Localization and expression profiles of gingival monocyte chemoattractant protein-1-induced protein-1 (MCPIP-1) and mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT-1).

OBJECTIVES: The purposes of this study were to localize monocyte chemoattractant protein-1-induced protein-1 (MCPIP-1) and its suppressor mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT-1) in gingival tissues and to profile their protein expression levels in relation to the clinical inflammation, Porphyromonas gingivalis colonization, and interleukin (IL)-8 levels. MATERIALS AND METHODS: Study samples were collected from two independent study populations: (1) Gingival tissues were collected from eight periodontally healthy individuals and eight periodontitis patients to localize MCPIP-1 and MALT-1 immunohistochemically, and (2) forty-one gingival tissue samples with marginal, mild, or moderate to severe inflammation were collected from 20 periodontitis patients to determine MCPIP-1 and MALT-1 levels using immunoblots, P. gingivalis levels with qPCR, P. gingivalis gingipain activities with fluorogenic substrates, and IL-8 levels with multiplex technique. RESULTS: MCPIP-1 was detectable in the epithelium and in connective tissue, being especially prominent around the blood vessel walls in healthy periodontal tissues. MALT-1 was observed at all layers of gingival epithelium and especially around the accumulated inflammatory cells in connective tissue. No difference in gingival tissue MCPIP-1 and MALT-1 levels was observed in relation to the severity of gingival inflammation. MALT-1 levels were elevated (p = 0.023) with the increase in tissue P. gingivalis levels, and there was an association between MALT-1 and IL-8 levels (ß = 0.054, p = 0.001). CONCLUSIONS: Interactions of MALT-1 levels with gingival tissue P. gingivalis counts and IL-8 levels suggest that activation of MALT-1 can take part in P. gingivalis-regulated host immune responses. CLINICAL RELEVANCE: Pharmacological targeting the crosstalk between immune response and MCPIP-1/MALT-1 may have benefits in periodontal treatment.

Ultrasonographic evaluation of the titanium-prepared platelet-rich fibrin effect in free gingival graft procedures.

BACKGROUND: Complications after free gingival graft (FGG) operations are generally related to the donor site. The titanium-prepared, platelet-rich fibrin (T-PRF) placement in the donor site accelerate the wound healing and prevent postoperative complications such as pain and hemorrhage. We aim to evaluate the effect of T-PRF regarding vascularization and tissue thickness and to report the advantages of the ultrasonography (US) in FGG. METHODS: Ten individuals were divided into two groups as T-PRF and control. While the T-PRF membrane was placed at the donor site in the T-PRF group, a gelatin sponge was placed in the control group. All patients underwent US examination in terms of vascularization and tissue thickness of left and right donor sites. The correlation between the right and left donor sites was analyzed with the Pearson correlation test. Tissue thicknesses and pulsatility index (PI) were analyzed with independent samples t-test. The results were evaluated statistically at the P <0.05 significance level. RESULTS: The T-PRF group showed increased vascularity which can be interpreted to improve healing in soft tissue. However, not a difference, but a positively very high correlation was observed between the right and left tissue thicknesses (P = 0,00; r = +0902). CONCLUSIONS: Evaluation of tissue thickness and vascularization density of donor sites with US not only increases clinical success rate but also reduces the risk of complications during surgery and postoperative pain in FGG. Studies evaluating T-PRF membrane as palatal dressing after FGG are only clinical, however, the efficiency of T-PRF was evaluated radiologically in this study for the first time.

Evaluation of Gene Polymorphism and Gingival Crevicular Fluid Levels of Matrix Metalloproteinase-3 in a Group of Turkish Periodontitis Patients.

INTRODUCTION: Periodontitis is characterized by the destruction of tooth-supporting tissues. Matrix metalloproteinases (MMPs) play a significant part in the degradation of collagen structure. The gingival crevicular fluid (GCF) levels of MMPs increase with the progression of periodontal inflammation. Polymorphisms can be responsible for high expression of MMPs and can exacerbate the breakdown of collagen structure. This study aims to investigate the effect of MMP-3 -1171 5A/6A polymorphism and the GCF levels of MMP-3 in a group of Turkish periodontitis patients. MATERIALS AND METHODS: Non-smoking, stage II grade A periodontitis (S II-Gr A) (n = 68) and stage II grade B periodontitis (S II-Gr C) (n = 64) patients were recruited. Healthy individuals (H) (n = 72) without signs of gingivitis or periodontitis served as the control. Venous blood was collected from participants to obtain DNA, and the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect polymorphism. GCF samples were taken to assess MMP-3 levels using an enzyme-linked immunosorbent assay (ELISA). RESULTS: The MMP-3 -1179 5A/6A distribution showed no significant difference between the groups (p > 0.05). However, the MMP-3 GCF levels of the S II-Gr C group were higher than those of both the S II-Gr A and H groups (p < 0.05), and elevated MMP-3 levels were detected in S II-Gr A compared to H (p < 0.05). CONCLUSION: The MMP-3 GCF levels showed an association with periodontal tissue destruction, although single nucleotide polymorphism was not associated with the S II-Gr C and S II-Gr A groups in the Turkish population.

Periodontitis and peri-implantitis tissue levels of Treponema denticola-CTLP and its MMP-8 activating ability.

BACKGROUND AND AIMS: Chymotrypsin-like-proteinase of Treponema denticola (Td-CTLP) can stimulate the protein expression and activation of matrix metalloproteinase (MMP)-8 (or collagenase-2), a potent tissue destructive enzyme from gingival cells in vitro. The aims of this study were 1) to demonstrate the proMMP-8 (or latent MMP-8) activation by Td-CTLP in vitro and 2) to detect Td-CTLP and MMP-8 protein levels in the tissue samples of peri-implantitis and periodontitis patients. MATERIALS AND METHODS: proMMP-8 activation by Td-CTLP was analyzed by immunoblots. Tissue specimens were collected from 38 systemically healthy and non-smoking patients; 14 of whom had moderate to severe periodontitis, 10 of whom were suffering from peri-implantitis, and finally 14 of whom showed no sign of periodontal inflammation nor radiological bone decay (control group). The immune-expression levels of MMP-8 and Td-CTLP in the epithelium and the connective tissue were analyzed immunohistochemically. A pixel color-intensity analyze was performed with ImageJ software (version 1.46c; Rasband WS, National Institutes of Health, Bethesda, MD, USA) to obtain a comparable numeral score for each patient's epithelium and connective tissue MMP-8 and Td-CTLP enzyme level. RESULTS: Td-CTLP activated proMMP-8 in vitro by converting the 70-75 kDa proMMP-8 to 65 kDa active MMP-8. Also, lower molecular size 25-50 kDa parts of MMP-8 were formed. There was no statistically significant difference between the study groups in terms of their MMP-8 and Td-CTLP levels in the epithelium or in the connective tissue. CONCLUSION: Regarding the limits of this study, it can thus be said that the Td-CTLP enzyme can activate the host proMMP-8 enzyme. Tissue protein levels of MMP-8 and Td-CTLP do not seem to be changed in peri-implantitis and in periodontitis.

A review with an additional case: amelanotic malignant melanoma at mandibular gingiva.

The purpose of this review with an additional case is to evaluate the clinical, ultrasonographic and histopathological features of a rare case of Amelanotic Malignant Melanoma (AMM) at mandibular gingiva and to compare our case with other published AMMs at mandibular gingiva. A 52-year-old male patient with no systemic diseases was referred to our clinic with a soft tissue lesion at mandibular gingiva. Ultrasonographic examination was performed and a lesion with malignant features was observed. A periapical radiograph was taken to investigate bone destruction and biopsy was planned. Histopathological examination revealed AMM and a literature search was performed to congregate reports which were indexed in PubMed, ScienceDirect, and ResearchGate. Three AMM cases at mandibular gingiva were found. Doppler Ultrasound examination suggested bone destruction and a 1.8 cm × 0.6 cm soft tissue mass with well-defined borders and increased vascularity. Due to its hypervascularity, depth of invasion and destruction at the bone, the lesion was prediagnosed as a malignancy. Lack of melanin pigmentation caused the large immunohistochemical panel study. The tumour cells showed HMB45 and S100 positivity and they were negative with SMA, Desmin, CK1.3, and CK20. Routine ultrasound examination of all soft tissue lesions is very important for assessing features such as vascularity, bone destruction and depth of invasion to detect malignancy. Melanocytic-associated immunohistochemical markers are crucial for AMM diagnosis.

The Prevalence of Periodontal Pathogenic Bacteria in Atherosclerotic Cardiovascular Disease.

BACKGROUND: A possible link between periodontal pathogenic bacteria and atherosclerosis may exist based on the inflammatory mechanisms initiated by bacteria found in periodontal lesions. Our aim was to investigate the presence of DNA originating from T. denticola, C. rectus, T. forsythia, and P. gingivalis in the vascular tissue specimens obtained from patients who underwent surgery for arteriosclerotic vascular disease in this study. METHODS: A total of 96 patients diagnosed with valvular heart disease due to atherosclerosis and 85 patients with advanced aortic valve stenosis due to rheumatic fever and had undergone aortic valve replacement were included as the study (PG) and the control groups (CG), respectively. Atheroma plaques and vascular tissue specimens were collected from PG and CG during cardiovascular surgical procedures. Revitalization of the lyophilized T. denticola, ATCC 35405; C. rectus, ATCC 33238; P. gingivalis, ATCC 33277 and T. forsythia, ATCC 43037 strains was performed according to the manufacturer's instructions. C. rectus, T. forsythia, and T. denticola DNA samples were analyzed using the one-step in-house PCR method. RESULTS: In one (1.04%) and three (3.13%) out of 96 atherosclerotic PG tissue specimens, P. gingivalis and T. for-sythia DNA were detected, respectively. No T. denticola or C. rectus DNA was found in the study specimens. Periodontal pathogenic bacteria were not observed in 85 CG tissue specimens. There was no statistically significant difference between PG and CG for the presence of P. gingivalis and T. forsythia DNA using Fischer's Exact test (p > 0.05). CONCLUSIONS: In conclusion, with the case-control studies on a small scale such as in our study, it is not possible to determine a causality relationship between periodontal pathogenic bacteria and formation of atherosclerosis. Periodontal pathogenic bacteria may not be the only factor that causes inflammatory diseases associated with atherosclerosis. Host response and inflammatory mechanisms may be affected by other factors such as ethnicity, dietary habits, nutritional availability, and lifestyle. Taken together, it is difficult to conclude a causal link between periodontal pathogenic bacteria and formation of atherosclerosis.

Preference of Suture Specifications in a Selected Periodontal and Implant Surgeries in Turkey.

OBJECTIVE: Various suture materials and needles are now available for use in the dental surgery. The aim of this study was to determine the preference of suture materials among Turkish dentists by a dental survey. MATERIALS AND METHODS: The survey was prepared and sent electronically to Turkish dentists through e-mail and/or Facebook. Dentists were asked to report their graduation year from dental school and their specialty if they have one. In addition, the type periodontal/implant operations and the frequency of those operations applied by them were questioned. The participants were to indicate their suture preferences for these procedures in a multiple-choice questionnaire. RESULTS: Fifty-seven regular dentists, 49 periodontists, 22 oral surgeons, and 8 other specialists completed a self-administered survey. The majority of clinicians worked in private practice (77.9%). Nonabsorbable sutures were the most preferred for all procedures except periodontal plastic surgery. In regenerative surgeries, monofilament, 5-0 diameter suture material on a reverse cutting, 3/8 circle needle was preferred. In addition, for mucogingival surgery, 5-0 diameter suture material on a reverse cutting and 3/8 circle needle was favored. For dental implants, 4-0 diameter suture material on a reverse cutting and 3/8 circle needle was preferred. Monofilament and braided sutures were selected almost equally for implant operations. CONCLUSIONS: In periodontal and implant surgeries, dentists highly preferred the use of nonabsorbable sutures. In addition, the shape and diameter of needle had an important role in the selection of suture material. The present study's results may serve as a guide for the future studies.

Regulatory effects of PRF and titanium surfaces on cellular adhesion, spread, and cytokine expressions of gingival keratinocytes.

Dental implant material has an impact on adhesion and spreading of oral mucosal cells on its surface. Platelet-rich fibrin (PRF), a second-generation platelet concentrate, can enhance cell proliferation and adhesion. The aim was to examine the regulatory effects of PRF and titanium surfaces on cellular adhesion, spread, and cytokine expressions of gingival keratinocytes. Human gingival keratinocytes were cultured on titanium grade 4, titanium grade 5 (Ti5), and HA discs at 37 °C in a CO2 incubator for 6 h and 24 h, using either elutes of titanium-PRF (T-PRF) or leukocyte and platelet-rich fibrin (L-PRF), or mammalian cell culture medium as growth media. Cell numbers were determined using a Cell Titer 96 assay. Interleukin (IL)-1ß, IL-1Ra, IL-8, monocyte chemoattractant protein (MCP)-1, and vascular endothelial growth factor (VEGF) expression levels were measured using the Luminex® xMAP™ technique, and cell adhesion and spread by scanning electron microscopy. Epithelial cell adhesion and spread was most prominent to Ti5 surfaces. L-PRF stimulated cell adhesion to HA surface. Both T-PRF and L-PRF activated the expressions of IL-1 ß, IL-8, IL-1Ra, MCP-1, and VEGF, T-PRF being the strongest activator. Titanium surface type has a regulatory role in epithelial cell adhesion and spread, while PRF type determines the cytokine response.

NFE2L2/NRF2, OGG1, and cytokine responses of human gingival keratinocytes against oxidative insults of various origin.

OBJECTIVE: Bacterial or tobacco-related insults induce oxidative stress in gingival keratinocytes. The aim of this study was to investigate anti-oxidative and cytokine responses of human gingival keratinocytes (HMK cells) against Porphyromonas gingivalis lipopolysaccharide (Pg LPS), nicotine, and 4-nitroquinoline N-oxide (4-NQO). MATERIALS AND METHODS: HMK cells were incubated with Pg LPS (1 µl/ml), nicotine (1.54 mM), and 4-NQO (1 µM) for 24 h. Intracellular and extracellular levels of interleukin (IL)-1ß, IL-1 receptor antagonist (IL-1Ra), IL-8, monocyte chemoattractant protein (MCP)-1, and vascular endothelial growth factor (VEGF) were measured with the Luminex® xMAP™ technique, and nuclear factor, erythroid 2 like 2 (NFE2L2/NRF2) and 8-oxoguanine DNA glycosylase (OGG1) with Western blots. Data were statistically analyzed by two-way ANOVA with Bonferroni correction. RESULTS: All tested oxidative stress inducers increased intracellular OGG1 levels, whereas only nicotine and 4-NQO induced NFE2L2/NRF2 levels. Nicotine, 4-NQO, and their combinational applications with Pg LPS induced the secretions of IL-1ß and IL-1Ra, while that of IL-8 was inhibited by the presence of Pg LPS. MCP-1 secretion was suppressed by nicotine, alone and together with Pg LPS, while 4-NQO activated its secretion. Treatment of HMK cells with Pg LPS, nicotine, 4-NQO, or their combinations did not affect VEGF levels. CONCLUSION: Pg LPS, nicotine, and 4-NQO induce oxidative stress and regulate anti-oxidative response and cytokine expressions in human gingival keratinocytes differently. These results may indicate that bacterial and tobacco-related insults regulate distinct pathways.

Pathogen profile and MMP-3 levels in areas with varied attachment loss in generalized aggressive and chronic periodontitis.

INTRODUCTION: The progression of periodontitis depends on the changes in bone and connective tissue homeostasis and the imbalance of the biofilm and the host immunoinflammatory response, particularly matrix metalloproteinases (MMP). AIM OF THE STUDY: To assess the probable relation between subgingival anaerobic flora and the expression of MMP-3 in patients with generalized aggressive periodontitis (AgP), chronic periodontitis (CP) and healthy subjects, and to evaluate these levels according to varied tissue loss severity. MATERIAL AND METHODS: The plaque index (PI), gingival index (GI), probing depth (PD) and clinical attachment levels (CAL) were evaluated. MMP levels obtained from gingival sulcus fluid (GCF) were measured with Enzyme Linked Immuno Assay (ELISA). The bacterial counts were determined with Parocheck®. RESULTS: Higher levels of MMP-3 in patients with AgP compared to subjects with CP and healthy individuals were observed. The microorganisms responsible of possible tissue destruction in both AgP and CP are red complex bacteria. T. denticola, T. forsythia, P. intermedia and F. nucleatum show positive correlation with MMP-3 levels. CONCLUSIONS: MMP-3 is a biomarker associated with AgP, and red complex bacteria levels are correlated with increasing periodontal tissue loss in both periodontitis forms. The diagnosis of aggressive periodontitis, or site-specific treatment strategies can be orchestrated based on the evaluation of MMP-3 and the bacterial counts in patients with periodontitis.

Elevated levels of 8-OHdG and PARK7/DJ-1 in peri-implantitis mucosa.

BACKGROUND: Reactive oxygen species contribute to periodontal tissue homeostasis under control of anti-oxidative responses. Disruption in this balance induces severe inflammation and extended tissue degradation. PURPOSE: Aim of this study was to identify the expression levels of nuclear factor, erythroid 2 like 2 (NFE2L2/NRF2), Parkinsonism associated deglycase (PARK7/DJ-1), kelch-like ECH associated protein 1 (KEAP1), and 8-hydroxy-deoxyguanosine (8-OHdG) in peri-implant mucosal tissues affected by peri-implantitis, and to compare the levels to those of periodontally diseased and healthy tissue samples. METHODS: Tissue biopsies were collected from systemically healthy, non-smoking 12 peri-implantitis patients, 13 periodontitis patients, and 13 periodontally healthy controls. Expression levels of NFE2L2/NRF2, PARK7/DJ-1, KEAP1, and 8-OHdG in tissue samples were analyzed immunohistochemically. Statistical analysis was performed by one-way ANOVA with Tukey's HSD test. RESULTS: Inflammatory cell infiltration in the connective tissue and loss of architecture in the spinous layer of the epithelium were prominent in peri-implantitis. Proportions of 8-OHdG and PARK7/DJ-1 expressing cells were elevated in both peri-implantitis (P = .025 for 8-OHdG and P = .014 for PARK7/DJ-1) and periodontitis (P = .038 for 8-OHdG and P = .012 for PARK7/DJ-1) groups in comparison with controls. Staining intensities of 8-OHdG and PARK7/DJ-1 were higher in the periodontitis and peri-implantitis groups than in the control (P < .01) groups. There was no difference in the expression levels of NFE2L2/NRF2 between the groups. KEAP1 was not observed in any tissue sample. CONCLUSIONS: Peri-implantitis is characterized by severe inflammation and architectural changes in the epithelium and connective tissue. The expressions of 8-OHdG and PARK7/DJ-1 are elevated in both peri-implantitis and periodontitis.