A simplified protocol for deep quantitative proteomic analysis of gingival crevicular fluid for skeletal maturity indicators.

Assessment of craniofacial skeletal maturity is of great importance in orthodontic diagnosis and treatment planning. Traditional radiographic methods suffer from clinician subjectivity and low reproducibility. Recent biochemical methods, such as the use of gingival crevicular fluid (GCF) protein biomarkers involved in bone metabolism, have provided new opportunities to assess skeletal maturity. However, mass spectrometry (MS)-based GCF proteomic analysis still faces significant challenges, including the interference of high abundance proteins, laborious sample prefractionation and relatively limited coverage of GCF proteome. To improve GCF sample processing and further discover novel biomarkers, we herein developed a single-pot, solid-phase-enhanced sample-preparation (SP3)-based high-field asymmetric waveform ion mobility spectrometry (FAIMS)-MS protocol for deep quantitative analysis of the GCF proteome for skeletal maturity indicators. SP3 combined with FAIMS could minimize sample loss and eliminate tedious and time-consuming offline fractionation, thereby simplifying GCF sample preparation and improving analytical coverage and reproducibility of the GCF proteome. A total of 5407 proteins were identified in GCF samples from prepubertal and circumpubertal groups, representing the largest dataset of human GCF proteome to date. Compared to the prepubertal group, 61 proteins were differentially expressed (31 up-regulated, 30 down-regulated) in the circumpubertal group. The six-protein marker panel, including ATP5D, CLTA, CLTB, DNM2, HSPA8 and NCK1, showed great potential to predict the circumpubertal stage (ROC-AUC 0.937), which provided new insights into skeletal maturity assessment.

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.

The influence of the tooth preparation finish line position on the expression of matrix metalloproteinase-9 and the presence of periodontopathogens in the gingival crevicular fluid.

OBJECTIVE: The objective of the study was to determine the concentration of matrix metalloproteinase 9 (MMP-9) and changes in the presence of periodontopathogens in the gingival crevicular fluid before and after tooth preparation with the subgingival and equigingival finish line position. PATIENTS AND METHODS: The clinical prospective study included 20 subjects with an indication for upper canine preparation, with the subgingival (group 1) and equigingival finish line (group 2). Samples were taken in four observation intervals: 5 minutes before (control samples), as well as 15 minutes, 24 and 72 hours after tooth preparation (experimental samples). Measurement of MMP-9 was done using Enzyme-linked Immunosorbent Assay (ELISA). The presence of bacteria in the gingival fluid was proven by the Polymerase chain reaction (PCR) analysis. RESULTS: The MMP-9 values did not differ statistically significantly between the groups (p=0.524). The MMP-9 values showed a statistically significant difference in the given observation period (p<0.001) with a significant linear increase in values (p<0.001). A significant quadratic trend recorded a decrease in the MMP-9 values 15 minutes after preparation, and an increase 24 hours after preparation, without a significant difference in the interaction between groups (p=0.392). After preparation, a significant difference in the presence of periodontopathogens was confirmed, i.e., a decrease in the presence of Prevotella intermedia (p=0.025) and Tannerella forsythia (p=0.016) in group 1, and an increase in the presence of Aggregatibacter actinomycetemcomitans in both groups (p=0.029, p=0.026). CONCLUSIONS: The study is a good basis for determining the influence of tooth preparation on gingival inflammation, with therapeutic (choice of preparation technique) and preventive significance regarding the protection of the periodontal tissue from possible iatrogenic damage.

Influence of Nutrition and Physical Activity on Local and Systemic Inflammatory Signs in Experimentally Induced Gingivitis.

Although numerous studies have been published investigating the relationship between various dietary components and inflammatory periodontal disease, it has not yet been possible to clearly distinguish between periodontally healthy and unhealthy diets. This clinical study aimed to assess the association of specific food ingredients and physical activity on local and systemic inflammatory signs in experimentally induced gingivitis. Thirty-nine non-smoking periodontally healthy volunteers (mean age 23.2 ± 3.8 years) refrained from oral hygiene in the right maxilla for 21 days to induce an experimental gingivitis. Clinical examination (baseline and day 21) included plaque index, bleeding on probing (BOP), gingival crevicular fluid volume and high sensitive C-reactive protein levels (blood sample). Accompanying the intervention, volunteers documented with validated questionnaires their physical activity converted into metabolic equivalent (MET) and their nutrition converted into the dietary inflammatory index (DII). Significantly lower BOP (p = 0.039) was found for subjects with a more anti-inflammatory DII than for those with a more pro-inflammatory DII; higher MET values were correlated with lower BOP at day 21 (correlation coefficient -0.36). The results show an influence of nutrition and physical activity on periodontal inflammation signs. The DII may be a suitable parameter to verify the relationship between nutrition and inflammatory periodontal diseases.

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.

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.

aMMP-8 point-of-care - diagnostic methods and treatment modalities in periodontitis and peri-implantitis.

INTRODUCTION: When collected in a standardized fashion, oral fluid analysis can refine the diagnosis of periodontal and peri-implant disease. In practice, dental professionals can perform active matrix metalloproteinase (aMMP-8) analysis chairside. AREAS COVERED: Periodontal tissues are mainly made up of type I collagen, and collagen breakdown is one of the main events in periodontal and peri-implantitis destructive lesions. In addition to traditional measurements, their diagnosis can be refined with tests utilizing oral fluids. The active matrix metalloproteinase-8 (aMMP-8) is possible to be determined from the gingival crevicular fluid (GCF), peri-implant sulcus fluid (PISF), and other oral fluids such as mouth rinse and saliva. We also investigated the applicability of aMMP-8 chair-side test kits in the evaluation of oral health benefits of different adjunctive host-modulating periodontal therapies including fermented lingonberry mouthwash (FLJ) and antibacterial photodynamic therapy (aPDT). EXPERT OPINION: The aMMP-8 levels can more reliably detect early activation of periodontal and peri-implant disease as compared to traditional diagnostic methods that assess the experienced health status or past disease, rather than the present or future pathology. Novel therapies like, fermented lingonberry juice as a mouthrinse or aPDT, are potential host-modulating adjunctive treatments to reduce the signs of oral inflammation and infection.

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.

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.

The role of extracellular vesicles in periodontitis: pathogenesis, diagnosis, and therapy.

Periodontitis is a prevalent disease and one of the leading causes of tooth loss. Biofilms are initiating factor of periodontitis, which can destroy periodontal tissue by producing virulence factors. The overactivated host immune response is the primary cause of periodontitis. The clinical examination of periodontal tissues and the patient's medical history are the mainstays of periodontitis diagnosis. However, there is a lack of molecular biomarkers that can be used to identify and predict periodontitis activity precisely. Non-surgical and surgical treatments are currently available for periodontitis, although both have drawbacks. In clinical practice, achieving the ideal therapeutic effect remains a challenge. Studies have revealed that bacteria produce extracellular vesicles (EVs) to export virulence proteins to host cells. Meanwhile, periodontal tissue cells and immune cells produce EVs that have pro- or anti-inflammatory effects. Accordingly, EVs play a critical role in the pathogenesis of periodontitis. Recent studies have also presented that the content and composition of EVs in saliva and gingival crevicular fluid (GCF) can serve as possible periodontitis diagnostic indicators. In addition, studies have indicated that stem cell EVs may encourage periodontal regeneration. In this article, we mainly review the role of EVs in the pathogenesis of periodontitis and discuss their diagnostic and therapeutic potential.

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.

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.

[Effect of ultrasonic curettage combined with sodium hyaluronate gel on chronic periodontitis and its effect on inflammatory factors hs-CRP, MCP-1 and MMP-13 in gingival crevicular fluid].

PURPOSE: To investigate the effect of ultrasonic curettage combined with sodium hyaluronate gel in the treatment of chronic periodontitis (CP) and the effect on inflammatory factor hypersensitive C-reactive protein (hs-CRP), monocyte chemotactic protein 1 (MCP-1) and matrix metalloproteinase 13 (MMP-13) in gingival crevicular fluid. METHODS: A total of 102 patients with CP from October 2021 to October 2022 were selected, divided into experimental group (n=51) and control group (n=51) by random number table method. The control group received ultrasonic subgingival curetage, and the experimental group received sodium hyaluronate gel adjuvant therapy on the basis of the control group. The periodontal rehabilitation indexes, clinical efficacy and the changes of gingival crevicular fluid hs-CRP, MCP-1 and MMP-13 were compared between the two groups. The periodontal pathogens, bone metabolism indexes and the occurrence of adverse events during treatment were compared between the two groups. The data were statistically analyzed using SPSS 22.0 software package. RESULTS: After treatment, the sulcus bleeding index (SBI), gingival index (GI), plaque index (PLI), periodontal pocket depth (PD) and attachment level (AL) of the two groups were significantly lower than before treatment (P＜0.05), and even significantly lower(P＜0.05) in the experimental group. Total effective rate of the experimental group was significantly higher than that of the control group(P＜0.05). hs-CRP, MCP-1 and MMP-13 in both groups after treatment were significantly lower than before treatment(P＜0.05), and hs-CRP, MCP-1 and MMP-13 in the experimental group after treatment were significantly lower than those in the control group(P＜0.05). The detection rates of Porphyromonas gingivalis, Forsetanella and Treponema dentalis were significantly lower in both groups after treatment than before treatment (P＜0.05), and the detection rates of the above indexes in the experimental group after treatment were significantly lower than those in the control group(P＜0.05). After treatment, the C-terminal peptide(CTX) of type Ⅰ collagen was significantly lower than that before treatment, and bone gla protein(BGP) was significantly higher than that before treatment(P＜0.05). The CTX and BGP of the experimental group were significantly lower than that of the control group and significantly higher than that of the control group(P＜0.05). There was no significant difference in the incidence of total adverse reactions between the two groups(P＞0.05). CONCLUSIONS：Ultrasonic curettage combined with sodium hyaluronate gel in the treatment of CP can promote periodontal tissue rehabilitation, enhance short-term efficacy, inhibit synthesis of inflammatory factors in gingival crevicular fluid, kill periodontal pathogens, regulate bone metabolism, and is safe and reliable.

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.

Impact of smoking on protein levels of beta-defensins in periodontal disease.

Antimicrobial peptides (AMPs) are important components of the host response against invading pathogens. In addition to their direct antimicrobial activity, they can also participate in the immune system modulation. However, the role of AMPs in the etiopathogenesis of periodontal disease and the risk factors that may influence their expression in the oral cavity are not fully understood. The aim of this study was to determine the impact of smoking on beta-defensin (hBD) 1 and 2 levels analyzing samples from periodontitis patients. Fifty patients with periodontitis, 25 smokers and 25 non-smokers, and 20 periodontally healthy patients were recruited. After periodontal clinical evaluation, gingival crevicular fluid (GCF) samples were collected from healthy sites of patients without periodontal disease and from healthy and diseased sites of patients with periodontitis. Peptides quantification was performed by sandwich ELISA technique. Smokers showed reduced GCF hBD 1 levels and increased hBD 2 levels compared to non-smokers in diseased sites (p <0.05). Higher levels of hBD 1 were observed in healthy sites of patients without periodontal disease than in healthy sites of patients with periodontitis (p<0.0001). Diseased sites of non-smokers presented higher levels of hBD 2 than healthy sites (p <0.05). These results reveal that protein levels of hBDs 1 and 2 can be impaired by cigarette smoking in the presence of periodontal disease.

Protective Effect of Iron Oxide Nanoparticles on Periodontal Injury in Rats by Inhibiting Collagenase-1 and Alkaline Phosphatase Expression.

This study intends to assess whether iron oxide nanoparticles affect periodontal injury and collagenase-1 (COL-1), and alkaline phosphatase (ALP) in rats. In this study, the ALP activity and Col-1 concentration in rats with periodontal injury were determined.We detected the periodontal histopathological changes and expression of periodontal pocket depth (PD) and attachment loss (AL) by Hematoxylin and eosin (HE) staining.We also detected Col-1 and ALP proteins in periodontal tissues by Western blot. Real-time reverse transcription-polymerase chain reaction (RT-PCR) detected Col-1 and ALP mRNA level in periodontal tissues of rats in each group, while ALP activity and Col-1 concentration in gingival crevicular fluid in model group increased compared to sham group (P < 0.05). After intervention by iron oxide nanoparticles, ALP activity and Col-1 concentration in the gingival crevicular fluid of model rats decreased greatly (P < 0.05). The gingival atrophy was more serious in model group, and many inflammatory cells infiltrated into the tissue and destroyed the alveolar tissue. Meanwhile, the periodontal tissue from rats in intervention group was greatly improved, and the degree of alveolar bone destruction was also significantly reduced, while the PD and AL periodontal indexes were significantly inhibited (P < 0.05). The protein and relative expression showed that the protein and mRNA expressions of ALP and Col-1 in periodontal tissue from model group were lower than those in sham group (P < 0.05). After intervention by iron oxide nanoparticles, the protein and mRNA expressions of ALP and Col-1 in the periodontal tissues in intervention group increased (P < 0.05). Iron oxide nanoparticles can thus inhibit the expression of ALP and COL-1 in periodontal injury rats, and improve the periodontal injury tissue.

Peri-implant crevicular fluid SIRT1 levels decrease in patients with peri-implant inflammatory: A prospective observational study.

BACKGROUND: A dental Implant is a prosthetic device made of alloplastic materials implanted into the bone to provide retention and support for a dental prosthesis. Sirtuin1 (SIRT1) molecule, a nicotinamide adenine dinucleotide (NAD)-dependent histone deacetylase, regulates a variety of physiological and pathological processes, including oxidative stress, metabolism, cell proliferation, cell differentiation, inflammatory, and apoptosis. We explored whether the expression of SIRT1 correlates in patients receiving implants with peri-implant mucositis (PIM) and peri-implantitis (PI) in comparison to patients with healthy peri-implant area (PIH). METHODS: A number of 198 patients with dentition defects were enrolled in the study after their implants were functional for at least 6 months. All 198 subjects were divided into 3 groups: 1) control patients with PIH healthy implants; 2) patients with PIM mucositis; and 3) patients with PI implantitis. To distinguish these three groups, peri-implant crevicular fluid (PICF) was collected by inserting a sterile paper strip into the gap around the implant and the levels of SIRT1 and cytokines were measured by the enzyme linked immunosorbent assay (ELISA). Demographic and clinical data included age, sex, Body Mass Index (BMI), probing depth (PD), plaque index (PLI), bleeding on probing (BOP), oral health impact profile (OHIP-14), history of periodontitis and the use time of implants. RESULTS: The PD, PLI, OHIP-14 evaluation scores in patients with periodontitis of PIM mucositis and PI implantitis were all significantly higher than in patients with PIH healthy implants. Overall, the SIRT1 levels in PICF of the PIM and PI patients were significantly lower than of the PIH patients. In comparison with PIM patients, SIRT1 levels of the PI patients were remarkably lower than the PIH patients. Pearson's analysis showed that SIRT1 levels were negatively correlated with levels of interleukin (IL)-6, C-reactive protein (CRP) and IL-1ß in patients with PIM and PI. We suggest that SIRT1 levels could serve as a potential diagnostic biomarker of PI or PIM. The PICF levels of SIRT1, CRP, IL-6, IL-1ß and the history of periodontitis were the risk factors for patients with peri-implant inflammatory process. CONCLUSION: The measurement of SIRT1 expression in PICF may serve as a biomarker for the ongoing inflammatory process in patients with dental implants. The low SIRT1 levels correlated with PI implantitis and PIM mucositis as well as the elevated levels of pro-inflammatory cytokines (CRP, IL-6 and IL-1ß).

Effects of a 980 nm Diode Laser as an Adjunct to Nonsurgical Periodontal Therapy on Periodontal Status and Inflammatory Markers in Patients After Myocardial Infarction: A Randomized Controlled Trial.

Objective: The aim of this study was to evaluate the efficacy of diode laser (DL) therapy as an adjunct to nonsurgical periodontal therapy in the treatment of periodontitis in patients after myocardial infarction (MI). Methods: After given permission by Ethics Commission of the Pomeranian Medical University (KB-0012/06/12), 36 patients <65 years of age (mean: 56.3 ± 7.9) with periodontitis, 6 weeks to 6 months after MI were enrolled for the study. The control group (n = 18) received nonsurgical periodontal therapy, whereas the test group (n = 18) received nonsurgical periodontal therapy followed by laser therapy of the periodontal pockets with 980 nm DL, 1 W, continuous wave mode, and 20 sec per tooth side. Procedures were repeated twice at 5-7 day intervals. Clinical periodontal parameters and inflammatory markers in gingival crevicular fluid (GCF) [elastase, aspartate transaminase (AST), alanine transaminase (ALT) and interleukin (IL)-6, proteins], bloodstream [fibrinogen, high-sensitivity CRP (hs-CRP), IL-6, AST and ALT], and lipid fractions (triglycerides, high-density lipoprotein, low-density lipoprotein, and total cholesterol) were measured before treatment, 2 weeks, and 3 months after treatment. Results: The difference between groups in the reduction of periodontal pocket depth (PPD) in pockets ≥7 mm was found to be significant in the test group (p < 0.05). There was also a statistically significant reduction in the volume of GCF and hs-CRP concentration in blood 2 weeks after the completion of treatment in the test group (p < 0.05). Conclusions: Within the limits of this study, it can be concluded that in the nonsurgical treatment of periodontitis with patients after MI, the additional use of DL enables greater reduction of PPD in pockets ≥7 mm. In addition, a faster reduction of GCF volume and hs-CRP was noted in the laser group.