Evaluation of Cortical Thicknesses and Bone Density Values of Mandibular Canal Borders and Coronal Site of Alveolar Crest.

Objectives: The objectives of this retrospective study are to measure the amount of the alveolar crest cortication and cortication around the mandibular canal, and to evaluate bone density values of alveolar crest, cortication around mandibular canal, and possible implant placement area for edentulous sites. Material and Methods: Six hundred forty-two cone-beam computed tomography scans from 642 subjects were evaluated in four centers. Cortical thicknesses of alveolar crest and mandibular canal cortical borders (buccal, lingual, apical, and coronal) in each mandibular posterior teeth region were measured. Bone density of alveolar crest and mandibular canal cortical borders (buccal, lingual, apical, and coronal) in each mandibular posterior teeth region were recorded. The correlations between numeric variables were investigated using Pearson's correlation test. Results: The largest cortical border of the canal was measured 1.1 (SD 0.71) mm at the left second molar area and in coronal side of the mandibular canal (MC). Left and right first premolar regions showed higher bone density values compared to the other sites in all bone density values evaluations. The buccal side of the canal at the right first premolar region showed the highest bone density values (832.32 [SD 350.01]) while the coronal side of the canal at the left second molar region showed the lowest (508.75 [SD 225.47]). The bone density of possible implant placement area at the both left (692.25 [SD 238.25]) and right (604.43 [SD 240.92]) edentulous first premolar showed the highest values. Positive correlations between the bone density values of alveolar crest and the coronal side of MC were found in molar and left second premolar regions (P < 0.05). Conclusions: Results may provide information about the amount of cortication and bone densities tooth by tooth for posterior mandible to surgeons for planning the treatment precisely.

Anti-Apoptotic and Pro-Apoptotic Bcl-2 Family Proteins in Peri-Implant Diseases.

OBJECTIVES: Intrinsic apoptosis, which is regulated by Bcl-2 family proteins, has an important role in chronic inflammatory diseases. The aim of the study was to identify the tissue levels and ratios of anti- and pro-apoptotic Bcl-2 family proteins in peri-implant diseases. MATERIALS AND METHODS: Twenty-three individuals with peri-implant mucositis, 25 individuals with peri-implantitis, and 24 controls were included. The following clinical parameters were recorded: keratinized mucosa width, modified bleeding index, probing depth, modified plaque index, modified gingival index, and keratinized tissue thickness. Marginal alveolar bone assessments were performed by a software program. Granulation tissues were collected during treatments of peri-implant diseases. The control tissue samples were collected during the second stage of implant surgery. The tissue levels of Bcl-2 family pro-apoptotic (Bak, Bax, active caspase-3) and anti-apoptotic (Bcl-2, Bcl-xL, Mcl-1) proteins were determined by multiplex immunoassay method. RESULTS: The pro-apoptotic proteins; Bak, Bax and anti-apoptotic proteins Bcl-2, Bcl-xL, Mcl-1 were detected significantly higher in controls compared with patients with peri-implant mucositis and peri-implantitis (p < .001), respectively. The higher active caspase-3 levels were also detected in controls in comparison with peri-implant mucositis (p = .018) and peri-implantitis (p = .005). Anti-apoptotic: pro-apoptotic protein ratios (Bcl-2:Bax, p < .001; Bcl-2:Bak, p = .01; Bcl-xL: Bax, p = .006, Bcl-xL:Bak, p = .011; Mcl-1:Bak, p < .001) were significantly increased in diseased groups. A positive correlation was demonstrated between clinical variables and anti-apoptotic: pro-apoptotic ratios. CONCLUSION: Our findings indicate dysregulation of the Bcl-2 family proteins in peri-implant diseases. This unregulated response may disturb the homeostasis of peri-implant tissue.

Human ß-Defensins in Diagnosis of Head and Neck Cancers.

Head and neck cancers are malignant growths with high death rates, which makes the early diagnosis of the affected patients of utmost importance. Over 90% of oral cavity cancers come from squamous cells, and the tongue, oral cavity, and salivary glands are the most common locations for oral squamous cell carcinoma lesions. Human ß-defensins (hBDs), which are mainly produced by epithelial cells, are cationic peptides with a wide antimicrobial spectrum. In addition to their role in antimicrobial defense, these peptides also take part in the regulation of the immune response. Recent studies produced evidence that these small antimicrobial peptides are related to the gene and protein expression profiles of tumors. While the suppression of hBDs is a common finding in head and neck cancer studies, opposite findings were also presented. In the present narrative review, the aim will be to discuss the changes in the hBD expression profile during the onset and progression of head and neck cancers. The final aim will be to discuss the use of hBDs as diagnostic markers of head and neck cancers.

Salivary and serum levels of neutrophil proteases in periodontitis and rheumatoid arthritis.

OBJECTIVE: The aim was to profile serum and salivary levels of active-matrix metalloproteinase (aMMP)-8, tissue inhibitor MMP (TIMP)-1, aMMP-8/TIMP-1 ratio, total MMP (tMMP)-9, tMMP-9/TIMP-1 ratio, myeloperoxidase (MPO), and human neutrophil elastase (HNE) in periodontitis and rheumatoid arthritis (RA). MATERIALS AND METHODS: Rheumatoid arthritis patients with periodontitis (RA + P, n = 26), periodontally healthy RA patients (RA, n = 23), systemically healthy periodontitis patients (P, n = 24), and controls (C, n = 24) were included. aMMP-8 levels were determined by a time-resolved immunofluorescence assay (IFMA), TIMP-1, tMMP-9, MPO, and HNE levels were measured by enzyme-linked immunosorbent (ELISA) assays. RESULTS: Higher salivary aMMP-8 (p < 0.001), aMMP-8/TIMP-1 ratio (p = 0.043), tMMP-9 (p = 0.011), tMMP-9/TIMP-1 ratio (p = 0.022), MPO (p = 0.026) and HNE (p < 0.001) levels were detected in P relative to the controls. Salivary TIMP-1 was increased in RA patients regardless of periodontal status (RA + P vs. P: p = 0.038; RA vs. C: p = 0.020). Serum neutrophil proteases were increased in RA groups (RA + P, RA) compared to systemically healthy groups (P, C) (p < 0.05). CONCLUSIONS: Serum levels of neutrophil proteases were increased in RA study groups; however rheumatologic status seemingly does not affect salivary levels of these proteins.

Salivary macrophage activation-related chemokines and mitogen-activated kinase kinase-degrading proteolytic activity in type 1 diabetes mellitus.

BACKGROUND: This cross-sectional study aimed to evaluate salivary concentrations of macrophage activation-related chemokines and mitogen-activated kinase kinase (MAPKK)-degrading proteolytic activity in children and adolescents with and without type 1 diabetes mellitus (T1DM). METHODS: A total of 122 children and adolescents (65 T1DM patients, 50.8% female, mean age:10.9 years; 57 systemically healthy controls, 36.8% female, mean age: 9.5 years) were included in the study. Salivary concentrations of interferon gamma inducible protein-10 (IP-10), monocyte chemoattractant protein (MCP)-1, MCP-2, MCP-3, MCP-4, macrophage-derived chemokine (MDC), macrophage migration inhibitory factor (MIF), monokine induced by interferon gamma (MIG), and macrophage inflammatory protein-1 alpha (MIP-1α) were quantified using a bead-based technique. MAPKK-degrading proteolytic activity was detected using fluorescent peptide substrates. RESULTS: The T1DM group had higher plaque index (PI%, p = 0.032) and bleeding on probing (BOP%, p = 0.045) scores, and lower decayed, missing, filled teeth (dmft/DMFT, p = 0.002) index scores compared to the healthy controls. Compared to the controls, salivary MCP-1 (p = 0.007), MCP-3 (p < 0.001), MIG (p = 0.007), and MIP-1α (p = 0.033) concentrations were elevated whereas MCP-4 concentrations decreased (p < 0.001) in the T1DM group. After adjusting for age, PI%, BOP%, and dmft/DMFT scores, significant differences in salivary concentrations of MIG (p = 0.033) and MIP-1α (p = 0.017) were observed between the groups. Moreover, protease activities directed to the cleavage sites of MEK23-18 (p = 0.001), MKK6b7-22 (p = 0.007), MKK451-66 (p = 0.005), MKK7b37-52 (p = 0.034), and MKK7b69-84 (p = 0.009) were elevated in the T1DM group. CONCLUSION: T1DM disrupts the salivary macrophage activation-related chemokine profile and dysregulates proteolytic MAPKK cleavage. These findings can be an outcome of the impaired systemic immune response in T1DM.

The Radiological Evaluation of Mandibular Canal Related Variables in Mandibular Third Molar Region: a Retrospective Multicenter Study.

Objectives: The aim of this retrospective study was to investigate anatomical structure of mandibular canal and the factors those increase the possibility of inferior alveolar nerve damage in mandibular third molar region of Turkish population. Material and Methods: Overall 320 participants with 436 mandibular third molars were included from four different study centers. Following variables were measured: type and depth of third molar impaction, position of mandibular canal in relation to third molars, morphology of mandibular canal, cortication status of mandibular canal, possible contact between the third molars and mandibular canal, thickness and density of superior, buccal, and lingual mandibular canal wall, bucco-lingual and apico-coronal mandibular canal diameters on cone-beam computed tomography scans. Results: Lingual mandibular canal wall density and thickness were decreased significantly as the impaction depth of mandibular third molar was increased (P = 0.045, P = 0.001 respectively). Highest buccal mandibular canal wall density and thickness were observed in lingual position of mandibular canal in relation to mandibular third molar (P = 0.021, P = 0.034 respectively). Mandibular canal with oval/round morphology had higher apico-coronal diameter in comparison to tear drop and dumbbell morphologies (P = 0.018). Additionally, mandibular canals with observed cortication border and no contact with mandibular third molar had denser and thicker lingual mandibular canal wall (P = 0.003, P = 0.001 respectively). Conclusions: Buccal and lingual mandibular canal wall density, thickness and mandibular canal diameter may be related with high-risk indicators of inferior alveolar nerve injury.

Salivary levels of hBDs in children and adolescents with type 1 diabetes mellitus and gingivitis.

OBJECTIVES: Type 1 diabetes mellitus (T1DM), a chronic autoimmune disease characterized by insulin deficiency, is related to periodontal diseases in children and adolescents. Our aim was to profile salivary human beta-defensin (hBD)-2 and hBD-3 concentrations in relation to periodontal and T1DM status in children and adolescent populations. MATERIAL AND METHODS: Unstimulated saliva samples were collected from 66 participants including periodontally healthy T1DM patients (T1DM + C; n = 18), T1DM patients with gingivitis (T1DM + G; n = 20), systemically and periodontally healthy individuals (SH + C: n = 15), and systemically healthy gingivitis patients (SH + G; n = 13). Full mouth plaque index (PI), bleeding on probing (BOP), probing pocket depth (PPD), and clinical attachment level (CAL) were recorded. Salivary hBD-2 and hBD-3 concentrations were evaluated by sandwich ELISA method. A p value of < 0.05 was considered statistically significant. RESULTS: Salivary hBD-3 concentrations were lower in T1DM groups in comparison to systemically healthy counterparts (SH + G vs. T1DM + G; p < 0.001 and SH + C vs. T1DM + C; p < 0.001). Salivary hBD-2 levels did not differ between related groups. The difference in hBD-3 concentrations between T1DM and control groups was still significant (p = 0.008) after being adjusted for PI%, BOP%, and age. CONCLUSION: In the limits of study, T1DM patients were found to have decreased salivary hBD-3 concentrations, regardless of their gingival inflammatory status. CLINICAL RELEVANCE: Altered salivary hBD-3 concentration can partly explain why diabetic children are more prone to periodontal diseases.

Evaluation of gingival microcirculation in patients with gestational diabetes mellitus.

OBJECTIVE: The aim of this study was to evaluate the possible microcircularity variations at periodontal mucous level in patients with gestational diabetes mellitus (GDM). MATERIAL AND METHODS: Overall 55 periodontally healthy and non-smoker participants were enrolled in the study by whom 30 were diagnosed with GDM (26 to 34 weeks pregnant) and 25 were systemically healthy unpregnant controls. The analysis was performed in the masticatory/gingival mucosa of maxillary anterior region and by the optical probe videocapillaroscopy technique equipped with 200× lenses. The following parameters were recorded: capillary loop visibility, capillary orientation to surface, microhemorrhages, capillary density and tortuosity. RESULTS: The average capillary density was significantly higher in participants with GDM (27 ± 5.46 no. loops/mm2) compared to controls (21.16 ± 3 no. loops/mm2) (P = 0.035) while increased tortuosity scores was observed in controls compared with the GDM group (P = 0.017). There was not any significantly difference between study groups among the other variables (P > 0.05). CONCLUSIONS: Capillary alterations including capillary density and tortuosity were demonstrated in gingival microcirculation of patients with GDM. These microcirculatory changes could provide us new understanding on the dynamics of the relationship between GDM and periodontal tissues.

Salivary and serum concentrations of monocyte chemoattractant protein-1, macrophage inhibitory factor, and fractalkine in relation to rheumatoid arthritis and periodontitis.

BACKGROUND: Monocyte chemoattractant protein-1 (MCP-1), macrophage migration inhibitory factor (MIF), and fractalkine are chemokines that are expressed by a variety of cell types to regulate macrophage inflammatory response. The aim of the study was to examine the effects of periodontitis and rheumatoid arthritis (RA) on their serum and salivary concentrations. METHODS: Adults with either periodontitis (P, n = 21), or with rheumatoid arthritis (RA, n = 23), or with both diseases (RA+P, n = 23) were included in the study. Systemically and periodontally healthy individuals (n = 22) served as controls. Saliva and serum samples were collected from all participants before the medical and periodontal examinations. Salivary and serum MCP-1, MIF, and fractalkine concentrations were measured by the Luminex technique. Total salivary protein levels were determined by the Bradford assay. RESULTS: Salivary MCP-1, MIF, and fractalkine concentrations were elevated in both RA groups (RA+P and RA) in comparison with systemically healthy controls. As related to total salivary protein levels, higher MCP-1 (P = 0.003) and fractalkine (P = 0.045) concentrations were found in controls compared with the P group. In serum, MCP-1 concentrations in the RA+P group were higher (P = 0.003) than those of group P. Elevated serum fractalkine concentrations were observed in both periodontitis groups (RA+P, P = 0.014; and P, P = 0.013) compared with controls. CONCLUSIONS: In RA, MCP-1, MIF, and fractalkine concentrations are elevated in saliva. These chemokines may disrupt oral macrophage responses and potentially take part in the interaction between periodontitis and RA.

Salivary human beta-defensins and cathelicidin levels in relation to periodontitis and type 2 diabetes mellitus.

Objective: Type 2 diabetes mellitus (T2DM) is a well-defined risk factor of periodontitis and it can affect expression of human beta-defensins (hBDs) and cathelicidin (LL-37) as well. The aim of the present study was to evaluate the impact of periodontitis and T2DM on salivary concentrations of these antimicrobial peptides.Material and methods: Unstimulated saliva samples, together with full-mouth periodontal recordings were collected from 92 individuals with periodontitis (63 with T2DM and 21 smokers) and 86 periodontally healthy controls (58 with T2DM and 21 smokers). Salivary hBD-1, -2, -3, LL-37, and advanced glycalization end products (AGE) concentrations were measured by enzyme-linked immunosorbent assay.Results: Among the periodontitis patients, T2DM group demonstrated lower levels of hBD-1 (p = .006), hBD-2 (p < .001) and hBD-3 (p < .001), and higher levels of LL-37 (p < .001) compared to systemically healthy controls. When only periodontally healthy controls were included into the analysis, higher hBD-1 (p = .002) and LL-37 (p < .001) levels were found in T2DM patients in comparison to systemically healthy controls. Salivary LL-37 levels were associated with HbA1c and periodontitis, while hBD-2, hBD-3 and levels associated only with HbA1c.Conclusion: In the limits of this study, hyperglycaemia can be proposed as a regulator of salivary hBD and cathelicidin levels. Periodontitis, on the other hand, affects only salivary LL-37 levels.

Gingival crevicular fluid levels of human beta-defensin-1 in type 2 diabetes mellitus and periodontitis.

OBJECTIVES: Human ß-defensin (hBD)-1 is an important gatekeeper of the gingiva against constant bacterial challenge, and glucose levels are involved in its optimal expression. The aims of the study were to investigate hBD-1 levels in gingival crevicular fluid (GCF) and to compare these levels between type 2 diabetics with or without periodontitis and healthy individuals. MATERIALS AND METHODS: Altogether, 81 subjects were included in the study: 21 subjects with type 2 diabetes mellitus (T2DM) suffering from generalized periodontitis (T2DM + GP), 18 systemically healthy generalized periodontitis patients (GP), 18 periodontally healthy T2DM subjects (T2DM + H), and 24 systemically and periodontally healthy subjects (control). Plaque index (PI), gingival index (GI), probing pocket depth (PPD), and clinical attachment level (CAL) were recorded, and GCF samples were collected. hBD-1 levels in GCF were measured using ELISA. RESULTS: hBD-1 levels were significantly reduced in the T2DM + GP and GP groups. Although PI and GI scores were similar in both periodontally healthy groups, hBD-1 levels were lower in the T2DM + H group. In the whole population, hBD-1 levels correlated negatively with all periodontal parameters. CONCLUSIONS: Both diabetes and periodontitis affect hBD-1 levels in GCF. CLINICAL RELEVANCE: The altered levels of hBD-1 in GCF of diabetics might be associated with the susceptibility of diabetics to periodontitis.

Analyses of Gingival Adhesion Molecules in Periodontitis: Theoretical In Silico, Comparative In Vivo, and Explanatory In Vitro Models.

BACKGROUND: A deeper understanding of periodontitis pathophysiology is central to future development of novel biomarkers and therapeutics. The following is reported here: 1) an in silico network model of interactions among cell adhesion molecules and a network-focused microarray analysis of the corresponding genes in periodontitis; 2) analysis of secretions of adhesion molecules in gingival tissue samples from patients with periodontitis and healthy controls; and 3) effect of the human neutrophilic peptide-1 (HNP-1) on epithelial adhesion molecules. METHODS: The network model identified 85 nodes in relation to the interactions of adhesion molecules. Subsequently, the relative gene expression was overlaid on the network model. Differential gene expression was analyzed, and false discovery rate control was performed for statistical assessment of the microarray data. Both tissue and cell culture samples were immunostained for desmocollin (DSC)2, occludin (OCLN), desmoglein (DSG)1, tight junction protein 2, and gap junction protein α. RESULTS: The differential gene expression analysis revealed that the epithelial adhesion molecules were significantly lower in abundance in individuals with periodontitis than controls. In contrast, the genes for leukocyte adhesion molecules showed a significant upregulation. Immunostainings revealed elevated secretions of both DSG1 and OCLN in periodontitis. An in vitro model suggested reduced DSC2 and OCLN secretions in the presence of HNP-1. CONCLUSIONS: Gene expression of gingival adhesion molecules in periodontitis is regulated by leukocyte transmigration, whereas the neutrophilic antimicrobial peptide HNP-1 is noted as a putative regulator of epithelial adhesion molecules. These observations contribute to the key mechanisms by which future biomarkers might be developed for periodontitis.

Overexpressions of hBD-2, hBD-3, and hCAP18/LL-37 in Gingiva of Diabetics with Periodontitis.

Antimicrobial peptides of the epithelium play a significant role in the innate immune response in the oral cavity, which is constantly exposed to microbes. Type 2 diabetes mellitus (T2DM) is a highly prevalent metabolic disease which is related to periodontal disease. To date, little is known about expressions of antimicrobial peptides in gingival epithelia of diabetics. Our aim was to examine the expression and localization of human beta-defensins (hBD)-2 and -3 and cathelicidin (hCAP18/LL-37) in diabetic subjects suffering from generalized periodontitis (GP). Gingival tissue sections were collected from three subject groups: 14 T2DM subjects with GP (T2DM+GP), 11 systemically healthy GP patients (GP), and 13 systemically and periodontally healthy subjects (control). Surgical incisions targeted the sulcular epithelium and/or the bottom of the selected periodontal pocket. Tissue specimens were fixed in paraformaldehyde and embedded in paraffin blocks. Immunohistochemistry stainings were performed for cytokeratin19, hBD-2, hBD-3 and hCAP18/LL-37. Stainings were examined under light microscope with 40× magnification. Results were statistically evaluated by the t-test. In controls, hBD-2 was localized at the superficial layers of the gingival epithelium, hBD-3 and hCAP18/LL-37 were at the basal layers, whereas in subjects with periodontitis both defensins were visible at all epithelial layers. hBD-2 was detected in the nucleus and cytoplasm, while hBD-3 and hCAP18/LL-37 were detected only in the cytoplasm of the cells. Expressions of hBD-2 (p=0.005), hBD-3 (p=0.007), and hCAP18/LL-37 (p=0.002) were elevated in subjects with T2DM+GP in comparison to controls. No statistically significant difference was found in the expression of hBD-2, -3, and hCAP18/LL-37 between the GP group and the control or T2DM+GP groups. Gingival antimicrobial peptides are overexpressed in T2DM. This outcome can be part of impaired immune response in diabetics, and underlying factors and mechanisms need to be elucidated.

Salivary Antimicrobial Peptides in Early Detection of Periodontitis.

In the pathogenesis of periodontitis, an infection-induced inflammatory disease of the tooth-supporting tissues, there is a complex interaction between the subgingival microbiota and host tissues. A periodontal diagnostic tool for detecting the initiation and progression of the disease, monitoring the response to therapy, or measuring the degree of susceptibility to future disease progression has been of interest for a long time. The value of various enzymes, proteins, and immunoglobulins, which are abundant constituents of saliva, as potential biomarkers has been recognized and extensively investigated for periodontal diseases. Gingival defensins and cathelicidins are small cationic antimicrobial peptides that play an important role in innate immune response. However, their applicability as salivary biomarkers is still under debate. The present review focuses on proteomic biomarkers and antimicrobial peptides, in particular, to be used at early phases of periodontitis.