Search Results (103)

Periodontitis is a prevalent chronic inflammatory disease with complex etiopathogenesis involving microbial dysbiosis, host immune response, environmental factors, and genetic susceptibility. Among the cytokines implicated in periodontal immunoregulation, interleukin-35 (IL-35) has emerged as a novel anti-inflammatory mediator with potential diagnostic and therapeutic relevance. This narrative review evaluates the role of IL-35 in periodontal disease by exploring its local and systemic expression, response to non-surgical periodontal therapy (NSPT), and association with clinical disease severity. Additionally, current evidence regarding IL-35 gene polymorphisms and their potential contribution to individual susceptibility and disease progression, as well as their relevance in related systemic conditions, is assessed. A comprehensive review and synthesis of recent clinical and experimental studies were conducted, focusing on IL-35 levels in saliva, serum, and gingival crevicular fluid (GCF) among patients with healthy periodontium, gingivitis, and various stages of periodontitis, both before and after NSPT. Emphasis was placed on longitudinal studies evaluating IL-35 dynamics in correlation with periodontal parameters, as well as genetic association studies investigating IL-12A and EBI3 gene polymorphisms. IL-35 levels were generally found to be higher in healthy individuals and reduced in periodontitis patients, indicating a possible protective role in maintaining periodontal homeostasis. Following NSPT, IL-35 levels significantly increased, corresponding with clinical improvement and reduced inflammatory burden. Genetic studies revealed variable associations between IL-35 polymorphisms and susceptibility to periodontitis and related systemic conditions, although further research is needed for validation. IL-35 appears to function as a modulator of immune resolution in periodontal disease, with potential utility as a non-invasive biomarker for disease activity and therapeutic response. Its upregulation during periodontal healing supports its role in promoting tissue stabilization. The integration of cytokine profiling and genetic screening may enhance personalized risk assessment and targeted interventions in periodontal care. Full article

Background/Objectives: Gingipains produced by P. gingivalis have been shown to be directly related to periodontal tissue degradation and are significant molecular targets in therapy of periodontitis. Blocking the activity of these enzymes should reduce survival of this pathogen and mitigate the effects of inflammation in periodontitis. Therefore, gingipains inhibitors and specific antibodies could be recommended in the treatment of periodontitis. Cysteine peptidase inhibitors can be obtained by chemical synthesis, or isolated from natural raw materials. This research has the following aims: 1. to analyze in vitro the inhibition of cysteine protease activity in the gingival crevicular fluid (GCF) and 2. to compare the toxicity of natural raw inhibitors (obtained from Fallopia japonica plant and egg white) with chlorhexidine (CHX) using an MTS viability test. Methods: Samples of GCF were collected from healthy (N = 17) individuals and (N = 65) periodontal patients. Cysteine peptidase activity was inhibited by adding a solution of cystatin from egg white (with 20% glycerol), or cystatin from knotweed, or low molecular weight inhibitors (MW < 3 kDa) from egg white and knotweed against Nα-Benzoyl-DL-arginine 4-nitroanilide hydrochloride. Results: There was a statistically significant difference between the inhibition means of cysteine protease activity for the five groups (p < 0.001). Means for the four groups of patients with periodontitis were not statistically significant different from each other (p = 0.320). The inhibition rates were higher in periodontitis patients. The toxicity of knotweed cystatin inhibitor was several times lower than the toxicity of E-64d, and of CHX. Conclusion: Cysteine protease inhibitors isolated from egg or plants were non-toxic, effectively inhibited the activity of cysteine proteases in GCF, and may be a promising alternative to more toxic standard antimicrobials (CHX) in preventing periodontal tissue breakdown. Full article

Background: This study explores the link between oral biofluids, microbial dysbiosis, and Alzheimer’s disease (AD), highlighting saliva and gingival crevicular fluid (GCF) as non-invasive diagnostic sources. AD onset and progression appear to be influenced not only by genetic and environmental factors but also by changes in the oral microbiome and related inflammatory and metabolic alterations. As global populations age, the incidence of AD is projected to rise significantly. Emerging evidence implicates the oral microbiome and salivary metabolites in neurodegenerative pathways, suggesting that oral health may mirror or influence brain pathology. Materials and Methods: A systematic review of recent multi-omics studies was performed, focusing on salivary and GCF analysis in patients with AD, those with mild cognitive impairment (MCI), and cognitively healthy individuals. Databases searched included PubMed, Web of Science, and Scopus, following PRISMA guidelines. Results: Across the 11 included studies, significant alterations were reported in both the salivary microbiome and metabolome in AD patients. Notable microbial shifts involved increased abundance of Veillonella parvula and Porphyromonas gingivalis, while key metabolites such as L-tyrosine, galactinol, and mannitol were consistently dysregulated. These biomarkers correlated with cognitive performance and systemic inflammation. Conclusions: Oral biofluids represent promising, accessible sources of biomarkers for early AD detection. Multi-omics integration reveals the oral–brain axis as a potential target for diagnosis, monitoring, and therapeutic strategies. Full article

Background: This study evaluates non-surgical therapy combined with injectable platelet-rich fibrin (i-PRF) on the clinical parameters and the levels of matrix metalloproteinase-8 (MMP-8) in the gingival crevicular fluid (GCF) in patients with periodontitis. Methods: Forty subjects diagnosed with periodontitis were randomly divided into two groups. In the test group, scaling and root planing (SRP) was performed with the subsequent application of i-PRF into periodontal pockets, while in the control group SRP was performed alone. Clinical examination was performed before and 1, 3 and 6 months after treatment. For MMP-8 level determination, the ELISA method was used. Results: In both groups, a statistically significant reduction in full mouth probing depth (FMPD), full mouth clinical attachment level (FMCAL), full mouth bleeding on probing (FMBOP), full mouth plaque index (FMPI) and full mouth marginal bleeding index FMMBI (p < 0.001) was observed. In the test group, the reduction in FMPD and FMBOP was statistically significantly greater than in the control group (p = 0.049 and p < 0.001, respectively). A significantly greater reduction of probing depth (PD) and clinical attachment level (CAL) in pockets > 5 mm between baseline and examination after 3 and 6 months was noted in the test group. The level of MMP-8 was statistically significantly reduced in both groups (p = 0.007 and p = 0.009). Conclusions: SRP significantly improves the clinical parameters and reduces MMP-8 levels in patients with periodontitis. Addition of i-PRF may further enhance the positive effects of periodontal treatment on clinical parameters, without significant influence on MMP-8 levels. The results of the research require confirmation in a more homogeneous group, taking into account the elimination of the specified limitations. Full article

This study applies multivariate data analysis to deconvolute the spectral profiles of the Amide III region in the infrared spectra of gingival crevicular fluid (GCF). This reveals the impact of major oral diseases, such as dental caries and periodontal diseases, on the transformation of the secondary structure of GCF proteins. A two-stage analytical approach was employed: first, principal component analysis (PCA) was performed to establish the main factors of variation in the data, followed by pairwise comparisons of the samples based on the results of the Amide III profile deconvolution. The analysis also accounted for comorbidities, such as oncological and gastrointestinal diseases. This approach allowed for the identification of subtle differences in the composition and conformation of the secondary structure of GCF proteins while accounting for the superposition of multiple influencing factors. This methodology was effective in identifying biomarkers of oral diseases in GCF. For the first time, it has been demonstrated that the relative content of the β-sheet-associated component in the spectral profile of the secondary structure element of the protein fraction of GCF serves as a statistically significant marker for dental caries, regardless of the presence or absence of other diseases. Additionally, a significant decrease in the relative content of α-helix structures was observed in GCF from patients with oncological diseases. The changes in the spectral profile of the Amide III band of GCF identified in this study have not been previously detected using molecular spectroscopy, correlated with the secondary structure of proteins, or analyzed using multivariate analysis methods. Full article

Background/Objectives: Periodontitis, a prevalent chronic inflammatory disease affecting tooth-supporting structures, has been increasingly linked to systemic conditions such as diabetes mellitus (DM) and cardiovascular diseases (CVDs). This study aimed to evaluate the periodontal status and levels of dipeptidyl peptidase-4 (DPP-4) and galectin-3 (Gal-3) in gingival crevicular fluid (GCF) of patients with periodontitis, heart failure (HF), and diabetes, exploring their potential as biomarkers for disease association. Methods: A cross-sectional study was conducted on 88 patients categorized into four groups: periodontally and systemically healthy (control, C); periodontitis (P); periodontitis and HF (P+HF); and periodontitis, HF, and diabetes (P+HF+D). Periodontal status was assessed using probing pocket depth (PPD) and Gingival Index (GI). GCF samples were collected and analyzed for DPP-4 and Gal-3 levels using ELISA. Statistical analyses were performed to assess differences between groups and potential correlations. Results: Results indicated significantly higher levels of DPP-4 in all test groups compared to controls (p < 0.0001), with the highest levels in the P+HF+D group. Similarly, Gal-3 levels were elevated in periodontitis patients, particularly in those with HF (p < 0.0001), and there was no significant difference between P+HF and P+HF+D groups. No significant differences were observed between smokers and non-smokers regarding these markers. Positive correlations were found between the periodontal parameters and the immunological markers in all test groups. Conclusions: The findings suggest that DPP-4 and Gal-3 may serve as potential biomarkers for periodontitis in association with heart failure and diabetes, with DPP-4 being more upregulated in the association with diabetes and Gal-3 with heart failure. Full article

Background: This study aimed to investigate endocan (ESM-1) levels in periodontitis patients before and after non-surgical periodontal treatment by analyzing the relationship between vascular endothelial growth factor A (VEGF-A) and tumor necrosis factor-alpha (TNF-α) in gingival crevicular fluid (GCF). Methods: This study included 26 periodontally healthy people as controls (Group 1) and 27 patients with Stage III-Grade B periodontitis (Group 2). Demographic and periodontal variables were assessed. GCF samples were collected from every subject both before and 6 weeks following non-surgical periodontal therapy (NSPT). Using an enzyme-linked immunosorbent test, biomarker levels were determined. Results: The periodontitis patients showed higher ESM-1 levels than the controls, although the difference was not significant (p > 0.005). The ESM-1 levels decreased significantly after treatment (p = 0.001). The VEGF-A levels did not differ significantly between the periodontitis patients and controls (p > 0.005) and decreased non-significantly following treatment (p > 0.005). The TNF-α levels were significantly higher in the periodontitis patients than the controls (p = 0.000) and decreased significantly after treatment (p = 0.000). A significant correlation was found between TNF-α and both probing depth (PD) and interproximal clinical attachment level (iCAL) in the control group (p < 0.05). In the periodontitis group, the VEGF levels were significantly correlated with the gingival index (GI) (p < 0.05). Significant correlations were identified between ESM-1 and VEGF-A and ESM-1 and TNF-α, as well as VEGF-A and TNF-α, in both the control group and following treatment (p < 0.05). Conclusions: ESM-1 and TNF-α levels decreased with non-surgical periodontal treatment in GCF. Within the limits of the study, the findings suggest that ESM-1 levels in periodontal tissues may be an indicator of periodontal disease. Full article

Background and Objectives: In recent years, numerous studies have investigated and analyzed the levels of molecular biomarkers of temporomandibular disorders (TMD) from various tissue samples and body fluids. However, no study has investigated gingival crevicular fluid (GCF) in TMD patients. The purpose of this study was to determine the concentrations of pro-inflammatory cytokines in GCF before and after stabilization splint (SS) therapy in patients with painful TMD, to investigate whether SS administration causes changes in the concentrations of pro-inflammatory cytokines. An additional aim was to investigate the relationship of GCF cytokine levels with chronic pain intensity and clinical parameters. Materials and Methods: This prospective cohort study included 36 patients who were diagnosed with painful TMD using the Diagnostic Criteria for TMD (DC/TMD). GCF samples were collected at baseline before SS treatment (T0) and at one month (T1) and three months (T2) after the start of therapy. Customized ProcartaPlex Multiplex assays from eBioscience (Invitrogen™, Thermo Fisher Scientific, Viena, Austria) were used for the quantitative analysis of pro-inflammatory cytokines (IL-1β, IL-6, IL-7, IL-8, IL-13, and TNF-α). Patients filled out Croatian versions of questionnaires for self-assessment from Axis II DK/TMP: Graded Chronic Pain Scale (v2) (GCPSv2) and Jaw Function Limitation Scale-20 (JFLS-20). Results: The results showed that the GCF levels of IL-7 (Friedman’s test, p = 0.008) and IL-13 (Friedman’s test, p = 0.003) were significantly decreased at T2. The GCF level of IL-13 was in negative correlation with chronic pain grade score at T2 (Rho = −0.333), while the GCF level of IL-8 was in positive correlation with mobility limitation (Rho = 0.382) at T1. Conclusions: The results indicate that SS therapy might have a role in reducing inflammation and that the GCF could be a valuable medium for assessing molecular biomarkers. Full article

The sensitive detection of inflammatory biomarkers in gingival crevicular fluid (GCF) is highly desirable for the evaluation of periodontal disease. Luminol-based electrochemiluminescence (ECL) immunosensors offer a promising approach for the fast and convenient detection of biomarkers. However, luminol’s low ECL efficiency under neutral conditions remains a challenge. This study developed an immunosensor by engineering an immunorecognition interface on the outer surface of mesoporous silica nanochannel film (SNF) and confining a Co₃O₄ nanocatalyst within the SNF nanochannels to improve the luminol ECL efficiency. The SNF was grown on an indium tin oxide (ITO) electrode using the simple Stöber solution growth method. A Co₃O₄ nanocatalyst was successfully confined within the SNF nanochannels through in situ electrodeposition, confirmed by X-ray photoelectron spectroscopy (XPS) and electrochemical measurements. The confined Co₃O₄ demonstrated excellent electrocatalytic activity, effectively enhancing luminol and H₂O₂ oxidation and boosting the ECL signal under neutral conditions. Using interleukin-6 (IL-6) as a proof-of-concept demonstration, the epoxy functionalization of the SNF outer surface enabled the covalent immobilization of capture antibodies, forming a specific immunorecognition interface. IL-6 binding induced immunocomplex formation, which reduced the ECL signal and allowed for quantitative detection. The immunosensor showed a linear detection range for IL-6 from 1 fg mL⁻¹ to 10 ng mL⁻¹, with a limit of detection (LOD) of 0.64 fg mL⁻¹. It also demonstrated good selectivity and anti-interference capabilities, enabling the successful detection of IL-6 in artificial GCF samples. Full article

Background and Objectives: This study aimed to evaluate the role of A-PRF (advanced platelet-rich fibrin) in the enhancement of wound healing and protecting the periodontal health of mandibular second molars after the extraction of mandibular third molars. Additionally, the study assessed the levels of pro-inflammatory cytokines in the gingival crevicular fluid (GCF) of mandibular second molars as markers of inflammation. Materials and Methods: Twenty-five systemically healthy adult patients with bilateral removal of impacted mandibular third molars were included. Each patient received A-PRF in one extraction site, while the contralateral site served as a control. Periodontal parameters of the adjacent second molar, including probing depth (PD) and clinical attachment level (CAL), were measured in distal–vestibular (DV) and distal–lingual (DL) sites. Pain, swelling, and overall healing were subjectively evaluated. Levels of tumor necrosis factor-alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6) in the GCF were analyzed. Evaluations occurred at baseline and three months post-surgery. Results: A-PRF significantly improved PD (from 4.69 ± 0.61 mm to 3.85 ± 0.34 mm in DV, and from 4.71 ± 0.65 mm to 3.79 ± 0.27 mm in DL, respectively) and CAL (from 2.41 ± 0.25 mm to 1.82 ± 0.21 mm in DV, and from 2.40 ± 0.36 mm to 1.75 ± 0.19 mm in DL, respectively) of the adjacent second molar, compared to control sites, three months post-surgery. Pain and swelling scores were notably lower on the 7th postoperative day in the A-PRF group. A-PRF also reduced pro-inflammatory cytokines in GCF, significantly more than in control sites, at three months post-surgery. Conclusions: A-PRF enhances the periodontal and inflammatory status of adjacent teeth and wound healing after the extraction of mandibular third molars. Full article

Background: Changes in the positions of teeth occur during orthodontic treatment due to the application of forces that cause restructuring of the periodontal tissue. In the last decade, substantial research has been conducted to detect different biomarkers in the gingival crevicular fluid (GCF) to obtain a better assessment of the periodontal status. Aim: The purpose of this review is to describe how the levels of certain biomarkers from the gingival fluid change during tissue remodeling throughout orthodontic treatment. Materials and methods: To carry out the purpose of this research, electronic databases were searched using specific keywords, leading to 387 articles, out of which 19 were used in writing this narrative review. A sampling period of the last 10 years was used in selecting the articles. Results: The results highlight that the origin of the gingival crevicular fluid is at the gingival blood vessels’ plexus. GCF has a complex composition with differences depending on the periodontal status and the tissue restructuring which takes place in the periodontium. The levels of inflammatory mediators, enzymes, and metabolic products of tissue remodeling in GCF change during orthodontic treatment. Being aware of their specific role, they can provide valuable information about bone remodeling during orthodontic tooth movement. Conclusions: By determining the biomarkers in GCF, as an investigative method, clinicians could easily monitor the orthodontic tooth movement, and, subsequently, the treatment period could be shortened and the adverse effects associated with it could be avoided. Full article

Evidence has shown the clear positive effects of nature-based products on biofilm control and improved gingival health. However, most studies have used in vitro models, have tested single natural components, or have not evaluated proteomic changes after treatment. This double-blind, parallel, randomized, and controlled clinical trial evaluated the benefits of a nature-based gel in controlling gingival inflammation and its effects on the proteomic gingival crevicular fluid (GCF) profile. Gingivitis patients were distributed into the following groups: (1) nature-based gel containing propolis, aloe vera, green tea, cranberry, and calendula (n = 10); (2) control—conventional toothpaste (n = 10). GCF was collected and evaluated by means of liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS). At 3 months, the groups showed similar clinical benefits (p < 0.05). A total of 480 proteins were identified across all groups. In a pooled comparison of both groups at both time points, exclusive proteins were identified in the nature-based gel (78) and the control (21) groups. The exclusive proteins identified for the toothpaste mainly acted in wound healing, and those for the nature-based gel mainly acted on immune system processes. The nature-based gel achieved similar clinical outcomes to conventional toothpaste. However, the nature-based gel markedly changed the proteomic profile of GCF after treatment, showing a profile associated with a host response. Full article

The objective of this study is to analyze the miRNA expression of oral fluids such as gingival crevicular fluid (GCF) in patients with periodontitis and Type 2 diabetes mellitus, and how these epigenetic biomarkers can influence the bidirectional relationship of these two inflammatory diseases. This review was conducted following the PRISMA criteria. PubMed, Scopus, Cochrane Library, Embase, and Web of Science databases were searched for clinical studies conducted on humans investigating, through GCF miRNA expression, the relationship between periodontal diseases and type 2 diabetes mellitus. In addition, the etiopathogenic pathways of the studied miRNAs were analyzed using the DIANA MIR path tool. A total of 1436 references were identified in the initial literature search, and seven articles were finally included in this review. Most of the articles included in this review were case–control studies and examined the expression of miRNAs in patients with periodontitis with or without diabetes. Due to their characteristics, miRNAs appear to be the ideal biomarkers for improving the understanding and knowledge of the etiopathogenic pathways that link both diseases. Among all the studied miRNAs, miR-146a, miR-155, miR-200b, miR-223, and miR-203 showed strong involvement in inflammatory and metabolic pathways, making them potential good diagnostic and prognostic biomarkers. Full article

Background and Objectives: This study aimed to investigate the effect of zinc phosphate (ZnP) cement, glass ionomer cement (GIC), and nano-integrated bio-ceramic (NIB) cement on mineralization when placed in contact with bone tissue-forming cells. Materials and Methods: ZnP cement, GIC, and NIB cement were divided into direct and indirect groups. A total of 72 cement pellets (24 pellets of each test sample) of 3 × 1 mm (width × height) were prepared using polytetrafluoroethylene molds. A total of 3 sample groups were demarcated using 96- cell well culture plates. In the control group, 24 wells were filled with mineralized osteoblasts and 1 µL of gingival crevicular fluid (GCF). In test group 1, to show a direct effect, 36 samples were plated with mineralized osteoblasts and 1 µL GCF for 24 h; the cells were directly exposed to cement pellets. A total of 36 samples were immersed in GCF for 24 h; later the supernatant was transferred to the mineralized osteoblasts to demonstrate an indirect effect in test group 2. To assess the mineralization, osteoblasts were stained with alizarin red and later observed under an inverted phase-contrast microscope. Data were analyzed using the statistical package for social sciences. An independent t-test compared the direct and indirect effects of the ZnP cement, GIC, NIB cement, and control groups on the mineralization of osteoblasts derived from hDPCs. Results: A statistically significant difference was observed between the ZnP cement, GIC, and NIB cement groups (p < 0.05). ZnP cement exhibited a moderate, NIB cement the least harmful effect, and GIC showed the most harmful effect on the mineralization of osteoblast cells. Conclusions: The biocompatibility of dental luting cements is an important aspect that clinicians should consider during their selection. Nano-integrated bio-ceramic cement showed the least negative effect on the mineralization of osteoblast cells which is beneficial for the cementation of cement-retained implant prostheses. However, further studies are needed to evaluate osteoblast and osteoclast activity in vivo. Full article

Gingival crevicular fluid (GCF) and oral fluid have emerged as promising diagnostic tools for detecting biomarkers. This review aimed to evaluate the existing literature on using oral fluids as a source of biomarkers for bone turnover diseases affecting the jawbone. A comprehensive search strategy was executed between August 2014 and August 2024 across five major databases (Web of Science, EBSCOhost Dentistry & Oral Sciences Source, Cochrane Library, Scopus, and PubMed) and grey literature sources. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) was applied. The screening was facilitated using Rayyan at rayyan.ai and Endnote X20 software tools, culminating in the evaluation of 14,965 citations from databases and 34 from grey literature. Following rigorous scrutiny, 37 articles were selected for inclusion in this review, encompassing diseases such as periodontitis, medication-related osteonecrosis of the jaw (MRONJ), and osteoporosis. The quality of the included observational studies was assessed using the Revised Risk of Bias Assessment Tool for Non-Randomized Studies (RoBANS 2). Interleukin-1 beta (IL-1β), sclerostin, osteoprotegerin (OPG), and interleukin-34 (IL-34) emerged as significant biomarkers in GCF, and they were mainly from periodontitis and osteoporosis. Osteocalcin (OC), IL-1β, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), OPG, and matrix metalloproteinase-9 (MMP-9) were significant in oral fluid or saliva, and they were from periodontitis, MRONJ, and osteoporosis. These findings underscore the potential use of oral fluids, which are regarded as non-invasive tools for biomarker identification in bone turnover. Many biomarkers overlap, and it is important to identify other specific biomarkers to enable accurate diagnosis of these conditions. Full article