Innovative i-PRF semisurgical method for gingival augmentation and root coverage in thin periodontal phenotypes: a preliminary study.

OBJECTIVES: The aim of the study was to evaluate the effect of injectable platelet-rich fibrin (i-PRF) on gingival thickness and gingival recession in individuals with thin periodontal phenotypes. METHOD AND MATERIALS: In this prospective study, i-PRF was applied via a semisurgical method to augment 53 tooth regions with thin periodontal phenotypes. In order to ensure that sufficient blood clot formed on the side of the gingiva facing the bone and that i-PRF reached the area, a minimal incision was made with the help of a scalpel in the apical region of the relevant region, and the periosteum was elevated with a microsurgical instrument. To ensure sustained exposure to angiogenetic growth factors and enhance the histoconductive properties, i-PRF injection was applied to the relevant areas in four sessions at 10-day intervals. RESULTS: An increase in gingival thickness was achieved in 92.5% of the areas treated with i-PRF, and the desired gingival thickness (0.8 mm) was achieved in 44.9% of these areas. In addition, significant reductions in the amount of recession were observed in 83.3% of the 12 gingival recession areas (P = .005). Moreover, complete coverage was achieved in 60% of these regions. CONCLUSION: With the new i-PRF semisurgical method, it was shown that gingival thickness can be increased in tooth regions with thin gingiva, and that areas of gingival recession can be covered. Further comprehensive studies are needed to fully understand the role of i-PRF in enhancing angiogenesis and the histoconductive properties of this fully autogenous blood concentrate.

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.

Salivary levels of BAFF, TWEAK, and soluble CD163 and salivary arginase activity before and after periodontal treatment.

OBJECTIVE: To monitor salivary B-cell activating factor (BAFF), tumor necrosis factor-like weak inducer of apoptosis (TWEAK), and soluble (s)CD163 levels and arginase activity in periodontitis patients following nonsurgical periodontal treatment. BACKGROUND: BAFF, TWEAK, and sCD163 and arginase are associated with activities of B cells and macrophages, which are important regulators of periodontal immune-inflammatory response and healing following treatment. Increased salivary BAFF and sCD163 levels and arginase activity in periodontitis have been demonstrated, but their changes following treatment have not been evaluated before. MATERIALS AND METHODS: Forty-four Stage III/IV periodontitis patients and 35 periodontally healthy controls were included in the study. Full-mouth periodontal measurements were recorded and unstimulated saliva was obtained from all participants at baseline. Sample collection and measurements were repeated in periodontitis patients at 2, 6, 12, and 24 weeks following full-mouth scaling and root debridement, whereas controls were only seen at baseline. BAFF, TWEAK, and sCD163 levels were analyzed with bead-based multiplexed immunoassay. Arginase activity was measured with Chinard's method. RESULTS: BAFF (p < .001) and sCD163 (p = .003) levels and arginase activity (p < .015) were higher in periodontitis patients compared to healthy controls. BAFF levels (p < .001) and arginase activity (p < .001) of periodontitis patients were reduced at 2 weeks posttreatment and continued to decrease up to 6 (p = .038) and 12 weeks (p = .024), respectively. The reduction of sCD163 levels became significant (p = .003) at 24 weeks posttreatment. CONCLUSIONS: The decrease in salivary BAFF levels 2 weeks after periodontal treatment indicates a change in cell signaling toward limited B-cell activation. Decreasing arginase activity similarly reflects a significant reduction in inflammatory response. The reduction in sCD163 levels that are observed at 24 weeks may reflect a longstanding anti-inflammatory macrophage activation, given their multiple functions in immune response, inflammation, and healing.

Baseline interleukin-10, CD163, and tumor necrosis factor-like weak inducer of apoptosis (TWEAK) gingival tissue levels in relation to clinical periodontal treatment outcomes: A 12-week follow-up study.

BACKGROUND: The aim of this study was to examine the relationship between healing response after non-surgical periodontal treatment and baseline gingival tissue levels of M2 macrophage activation-related proteins CD163, interleukin (IL)-10, interferon (IFN)-γ, and tumor necrosis factor-like weak inducer of apoptosis (TWEAK), and the CD163/TWEAK ratio. METHODS: Eighty-eight gingival tissue samples from 44 Stage III/IV, Grade C periodontitis patients (18 smokers) and 41 tissue samples from 41 periodontally healthy participants (18 smokers) were evaluated. Clinical parameters were recorded in periodontally healthy individuals at baseline and in periodontitis patients at pre-treatment and 2, 6, and 12 weeks following therapy. IL-10, IFN-γ, CD163, and TWEAK levels were analyzed with Luminex technique. RESULTS: Tissue levels (median, 1st -3rd quartile) of IL-10 (pg/ng protein), CD163 (pg/µg protein) and TWEAK (pg/µg protein) were as follows: IL-10 periodontitis: 2.08, 0.86-5.32 and periodontally healthy: 5.22, 3.20-10.25; CD163 periodontitis: 8.85, 4.92-14.06 and periodontally healthy: 18.36, 12.51-34.02; TWEAK periodontitis: 0.08, 0.05-0.11 and periodontally healthy: 0.16, 0.12-0.21. IL-10, CD163, and TWEAK levels were higher (P < 0.001) in periodontally healthy tissues than in periodontitis tissues. Pocket closure at 12 weeks was associated with elevated baseline gingival CD163 levels (P = 0.047) and CD163/TWEAK ratio (P = 0.001). Elevated baseline gingival CD163/TWEAK ratio was associated with pocket reduction at 6 (P = 0.022) and 12 weeks (P = 0.002). CONCLUSION: Associations of pocket closure with pre-treatment gingival tissue CD163 levels and CD163/TWEAK ratio indicate that baseline M2 macrophage activation profile may play a role in periodontal wound healing.

Regulation of gingival keratinocyte monocyte chemoattractant protein-1-induced protein (MCPIP)-1 and mucosa-associated lymphoid tissue lymphoma translocation protein (MALT)-1 expressions by periodontal bacteria, lipopolysaccharide, and interleukin-1ß.

BACKGROUND: The aim of this study was to evaluate oral bacteria- and interleukin (IL)-1ß-induced protein and mRNA expression profiles of monocyte chemoattractant protein-1-induced protein (MCPIP)-1 and mucosa-associated lymphoid tissue lymphoma translocation protein (MALT)-1 in human gingival keratinocyte monolayers and organotypic oral mucosal models. METHODS: Human gingival keratinocyte (HMK) monolayers were incubated with Porphyromonas gingivalis, Fusobacterium nucleatum, P. gingivalis lipopolysaccharide (LPS) and IL-1ß. The protein levels of MCPIP-1 and MALT-1 were examined by immunoblots and mRNA levels by qPCR. MCPIP-1 and MALT-1 protein expression levels were also analyzed immunohistochemically using an organotypic oral mucosal model. One-way analysis of variance followed by Tukey correction was used in statistical analyses. RESULTS: In keratinocyte monolayers, MCPIP-1 protein expression was suppressed by F. nucleatum and MALT-1 protein expression was suppressed by F. nucleatum, P. gingivalis LPS and IL-1ß. P. gingivalis seemed to degrade MCPIP-1 and MALT-1 at all tested time points and degradation was inhibited when P. gingivalis was heat-killed. MCPIP-1 mRNA levels were increased by P. gingivalis, F. nucleatum, and IL-1ß, however, no changes were observed in MALT-1 mRNA levels. CONCLUSION: Gingival keratinocyte MCPIP-1 and MALT-1 mRNA and protein expression responses are regulated by infection and inflammatory mediators. These findings suggest that periodontitis-associated bacteria-induced modifications in MCPIP-1 and MALT-1 responses can be a part of periodontal disease pathogenesis.

Tissue Levels of CD80, CD163 and CD206 and Their Ratios in Periodontal and Peri-Implant Health and Disease.

This study aimed to compare tissue levels of CD80 (pro-inflammatory macrophage-related surface marker), CD163, and CD206 (anti-inflammatory macrophage-related surface markers), and their ratios in periodontal and peri-implant health and disease. Altogether, 36 tissue samples were obtained from 36 participants with clinically healthy gingiva (n = 10), healthy peri-implant mucosa (n = 8), periodontitis lesions (n = 9), and peri-implantitis lesions (n = 9). CD80, CD163, and CD206 levels were assessed with immunoblotting. CD163 levels were found to be decreased (p = 0.004), and the CD80/CD163 ratio was found to be elevated (p = 0.002) in periodontitis lesions compared to healthy gingiva. Peri-implantitis lesions showed a tendency towards a higher CD80/CD163 ratio than in healthy peri-implant mucosa with a borderline difference (p = 0.054). No statistically significant difference was detected in CD80, CD163, and CD206 levels of periodontitis lesions when compared to peri-implantitis, and in healthy gingiva when compared to healthy peri-implant mucosa. A disruption in CD80/CD163 balance seems to be related to the pathogenesis of periodontitis and peri-implantitis, being less prominent in the latter. The reason behind this phenomenon may be either suppressed CD163 expression or reduced CD163+ anti-inflammatory macrophage abundance.

Analysis of Biomarkers and Marginal Bone Loss in Platform-Switched and Nonplatform-Switched Implants: A Randomized Clinical Trial.

Objectives: To compare the peri-implant crevicular fluid (PICF) biomarker levels, peri-implant status, and marginal bone level (MBL) differences of implants restored with randomly assigned nonplatform-switched (NPS) or platform-switched (PS) abutments. Methods: Ninety-four implants in 27 subjects were included in this study. Receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), interleukin-1ß (IL-1ß), monocyte chemotactic protein-1 (MCP-1) levels in PICF, peri-implant health, and the change in the MBL were evaluated at the time of restoration (T 1) and after 12 months (T 2). Results: The IL-1ß levels decreased and the RANKL, OPG, and MCP-1 levels increased from T 1 to T 2 (P < 0.05) in both groups. RANKL/OPG ratio at T 1, MCP-1 levels at T 2, and the MCP-1 change from T 1 to T 2 were lower in the PS group than in the NPS group (P < 0.05). MBL change was lower (0.51 ± 0.31 mm) in the PS group than that (0.75 ± 0.29 mm) in the NPS group at T 2 (P < 0.001). Peri-implant health status between the study groups was negligible. Conclusion: PS was superior to NPS regarding the preservation of MBL. Higher MCP-1 levels, altered RANKL/OPG ratio, and lower OPG levels in the NPS group could be associated with subclinical peri-implant bone remodeling.

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.

Resolvin E1 Regulates Th17 Function and T Cell Activation.

Resolvin E1 (RvE1) is a specialized pro-resolving lipid mediator derived from eicosapentaenoic acid and plays a critical role in resolving inflammation and tissue homeostasis. Th17 cells are a distinct group of T helper (Th) cells with tissue-destructive functions in autoimmune and chronic inflammatory diseases via the secretion of IL-17. Dendritic cell (DC)-mediated antigen presentation regulates the Th17-induced progression of inflammation and tissue destruction. In this study, we hypothesized that the RvE1 would restore homeostatic balance and inflammation by targeting the Th17 function. We designed three experiments to investigate the impact of RvE1 on different phases of Th17 response and the potential role of DCs: First CD4+ T cells were induced by IL-6/TGFß to measure the effect of RvE1 on Th17 differentiation in an inflammatory milieu. Second, we measured the impact of RvE1 on DC-stimulated Th17 differentiation in a co-culture model. Third, we measured the effect of RvE1 on DC maturation. RvE1 blocked the CD25, CCR6 and IL-17 expression; IL-17, IL-21, IL-10, and IL-2 production, suggesting inhibition of T cell activation, Th17 stimulation and chemoattraction. RvE1 also suppressed the activation of DCs by limiting their pro-inflammatory cytokine production. Our findings collectively demonstrated that the RvE1 targeted the Th17 activation and the DC function as a potential mechanism for inflammatory resolution and acquired immune response.

Periodontitis Prevalence, Severity, and Risk Factors: A Comparison of the AAP/CDC Case Definition and the EFP/AAP Classification.

BACKGROUND: This cross-sectional study evaluated the utility of the 2018 European Federation of Periodontology/American Academy of Periodontology (EFP/AAP) classifications of epidemiological studies in terms of periodontitis severity, prevalence and associated risk factors and the 2012 American Academy of Periodontology/Centers for Disease Control and Prevention (AAP/CDC) case definitions. METHODS: We included 488 participants aged 35-74 years. Measurements were recorded at six sites per tooth by two qualified examiners. The evaluated parameters included pocket depth (PD), clinical attachment loss (CAL) and bleeding on probing (BOP). Periodontitis prevalence and severity were reported using the 2018 EFP/AAP classification and the AAP/CDC case definitions. The data were stratified by recognized risk factors (age, diabetes and smoking status). RESULTS: The 2018 EFP/AAP classification indicated that all patients suffered from periodontitis. When CAL served as the main criterion, the frequency of patients with severe (Stages III-IV) periodontitis was 54%. When the AAP/CDC case definitions were applied, the prevalence of periodontitis was 61.9% and that of severe periodontitis 16.8%. Age was the most significant risk factor, regardless of the chosen case definition. CONCLUSION: It is essential to employ a globalized standard case definition when monitoring periodontitis and associated risk factors.

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.

Wetting properties of blood lipid fractions on different titanium surfaces.

BACKGROUND: Blood is the first tissue contacting the implant surface and starting the biological interactions to enhance osseointegration and stimulate bone formation with the progenitor cytokines, chemokines, and growth factors. The coagulation cascade initiates the first step of osseointegration between implant and neighboring tissues. The wound healing may be inadequate unless the blood wets the implant surface properly. Wettability is one of the most important features of the implant surface while lipid level constitutes a milestone that may change the energy of blood, which determines its distribution on implant material. Thus, the aim of this study was to evaluate the effect of lipid component of blood as cholesterol and its treatment on their wetting behavior of titanium surfaces. METHODS: Five surface groups were formed including grade 4 titanium-machined, grade 4 titanium-SLA, grade 4 titanium-SLActive, Roxolid-SLA, and Roxolid-SLActive. In healthy, hyperlipidemic, and treatment situations, blood was taken from eight rabbits and dropped to the disc surfaces. Contact angles were measured between the blood samples and disc surfaces. RESULTS: A significant difference was found between both machined and SLActive surfaces, SLA and SLActive surfaces in the hyperlipidemic period, and only Roxolid-SLA and SLActive surfaces during the treatment period. When evaluated according to time, only grade 4-machined and Grade 4-SLA surfaces showed a significant difference. CONCLUSIONS: Our findings indicated that each period has its own characteristics and showed the importance of cholesterol in blood structure on applicability of implant surfaces.

Investigation of the effects of semaphorin 3A on new bone formation in a rat calvarial defect model.

PURPOSE: This study investigates the effects of semaphorin 3A on new bone formation in an experimental rat model. MATERIALS AND METHODS: Cortical bone defects, 5 mm, were created in the calvaria of 40 Wistar rats, which were then separated into three groups: empty defect (control) group, collagen group, collagen + semaphorin 3A group. The bone blocks were harvested after 4 and 8 weeks. New bone formation was assessed by micro-computed tomography (micro-CT), histology, histomorphometry, transmission electron microscope (TEM) and immunohistochemistry. RESULTS: Increased bone formation was observed in collagen + semaphorin 3A groups both histologically and with micro-CT. In the histomorphometic analysis, the control group had significantly less bone formation compared to both the collagen and collagen + semaphorin 3A group at 4 weeks (p = 0.0001) and 8 weeks (p = 0.0001). The collagen group had significantly less bone formation compared to collagen + semaphorin 3A group both at 4 weeks (p = 0.002) and 8 weeks (p = 0.005). Immunohistochemical analysis revealed that semaphorin 3A inhibited receptor activator of nuclear factor-kB ligand (RANKL) expression and increased the expressions of osteoblastic bone markers at 4 weeks. In TEM analysis, the collagen + semaphorin 3A group had an increased proliferation and bone formation rate at 4 weeks, whereas bone quantity and maturation were enhanced at 8 weeks. CONCLUSION: Locally applied semaphorin 3A increases callus formation at 4 weeks and bone formation at 8 weeks. Semaphorin 3A prevents bone resorption by inhibiting osteoclasts and increases bone formation by inducing osteoblasts.

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.

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.

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.

The effect of low-level laser therapy on tooth movement during canine distalization.

The aim of the study is to determine the effects of low level laser therapy on tooth movement during canine distalization by evaluating IL-1ß, TGF-ß1 levels in gingival crevicular fluid. Maxillary first premolars of the 15 Angle Class II division I patients (12-19 years old) were extracted. Right maxillary canines were distalized by standard protocol as control group whereas the left maxillary canines distalized by laser application. A gallium-aluminum-arsenide diode laser with an output power of 20 mW was applied as five doses from the buccal and the palatal side on the day 0, and the 3rd, 7th, 14th, 21th 30th, 33st, 37th, 60th, 63th, and 67th days. Gingival crevicular fluid samples were obtained with filtration paper at the initial, 7th, 14th, and 21th days, and the IL-1ß and TGF-ß1 cytokine levels were analyzed. Orthodontic models and periodontal indices were taken initially and on the days 30th, 60th, and 90th of canine distalization period. Tooth movement was assessed by scanning models (3Shape). The amount of tooth movement in the laser group was 40% more than the control group. First day IL-1ß levels were statistically higher than initial and 21st day levels (P= 0.003, P = 0.012). The rise in IL-1ß levels caused the negative correlations between 7th day IL-1ß and 21st day TGF-ß1 levels describes the tissue effects of laser application. Periodontal indices showed no sign of gingival inflammation during canine distalization period. As conclusion, laser does accelerate tooth movement and could shorten the whole treatment duration.

Prolonged pharmacological inhibition of cathepsin C results in elimination of neutrophil serine proteases.

Cathepsin C (CatC) is a tetrameric cysteine dipeptidyl aminopeptidase that plays a key role in activation of pro-inflammatory serine protease zymogens by removal of a N-terminal pro-dipeptide sequence. Loss of function mutations in the CatC gene is associated with lack of immune cell serine protease activities and cause Papillon-Lefèvre syndrome (PLS). Also, only very low levels of elastase-like protease zymogens are detected by proteome analysis of neutrophils from PLS patients. Thus, CatC inhibitors represent new alternatives for the treatment of neutrophil protease-driven inflammatory or autoimmune diseases. We aimed to experimentally inactivate and lower neutrophil elastase-like proteases by pharmacological blocking of CatC-dependent maturation in cell-based assays and in vivo. Isolated, immature bone marrow cells from healthy donors pulse-chased in the presence of a new cell permeable cyclopropyl nitrile CatC inhibitor almost totally lack elastase. We confirmed the elimination of neutrophil elastase-like proteases by prolonged inhibition of CatC in a non-human primate. We also showed that neutrophils lacking elastase-like protease activities were still recruited to inflammatory sites. These preclinical results demonstrate that the disappearance of neutrophil elastase-like proteases as observed in PLS patients can be achieved by pharmacological inhibition of bone marrow CatC. Such a transitory inhibition of CatC might thus help to rebalance the protease load during chronic inflammatory diseases, which opens new perspectives for therapeutic applications in humans.