BACKGROUND: The presence of a polymicrobial dysbiotic film in direct and constant contact with periodontal tissues initiates the host immune response. Interleukin 18 (IL-18) triggers up-regulates the production of other proinflammatory cytokines (TNF-α, IL-1β, IL-6), creating a vicious cycle that expands the inflammatory and destructive process in the periodontal tissue. A systematic review and meta-analysis was carried out with the main propose to investigate IL-18 expression in different biological samples from subjects with chronic periodontitis.
METHODS: The protocol followed PRISMA guidelines and was registered in Open Science Framework (OSF): https://doi.org/10.17605/OSF.IO/BS9GM . A digital search was conducted in the databases PubMed, ScienceDirect, Google Scholar, Web of Science and Dentistry & Oral Sciences Source databases were consulted from March 15th, 2005 to February 10th, 2023. Study quality was assessed using the JBI tool for cross-sectional studies and clinical trials. A meta-analysis was performed using a random/fixed effects model to evaluate the concentration of IL-18 in serum, plasma, saliva, gingival tissue and GCF of exposure group compared to control group.
RESULTS: The search strategy provided a total of 3,156 articles, of which 18 investigations met the inclusion criteria and 15 articles were quantitatively analyzed. The total number of patients studied was 1,275 (682 cases and 593 controls). The meta-analysis revealed significantly elevated IL-18 levels of serum, saliva and GCF of subjects with chronic periodontitis compared to healthy subjects (Serum: SMD = 62.73, 95%CI: 25.43-100.03, Z = 3.29, p = 0.001*; Saliva: SMD = 243.63, 95%CI: 8.68-478.59, Z = 2.03, p = 0.042*; GCF: SMD = 150.26, 95%CI: 56.86-243.66, Z = 3.15, p = 0.02*).
CONCLUSIONS: IL-18 levels in serum, saliva and GCF could have the potential to be used as complementary diagnostic tools to the clinical and radiographic parameters in subjects with periodontitis.

This study aims to histologically and immunohistochemically evaluate the effect recombinant human bone morphogenetic protein (rh-BMP2) injected in gingival tissue has on the acceleration of the epithelial migration from the wound edges and epithelial cell proliferation after implant surgery.
METHODS: The study includes 20 patients who underwent bilateral implant surgeries in the premolar-molar region of the mandible, followed by guided bone regeneration. Each patient received an implant in both locations, but rh-BMP2 was only on the right side. At 9 days from the surgery, a gingival biopsy was performed 3 mm distally to the last implant. In total, 20 samples were collected from the left side (control group #1) and 20 from right (test group #1). This was repeated at a 4-month interval during healing abutment placements. Tissues were processed and stained with hematoxylin-eosin and then immunohistochemically for the expression of Ki-67 and further histological examination.
RESULTS: Complete closure of the epithelium with new cell formation was observed in the 55% test group and 20% control group after 9 days. At 4 months, although 100% samples of all groups had complete epithelial closure, the test group showed that the epithelial cells were more organized and mature due to the increased number of blood vessels. The average number of new epithelial cells was 17.15 ± 7.545 and 16.12 ± 7.683 cells per mm in test group, respectively, at 9 days and 4 months and 10.99 ± 5.660 and 10.95 ± 5.768 in control groups.
CONCLUSIONS: Evident from histological observations, rh-BMP-2 can accelerate the closure of gingival wounds, the healing process of epithelial gingival tissue, and the formation of epithelial cells in patients undergoing dental implant treatment.

UNASSIGNED: Three-dimensional (3D) tissue models bridge the gap between conventional two-dimensional cell cultures and animal models. The aim of this study was to develop an organotypic 3D gingival (OTG) model to provide a tool to investigate bacterial and viral pathogens in periodontitis.
UNASSIGNED: The OTG model composed of gingival fibroblasts (GFs) and telomerase-immortalized gingival keratinocytes (TIGKs) was constructed and applied to study infections by Porphyromonas gingivalis and herpes simplex virus 1 (HSV-1). Immunohistochemical staining, confocal microscopy, qPCR, titration techniques, and colony-forming unit counts were applied to interrogate epithelial markers expression, monitor P. gingivalis and HSV-1 presence, and evaluate the immune response along with the efficiency of antimicrobial drugs.
UNASSIGNED: The OTG model resembled the morphology of the human gingiva. During infection, both pathogens penetrated deep into the tissue and persisted for a few days with P. gingivalis also forming a biofilm on the cell surface. The infection triggered the expression of inflammatory mediators in cells and both pathogens were efficiently eliminated by specific antimicrobials.
UNASSIGNED: Presented OTG model constitutes a simple and convenient tool to study the interaction between bacterial and viral pathogens within the gingival tissue, including penetration, persistence and biofilm formation. It is also suitable to examine the efficiency of antimicrobial drugs.

OBJECTIVE: To investigate the proportional variation of macrophage and T-lymphocytes subpopulations in acute coronary syndrome (ACS) patients, its association with periodontitis (P), and to compare with control individuals.
METHODS: Three groups of subjects participated: one group consisted of 17 ACS patients with P (ACS + P), another group consisted of 22 no ACS + P patients, and a control group consisted of 23 participants with gingivitis (no ACS + G). Macrophage, CD4 + , and CD8 + T-lymphocytes and CD4 + /CD8 + ratio values in gingival tissue were determined histometrically.
RESULTS: Significant differences were found among three groups regarding the mean number of macrophage (no ACS + P > ACS + P > no ACS + G; p < 0.05) and CD8 + T-lymphocytes (no ACS + P > ACS + P > no ACS + G; p < 0.05). Significant variations were observed between the groups both CD4 + T-lymphocytes densities (ACS + P > no ACS + P and ACS + P > no ACS + G; p < 0.05) and CD4 + / CD8 + ratio (no ACS + P < no ACS + G and ACS + P < no ACS + G; p < 0.05).
CONCLUSIONS: The increased number of CD8 + T-lymphocytes in both group ACS + P and group no ACS + P resulted in a reduction of the CD4 + /CD8 + ratio in gingival tissue when compared with no ACS + G group.
CONCLUSIONS: The decrease of CD4 + /CD8 + ratio in gingival tissue reflects periodontitis and may be associated with severe adverse outcomes in people with ACS.

The aim of this study is to evaluate the effect on the initiation of new blood vessel formation of rh-BMP-2 administration in the human gingival tissue during bone regeneration surgery.
METHODS: The randomized controlled clinical trial included twenty patients with bilateral partial edentulous of the mandibular premolar and molar region. Each patient received one implants on each side. Only one side received a 0.25 µg injection of rhBMP-2 into the gingival flap and grafted material during guided bone regeneration (GBR) for dental implantation. And the other side received GBR without injection. Three samples were collected from each patient as follows: one from the anterior area of the mandible (control group #1) collected at the time of all implant surgeries, and the two other samples during the placement of healing abutments at 4 months of follow-up, from treated side with rh-BMP-2 (test group) and untreated ones (control group #2). A total of 60 gingival samples were collected. Samples were stained with hematoxylin-eosin, and immunohistochemistry was performed with a vascular endothelial growth factor marker. The number of new vessels in each sample was counted.
RESULTS: Statistical analyses showed a significantly higher number of new vessels in the gingival tissue of the test group.
CONCLUSIONS: Rh-BMP-2 injections into the gingival flap significantly improved new blood vessel formation.

This study aimed to analyse the immunohistochemical profile of inflammatory infiltrates in the gingival tissue of patients undergoing orthodontic treatment in relation to patients’ titanium and/or nickel allergy status. Patients with gingival enlargement received initial periodontal therapy, followed by external gingivectomy in the case of persistent gingival enlargement. The sample included 44 patients (22 had metal allergic sensitisation). Histopathological changes were assessed, and an immunohistochemical analysis was performed on formalin-fixed and paraffin-embedded gingival samples using antibodies against CD1a, CD3, CD4, CD8, CD20, CD68, and CD138. Computer-assisted image analysis was performed to evaluate the positive cell count in the gingival tissue. The gingiva of the sensitised patients was characterised by the absence of multifocal inflammatory infiltrates (p < 0.05), while pronounced exocytosis and band-like inflammatory infiltrates were more frequently observed in sensitised patients. In addition, there was an increase in Langerhans cells and T-helper lymphocytes and a decrease in naïve T-lymphocytes, cytotoxic T-lymphocytes, macrophages, and plasma cells in the sensitised subjects compared to non-sensitised. However, the differences were only statistically significant for macrophages, with a moderate effect size (82.8 vs. 133.9; p = 0.041; r = 0.308). The absence of multifocal inflammation appears to be the most characteristic histopathological feature of the gingiva of sensitised patients. Although their gingiva presented certain characteristics of late hypersensitivity immune reactions the observed changes imply dominant irritative effect e.

UNASSIGNED: Although some studies have taken an interest in the participation of platelets in periodontitis, so far, we know very little about the roles of platelets in periodontitis. The objective of this study is to explore the involvement of platelets in the development of experimental periodontitis in mice.
UNASSIGNED: Twenty C57BL/6 male mice were used for this study. Experimental periodontitis models of mice were constructed by ligating for 1, 3, 7, and 14 days, respectively. Morphological changes in the alveolar bone were assessed by micro-computed tomography (Micro-CT). The gingival crevicular fluid samples of ligation sites were collected and stained by immunocytochemistry. Immunohistochemistry was used to detect platelets infiltration in gingival tissues of mice.
UNASSIGNED: The results of Micro-CT showed that with the extension of ligation time, alveolar bone resorption increased, suggesting that the experimental periodontitis models were established. Immunochemical staining showed that there were almost no platelets in the gingival crevicular fluid of mice ligated for 1 and 3 days. And at 7 and 14 days of ligation, a large number of platelets were present in the gingival crevicular fluid and formed complexes with neutrophils. And with the extension of ligation time, the extent of platelet infiltration increased in mice gingival tissues.
UNASSIGNED: Platelets were infiltrated increasedly in the gingival sulcus and gingival tissues following the experimental time, and may participate in the development of mouse experimental periodontitis.

There is a shortage of suitable tissue-engineered solutions for gingival recession, a soft tissue defect of the oral cavity. Autologous tissue grafts lead to an increase in morbidity due to complications at the donor site. Although material substitutes are available on the market, their development is early, and work to produce more functional material substitutes is underway. The latter materials along with newly conceived tissue-engineered substitutes must maintain volumetric form over time and have advantageous mechanical and biological characteristics facilitating the regeneration of functional gingival tissue. This review conveys a comprehensive and timely perspective to provide insight towards future work in the field, by linking the structure (specifically multilayered systems) and function of electrospun material-based approaches for gingival tissue engineering and regeneration. Electrospun material composites are reviewed alongside existing commercial material substitutes\', looking at current advantages and disadvantages. The importance of implementing physiologically relevant degradation profiles and mechanical properties into the design of material substitutes is presented and discussed. Further, given that the broader tissue engineering field has moved towards the use of pre-seeded scaffolds, a review of promising cell options, for generating tissue-engineered autologous gingival grafts from electrospun scaffolds is presented and their potential utility and limitations are discussed.

Objective: Chronic periodontitis is a bone-destructive disease affecting periodontal support structures. Although leptin has a protective effect against periodontitis, the underlying mechanism remains unclear. Therefore, this study aimed to investigate the possible role of leptin by examining its relationship with OPG and RANKL in human gingival tissues obtained from patients with chronic periodontitis. Method: Twenty-two patients with chronic periodontitis were enrolled (10 with moderate periodontitis and 12 with severe periodontitis) in the experimental group, and 12 healthy individuals were enrolled in the control group. Gingival tissue samples were collected, and the protein levels and localization of leptin, OPG, and RANKL were studied using immunohistochemistry (IHC). The staining intensities of leptin, OPG, and RANKL were correlated with the periodontal clinical index. Moreover, real-time quantitative PCR (RT-qPCR) was used to determine OPG and RANKL mRNA levels in gingival fibroblasts stimulated with gradient concentrations of leptin protein in vitro. Result: Leptin, OPG, and RANKL were located in the cytoplasm of gingival epithelial cells and the connective tissue. Leptin was widely and significantly expressed in the control group, whereas it was lightly stained in the severe group. RANKL was lightly stained in the control group, whereas it was widely and significantly expressed in the severe group. The control and the moderate groups had similar OPG levels, which were significantly higher than that in the severe group. Leptin was positively correlated with OPG(r = 0.905, p < 0.01) and negatively correlated with RANKL (r = -0.635, p < 0.01). In vitro low concentrations of leptin led to an increased OPG/RANKL mRNA ratio, whereas the opposite effect was observed at high concentrations. Conclusion: Leptin can regulate OPG and RANKL expression in gingival fibroblasts and may thus play a role in the development of chronic periodontitis by modulating the OPG/RANKL ratio.

UNASSIGNED: Cigarette smoking has been recognized as an important risk factor in periodontal diseases. One of the suggested mechanisms behind this association is that nicotine alters the microcirculation and causes vasoconstriction and reduced blood flow through the periodontal tissues. Scarce information is currently available relative to the microvascular alterations associated with smoking and the distribution of capillaries through the various areas of the gingival tissues. The aims of this study were to assess, in human interproximal gingival biopsies, the number and diameter of gingival capillaries in periodontally affected smokers and nonsmokers using the CD34 immunohistochemical staining method. The pattern of distribution of vessels in the different areas of the gingival tissues was also assessed.
UNASSIGNED: Systemically healthy patients with moderate chronic periodontitis and ranging in age between 30 and 60 years were recruited for the study from the patient population attending the Periodontology Department of the Faculty of Dental Medicine at the Lebanese University of Beirut. The patients were selected to have a group of 10 patients (Group SP) of smokers (>10 cigarettes/day for the last 10 years) and a second group (Group NP) consisting of nonsmoking periodontally affected patients. Three to four weeks following initial preparation, one interproximal gingival biopsy was obtained from each patient. Immunohistochemical staining with CD34 mouse monoclonal antibody was used to identify the endothelial cells of the blood vessels within each sample. Twelve biopsy samples (five in Group NP and seven in Group SP) were chosen for the measurement of the number and diameter of vessels in three regions of the connective tissue of the biopsy under a blinded protocol.
UNASSIGNED: In the two groups, the quantitative distribution of small, medium, and large vessels followed a similar trend with the number of small vessels being significantly greater than both medium and large vessels. Small vessels prevailed in the peripheral regions, whereas large vessels were more abundant in the deeper connective tissue areas. The total number of vessels seemed unaffected by chronic cigarette smoking in both groups in the entire biopsy area and in the separate connective tissue regions. Quantitative alteration in the total number of gingival capillaries was not observed in chronic smokers. A redistribution of small and large vessels in the superficial and deeper connective tissue areas of the gingival papilla was noted as a result of smoking in periodontal patients.
UNASSIGNED: The quantitative distribution of small, medium, and large vessels follows a similar trend with the content in small vessels being significantly more important than both medium and large vessels. Smoking and periodontitis result in a redistribution of small and large vessels in the superficial and deeper connective tissue areas of the gingival papilla compared to nonsmoking periodontal patients. The significance and clinical implications of such rearrangement of vasculature within the gingival tissue need to be further investigated.