Periodontitis Patients Research Articles

Immunoaffinity-enriched salivary small extracellular vesicles in periodontitis

Aim: Saliva extracellular vesicles (EVs) serve as a significant reservoir of biomarkers that may be of clinical use in disease diagnosis. Saliva, however, contains EVs of both host- and bacterial- origin. Identifying suitable EVs for disease diagnosis involves enriching host EVs and limiting non-host contamination with effective isolation methods. The objectives of this research were: (1) to evaluate the salivary EVs enrichment in 12 periodontally healthy patients by two different methods: size exclusion chromatography (SEC) and bead-based immunoaffinity capture (EXO-NET®); (2) to analyze the variance expression of inflammatory cytokines in EXO-NET-enriched EVs, comparing individuals with periodontitis (n = 20) to non-periodontitis (n = 12). Methods: Whole unstimulated saliva samples were collected from 12 periodontally healthy and 20 periodontitis patients. EVs were isolated from the 12 non-periodontitis patients using SEC (referred to as SEC-EVs) and EXO-NET (referred to as EXO-NET EVs), after which their total protein content, 37 EV surface markers, and bacterial pathogens expression were compared. Subsequently, the inflammatory cytokines expression levels (interleukin-IL-6, IL-1β, IL-8, and IL-10) in EXO-NET EVs were measured for non-periodontitis and periodontitis. Results: EXO-NET EVs contained more EV-specific protein and substantially higher expression of EV surface markers (CD9, CD81, CD63), but less pathogenic DNA was detected compared to that in SEC-EVs. Additionally, EXO-NET EVs from periodontitis patients contained higher amounts of IL-6 and IL-8, and decreased IL-10, compared to those from non-periodontitis patients. Conclusion: The findings suggest that immunoaffinity capture (EXO-NET) is a dependable method for salivary EVs enrichment, resulting in a higher yield of host EVs with reduced bacterial DNA detection compared to SEC. Furthermore, the research proposes that immunoaffinity capture enriched EVs can function as biomarkers for periodontitis, demonstrated by an increased expression of proinflammatory cytokines from periodontitis patients.

Osteocalcin levels in gingival crevicular fluid periodontitis patient with and without type 2 Diabetes Mellitus

Background: Diabetes Mellitus (DM) is a systemic disease with a high prevalence in Indonesia. Periodontitis is a complication of DM with frequent occurrences. Periodontitis with DM causes greater bone damage than non-DM periodontitis because its conditions can affect osteoblast and osteoclast activity. Osteocalcin (OC), mostly synthesized by osteoblasts, regulates the activity of bone formation and turnover so that it can be a marker of bone formation and turnover to see the severity of the disease. OC can be found in gingival crevicular fluid (GCF). This study aimed to determine osteocalcin levels in gingival crevicular fluid periodontitis patients with uncontrolled type 2 Diabetes Mellitus and non-Diabetes Mellitus.Method: Samples used in this study were gingival crevicular fluid taken from 20 samples of periodontitis patients with DM type 2 (DMT2P) and 20 samples of periodontitis non-DM patients. The OC levels were analyzed using ELISA Kit Osteocalcin then statistical tests were carried out.Result: The results showed that the OC level of the DMT2P group was 0.369 ± 0.140 while the periodontitis non-DM group was 0.664 ± 0.141.Conclusion: The OC levels of periodontitis patients with DM are lower than periodontitis patients non-DM.

Relationship Between Staging and Grading of Periodontitis and Periimplantitis: A Retrospective Study

Objective: The purpose of this retrospective study was to determine the relationship between peri-implantitis and stage/grade of periodontitis. 
 Methods: Records of 171 periodontitis patients with 318 dental implants were screened. Classification of diagnosed patients with periodontitis were done by both stage (1, 2, 3 and 4) and grade (A, B and C). Peri-implant health status was evaluated as no peri-implantitis or with peri-implantitis. Marginal bone loss severity of implants diagnosed with peri-implantitis, prosthesis type (single, bridge), location of dental implants (maxilla, mandibula, anterior and posterior), smoking (yes/no) and diabetes history (yes/no) of the patients were also evaluated. Analysis was done at implant level.
 Results: A total of 203 (63.8%) dental implants were diagnosed with peri-implantitis. There were statistical differences in the stage and grade of periodontitis between implants diagnosed with no peri-implantitis and peri-implantitis (p

Salivary Protein Quantification among Periodontitis Patients with Different Disease Severity: A Pilot Study

The salivary protein quantification reflecting the periodontal pathophysiology among patients with different periodontitis severity has not been well explored. This quantification is fundamental as the changes in the quantity of proteins in different severities of periodontitis can reflect the periodontal complex alterations. This pilot study aimed to quantify and compare the salivary proteins by collecting unstimulated saliva from periodontitis patients and healthy individuals. The radioimmunoprecipitation assay (RIPA) protocol was used to extract protein from patients’ saliva and quantified using the Nanodrop Spectrophotometer 2000/2000c (A280) set at 280 nm wavelength. The mean concentration of salivary proteins was shown to be 26.48 + 2.95 mg/mL in healthy patients, 22.00 + 0.38 mg/mL in Stage 1 periodontitis patients, 27.34 + 1.61 mg/mL in Stage 2 periodontitis patients, 27.11 + 0.66 mg/mL in Stage 3 periodontitis patients and 24.89 + 1.91 mg/mL in Stage 4 periodontitis patients. However, the mean difference between all groups was not statistically significant. Within the limitations of this study, there was no difference in the salivary protein quantity among patients with different stages of periodontitis.

Gingival crevicular fluid levels of apelin correlates with clinical periodontal diagnosis.

Limitations of clinical periodontal measurements have led to the search for reliable biomarkers that can be used in diagnosis and monitoring of periodontal diseases. Considering the relationship of adipokines with periodontal disease, diabetes, and obesity, apelin may be a biomarker for periodontal diseases due to its modulating effects on inflammation. The present study was conducted to determine gingival crevicular fluid (GCF) apelin levels in systemically healthy individuals and to evaluate the potential of apelin as a biomarker for periodontal diagnosis. Ten individuals with clinically healthy periodontal tissues, 10 patients diagnosed with gingivitis, and 10 patients with periodontitis were included in the present study. Whole mouth clinical periodontal measurements were recorded and GCF samples were obtained from the buccal approximal regions of single-rooted teeth with features that would represent clinical periodontal diagnosis. Apelin level in the samples was determined by ELISA. Clinical and biochemical findings were statistically analyzed. Possible relationship between the variables was evaluated with Pearson correlation analysis. Apelin level in the gingivitis group was higher than that in the clinically healthy group (p = 0.000) and lower than that in the periodontitis group (p = 0.000). A positive correlation was found between GCF apelin concentration and plaque score, bleeding on probing, and probing depth (p = 0.000). Within the limits of this study, it can be suggested that GCF apelin concentration may be a biomarker that can distinguish between healthy periodontal tissues, gingivitis, and periodontitis patients. Apelin concentration in the gingival crevicular fluid may aid in the diagnosis of periodontal disease.

High incidence of periodontitis in patients with ascitic decompensated cirrhosis.

Periodontitis has been associated with various liver diseases. However, the relevance of periodontitis in the progression of decompensated cirrhosis remains inconclusive. In particular, it is unclear whether the common periodontitis pathogens, Porphyromonas gingivalis (P. gingivalis) and Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans), can be detected not only in the oral mucosa but also in ascites and stool. To investigate the significance of periodontitis, P. gingivalis, and A. actinomycetemcomitans in cirrhosis patients with ascitic decompensation. This prospective study was conducted at the University Hospital Hamburg-Eppendorf, a tertiary center in Northern Germany. A cohort of 27 patients with ascitic decompensated liver cirrhosis underwent dental examinations to assess the association between periodontitis and various clinical parameters of cirrhosis, as well as patient outcomes. PCR was used to test gingival samples, ascites, and stool for the presence of P. gingivalis and A. actinomycetemcomitans. Gingival samples were collected by probing the deepest gum pocket of a sextant and wiping them on a cotton swab. Periodontitis was diagnosed in 22 out of 27 (82%) ascite patients, which is significantly more common than in a control cohort of 100 unselected patients (59%, P = 0.04). P. gingivalis was detected in the gingiva of six patients, and one of them also had P. gingivalis in their stool. However, P. gingivalis was not found in the ascites of any patient. Five out of six patients with P. gingivalis had periodontitis (83%). A. actinomycetemcomitans was not detected in any sample. Patients without periodontitis had a significantly higher mortality rate compared to those with periodontitis, and survival (Kaplan-Meier analysis) was longer in patients with periodontitis (P = 0.02). Transplant-free survival was also more common in patients with periodontitis compared to those without (63% vs 0%, P = 0.02). Decompensated cirrhotic patients frequently suffer from periodontitis. However, there was no evidence of the translocation of P. gingivalis or A. actinomycetemcomitans into ascites. The survival of cirrhotic patients with periodontitis was not reduced.

High-Throughput Combined Analysis of Saliva Microbiota and Metabolomic Profile in Chinese Periodontitis Patients: A Pilot Study.

The onset and progression of periodontitis involves complicated interactions between the dysbiotic oral microbiota and disrupted host immune-inflammatory response, which can be mirrored by the changes in salivary metabolites profile. This pilot study sought to examine the saliva microbiome and metabolome in the Chinese population by the combined approach of 16s rRNA sequencing and high-throughput targeted metabolomics to discover potential cues for host-microbe metabolic interactions. Unstimulated whole saliva samples were collected from eighteen Stage III and IV periodontitis patients and thirteen healthy subjects. Full-mouth periodontal parameters were recorded. The taxonomic composition of microbiota was obtained by 16s rRNA sequencing, and the metabolites were identified and measured by ultra-high performance liquid chromatography and mass spectrometry-based metabolomic analysis. The oral microbiota composition displayed marked changes where the abundance of 93 microbial taxa differed significantly between the periodontitis and healthy group. Targeted metabolomics identified 103 differential metabolites between the patients and healthy individuals. Functional enrichment analysis demonstrated the upregulation of protein digestion and absorption, histidine metabolism, and nicotinate and nicotinamide metabolism pathways in the dysbiotic microbiota, while the ferroptosis, tryptophan metabolism, glutathione metabolism, and carbon metabolism pathways were upregulated in the patients. Correlation analysis confirmed positive relationships between the clinical parameters, pathogen abundances, and disease-related metabolite levels. The integral analysis of the saliva microbiome and metabolome yielded an accurate presentation of the dysbiotic oral microbiome and functional alterations in host-microbe metabolism. The microbial and metabolic profiling of the saliva could be a potential tool in the diagnosis, prognosis evaluation, and pathogenesis study of periodontitis.

A genome-wide association study meta-analysis in a European sample of stage III/IV grade C periodontitis patients ≤35 years of age identifies new risk loci.

Few genome-wide association studies (GWAS) have been conducted for severe forms of periodontitis (stage III/IV grade C), and the number of known risk genes is scarce. To identify further genetic risk variants to improve the understanding of the disease aetiology, a GWAS meta-analysis in cases with a diagnosis at ≤35 years of age was performed. Genotypes from German, Dutch and Spanish GWAS studies of III/IV-C periodontitis diagnosed at age ≤35 years were imputed using TopMed. After quality control, a meta-analysis was conducted on 8,666,460 variants in 1306 cases and 7817 controls with METAL. Variants were prioritized using FUMA for gene-based tests, functional annotation and a transcriptome-wide association study integrating eQTL data. The study identified a novel genome-wide significant association in the FCER1G gene (p = 1.0 × 10-9 ), which was previously suggestively associated with III/IV-C periodontitis. Six additional genes showed suggestive association with p < 10-5 , including the known risk gene SIGLEC5. HMCN2 showed the second strongest association in this study (p = 6.1 × 10-8 ). This study expands the set of known genetic loci for severe periodontitis with an age of onset ≤35 years. The putative functions ascribed to the associated genes highlight the significance of oral barrier tissue stability, wound healing and tissue regeneration in the aetiology of these periodontitis forms and suggest the importance of tissue regeneration in maintaining oral health.

Prevalence of inflammatory periodontal disease in patients attended in a university dental clinic – a retrospective analysis

OBJECTIVE: To estimate the prevalence of inflammatory periodontal disease (IPD) in individuals treated in a university dental clinic located in the Center-West of Brazil. METHODS: We performed a retrospective cross-sectional study with census sample. Previously collected data from dental records were tabulated and analyzed descriptively. RESULTS: 151 patients were included on the research. Approximately 60% presented gingivitis and 32.4% periodontitis. 85.4% showed high plaque index (&gt;10%) and 31.7% declared not to use dental floss. 12% of the participants reported being diabetic and 17.2% were smokers. The prevalence of deep pockets was the worst periodontal condition found (15.2%). CONCLUSION: The prevalence of ILD found in this population was higher than the Brazilian national average. The results can be useful to support buccal and general health promotion strategies on public policies.

Alterations in oxidative stress biomarkers and helper T-cell subgroups in patients with periodontitis and IgA nephropathy.

Investigating the changes in the oxidative stress levels and helper T lymphocyte (Th) subsets in patients with periodontitis and IgA nephropathy (IgAN) to determine their relationship. IgAN has a high prevalence, poor prognosis, and no effective cure. Accumulating evidence has implicated a close relationship between periodontitis and chronic kidney diseases, in which both IgAN and chronic periodontitis show chronic inflammation and abnormal metabolism. However, few studies have been conducted on the relationship between the two diseases from this perspective. We divided 86 IgAN patients into patients with healthy periodontium (IgAN-H, n = 34) and patients with periodontitis IgAN (IgAN-P, n = 52); moreover, we divided 72 systemically healthy participants into patients with periodontitis (H-P, n = 35) and participants with healthy periodontium (H-H, n = 37). The proportions of Th subsets in peripheral blood were estimated using flow cytometry. Cytokine levels in plasma were assessed using cytokine assay kits. Enzyme-linked immunosorbent assay was used to evaluate the plasma levels of oxidative stress. Our results from analyzing the Th cell subsets indicated that Th2 cell counts in the IgAN-P group were significantly lower than those in the IgAN-H group, while Th17 cell counts were increased (p < 0.05). Moreover, the Th1/Th2 ratio and interleukin-6 levels in the IgAN-P group were significantly higher than those in the H-H group (p < 0.01). Compared with that in the H-H group, in the remaining three groups, plasma total oxidation state (TOS) levels were increased (p < 0.01), while plasma total antioxidant state (TAS) levels were decreased (p < 0.05). Furthermore, estimated glomerular filtration rate was negatively correlated with the probing depth and gingival bleeding index. IgAN was a risk factor for periodontitis, while TAS was a protective factor for periodontitis. The oxidative stress index (OSI) might be valuable for distinguishing periodontitis patients from healthy controls (area under the receiver operator characteristic curve = 0.951). IgAN is an independent risk factor of periodontitis, and the Th17 cell-mediated inflammatory response might be associated with the occurrence of periodontitis in patients with IgAN. Patients with coexisting IgAN and periodontitis exhibit increased oxidative stress, in which TOS and OSI are potential biomarkers for diagnosing periodontitis.

Modulating the sEH/EETs Axis Restrains Specialized Proresolving Mediator Impairment and Regulates T Cell Imbalance in Experimental Periodontitis.

Epoxyeicosatrienoic acids (EETs) and other epoxy fatty acids are short-acting lipids involved in resolution of inflammation. Their short half-life, due to its metabolism by soluble epoxide hydrolase (sEH), limits their effects. Specialized proresolving mediators (SPMs) are endogenous regulatory lipids insufficiently synthesized in uncontrolled and chronic inflammation. Using an experimental periodontitis model, we pharmacologically inhibited sEH, examining its impact on T cell activation and systemic SPM production. In humans, we analyzed sEH in the gingival tissue of periodontitis patients. Mice were treated with sEH inhibitor (sEHi) and/or EETs before ligature placement and treated for 14 d. Bone parameters were assessed by microcomputed tomography and methylene blue staining. Blood plasma metabololipidomics were carried out to quantify SPM levels. We also determined T cell activation by reverse transcription-quantitative PCR and flow cytometry in cervical lymph nodes. Human gingival samples were collected to analyze sEH using ELISA and electrophoresis. Data reveal that pharmacological sEHi abrogated bone resorption and preserved bone architecture. Metabololipidomics revealed that sEHi enhances lipoxin A4, lipoxin B4, resolvin E2, and resolvin D6. An increased percentage of regulatory T cells over Th17 was noted in sEHi-treated mice. Lastly, inflamed human gingival tissues presented higher levels and expression of sEH than did healthy gingivae, being positively correlated with periodontitis severity. Our findings indicate that sEHi preserves bone architecture and stimulates SPM production, associated with regulatory actions on T cells favoring resolution of inflammation. Because sEH is enhanced in human gingivae from patients with periodontitis and connected with disease severity, inhibition may prove to be an attractive target for managing osteolytic inflammatory diseases.

A novel clinical protocol for dentin hypersensitivity management based on regenerative dental gel associated with calcium: A case study in a patient with periodontal disease

Introduction: Dentin hypersensitivity (DH) is defined as pain caused by dentin exposure in response to various stimuli. Patients with periodontal disease (PD) frequently report DH due to root surface exposure or after periodontal treatment. Bioactive agents derived from calcium and/or silicon-based systems, such as REFIX technology, have been used to promote tooth regeneration, with focus on tooth sensitivity treatment. Case Report: Based on this, the efficacy of REFIX gel combined with calcium on the treatment of a DH was evaluated in a PD 45-year-old male patient. The patient’s right and left sides were randomly selected to use one of the prophylaxis dental protocols: REFIX dental gel/calcium (RDGCa) (experimental: 13, 14, 15 teeth) or Pumice Stone (PS) (control: 23, 24, 25 teeth). The products were applied using dental polishing brush during 10 seconds in each tooth surface. Schiff Air Index (SAI) for sensitive teeth (0–3) were performed by applying air jet to each tooth’s vestibular surface for 5 seconds after and before the prophylaxis. The patient reported pain and interruption of the stimulus before and after PS prophylaxis (SAI 3 and 3 respectively). There was a reduction in pain when RDGCa protocol was used (SAI 3—before/SAI 1—after). In this instance, the patient felt the stimulus but did not request to be interrupted. This contrasted with the outcomes of the initial stimulus as well as PS. Conclusion: The results showed that the RDGCa strategy was successful in lowering DH in PD patients.

Establishment of models to predict factors influencing periodontitis in patients with type 2 diabetes mellitus.

Type 2 diabetes mellitus (T2DM) is associated with periodontitis. Currently, there are few studies proposing predictive models for periodontitis in patients with T2DM. To determine the factors influencing periodontitis in patients with T2DM by constructing logistic regression and random forest models. In this a retrospective study, 300 patients with T2DM who were hospitalized at the First People's Hospital of Wenling from January 2022 to June 2022 were selected for inclusion, and their data were collected from hospital records. We used logistic regression to analyze factors associated with periodontitis in patients with T2DM, and random forest and logistic regression prediction models were established. The prediction efficiency of the models was compared using the area under the receiver operating characteristic curve (AUC). Of 300 patients with T2DM, 224 had periodontitis, with an incidence of 74.67%. Logistic regression analysis showed that age [odds ratio (OR) = 1.047, 95% confidence interval (CI): 1.017-1.078], teeth brushing frequency (OR = 4.303, 95%CI: 2.154-8.599), education level (OR = 0.528, 95%CI: 0.348-0.800), glycosylated hemoglobin (HbA1c) (OR = 2.545, 95%CI: 1.770-3.661), total cholesterol (TC) (OR = 2.872, 95%CI: 1.725-4.781), and triglyceride (TG) (OR = 3.306, 95%CI: 1.019-10.723) influenced the occurrence of periodontitis (P < 0.05). The random forest model showed that the most influential variable was HbA1c followed by age, TC, TG, education level, brushing frequency, and sex. Comparison of the prediction effects of the two models showed that in the training dataset, the AUC of the random forest model was higher than that of the logistic regression model (AUC = 1.000 vs AUC = 0.851; P < 0.05). In the validation dataset, there was no significant difference in AUC between the random forest and logistic regression models (AUC = 0.946 vs AUC = 0.915; P > 0.05). Both random forest and logistic regression models have good predictive value and can accurately predict the risk of periodontitis in patients with T2DM.

The spatial transcriptomic landscape of human gingiva in health and periodontitis.

The gingiva is a key oral barrier that protects oral tissues from various stimuli. A loss of gingival tissue homeostasis causes periodontitis, one of the most prevalent inflammatory diseases in humans. The human gingiva exists as a complex cell network comprising specialized structures. To understand the tissue-specific pathophysiology of the gingiva, we applied a recently developed spatial enhanced resolution omics-sequencing (Stereo-seq) technique to obtain a spatial transcriptome (ST) atlas of the gingiva in healthy individuals and periodontitis patients. By utilizing Stereo-seq, we identified the major cell types present in the gingiva, which included epithelial cells, fibroblasts, endothelial cells, and immune cells, as well as subgroups of epithelial cells and immune cells. We further observed that inflammation-related signalling pathways, such as the JAK-STAT and NF-κB signalling pathways, were significantly upregulated in the endothelial cells of the gingiva of periodontitis patients compared with those of healthy individuals. Additionally, we characterized the spatial distribution of periodontitis risk genes in the gingiva and found that the expression of IFI16 was significantly increased in endothelial cells of inflamed gingiva. In conclusion, our Stereo-seq findings may facilitate the development of innovative therapeutic strategies for periodontitis by mapping periodontitis-relevant genes and pathways and effector cells.

Salivary Proteome and Intact N-Glycopeptides Analysis Reveal Specific Signatures in Periodontitis.

Periodontitis is a prevalent oral inflammatory disease that can result in tooth loss and is closely linked to type 2 diabetes (T2D). In this study, we analyzed the salivary proteome and intact N-glycopeptides (IGPs) of individuals with mild-moderate, severe, aggressive periodontitis, and periodontitis with T2D, including those treated with antidiabetic drugs, to identify specific signatures associated with the disease. Our results revealed that salivary proteins and glycoproteins were altered in all periodontitis groups (PRIDE ID: 1-20230612-72345), with fucose- and sialic acid-containing N-glycans showing the greatest increase. Additionally, differentially expressed proteins were classified into 9 clusters, including those that were increased in all periodontitis groups and those that were only altered in certain types of periodontitis. Interestingly, treatment with antidiabetic drugs reversed many of the changes observed in the salivary proteome and IGPs in T2D-related periodontitis, suggesting a potential therapeutic approach for managing periodontitis in patients with T2D. Consistent with MS/MS results, the expression of salivary IGHA2 and Fucα1-3/6GlcNAc (AAL) was significantly increased in MP. These findings provide new insights into the pathogenesis of periodontitis and highlight the potential of salivary biomarkers for diagnosis, prognosis, and monitoring of disease progression and treatment response.

Human gingival fibroblasts are a source of B cell-activating factor during periodontal inflammation.

Host-modulating therapy is a possible treatment for individuals that respond poorly to conventional periodontal therapy. B cells, abundant in periodontitis lesions, require the cytokines B cell-activating factor (BAFF) and A proliferation-inducing ligand (APRIL) for survival and maturation. Although mRNA levels of BAFF and APRIL are increased in tissue from periodontitis lesions, it is unknown if periodontal resident cells express BAFF and/or APRIL during periodontal inflammation. In this study, we aim to analyze the expression of BAFF and APRIL in human gingival fibroblasts after stimulation with proinflammatory cytokines. Furthermore, we perform protein analysis in tissues and serum from periodontitis patients and healthy controls. Human gingival fibroblasts were cultured and stimulated with the proinflammatory cytokines' tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β). The mRNA expression of BAFF and APRIL was analyzed by real-time quantitative polymerase chain reaction (qPCR), and the protein was detected in tissue sections using immune staining. Serum levels of BAFF were analyzed with enzyme-linked immunosorbent assay (ELISA). In gingival fibroblasts, TNF-α upregulated BAFF mRNA, but APRIL was unaffected. IL-1β affected neither BAFF nor APRIL expression. BAFF protein was detected in the oral epithelium and in cells of the underlying connective tissue in periodontitis tissue, and BAFF protein was increased in the serum of periodontitis patients. Periodontal resident cells express BAFF during periodontal inflammation and participate in providing a favorable milieu for the survival and action of B cells.

Assessment of Salivary MMP-8 and IL-1β for the Diagnosis of Periodontal Diseases in Pakistani Population.

Clinical methods use the subjective diagnosis of periodontal diseases by visual observation that could result in differences and variability of diagnosis. The addition of specific markers could aid in the accurate diagnosis of the local population. The objective of the study was to target two of the major proteins for possible significance in such an approach. Unstimulated saliva samples were collected from 60 participants aged between 18 and 70 years. Three groups each with twenty participants were recruited into periodontitis, gingivitis, and healthy control. The samples were analyzed using human enzyme-linked immunosorbent assay kits for matrix metalloproteinase-8 (MMP-8) and interleukin-1β (IL-1β). SPSS version 20 was used to analyze the result. Posthoc analysis by Tukey's test revealed that MMP-8 levels were higher in gingivitis and periodontitis groups as compared with healthy controls. The test also revealed that IL-1β levels were higher in the periodontitis group compared with the healthy control and gingivitis group. Additionally, one-way analysis of variance analysis showed a significant effect on probing depth in gingivitis and periodontitis patients. The mean age of periodontitis group was significantly higher than other groups. Salivary biomarkers may provide useful diagnostic information and could be utilized as tests for periodontal disease screening, prognosis, and prediction.

CCL5's Role in Periodontal Disease: A Narrative Review.

Persistent host inflammatory and immune responses to biofilm play a critical role in the mechanisms that govern soft and hard tissue destruction in periodontal disease. Among the less explored facets of these mechanisms are chemokines, including CCL5 (C-C motif chemokine ligand 5), also known as RANTES (regulated on activation, normal T cell expressed and secreted), a proinflammatory CC subfamily chemokine synthesized by T lymphocytes. Despite its importance, there is currently no comprehensive review of the role of CCL5 in periodontitis in the literature. Therefore, this paper aims to fill this gap by summarizing the existing knowledge on the involvement of CCL5 in the onset and progression of periodontitis. In addition, we aim to stimulate interest in this relatively overlooked factor among periodontitis researchers, potentially accelerating the development of drugs targeting CCL5 or its receptors. The review examines the association of CCL5 with periodontitis risk factors, including aging, cigarette smoking, diabetes, and obesity. It discusses the involvement of CCL5 in pathological processes during periodontitis, such as connective tissue and bone destruction. The data show that CCL5 expression is observed in affected gums and gingival crevicular fluid of periodontitis patients, with bacterial activity contributing significantly to this increase, but the reviewed studies of the association between CCL5 expression and periodontal disease have yielded inconclusive results. Although CCL5 has been implicated in the pathomechanism of periodontitis, a comprehensive understanding of its molecular mechanisms and significance remains elusive, hindering the development of drugs targeting this chemokine or its receptors.

Retinol-Binding Protein 4 and Visfatin Levels in Patients with Periodontitis and Obesity/Overweight: A Systematic Review and Meta-Analysis.

Prior studies demonstrated an equivocal conclusion about the association between the level of retinol-binding protein 4 (RBP4)/visfatin and periodontitis patients with obesity. The aim of our study (Prospero ID: CRD42023469058) was to systematically review the available articles linking the biofluid levels of RBP4/visfatin to the comorbidity of periodontitis and obesity. Clinical trials were screened in accordance with specific inclusion criteria from seven databases up to November 2023. A quality assessment was performed with the Newcastle-Ottawa Scale and ROBINS-I tools for observational and interventional trials, respectively. The standard mean difference (SMD) with a 95% confidence interval (CI) related to the RBP4 level was recorded; the other indicators related to the visfatin level were measured via the mean difference (MD) with the corresponding 95% CI, and Fisher's Z transformation was measured to reveal the association using Review Manager 5.4. The current evidence was based on five observational studies and two interventional studies. All of them were included in the systematic review, and six of them were in the meta-analysis. Statistical analysis indicated that there was no significant difference in the circulating levels of RBP4 in the periodontitis patients with obesity or without, who were labeled as OP or NP, respectively (155 OP-107 NP: SMD = 1.38; 95% CI: -0.18-2.94, p = 0.08), as well as the periodontal healthy patients with a normal weight, who were labelled as NnP (116 OP-79 NnP: SMD = 6.76; 95% CI: -5.34-18.87, p = 0.27). Meanwhile, a significant higher level of serum visfatin was found in the OP patients than that of the NP (86 OP-45 NP: MD = 4.21; 95% CI: 2.65-5.77, p < 0.00001)/NnP (164 OP-88 NnP: MD = 13.02; 95% CI: 7.34-18.70, p < 0.00001) group. In addition, a positive association was observed between the serum RBP4 and body mass index/clinical attachment loss (CAL). And, then, there was a positive association between the serum visfatin and periodontal parameters, including the probing depth, CAL, and plaque index, as well as metabolic parameters, including the total cholesterol, triglycerides, fasting blood glucose, and low-density lipoprotein cholesterol. Here, the circulating RBP4 level was not independently related to the comorbidity of periodontitis and obesity, while serum visfatin was significantly associated with periodontitis and obesity. Notably, the positive association between circulating RBP4/visfatin and the periodontal parameters/metabolic parameters firmly suggested that the higher severity of the obese or periodontal status was associated with an elevated level of serum visfatin or RBP4 in the OP group. With more rigorous longitudinal research, the exact causations between RBP4/visfatin and the patients affected by obesity and periodontitis could be disentangled. RBP4 and visfatin might be novel, enlightening prospective bio-indexes for the targeted treatment of comorbidities.

Evaluation of a biomarker (NO) dynamics in inflammatory zebrafish and periodontitis saliva samples via a fast-response and sensitive fluorescent probe

Many pathological processes include nitric oxide (NO), a signaling transduction molecule. Tumors, cardiovascular, cerebrovascular, neurodegenerative, and other illnesses are linked to abnormal NO levels. Thus, evaluating NO levels in vitro and in vivo is crucial for studying chemical biology process of associated disorders. This work devised and manufactured a coumarin-based fluorescent probe ZPS-NO to detect nitric oxide (NO). The reaction between ZPS-NO and NO produced a highly selective and sensitive optical response that caused a powerful fluorescence “turn-on” effect with a ultra-low NO detection limit of 14.5 nM. Furthermore, the probe was applied to sense and image NO in living cells and inflammatory model of zebrafish, as well as to detect NO in periodontitis patients’ saliva samples. We anticipate that probe ZPS-NO will serve as a practical and effective tool for assessing the interactions and evaluation of periodontitis development.