Periostin in the relation between periodontal disease and atherosclerotic coronary artery disease: A pilot randomized clinical study.

OBJECTIVE: The aim of this study was to analyze the effects of periodontal treatment on markers of atherosclerotic coronary artery disease and circulating levels of periostin. BACKGROUND: Periostin is necessary for periodontal stability, but it is highly present in atherosclerotic plaques. Treatment of periodontal disease, with low levels of local periostin, is thought to reduce systemic levels of periostin. Thus, this may contribute to cardiovascular health. METHODS: A pilot randomized controlled clinical trial was designed to include patients with severe periodontal disease and history of atherosclerotic coronary artery disease. Samples of gingival crevicular fluid (GCF) and serum were collected before and after periodontal treatment by periodontal surgery or non-surgical therapy. The levels of several markers of inflammation and cardiovascular damage were evaluated including CRP, IFN-γ, IL-1ß, IL-10, MIP-1α, periostin, and TNF-α in GCF and CRP, Fibrinogen, IFN-γ, IL-1ß, IL-6, IL-10, L-Selectin, MIP-1α, Periostin, TNF-α, and vWF in serum. RESULTS: A total of 22 patients with an average of 56 years old were recruited for participating in this study. Twenty of them were male. Most of them (82%) had suffered an acute myocardial event and underwent surgery for placing 1, 2, or 3 stents in the coronary arteries more than 6 months ago but less than 1 year. The treatment of periodontal disease resulted in an overall improvement of all periodontal parameters. Regarding the evaluation of GCF and serum, a significant increase of periostin in the GCF was observed after periodontal surgery. In contrast, although other markers in GCF and serum improved, no significant correlations were found. CONCLUSION: Treatment of periodontal disease through periodontal surgery induces a local and transient increase in the levels of periostin in the gingival crevicular fluid. The effects on systemic markers of inflammation and cardiovascular function have not been confirmed.

miR-221-3p as a potential biomarker of chronic periodontitis and its regulatory effect on inflammatory response.

PURPOSE: To explore the function of miR-221-3p in the development and course of chronic periodontitis (CP) and offer a fresh avenue for CP diagnosis and management. METHODS: miR-221-3p expression was detected by RT-qPCR. The clinical diagnostic value of miR-221-3p in CP patients was analyzed by receiver operating characteristic (ROC). ELISA was used to determine the IL-1ß and IL-6 in CP subjects and healthy controls. Pearson correlation analysis was performed with miR-221-3p. PDLCs were induced by LPS, transfected with miR-221-3p mimics, and their expression was analyzed for the effects of IL-1ß, and IL-6. RESULTS: The miR-221-3p expression was lower in the gingival sulcus fluid GCF of CP subjects compared to healthy controls. miR-221-3p showed high potential for clinical diagnosis in CP patients by ROC analysis, with high specificity and sensitivity. miR-221-3p was negatively correlated with Probing pocket depth (PD), Attachment loss (AL), Plaque index (PI), and Bleeding index (BI), and negatively correlated with inflammatory factors IL-1ß and IL-6. In LPS-induced PDLCs, IL-1ß and IL-6 were significantly increased, whereas miR-221-3p was significantly downregulated. Overexpression of miR-221-3p inhibited the production of inflammatory factors IL-1ß and IL-6 in LPS-induced PDLCs. CLINICAL SIGNIFICANCE: miR-221-3p expression may be a potential biological marker for the diagnosis of chronic periodontitis and provide a new direction for its treatment of chronic periodontitis.

Clinical Efficacy of Extracellular Vesicle Therapy in Periodontitis: Reduced Inflammation and Enhanced Regeneration.

Periodontitis, a prevalent inflammatory condition, affects the supporting structures of teeth, leading to significant oral health challenges. Traditional treatments have primarily focused on mechanical debridement, antimicrobial therapy, and surgery, which often fail to restore lost periodontal structures. Emerging as a novel approach in regenerative medicine, extracellular vesicle (EV) therapy, including exosomes, leverages nano-sized vesicles known for facilitating intercellular communication and modulating physiological and pathological processes. This study is a proof-of-concept type that evaluates the clinical efficacy of EV therapy as a non-surgical treatment for stage I-III periodontitis, focusing on its anti-inflammatory and regenerative potential. The research involved seven patients undergoing the therapy, and seven healthy individuals. Clinical parameters, including the plaque index, bleeding on probing, probing depth, and attachment level, were assessed alongside cytokine levels in the gingival crevicular fluid. The study found significant improvements in clinical parameters, and a marked reduction in pro-inflammatory cytokines post-treatment, matching the levels of healthy subjects, underscoring the therapy's ability to not only attenuate inflammation and enhance tissue regeneration, but also highlighting its potential in restoring periodontal health. This investigation illuminates the promising role of EV therapy in periodontal treatment, advocating for a shift towards therapies that halt disease progression and promote structural and functional restoration of periodontal tissues.

A Study of the Association between Primary Oral Pathologies (Dental Caries and Periodontal Diseases) Using Synchrotron Molecular FTIR Spectroscopy in View of the Patient's Personalized Clinical Picture (Demographics and Anamnesis).

In this exploratory study, we searched for associations between the two most common diseases of the oral cavity-dental caries and periodontal diseases-taking into account additional factors, such as personalized clinical pictures (the individual risk factors of the patient), based on the method of a multivariate data analysis of the molecular changes in the composition of human gingival crevicular fluid (GCF). For this purpose, a set of synchrotron Fourier-transform infrared spectroscopy (FTIR) spectra of gingival crevicular fluid samples from patients with different demographics, levels of dental caries development and periodontal diseases, and the presence/absence of concomitant chronic diseases were obtained and analyzed. Using a set of techniques (v-, F-, Chi-square tests; a principal component analysis (PCA); and the hierarchical clustering of principal components (HCPCs)) implemented in the R package FactoMineR allowed us to assess the relationship between the principal components (PCs) and characteristics of the respondents. By identifying the features (vibrational modes in the FTIR spectra) that contribute most to the differentiation of the spectral dataset, and by taking into account the interrelationships between the patients' characteristics, we were able to match specific biological markers (specific molecular groups) to the two factors of interest-two types of oral pathologies. The results obtained show that the observed changes in the quantitative and qualitative composition of the modes in the infrared (IR) spectra of the GCF samples from patients with different dental caries developments and periodontal diseases present confirm the difficulty of identifying patient-specific spectral information. At the same time, different periodontal pathologies are more closely associated with other characteristics of the patients than the level of their caries development. The multivariate analysis performed on the spectral dataset indicates the need to take into account not only the co-occurrence of oral diseases, but also some other factors. The lack of this consideration (typical in lots of studies in this area) may lead to misinterpretations and consequently to a loss of data when searching for biological markers of certain oral diseases.

Impact of Inflammatory Markers and Senescence-Associated Secretory Phenotype in the Gingival Crevicular Fluid on the Outcomes of Periodontal Regeneration.

The aim of this study was to test the molecular expression profile (senescence-associated secretory phenotype; SASP) in gingival crevicular fluid (GCF) prior to surgery in relation to the distribution of clinical success of periodontal regeneration. Forty consecutive patients presenting sites with residual probing pocket depth (PPD) ≥ 6 mm and intrabony defects ≥ 3 mm were treated through a minimally invasive surgical technique. Pre-operatively, GCF was sampled for inflammatory biomarker analysis related to SASP [interleukin (IL)-1ß, IL-6, and IL-12; matrix-metalloproteinases (MMP)-8 and -9]. Better or worse responders were classified depending on the achievement of a composite outcome measure at 1-year [COM; PPD ≤ 4 mm and clinical attachment gain (CAL) gain ≥ 3 mm]. Correlation analyses and logistic regression models were performed. Periodontal regeneration led to significant improvements in mean clinical and radiographic parameters. Teeth achieving COM presented significantly lower amounts of SASP factors compared with non-successful teeth. Higher CAL gain, PPD reduction, and radiographic bone fill were negatively correlated with IL-1ß and MMP-8 and -9 (p < 0.001), while IL-12 showed a direct relationship with CAL gain (p = 0.005) and PPD reduction (p = 0.038). Sites expressing higher SASP expression in the GCF before periodontal regeneration achieved worse clinical and radiographic outcomes.

Influence of locally delivered doxycycline on the clinical and molecular inflammatory status of intrabony defects prior to periodontal regeneration: A double-blind randomized controlled trial.

OBJECTIVES: To test the effect of locally delivered doxycycline (DOX) administered 2 weeks prior to minimally invasive periodontal regeneration in terms of presurgical inflammatory status and cytokine expression profile in the gingival crevicular fluid (GCF). Secondary aim was to assess the early wound healing index (EHI) at 2 weeks after surgery. BACKGROUND: It is hypothesized that healing after periodontal regeneration is dependent on preoperative soft tissue condition, and that local antibiotics may improve the site-specific inflammatory status at short time. METHODS: Sites associated with periodontal intrabony defects requiring regenerative surgery and showing bleeding on probing (BoP) were included. At T0, experimental sites were randomly treated with subgingival instrumentation with or without topic DOX application. After 2 weeks (T1), defects were approached by means of minimally invasive surgical technique. GCF was sampled at both T0 and T1 for inflammatory biomarker analysis. Two weeks after surgery, the EHI was evaluated (T2). RESULTS: Forty-four patients were included. At T1, the number of BoP+ sites was statistically significantly less in the test group (27.3% vs. 72.7%; p < .01). The total amount of interleukin (IL)-1ß (p < .001), matrix-metalloproteinases (MMP)-8 (p < .001), and MMP-9 (p = .010) in the GCF significantly decreased in the test group at T1, with relevant differences compared to controls. At T2, the EHI had an average value of 1.45 ± 0.86 in the test group while in the control, it was 2.31 ± 1.43 (p = .027). A statistically significantly positive correlation was observed between the amount of IL-1ß and MMP-9 and EHI scores. CONCLUSIONS: Within the limitations of this study, sites treated with DOX showed improved clinical and molecular inflammatory parameters before surgery, as well as soft tissue healing 2 weeks after surgery.

Effects of Kangfuxinye on NF-κB and Inflammatory Cytokines in Gingival Crevicular Fluid of Patients with Orthodontic Gingivitis Caused by Orthodontic Treatment.

Explore the Kangfuxinye effection on the expressions of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and inflammatory cytokines (IC) in the gingival crevicular fluid of patients with orthodontic gingivitis caused by orthodontic treatment. 98 patients with orthodontic gingivitis in Qingdao Stomatological Hospital caused by orthodontic treatment were divided into two groups, namely, the control treatment group and the Kangfuxinye treatment group. In this study, the expressions of those proteins and IC in gingival crevicular fluid before and after treatment were analyzed at first, and the correlations of the NF-κB p65 expression with IC were explored. Then the differences in the expressions of those proteins and IC and the efficacy between the control treatment group and the Kangfuxinye treatment group were analyzed. Compared with those before treatment, the expressions of NF-κB-related proteins and IC interleukin-1ß (IL-1ß), tumor necrosis factor-alpha (TNF-α) and vascular endothelial growth factor (VEGF)] were significantly decreased after treatment (p<0.05). After treatment, the expression of NF-κB p65 was positively correlated with IL-1ß, TNF-α and VEGF, but negatively related to IL-4 and IL-10. In addition, compared with the control treatment, Kangfuxinye significantly reduced the expressions of those proteins and their messenger ribonucleic acids (mRNAs) (p<0.05), decreased the expressions of IL-1ß, TNF-α and VEGF (p<0.05) but improved the total effective rate of treatment. Kangfuxinye can reduce the NF-κB expressions and IC in the gingival crevicular fluid of patients with orthodontic gingivitis caused by orthodontic treatment and enhance the efficacy.

Chemerin contributes to inflammatory responses and suppresses osteogenic differentiation in chronic periodontitis.

BACKGROUND: Chronic periodontitis (CP) is a common disease of oral cavity, and approximately 35% of adults suffered from CP. Therefore, its underlying mechanism needs to be explored for new therapeutic approaches. Chemerin, as a multifunctional adipokine, is found to be highly expressed in the gingival crevicular fluid (GCF), gingival tissues and the plasma of periodontitis patients. Thus, we aimed to uncover the underlying mechanism of chemerin in CP. METHODS: Thirty six CP patients and 25 healthy volunteers were enrolled. Periodontal ligament stem cells (PDLSCs) were isolated from CP patients and healthy ones, respectively. Then, normal PDLSCs or PDLSCs-differentiated osteoblasts were treated with different doses of recombinant human chemerin. RESULTS: Chemerin and inflammatory cytokines, including interleukin-1ß, interleukin-6, and tumor necrosis factor-α, were higher in the GCF and serum of CP patients than healthy ones. Moreover, chemerin was positively correlated with these three inflammatory cytokines, respectively, in CP patients. PDLSCs isolated from CP patients had higher expressions of chemerin and these cytokines than the ones isolated from normal individuals. Furthermore, chemerin dose-dependently increased inflammatory responses and inhibited osteogenic differentiation of PDLSCs. CONCLUSION: Chemerin accelerated inflammatory responses and suppressed osteogenic differentiation of PDLSCs, thus chemerin might sever as a therapeutic target of CP.

Biomarkers from Peri-Implant Crevicular Fluid (PICF) as Predictors of Peri-Implant Bone Loss: A Systematic Review.

The aim of the present systematic review is to summarize current knowledge regarding the analysis of biomarkers extracted from peri-implant crevicular fluid (PICF) as predictors of peri-implant bone loss (BL). An electronic search was conducted on three databases, PubMed/MEDLINE, Cochrane Library, and Google Scholar, to find clinical trials published until 1 December 2022 suitable to answer the following focused question: in patients with dental implants, are biomarkers harvested from PICF predictive of peri-implant BL? The initial search yielded a total of 158 entries. After a full-text review and application of the eligibility criteria, the final selection consisted of nine articles. The risk of bias in included studies was assessed using the Joanna Briggs Institute Critical Appraisal tools (JBI). According to the present systematic review, some inflammatory biomarkers harvested from PICF (collagenase-2, collagenase-3, ALP, EA, gelatinase b, NTx, procalcitonin, IL-1ß, and several miRNAs) seem to be correlated with peri-implant BL and may assist in the early diagnosis of pathological BL, that characterizes peri-implantitis. MiRNA expression demonstrated a predictive potential of peri-implant BL that could be useful for host-targeted preventive and therapeutic purposes. PICF sampling may represent a promising, noninvasive, and repeatable form of liquid biopsy in implant dentistry.

Effects of Candidalysin Derived from Candida albicans on the Expression of Pro-Inflammatory Mediators in Human Gingival Fibroblasts.

Candida albicans (Ca) is frequently detected in the peri-implant sulcus with peri-implantitis, a major postoperative complication after oral implant therapy. However, the involvement of Ca in the pathogenesis of peri-implantitis remains unclear. In this study, we aimed to clarify Ca prevalence in the peri-implant sulcus and investigated the effects of candidalysin (Clys), a toxin produced by Ca, on human gingival fibroblasts (HGFs). Peri-implant crevicular fluid (PICF) was cultured using CHROMagar and Ca colonization rate and colony numbers were calculated. The levels of interleukin (IL)-1ß and soluble IL-6 receptor (sIL-6R) in PICF were quantified by enzyme-linked immunosorbent assay (ELISA). Pro-inflammatory mediator production and intracellular signaling pathway (MAPK) activation in HGFs were measured by ELISA and Western blotting, respectively. The Ca colonization rate and the average number of colonies in the peri-implantitis group tended to be higher than those in the healthy group. IL-1ß and sIL-6R levels in the PICF were significantly higher in the peri-implantitis group than in the healthy group. Clys significantly induced IL-6 and pro-matrix metalloproteinase (MMP)-1 productions in HGFs, and co-stimulation with Clys and sIL-6R increased IL-6, pro-MMP-1, and IL-8 production levels in HGFs compared with Clys stimulation alone. These findings suggest that Clys from Ca plays a role in the pathogenesis of peri-implantitis by inducing pro-inflammatory mediators.

Periostin level in gingival crevicular fluid in periodontal disease: a systematic review and meta-analysis.

BACKGROUND: Periostin, a secreted adhesion molecule, is a matricellular protein secreted most in periodontal ligament and periosteum. Periostin is also needed for integrity and maturation of periodontal tissue. This meta-analysis was conducted to compare the gingival crevicular fluid (GCF) periostin levels in subjects having periodontal disease and healthy periodontium. METHODS: In this meta-analysis, three international database including PubMed, Scopus and Web of Science were searched and 207 studies retrieved. Also, the Google Scholar was searched to find more related studies (two studies were found). To assess the risk of bias of included studies, the Newcastle-Ottawa assessment scale adapted for case-control was used. Finally, required data was extracted and included into analysis. All statistical analysis were done using Stata software. RESULTS: Eight studies were included in this meta-analysis. Results showed that GCF periostin level is significant lower in chronic periodontitis group compare to healthy people (the standardized mean difference (SMD) = -3.15, 95% CI = -4.45, -1.85, p < 0.001). The syntheses of studies shown a significant decrease in the periostin level of chronic periodontitis patients compared to the gingivitis patients (SMD = -1.50, 95%CI = -2.52, -0.49, P = 0.003), while the mean level of periostin between the gingivitis patients and healthy group has no significant difference (SMD = -0.88, 95%CI = -2.14, 0.38, P = 0.173). CONCLUSION: The mean concentration of GCF periostin in people with chronic periodontitis significantly decreased compared to people with gingivitis and also compared to healthy people, while no significant difference was observed between the two groups with gingivitis and healthy people. Therefore, this marker may be used as a diagnostic criterion for the disease, which requires further studies.

Diagnostic potential of miR-200 family members in gingival crevicular fluid for chronic periodontitis: correlation with clinical parameters and therapeutic implications.

OBJECTIVE: The purpose of this study was to evaluate the potential of miR-200 family members in gingival crevicular fluid (GCF) as diagnostic biomarkers for chronic periodontitis (CP), aiming to provide valuable insights for the early detection and management of the disease. METHODS: GSE89081 dataset profiled miRNAs in GCF derived from 5 healthy and 5 periodontitis was analyzed by GEO2R. Quantitative real-time PCR was used to quantify the expression levels of miR-200 family members (miR-200a-3p, miR-200a-5p, miR-200b-3p, miR-200b-5p, miR-200c-3p, miR-200c-5p, miR-141-3p, miR-141-5p, and miR-429) in the GCF samples from 103 CP patients and 113 healthy controls. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic potential of miR-200 family members in differentiating CP patients from healthy controls. RESULTS: By analyzing the GSE89081 dataset, miR-200a-5p, miR-200b-5p and miR-200c-5p were significantly upregulated in GCF of the CP patients compared to the healthy control. In this study, miR-200a-3p, miR-200a-5p, miR-200b-3p, miR-200b-5p, miR-200c-3p, miR-200c-5p were significantly increased in GCF of CP patients compared to the healthy control, while miR-141 and miR-429 did not show significant differences. MiR-200a, -200b and 200c had good diagnostic value, and when these miRNAs were combined, they demonstrated excellent diagnostic value for CP with an AUC of 0.997, sensitivity of 99.03%, and specificity of 98.23%. MiR-200a, -200b and 200c in GCF showed significant and positive correlation with plaque index (PI), gingival index (GI), bleeding on probing (BOP), clinical attachment level (CAL), and probing pocket depth (PPD). CONCLUSION: MiR-200a, -200b and 200c in GCF may serve as potential biomarkers for the early diagnosis of CP, which was correlated with clinical parameters, being therapeutic targets for CP.

Levels of Inflammatory and Bone Metabolic Markers in the Gingival Crevicular Fluid of Individuals Undergoing Fixed Orthodontic Treatment in Comparison to Those Utilizing Invisalign.

Background and Objectives: Evaluation of the levels of cytokine and bone metabolic biomarkers (BMBs) in patients receiving fixed orthodontic therapy (FOT) and Invisalign. Materials and Methods: Sixty participants were enrolled after meeting the predefined inclusion criteria. Patients then underwent either FOT or Invisalign by allocating them randomly to each group (n = 30). The basic periodontal assessment was performed, including the plaque index (PI), gingival index (GI), and bleeding on probing (BoP), at baseline and again after 4 weeks. Gingival crevicular fluid (GCF) samples were taken from each individual at baseline and after 4 weeks. An enzyme-linked immunosorbent assay (ELISA) technique was used to determine the cytokine and BMB levels. An unpaired t-test compared the FOT and Invisalign group's means and SDs. Paired t-tests examined the difference between T0 baseline and T1. Results: Patients treated with either FOT or Invisalign presented no statistically significant difference in terms of periodontal parameters such as PI, GI, and BoP (p > 0.05). The levels of IL-6 were significantly higher in patients treated with FOT as compared to Invisalign at T1 (p < 0.05) The other tested cytokines, IL-10, 13, 17, and GM-CSF, were not significantly different in either the FOT or Invisalign group at baseline and 4 weeks follow-up (p > 0.05). Regarding BMBs, it was detected that NTx and OC levels in both of the investigated groups were not significantly different at baseline and after 4 weeks (p > 0.05). However, NTx levels rose significantly (p < 0.05) and OC levels fell from T0 to T1. Conclusions: FOT and Invisalign displayed comparable outcomes in terms of cytokine and BMB levels. However, only IL-6 and NTx were significantly different at week 4 from baseline.

Obesity Is Associated with a Weakened Gingival Inflammatory Cytokine Response.

Background and Objectives: An obesity-related elevated body mass index (BMI) across life is associated with chronic low-grade inflammation and increased levels of C-reactive protein (CRP) in blood. CRP is a marker and promoter of inflammation. The objectives of this study were to examine the effect of obesity on the relationship between peripheral and gingival CRP levels and to examine the effects of gingival CRP levels on gingival fluid inflammatory cytokines in periodontitis-resistant obese individuals. Materials and Methods: Thirty-nine participants in good periodontal health were recruited. Twenty patients were classified as lean and nineteen as obese based on their BMI levels. A thorough periodontal assessment was carried out. Gingival crevicular fluid (GCF) and blood samples were collected. Both GCF and blood samples were analyzed for interleukin-1ß (IL-1ß), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), interleukin-17A (IL-17A), and CRP. Results: GCF CRP levels were significantly higher in the obese than in the lean individuals. No statistically significant differences were noted between the two groups in either GCF or blood in terms of any of the inflammatory cytokine levels. IL-17A was not detected in the GCF of most subjects in both groups. GCF CRP levels were positively associated with blood CRP levels, and the association tended to be stronger in the obese individuals. GCF CRP showed no associations with GCF IL-10 in both groups. Although GCF CRP levels were positively associated with multiple GCF inflammatory cytokines (e.g., IL-1ß, IL-6, IL-8, and TNF-α) in all subjects, the associations tended to be weaker in the obese individuals (e.g., IL-1ß, IL-6, and TNF-α). Furthermore, the levels of the GCF inflammatory cytokines IL-6 and TNF-α were decreased in the obese individuals. Conclusions: Obesity unfavorably influences the relationship between blood and GCF CRP levels and promotes increased CRP levels in GCF. Collectively, the findings suggest a weakened inflammatory cytokine response in the gingival tissues of obese individuals.

Expression of Wnt signaling agonists and antagonists in periodontitis and healthy subjects, before and after non-surgical periodontal treatment: A systematic review.

Periodontitis is a preventable and treatable multifactorial chronic inflammatory disease that can lead to irreversible periodontal destruction and tooth loss. Wnt signaling and its regulators play an important role in periodontal inflammation, destruction, regeneration, and reconstruction. This systematic review aimed at investigating the involvement of Wnt signaling agonists and antagonists in periodontitis and healthy subjects, before and after periodontal treatment. Electronic searches were carried out using MEDLINE/PubMed, EMBASE, and Cochrane Library databases in addition to hand searches. Studies having different designs assessing the levels of Wnt signaling antagonist and agonist levels in gingival crevicular fluid, serum, and tissue in patients diagnosed with periodontitis or gingivitis, compared with healthy individuals were included. In addition, studies compared these levels in periodontitis patients before and after non-surgical periodontal therapy were also eligible. Sixteen studies met the eligibility criteria. Sclerostin (SOST) has been mainly investigated in the literature (8 publications). Sclerostin (5 studies), Wnt-5a (2 studies), secreted frizzled-related protein 1 (SFRP1) (3 studies), and ß-catenin (3 studies) show increased levels in periodontitis compared with periodontal health. Strong correlations between marker levels and periodontal clinical parameters were identified for SOST (5 studies), SFRP1 (2 studies), and ß-catenin (2 studies). SOST (3 studies) and SFRP1 (1 study) levels significantly decrease following non-surgical periodontal treatment. The present systematic review demonstrated an association between Wnt signaling agonist and antagonist levels and periodontitis. Wnt agonists and antagonists may serve as valuable diagnostic and prognostic markers for periodontitis onset and progression. Further case-control and longitudinal studies should be conducted for different Wnt signaling agonists and antagonists.

Effects of periodontal and bisphosphonate treatment on the gingival crevicular levels of sclerostin and dickkopf-1 in postmenopausal osteoporosis with and without periodontitis.

OBJECTIVE AND BACKGROUND: Both periodontitis and osteoporosis are associated with osteoclast-related bone resorption. Bone metabolism is regulated by wingless-type MMTV integration site family (WNT), and WNT/ß-catenin signals are controlled by physiological antagonists including dickkopf-1 (DKK-1) and sclerostin (SOST). This study examined the effects of periodontal and bisphosphonate (BP) treatment on the gingival crevicular fluid (GCF) sclerostin (SOST) and dickkopf-related protein-1 (DKK-1) levels in osteoporotic and systemically healthy postmenopausal women with and without periodontitis. MATERIALS AND METHODS: A total of 48 postmenopausal women were divided into 4 groups (n = 12) according to periodontal health and osteoporosis status, as follows: Group OP/P: subjects with both osteoporosis and periodontitis; Group P: systemically healthy subjects with periodontitis; Group OP: periodontally healthy subjects with osteoporosis; Group H: systemically and periodontally healthy controls. Clinical data and GCF SOST and DKK-1 levels of the participants were collected at baseline and at 6 and 12 months following the initiation of periodontal and/or BP treatment in the experimental groups. GCF SOST and DKK-1 data were obtained by ELISA. RESULTS: Clinical improvements were observed in all experimental groups. GCF SOST and DKK1 baseline levels varied significantly between groups due to periodontal disease (p < .001). Following treatment, significant increases in SOST and DKK-1 concentrations and significant decreases in total amounts of SOST were observed in both periodontitis groups (OP/P, P). However, while total amounts of DKK-1 decreased in Group OP/P, in Group P, these amounts had significantly increased at 12 months post-treatment (p < .05). At both 6 and 12 months post-treatment, SOST and DDK1 total amounts in Groups OP/P, OP, and H were similar (p > .05), whereas significant differences were observed between Groups H and P, indicating a deviation from periodontal health in Group P (p < .01). CONCLUSIONS: Significant changes in GCF SOST and DKK-1 levels were observed among women with osteoporosis who received both periodontal and BP treatment. A more detailed examination of how these treatment protocols can be combined may lead to new therapeutic approaches towards periodontal disease.

The Role of Matrix Metalloproteinases (MMP-8, MMP-9, MMP-13) in Periodontal and Peri-Implant Pathological Processes.

Severe periodontitis, a destructive inflammatory disease of the supporting tissues of the teeth, ranks sixth in terms of global spread, affecting about 11% of the population. Metalloproteinases (MMPs) are extracellular matrix (ECM) macromolecules that are important in cellular development and morphogenesis, and they are capable of activating growth factors in their proximity, cell surface receptors, and adhesion molecules. MMPs are part of a major family of zinc-dependent endopeptidases, and their activity is modulated and regulated by certain inhibitors known as tissue metalloproteinase inhibitors (TIMPs). Because type I collagen is the major component of the periodontal extracellular matrix, special attention has been paid to the role of collagenases, especially MMP-8 and MMP-13 and gelatinases, MMP-2 and MMP-9, in periodontal diseases. In fact, MMP-8 (or collagenase 2) is currently one of the most promising biomarkers for periodontitis in oral fluids. Among them, salivary MMP-9 has been shown to be a more sensitive marker for periodontal inflammation during orthodontic treatment, which opens new perspectives in reducing periodontal hazards during such treatments. Both MMP-8 and MMP-9 are extremely valuable diagnostic tools in treating periodontitis, and future studies and healthcare policies should focus on implementing more accessible methods of chairside testing in order to reduce the prevalence of this disease.

The expression and clinical significance of miR-30b-3p and miR-125b-1-3p in patients with periodontitis.

OBJECTIVE: Periodontitis is a chronic inflammatory infectious disease caused by the deposition of dental plaque on the tooth surface, leading to adverse systemic consequences. Accumulating evidence shows that dysregulated microRNAs (miRNAs) are associated with the disease severity of periodontitis. Herein, we report two novel miRNAs, miR-30b-3p and miR-125b-1-3p, in the context of periodontitis and their relationships with disease severity of periodontitis. METHODS: The miRNA profiles of gingival crevicular fluid (GCF) samples were used to screen differentially expressed miRNAs (DEmiRNAs) between periodontitis patients and periodontally healthy individuals. Clinical human GCF samples were collected from 80 patients diagnosed with periodontitis (PD +) for the first time and 100 periodontally healthy individuals (PD-). The severity of periodontitis was categorized into mild/moderate (MPD) and severe (SPD) groups. The expressions of miR-30b-3p and miR-125b-1-3p were determined by quantitative real-time PCR. The levels of IL-1ß, IL-6, and TNF-α were determined by ELISA methods. RESULTS: We applied GEO2R bioinformatics tool to analyze the raw data of the GSE89081 dataset and identified miR-30b-3p (|logFC|= 1.987) and miR-125b-1-3p (|logFC|= 1.878) between periodontitis patients and periodontally healthy individuals. It was found that PPD, CAL, BOP, and the relative expression levels of miR-30b-3p and miR-125b-1-3p were all higher in the PD + group than the PD- group, in the SPD group than the MPD group (P < 0.05). The periodontitis patients with high-miR-30b-3p expression exhibited increased PPD, CAL, and BOP compared to those low-miR-30b-3p expression, while high-miR-125b-1-3p expression group showed significant differences on PPD and BOP from low-miR-125b-1-3p expression group (P < 0.05). Pearson correlation analysis demonstrated a significantly positive correlation between the levels of inflammatory cytokines, miR-30b-3p expression, and miR-125b-1-3p expression (P < 0.001). Results of ROC curves showed AUC of 0.878 and 0.927, sensitivity of 0.843 and 0.855, and specificity of 0.791 and 0.801, respectively, when miR-30b-3p and miR-125b-1-3p expression levels were used to diagnose periodontitis. CONCLUSION: These data unveiled that miR-30b-3p and miR-125b-1-3p expressions may be associated with the pathogenesis of periodontitis.

Profiling of cytokines, chemokines and growth factors in saliva and gingival crevicular fluid.

In saliva and gingival crevicular fluid (GCF) soluble factors such as cytokines, chemokines and growth factors have shown a great potential serving as biomarkers for early detection and/or diagnosis of oral and systemic diseases. However, GCF and saliva, which one is a better source is still under debate. This study aimed to gain an overview of cytokines, chemokines and growth factors in saliva and GCF to pave the way for selecting suitable oral fluids for oral and systemic diseases. Multiplex cytokine assay was conducted to determine concentrations of cytokines, chemokines and growth factors in saliva and GCF samples from healthy subjects. The protocol for sample collection was carefully optimized. Stabilization, repeatability, and donor variation of the profiles were analyzed. We found that for different donors, cytokine and chemokine profiles showed unique patterns in saliva but similar patterns in GCF. In terms of growth factors, the profiles were individualized in saliva and GCF. All profiles stayed stable for the same healthy individual. In saliva, profiles of cytokines, chemokines and growth factors are individualized for different donors. In GCF, profiles of cytokines and chemokines are similar. Other factors, such as growth factors and T helper-related cytokines, are highly variable in donors. Profiles of soluble factors are not correlated in saliva and GCF. The comprehensive cytokine profiles in saliva and GCF reported in this work would serve as a good base for choosing promising cytokines for developing biomarkers in oral fluids.

Emerging roles of Interleukin-34 together with receptor activator of nuclear factor-kB ligand and osteoprotegerin levels in periodontal disease.

OBJECTIVE: The dependence between gingival crevicular fluid (GCF) of Interleukin-34 (IL-34) level and Receptor activator of nuclear factor -kB ligand/ osteoprotegerin (RANKL/OPG) ratio in the severity of periodontitis might reveal an unknown pathway of diseases with bone destruction. There is no study about the evaluation of IL-34 levels together with GCF RANKL and OPG levels in periodontitis patients before and after non-surgical periodontal treatment (NSPT). The objectives of this research were to investigate changes in the levels and relative ratios of IL-34, OPG, and RANKL in the GCF of patients with periodontitis before and after NSPT. MATERIALS AND METHODS: 20 healthy participants (CTRL), 20 patients with stage 3-grade B periodontitis and 20 with stage 3-grade C periodontitis were recruited. GCF and clinical periodontal recordings were investigated at the baseline and 6 weeks after NSPT. Enzyme-linked immunosorbent assay (ELISA) were used for quantifying of GCF IL-34, RANKL and OPG levels and their relative ratios were calculated. RESULTS: Greater values for GCF IL-34 and RANKL levels were found in the both of periodontitis groups than in CTRL group at baseline, whereas GCF OPG levels were statistically lower at baseline (P < 0.05). GCF IL-34 and RANKL levels decreased in the 6th week after NSPT in the both periodontitis groups, while the concentration OPG levels statistically increased (P < 0.05). Significantly positive correlations among the IL-34 with RANKL, sampled-site clinical attachment level (CAL), and gingival index (GI), whereas negative correlation with OPG were reported (P < 0.05). CONCLUSIONS: GCF IL-34 levels was high in patients with periodontitis and decreased after NSPT and its levels showed positive correlations with RANKL/OPG ratio levels CAL and GI. Determining of IL-34 levels together with RANKL/OPG ratio in GCF may therefore be valuable in detecting high risk individuals with periodontitis patients.