Behçet's disease (BD) pathogenesis involves severe outcomes such as blindness, central nervous system manifestations, and deep venous thrombosis that impacts systemic and local inflammatory changes. We tested the hypothesis that BD negatively affects gingival health and increases the severity of gingivitis. The study included 37 BD patients with gingivitis without any sign of periodontitis. Systemically healthy 19 patients with gingivitis (G) and 20 periodontally and systemically healthy individuals (C) were recruited as controls. BD patients were further grouped as stable and unstable based on their responses to BD treatment. Clinical periodontal parameters were measured to determine the impact of BD on gingival health. Serum and saliva levels of ELA-2 (neutrophil elastase-2), SLPI (secretory leukocyte protease inhibitor), α1-AT (alpha1-anti-trypsin), VEGF (vascular endothelial growth factor), IL-6 (interleukin-6), IL-8 (interleukin-8), and TNF-α (tumor necrosis factor alpha) were analyzed using multiplex immunoassay to measure the systemic and local inflammatory impact of BD. Plaque index (PI), probing pocket depth (PPD), and bleeding on probing (BOP) were significantly higher in the BD group than in the controls (p<0.05). IL-6 was higher in both serum and saliva in the BD group than in the G group (p<0.05). ELA-2 levels in saliva were higher in the stable BD group than in the controls, while TNF-α and SLPI were statistically significantly higher in BD than in the control (p<0.05). Salivary α1-AT level was statistically lower in the BD group compared to the control group. Our study suggested that the gingival inflammatory profile was impaired in patients with BD.

Few investigations evaluated smoking's impact on the periodontal proteome. Therefore, this study aimed to analyse the influence of tobacco on the overall periodontal proteome and the differential expression of gingival crevicular fluid (GCF) proteins using sequential window acquisition of all theoretical mass spectra (SWATH-MS). GCF samples were collected from 40 periodontitis subjects (stages III-IV). These were separated based on smoking status into smokers (17), ex-smokers (10), and non-smokers (13). Samples were analysed using SWATH-MS, and proteins were identified using the UniProt human-specific database. Data are available via ProteomeXchange with the identifier PXD043474. Principal component analysis (PCA) was employed to examine the spectral mass distribution of the proteome. Protein expression was different for a p-value<0.05 and a log2 fold change ≥0.3 (upregulated) or ≤-0.3 (downregulated). The distribution of overall proteome did not differ between non-smokers, smokers, and ex-smokers. Considering protein expression, 23 were differentially expressed in smokers vs. non-smokers (16 upregulated and 7 downregulated), 17 in ex-smokers vs. non-smokers (2 upregulated and 15 downregulated), and only 8 in smokers vs. ex-smokers (7 upregulated and 1 downregulated). Smoking increased the expression of proteins related to epithelial hyperkeratinization (keratins type II cytoskeletal 4, type I cytoskeletal 13 and type I cytoskeletal 19, cornulin, and fatty acid-binding protein 5). However, multiple immunoglobulins were underexpressed when comparing smokers and ex-smokers to non-smokers. Although smoking does not significantly modify the overall GCF proteome associated with periodontitis, it alters the expression of several proteins compared to never-smokers and ex-smokers. Smoking is a critical risk factor for the development and progression of periodontitis. However, evidence of the effect of smoking on the subgingival proteome is scarce. Therefore, this study aimed to determine the impact of smoking on the overall proteome and differential expression of gingival crevicular fluid (GCF) proteins using the sequential window acquisition of all theoretical mass spectra (SWATH-MS) proteomic technique. For this purpose, GCF samples were collected from 40 subjects with periodontitis, of which 17 were smokers, 10 were ex-smokers, and 13 were non-smokers. These samples were analysed by SWATH-MS, and proteins were identified using the UniProt human-specific database. Analysis of the overall proteome showed that its distribution was not significantly different between smokers, ex-smokers, and non-smokers. However, several proteins were found to be differentially expressed according to the smoking status. Smoking can increase the expression of several keratins and proteins related to hyperkeratinization of the epithelium. However, in ex-smokers, these proteins return to similar levels to those of non-smokers. Moreover, smoking may induce a lower expression of proteins related to adaptive immunity, such as immunoglobulins. This immunosuppressive effect may persist in ex-smokers.

The polarization of macrophages into an anti-inflammatory phenotype is crucial for resolving periodontal inflammation. It has been reported that B10 cells can regulate the immune response of macrophages during inflammation and are also able to regulate inflammation in periodontitis. However, whether B10 cells' regulation function in periodontitis is related to macrophage polarization remains unclear. This study aims to investigate whether B10 cells can regulate macrophage polarization in periodontitis. Macrophages were cocultured with B10 cells in vitro for 5days. After coculture, macrophages were obtained for analysis directly or followed by stimulation with Pg-LPS/IFN-γ or IL-4/IL-13. Flow cytometry and/or reverse transcriptase-polymerase chain reaction (RT-PCR) were employed to detect the expression of IL-1β, iNOS, TNF-α, CD206, and ARG-1 in macrophages. B10 cells were transferred on the 5th day after ligation in wild or macrophage-depletion mice. Toluidine blue and TRAP staining were used to evaluate alveolar bone resorption and osteoclast activation. Immunohistochemistry was employed to detect the expression of CD68, IL-1β, TNF-α, iNOS, ARG-1, and IL-10. Immunofluorescence was used to detect the expression of CD68+CD86+M1 macrophages and CD68+CD206+M2 macrophages. In vitro, B10 cells inhibit the expression of IL-1β, iNOS, and TNF-α in macrophages while increasing the expression of CD206 and ARG-1. In experimental periodontitis, B10 cells inhibit the polarization of CD68+CD86+M1 macrophages and iNOS expression but enhance the polarization of CD68+CD206+M2 macrophages and ARG-1 expression. Importantly, the depletion of macrophages partially weakened the regulation function of B10 cells in periodontitis. B10 cells promote M2 macrophage polarization, inhibit M1 macrophage polarization in periodontitis, and alleviate periodontitis partially by regulating macrophage polarization.

Lipocalin-2 (LCN-2) is an osteokine that suppresses appetite, stimulates insulin secretion, regulates bone remodeling, and is induced by proinflammatory cytokines. The aim of this work was to investigate the participation of LCN-2 in periodontitis associated with type 2 diabetes (T2D) by evaluating alveolar bone loss, glycemic control, inflammation, and femur fragility. A murine model of periodontitis with T2D and elevated LCN-2 concentration was used. Functional LCN-2 inhibition was achieved using an anti-LCN-2 polyclonal antibody, and isotype immunoglobulin G was used as a control. The alveolar bone and femur were evaluated by micro-CT. Glucose metabolism was determined. Tumor necrosis factor (TNF-α) and receptor activator of nuclear factor kappa-B ligand (RANKL) levels in alveolar bone lysates were quantified using ELISA, and serum cytokines were quantified using flow cytometry. A three-point bending test was performed in the femur, and RANKL levels were measured in femur lysates using ELISA. Functional inhibition of LCN-2 in T2D-periodontitis mice decreased alveolar bone loss in buccal and palatal surfaces and preserved the microarchitecture of the remaining bone, decreased TNF-α and RANKL in alveolar bone, reduced hyperglycemia, glucose intolerance, and insulin resistance, and increased insulin production through improving the functionality of pancreatic β cells. Furthermore, this inhibition increased serum free-glycerol levels, decreased serum interleukin (IL)-6, increased serum IL-4, and reduced femur fragility and RANKL expression in the femur. LCN-2 participates in periodontitis associated with T2D. Inhibiting its function in mice with T2D and periodontitis improves pancreatic β-cell function, and glucose metabolism and decreases inflammatory cytokines and bone-RANKL levels, which results in the preservation of femoral and alveolar bone microarchitecture. In this study, we explored the role of a bone protein known as lipocalin-2 (LCN-2) in the connection between periodontitis and type 2 diabetes (T2D). Periodontitis is a destructive gum and alveolar bone disease. LCN-2 levels are increased in both T2D and periodontitis. Using a mouse model of T2D with periodontitis, we examined how blocking LCN-2 function affected various aspects of these two diseases. We found that this inhibition led to significant improvements. First, it reduced alveolar bone loss and preserved bone structure by decreasing local inflammation and bone resorption. Second, it improved glucose and lipid metabolism, leading to better blood-sugar control and decreased insulin resistance. Blocking the functions of LCN-2 also decreased systemic inflammation throughout the body and strengthened bone integrity. Overall, our results suggest that LCN-2 plays a crucial role in the periodontitis associated with T2D. By inhibiting LCN-2 function, we were able to improve pancreatic function, improve glucose metabolism, reduce inflammation, and enhance bone health. Targeting LCN-2 could be a promising strategy for the harmful effects of T2D and periodontitis.

This study investigated the effect of a 4-week free-sugar avoidance on periodontal parameters during periodontal therapy. Twenty-one patients with untreated periodontitis and daily free-sugar intake were allocated to a sugar avoidance group (SAG) and a control group (CG). The SAG received a 45-min dietary consultation and was instructed to avoid free sugars during the following 4 weeks after subgingival instrumentation, while the CG continued with their regular diet. Bleeding on probing (BOP), plaque control record, body weight (BW), visceral fat (FATv), and a food frequency questionnaire (FFQ) were collected at baseline (T1), 4 weeks (T2), and 8 weeks (T3) after subgingival instrumentation. The main outcome parameter BOP was significantly reduced at T2 by 40.3% ± 15.54 in the SAG and 34% ± 12.47 in the CG (intra-p value both <0.001, inter-p value 0.361). A linear regression analysis of changes at patient level adjusted for age and FATv revealed a significant group difference for BOP (regression coefficient = -6.8; p = 0.019). Significant reductions were observed in BW, FATv and mean daily intake of free sugars (-14.4g/day), and a significant increase of vitamin C derived from fruits (75.89mg/day) at T2 in the SAG only. This study may indicate additional beneficial effects of a sugar avoidance on periodontal and metabolic parameters, and nutritional intake during periodontal therapy. German Clinical Trials Register (DRKS00026699). The current widespread free-sugar consumption is linked to an increasing incidence of chronic non-communicable diseases. Data indicate a relationship between sugar intake and a higher prevalence of periodontitis and increased gingival inflammation. This study showed that free-sugar avoidance after periodontal therapy had additional beneficial effects on periodontal and metabolic parameters in 10 test and 11 control patients. After 4 weeks of avoiding free sugars like sweets, processed white flour, juice, and so forth, periodontal bleeding was significantly reduced in both groups (-40.3% test group, -34% control group). Further regression analysis revealed a significant difference between groups favoring the intervention. Additionally, body weight and visceral fat were significantly reduced in the intervention group, only. To avoid sugar, patients were allowed to replace it with whole fruit, which led to increased levels of micronutrients such as vitamin C. Therefore, free-sugar avoidance may be of therapeutic benefit in addition to periodontal therapy. Further research is needed to investigate this effect in larger cohorts.

To compare the efficacy of combined treatment of Er:YAG laser (ERL) and low-level laser therapy (LLLT) with single laser applications, and scaling and root planing (SRP) for non-surgical periodontal treatment. In a randomized controlled trial, 25 non-smoking Stage II or Stage III periodontitis patients were recruited. The four intraoral quadrants were randomly assigned to four different treatments: (1) combined application with ERL plus SRP plus LLLT; (2) ERL plus SRP; (3) SRP plus LLLT; and (4) SRP. We assessed periodontal indexes, including probing depth (PD), clinical attachment level (CAL), bleeding index (BI), and plaque index (PLI), along with three cytokines (IL-1β, TNF-α, IL-10) from gingival crevicular fluid and red complex pathogens from subgingival dental plaque at baseline, 3 months, and 6 months. For initial moderate pockets (4mm≤PD≤6mm), quadrants treated with ERL+SRP+LLLT, ERL+SRP, and SRP+LLLT exhibited greater PD improvement compared to the control (SRP) quadrants at the 3-month follow-up (1.25±1.06, 1.23±1.12, 1.00±1.21vs. 0.98±1.21mm) and the 6-month follow-up (1.35±1.06, 1.23±1.17, 1.35±0.98vs. 0.98±1.23mm) (p=0.002). Quadrants treated with ERL+SRP+LLLT and SRP+LLLT showed more CAL gain means than the control quadrants at the 3-month follow-up (0.96±1.42, 0.61±1.39vs. 0.55±1.57mm) and the 6-month follow-up (0.84±1.54, 0.89±1.49vs. 0.48±1.68mm) (p=0.008). For initial deep pockets (PD≥7mm), the ERL+SRP+LLLT quadrants had more PD improvement and CAL gain compared to the control quadrants at follow-up. There were no significant differences in BI, PLI, inflammatory cytokines, and periodontal pathogens among the four groups. The combined application of ERL and LLLT demonstrated potential efficacy in reducing PD, particularly for deep pockets. To compare the therapy effect of combined use of Er:YAG laser (ERL) and low level laser therapy (LLLT) with single laser applications, and traditional periodontal treatment (SRP). A total of 25 nonsmoking patients with periodontitis were involved, and their mouths were divided into four sections, each receiving a different treatment: ERL+SRP+LLLT, ERL+SRP, SRP+LLLT, and SRP. Clinical indexes and laboratory indicators were assessed at baseline, 3 months, and 6 months. After six months, for initial moderate pockets, combined laser group and single laser group showed better improvements than traditional group in reducing the depth of periodontal pockets and increasing attachment levels. But for initial deep pockets, only combined laser group showed better improvement than traditional group. There were no significant differences in bleeding, plaque, inflammation, or harmful bacterial levels among the groups. These findings suggest that the integration of Er:YAG laser and lowlevel laser therapy into standard periodontal treatment may enhance the treatment's benefits in reducing pocket depth, especially for severe conditions.

This study aimed to analyze the influence of concave and cylindrical abutments on peri-implant soft tissue. Dimensions, collagen fiber orientation, and immunohistochemical data were assessed. A multicenter, split-mouth, double-blind randomized clinical trial was conducted. Two groups were analyzed: cylindrical abutments and concave abutments. After a 12-week healing period, peri-implant soft tissue samples were collected, processed, and evaluated for dimensions, collagen fiber orientation, and immunohistochemical data. Inflammatory infiltration and vascularization were assessed, and the abutment surfaces were analyzed using scanning electron microscopy. The statistical analysis was performed using the SPSS version 20.0 statistical package. A total of 74 samples in 37 patients were evaluated. Histological evaluation of peri-implant soft tissue dimensions revealed significant differences between concave and cylindrical abutments. Concave abutments exhibited greater total height (concave: 3.57±0.28 - cylindrical: 2.95±0.27) and barrier epithelium extension (concave: 2.46±0.17 - cylindrical: 1.89±0.21) (p<0.05), while the supracrestal connective tissue extension (concave: 1.11±0.17 - cylindrical: 1.03±0.16) was slightly greater (p>0.05). Collagen fiber orientation favored concave abutments (23.76±5.86), with significantly more transverse/perpendicular fibers than for cylindrical abutments (15.68±4.57). The immunohistochemical analysis evidenced greater inflammatory and vascular intensity in the lower portion for both abutments, though concave abutments showed lower overall intensity (concave: 1.05±0.78 - cylindrical: 1.97±0.68) (p<0.05). The abutment surface analysis demonstrated a higher percentage of tissue remnants on concave abutments (42.47±1.32; 45.12±3.03) (p<0.05). Within the limitations of this study, concave abutments presented significantly greater peri-implant tissue height, linked to an extended barrier epithelium, versus cylindrical abutments in thick tissue phenotype. This enhanced soft tissue sealing, favoring a greater percentage of transversely oriented collagen fibers. The concave design reduced chronic inflammatory exudation with T and B cells, thus minimizing the risk of chronic inflammation. This study looked at how 2 different shapes of dental implant abutments (the parts that connect the implant to the crown), specifically concave and cylindrical, affect the soft tissue around the implants. We wanted to see how these shapes influenced the tissue's size, structure, and health. We conducted a clinical trial with 37 patients, comparing the 2 types of abutments in the same mouth over 12weeks. Our findings showed that the concave abutments led to a taller and more extensive layer of protective tissue around the implant compared to the cylindrical ones. This protective tissue had more favorable collagen fiber orientation, which is important for the strength and health of the tissue. Additionally, the concave abutments resulted in less inflammation and better tissue integration. In conclusion, concave abutments may provide better support and health for the soft tissue around dental implants, reducing the risk of chronic inflammation and potentially leading to better long-term outcomes for patients with dental implants.

To assess how the diagnostic reproducibility of the 2018 Classification of Gingival Recession Defects (GRD) could be applied when comparing in-person chairside measurements with photographic measurements. Thirty-four GRD were photographed and evaluated by 4 masked operators. For each case, the operators measured twice recession type (RT), recession depth (RD), keratinized tissue width (KTW), gingival thickness (GT), detectability of the cemento-enamel junction (CEJ), and presence of root steps (RSs), chairside, and on photographs. Intraclass correlation coefficient (ICC) with 95% confidence intervals (CI) was calculated for RD and KTW; Kappa with 95% CI was used for GT, CEJ, and RS; quadratic weighted Kappa with 95% CI was used for RT. RD, KTW, and RT showed excellent overall intra-operator agreement (>0.93), and from good to excellent overall inter-operator agreement (>0.80), for both clinical and photographic measurements. Agreements were lower for GT, CEJ, and RS. Overall clinical and photographic agreements were within 0.1 difference for every variable, except for inter-operator agreement for RS which was 0.72 for clinical measurements and 0.45 for photographic measurements. The lowest overall agreement between clinical versus photographic measurements existed for CEJ (0.28) and RS (0.35). Variables composing the 2018 Classification of GRD are reproducible, both clinically and on photographs, with comparable agreements. The overall agreement was higher for KTW, RD, and RT, and lower for GT, CEJ, and RS, for both clinical and photographic measurements. The comparison between chairside and photographic evaluations indicated fair to excellent agreement for most variables, with CEJ and RS showing fair agreement. As digital diagnostics evolve to facilitate clinical diagnostic measurement, we aimed to assess the effectiveness of intraoral photography for diagnosing gingival recession defects (GRD) according to the 2018 Classification of GRD, compared to traditional clinical examination. Standardized photographs of thirty-four GRD cases were captured. Four masked operators evaluated the same gingival recessions twice in a clinical setting and twice using photographs. Measurement repeatability within and between operators was calculated for both clinical and photographic settings, and the two settings were compared. Continuous measurements such as recession depth and keratinized tissue width, as well as the evaluation of interproximal attachment height (recession type), showed excellent agreement both clinically and photographically. Agreement was lower for gingival thickness and the detectability of tooth anatomical landmarks, such as the cemento-enamel junction and the presence of root steps. Overall, the agreement between chairside and photographic evaluations was generally good, but lower when evaluating tooth anatomical landmarks. The variables composing the 2018 Classification of GRD are reproducible in both clinical and photographic settings, with comparable levels of agreement. However, there was consistently worse agreement for gingival thickness and when evaluating tooth anatomical landmarks.

It is unclear whether an intact buccal bony plate is a prerequisite for immediate implant placement in postextraction sockets. The aim of this 10-year randomized controlled trial was to compare peri-implant soft and hard tissue parameters, esthetic ratings of, and patient-reported satisfaction with immediate implant placement in postextraction sockets with buccal bony defects ≥5mm in the esthetic zone versus delayed implant placement after alveolar ridge preservation. Patients presenting a failing tooth in the esthetic region and a buccal bony defect ≥5mm after an extraction were randomly assigned to immediate (Immediate Group, n=20) or delayed (Delayed Group, n=20) implant placement. The second-stage surgery and provisional restoration placement occurred 3months after implant placement in both groups, followed by definitive restorations 3months thereafter. During a 10-year follow-up period, marginal bone levels (primary outcome), buccal bone thickness, soft tissue parameters, esthetics, and patient-reported satisfaction were recorded. The mean marginal bone level change was -0.71±0.59mm and -0.36±0.39mm in the Immediate Group and the Delayed Group after 10years (p=0.063), respectively. The secondary outcomes were not significantly different between both groups. Marginal bone level changes, buccal bone thickness, clinical outcomes, esthetics, and patients' satisfaction following immediate implant placement, in combination with bone augmentation in postextraction sockets with buccal bony defects ≥5mm, were not statistically different to those following delayed implant placement after ridge preservation in the esthetic zone. Immediate implant placement in case of a failing tooth is a favorable treatment option for patients because it considerably shortens treatment time and the number of surgical treatments. The question is if an intact buccal bone wall is necessary for immediate implant placement. A 10-year study was performed in which 20 patients with a failing tooth in the frontal region of the upper jaw were treated with immediate implant placement and were compared with 20 patients in whom a more conventional treatment strategy was followed in which the failing tooth was first removed and the bone gap restored and the implant placed in a second step. After a 10-year follow-up period, it appeared that the bone around the implant was very stable, gums were healthy, and patients were very satisfied with the result. There was no difference between the two treatment procedures. Such results mean that professionals can discuss the procedure with the patient and apply the individual's preference.