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Objective: To elucidate the molecular healing of intrabony defects following non‐surgical periodontal therapy (NSPT) using gingival crevicular fluid (GCF). Background Data: Currently limited information is available regarding the GCF of intrabony defects and the change in biomarker levels in the GCF at early time points following treatment interventions. Methods: Twenty‐one patients (Periodontitis Stage III or IV) who have received NSPT, contributing one intrabony defect and one healthy site were included in this study. GCF sampling was performed at baseline, 1 day, 5 days and 3 months after NSPT. Multiplex bead immunoassays allowed the profiling of GCF for 27 markers, associated with inflammation and repair/regeneration. A mixed effects model with Bonferroni correction for multiple comparisons was employed to compare the changes in the levels of GCF markers over time. Results: Following NSPT, changes were observed for several GCF markers, marked by significant increases 1 day post‐intervention, before returning to baseline levels by 3 months. Specifically, GCF concentrations of IL‐2, IL‐4, IL‐6, IL‐8, MMP‐1, MMP‐3, TIMP‐1 and FGFb significantly increased 1 day after NSPT. Signs of activation of cellular senescence were observed 1 day following treatment of intrabony defects, rapidly regressing by 5 days. Conclusion: Significant molecular changes are observed as early as 1 day following NSPT in intrabony defects, along with activation of cellular senescence. [ABSTRACT FROM AUTHOR]

Adrenomedullin (AM) and corticotrophin (ACTH) are both vasoactive peptides produced by a variety of cell types, including endothelial cells. Although AM and ACTH are considered to be important in the control of blood pressure and the response to stress, respectively, their role in inflammation and the immune response has not been clarified. This study shows, with the use of a cell-based ELISA, that AM and ACTH induce cell surface expression of the adhesion molecules E-selectin, VCAM-1, and ICAM-1 on human umbilical vein endothelial cells (HUVEC). Furthermore, this effect appears to be mediated in part via elevation of cAMP, given that both peptides elevate cAMP, the cell-permeable cAMP analog dibutyryl cAMP is able to mimic induction of all three cell adhesion molecules and the effect of AM and ACTH is inhibited by the adenylyl cyclase inhibitor SQ-22536. These findings demonstrate a role for AM and ACTH in the regulation of the immune and inflammatory response. E-selectin; intercellular adhesion molecule-1; vascular cell adhesion molecule-1; adrenomedullin; adrenocorticotropic hormone; human umbilical vein endothelial cells

Aim: To profile, for the first time, the gingival crevicular fluid (GCF) of intrabony defects against a wide array of inflammatory and regenerative markers. Materials and methods: Twenty‐one patients contributed one intrabony defect and one periodontally healthy site. Clinical and radiographic measures were obtained. GCF samples were analyzed with multiplex bead immunoassays over 27 markers previously identified by our group. Comparisons were performed using Wilcoxon matched‐pairs signed‐ranks tests, using a Bonferroni corrected α = 0.05/27 = 0.0019. Results: Intrabony defect sites presented significantly increased GCF volume and disease‐associated clinical and radiographic characteristics (p <.05). Intrabony defect sites presented significantly increased IL‐1α, IL‐1β, IL‐6, IFN‐γ, and MMP‐8 levels compared with periodontally healthy sites (p <.0019). For regeneration markers, significantly higher FGF basic and VEGF levels were observed (p <.0019). Notably, traits of cell senescence were identified for the first time in the GCF. Conclusions: The differentiation of intrabony defects from periodontally healthy control sites can be based on clinical and radiographic measures and on a differentiated GCF profile that is site‐specific. Alongside catabolic processes, through significant up‐regulation of inflammation and connective tissue remodeling, unique molecular characteristics of intrabony defects may render them a microenvironment amenable to regeneration. Traits of the senescence‐associated secretory phenotype may suggest the existence of senescent cells during periodontal inflammation in intrabony defects. [ABSTRACT FROM AUTHOR]

TLRs are PRRs that play a pivotal role in sensing exogenous pathogens and endogenous danger signals. Their role in the pathogenesis of inflammatory and immune-related diseases is gradually being unravelled. TLR2 and TLR4 are capable of sensing the oral microbial community, which is considered a potential trigger for Behçet’s disease (BD). This study aimed to investigate the expression and function of TLR2 and TLR4 in the oral mucosa of BD. A total of 87 patients was included: 55 BD, 24 healthy controls and eight recurrent aphthous stomatitis. Total RNA was purified from non-lesional oral mucosal brush biopsies and analysed for the presence of TLR2 and TLR4 mRNA, along with their splice variants. The response of peripheral blood mononuclear cells to classical TLR2 and TLR4 agonists was also investigated. TLR2b, TLR2d, TLR2e, TLR4.3 and TLR4.4 were significantly elevated in relapsed BD. A significant defect in the response to cognate agonists of TLR1/2 heterodimer and TLR4 was also observed in BD. The expression of unusual splice variants of TLR2 and TLR4 might explain the observed defect in these receptors’ function in BD. [ABSTRACT FROM AUTHOR]

Adrenomedullin is a peptide found in a variety of cells and tissues and involved in a multitude of biological processes. Recently, adrenomedullin has been identified as a host defense peptide and as such it plays a role in the inflammatory response. The transcription factor NF-kappaB is a major regulator of genes involved in the inflammatory response and the aim of this study was to determine whether NF-kappaB played a role in the inflammatory process triggered by adrenomedullin. Skin epithelial cells (HaCaTs) were used as our model in vitro. Western blot analysis from adrenomedullin-stimulated HaCaT cells revealed a rapid degradation of NF-kappaB inhibitor alpha and beta followed by the translocation of free NF-kappaB to the nucleus, where it was detected by Texas Red immunostaining after incubation with adrenomedullin for 15 min. Electromobility shift assay showed that NF-kappaB present in the nucleus was active, since it bound to a probe containing an NF-kappaB binding site. Supershift assays indicated that p50 and p65, members of the NF-kappaB family, were both part of the NF-kappaB dimmers involved in adrenomedullin cell signaling. HaCaTs secreted interleukin-6 in response to AM, which was significantly attenuated by the NF-kappaB inhibitor SN-50. Taken together, the data lend support for an immunoregulatory role for AM.