOST, WNT-5a, and TNF-α in GCF were similar in periodontitis and non-periodontitis patients (P>0.05). Sclerostin and WNT-5a gingival protein levels demonstrated a high diagnostic value for generalized moderate and severe chronic periodontitis, while a low accuracy was detected for localized chronic periodontitis.

Increased levels of soluble CD163 in periodontitis patients.

Soluble CD163 (sCD163) has been implicated as a new biomarker in inflammatory conditions. The aim of this cross-sectional study was to assess CD163 levels systemically and locally in patients with chronic periodontitis. sCD163 levels were measured by ELISA in serum samples from 70 periodontitis and 70 periodontally healthy subjects, and in saliva samples in a subset of the population. Two gingival biopsies were harvested per subject from 20 periodontitis patients: one from a periodontally affected site, the other from a healthy site, and the relative expression of CD163 mRNA was assessed by real-time PCR. Serum sCD163 was significantly higher in periodontitis patients compared to periodontally healthy subjects (720.0±330.6ng/ml versus 510.7±219.6ng/ml, respectively; p<.001). Similarly, sCD163 levels in saliva were significantly increased in periodontitis compared to healthy subjects (3.01±5.07ng/ml versus 1.98±4.95ng/ml, respectively; p=.009). Serum and saliva sCD163 levels showed a positive correlation (Kendall's tau .27, p=.018). Importantly, CD163 gene expression was significantly higher in affected sites compared to unaffected sites in periodontitis patients, with a mean fold upregulation of 9.9 (STD: 15.3, p=.010). Our findings suggest that CD163 may be involved in the pathogenesis of periodontitis and its link with systemic conditions.

LL‐37 in periodontal health and disease and its susceptibility to degradation by proteinases present in gingival crevicular fluid

To determine the levels of LL-37 in and its susceptibility to degradation by components of gingival crevicular fluid (GCF) in periodontal health and disease. Levels of LL-37 in GCF from periodontitis patients and periodontally healthy subjects were determined by ELISA. In addition, degradation of synthetic/exogenous LL-37 by components of GCF in the presence and absence of inhibitors was determined by matrix-assisted laser desorption/ionization time of flight mass spectrometry. The concentration of native LL-37 in GCF from Porphyromonas gingivalis positive (Pg+) and P. gingivalis negative (Pg-) sites in periodontitis patients was significantly higher than in GCF from healthy subjects. When synthetic LL-37 was added to healthy GCF, the peptide was not degraded. Conversely, GCF from Pg+ sites rapidly degraded synthetic LL-37 which was prevented in the presence of Arg- and Lys- gingipain inhibitors. Synthetic LL-37 was degraded more slowly by GCF from Pg- sites. LL-37 is detectable in GCF in periodontal health and disease. The rapid degradation of synthetic LL-37 in periodontitis GCF, particularly in Pg+ sites, limits its role as a potential therapeutic in the gingival crevice. These results highlight the need to design stable peptide mimetics of LL-37 as future therapeutics in periodontitis.

Porphyromonas gingivalis, Gamma Interferon, and a Proapoptotic Fibronectin Matrix Form a Synergistic Trio That Induces c-Jun N-Terminal Kinase 1-Mediated Nitric Oxide Generation and Cell Death

During infection and inflammation, bacterial and inflammatory proteases break down extracellular matrices into macromolecular fragments. Fibronectin fragments are associated with disease severity in arthritis and periodontitis. The mechanisms by which these fragments contribute to disease pathogenesis are unclear. One likely mechanism is that fibronectin fragments induce apoptosis of resident cells, which can be further modulated by nitric oxide. Nitric oxide levels are increased at inflammatory sites in periodontitis patients. The aim of this study was to examine whether a proapoptotic fibronectin matrix (AFn) exerts its action by inducing nitric oxide and whether priming by bacterial and inflammatory components exacerbates this mechanism. Our data demonstrate that AFn increased the levels of nitric oxide and inducible nitric oxide synthase (iNOS) dose and time dependently in periodontal ligament (PDL) cells. These effects and apoptosis were inhibited by iNOS suppression and enhanced by iNOS overexpression. Nitric oxide and iNOS induction were paralleled by increased c-Jun N-terminal kinase 1 (JNK-1) phosphorylation. JNK-1 overexpression enhanced the expression of nitric oxide and iNOS, whereas inhibiting JNK-1 by small interfering RNA or a kinase mutant reversed these findings. Priming PDL cells with Porphyromonas gingivalis, its lipopolysaccharide (LPS), or gamma interferon (IFN-gamma) further increased nitric oxide levels and apoptosis. Escherichia coli and Streptococcus mutans induced lesser effects. Gingival fibroblasts and neutrophils responded to a lesser degree to these stimuli, whereas keratinocytes were resistant to apoptosis. Thus, proapoptotic matrices trigger nitric oxide release via JNK-1, promoting further apoptosis in host cells. LPS and IFN-gamma accentuate this mechanism, suggesting that during inflammation, the affected matrices and bacterial and inflammatory components combined exert a greater pathogenic effect on host cells.

Gingival Crevicular Fluid Leptin Levels in Periodontitis Patients With Long‐Term and Heavy Smoking

The aim of the present study was to evaluate gingival crevicular fluid (GCF) leptin levels and the influence of long-term and heavy smoking on GCF leptin levels in patients with chronic periodontitis. In this study, 143 individuals were divided into three groups: non-smokers (NS), smokers (S), and control (C). Three subgroups of NS and S were grouped as follows: a) probing depth (PD) <or=3 mm; b) PD = 4 to 5 mm; and c) PD >5 mm. For each patient, PD, gingival index (GI), plaque index (PI), gingival bleeding time index (GBTI), and clinical attachment level (CAL) values were recorded. The GCF leptin levels obtained from sampling sites were determined by using enzyme-linked immunosorbent assay kits. The GCF leptin levels were found significantly lower in the a and b subgroups in the S group than those in the NS group (P <0.05). The inflammatory markers GI and GBTI showed significant correlations with leptin in NS (P <0.05). Our results suggest that higher leptin GCF levels in healthy sites in periodontitis patients may play a protective role in periodontal disease. Further studies are needed to determine the cellular origin of the leptin in the gingiva and the effect of plasma leptin levels on GCF leptin concentrations.

Capnocytophaga spp. in Periodontitis Patients Manifesting Diabetes Mellitus

The subgingival microflora in patients presenting concurrently with periodontitis and diabetes mellitus (DM) are poorly understood. While traditional putative periodontal pathogens are implicated, research involving other oral organisms; e.g., Capnocytophaga spp., is lacking. These organisms produce a range of bacterial enzymes relevant to periodontal breakdown. It is inferred that periodontal bacteria acquire systemic access through the ulcerated periodontal pocket surface; conclusive evidence supporting this notion is limited. The aims of this investigation were to: 1) quantify and identify Capnocytophaga spp. present in healthy and diseased sites in periodontitis patients with and without DM, and 2) isolate periodontal pathogens from these patients' blood. Twenty-one DM-periodontitis and 25 periodontitis patients were recruited. Subgingival plaque was collected from three healthy and three diseased sites per subject. Capnocytophaga spp. and total (facultative and obligate) anaerobic counts from each site were estimated. Capnocytophaga spp. were identified using 16S rRNA polymerase chain reaction (PCR) restriction fragment length polymorphism (RFLP). Statistical analyses were performed using multilevel modeling. Blood samples were subjected to HbA(1c) estimation and bacterial culture. A total of 848 Capnocytophaga spp. were isolated and identified. Significantly higher numbers of Capnocytophaga spp. (P <0.001) and anaerobes (P <0.001) were present in diseased sites in DM-periodontitis subjects compared to healthy sites in non-DM-periodontitis and DM-periodontitis subjects. C. ochracea (and variant) and C. granulosa were the most prevalent species. Blood samples were negative for Capnocytophaga spp. Total mean counts for Capnocytophaga spp. were significantly higher in DM-periodontitis subjects versus non-DM-periodontitis (P = 0.025) and at diseased sites versus healthy sites (P <0.001). Analysis of individual species revealed that the outcome varied with site status and DM status.

Elastase and lactoferrin in gingival crevicular fluid: possible indicators of a granulocyte‐associated specific host response

Periodontitis affects a limited number of susceptible humans. The aim of this study was to determine whether there is a difference in the inflammatory reaction between patients with gingivitis and those with periodontitis. For this purpose the levels of elastase and lactoferrin were measured in gingival crevicular fluid (GCF) from three types of sites: i) inflamed sites in patients with gingivitis alone, inflamed sites both ii) with and iii) without tissue destruction in patients with periodontitis. Elastase activity, measured with a chromogenic substrate was significantly higher in the two types of sites in periodontitis patients. Lactoferrin levels, measured with ELISA were the same in the three types of sites. In vitro activation of granulocytes from healthy volunteers with Fc-receptor stimulation showed that the entire release of lactoferrin occurred immediately. In contrast, elastase release was time-dependent and continued throughout the experiment. Thus, the degranulation of the specific (lactoferrin) and azurophil granule (elastase) are under separate control and the two parameters can be combined in a ratio in order to characterize the granulocytes of a given patient. Assuming an immediate release of lactoferrin from the activated granulocytes in vivo, similar amounts of lactoferrin in the three types of sites can be regarded as reflecting similar numbers of granulocytes in the three types of sites. Consequently, a higher elastase activity in GCF from patients with periodontitis indicates a higher rate of release from the cells per se and a granulocyte-associated specific host response.

The polyamines putrescine, spermidine and spermine in human gingival crevicular fluid

In human periodontal disease, there may be periods of exacerbation and remission. Definition of the homeostatic mechanisms in the periodontium may therefore be important in understanding the natural history of this disorder. The polyamines are biologically active amines involved in the regulation of cell growth, regeneration of tissue and modulation of inflammation. Their occurrence was examined in gingival crevicular fluid (GCF). Fifteen sites were evaluated in four patients with moderately advanced periodontitis before and after root planing and scaling, and 15 sites were evaluated in four patients with mild inflammatory gingivitis and no attachment loss. Polyamine analysis was by high-performance liquid chromatography. GCF from untreated sites in periodontitis patients contained the highest concentration of putrescine (10 4 greater than serum). This polyamine was detected in all periodontitis samples and 12 of 15 gingivitis samples. Significant differences were seen when the amount of putrescine/30 s sample was compared: periodontitis sites before treatment 1005.7 ± 106.1 pmol; periodontitis sites after treatment 504.7 ± 89.2 pmol; gingivitis sites 186.7 ± 40.1 pmol. In contrast, spermidine and spermine were detected only occasionally. Thus putrescine may play an important homeostatic role in the periodontium.