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Supplemental Material, sj-pdf-1-cmt-10.1177_19433875211027694 for Current Practice for Transverse Mandibular and Maxillary Discrepancies in the Netherlands: A Web-Based Survey Among Orthodontists and Oral and Maxillofacial Surgeons by Atilla Gül, Stephen T. H. Tjoa, Jan P. de Gijt, Justin T. van der Tas, Hadi Sutedja, Eppo B. Wolvius, Karel G. H. van der Wal and Maarten J. Koudstaal in Craniomaxillofacial Trauma & Reconstruction

Objectives To assess validity and reliability of Turkish oral hygiene-related self-efficacy (OHSE) scale, and the scores of general self-efficacy (GSE) and OHSE in the patients with gingivitis and chronic periodontitis (CP). Additionally, the effect of motivation and oral hygiene instructions on the OHSE scores and plaque index (PI) was also aimed. Materials and methods GSE and OHSE scores were obtained from 210 patients with gingivitis and CP (Group A). After determination of "cut-off" values on OHSE scores, patients with OHSE scores = 50 (OHSE-high) were included in Group B (gingivitis, N = 25; CP, N = 27). PI values were determined, and motivation and oral hygiene instructions were performed in Group B at baseline. PI measurements and OHSE questionnaire were repeated at 1 and 3 months. Results the intraclass reliability coefficient and Cronbach's alpha for Turkish translation of OHSE scale were 0.920 and 0.911. OHSE scores significantly correlated with GSE scores and smokers had lower OHSE scores than those of non-smokers in patients (n = 210). in both gingivitis and CP groups, OHSE scores increased and PI values decreased at 3 months. PI values were lower in gingivitis and CP groups with OHSE-high scores compared to patients with OHSE-low scores at 3 months. Conclusions Within the limitations of this study, it seems that Turkish version of OHSE questionnaire is reliable and valid. Patients with OHSE-low scores in both gingivitis and CP groups were less successful in performing daily plaque control, compared to patients with OHSE-high scores., Ege University Research FoundationEge University [2015-DIS-004], This study was supported by a grant from Ege University Research Foundation (2015-DIS-004).

Objectives: hCAP18/LL-37 is an endogenous antibiotic having a role in innate immunity. The aim of the present study was to evaluate serum and gingival crevicular fluid (GCF) hCAP18/LL-37 levels in patients with generalized aggressive periodontitis (G-AgP). Materials and methods: Twenty-six G-AgP patients, 24 gingivitis patients, and 25 healthy subjects were included in this study. Periodontal parameters including probing depth, clinical attachment level, plaque index, and papilla bleeding index were recorded. GCF and serum hCAP18/LL-37 levels were analyzed by enzyme-linked immunosorbent assay. Results: GCF hCAP18/LL-37 level was significantly higher in G-AgP compared to others (p = 0.038, p 0.05). Conclusion: Increased levels of GCF hCAP18/LL-37 in G-AgP might show that it is abundantly expressed in the presence of periodontal tissue destruction. Serum hCAP18/LL-37 levels do not seem to be related with the presence of G-AgP. Clinical relevance: hCAP18/LL-37 antimicrobial peptide might be associated with periodontal tissue destruction in the presence of aggressive periodontitis. © 2016, Springer-Verlag Berlin Heidelberg.

Platelet-rich fibrin (PRF) has a controlled release of growth factors due to the fibrin matrix structure. Different centrifugation protocols were suggested for PRF preparation. Since the derivation method of PRF can alter its contents, in the present study it is aimed to investigate the cell contents and transforming growth factor beta-1 (TGF-β1), platelet-derived growth factor (PDGF-AB), vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-1 and-8 release from experimental PRF-type membranes obtained with different centrifugation times at 400 gravity. Three blood samples were collected from 20 healthy non-smoker volunteers. One tube was used for whole blood analyses. The other two tubes were centrifuged at 400 g for 10 minutes (group A) or 12 minutes (group B). Each experimental PRF-type membrane was placed in Dulbecco's Modified Eagle's Medium (DMEM)and at 1, 24 and 72 hours, TGF-β1, PDGF-AB, VEGF, MMP-1 and -8 release amounts were analysed by enzyme-linked immunosorbent assay (ELISA). The blood cell count of membranes was determined by subtracting plasma supernatant and red blood cell (RBC) mixture from the whole blood cell counts. At 72 hours, the VEGF level of group B was statistically higher than that of group A (p = 0.040). The centrifugation time was not found to influence the release of other growth factors, enzymes and cell counts. Within the limits of the present study, it might be suggested that centrifugation time at a constant gravity has a significant effect on the VEGF levels released from experimental PRF-type membrane. It can be concluded that due to the importance of VEGF in the tissue healing process, membranes obtained at 12-minute centrifugation time may show a superior potential in wound healing.

LL-37 contributes to maintaining the balance between health and disease. Smoking is a risk factor for periodontitis that impairs neutrophil functions. The aim of the present study was to comparatively evaluate gingival crevicular fluid (GCF) LL-37 levels in smoker and non-smoker chronic periodontitis (CP) patients and controls, as well as the effect of non-surgical periodontal treatment on GCF LL-37 levels.Thirty-one CP patients (16 smokers, 15 non-smokers) and thirty-one controls (16 smokers, 15 non-smokers) were included in the study. CP patients received non-surgical treatment. GCF LL-37 levels and periodontal parameters were assessed at baseline, 1 and 3 months after completion of non-surgical periodontal treatment. GCF LL-37 levels were analyzed by ELISA.No significant difference was observed in GCF LL-37 levels between smoker and non-smoker controls (p0.05). Smoker CP group had significantly lower GCF LL-37 level than non-smoker CP group at baseline (p0.05). GCF LL-37 levels significantly decreased in non-smoker CP group at first week, 1 and 3 months after completion of non-surgical periodontal treatment (p0.05) although no significant decrease in GCF LL-37 levels was observed in smoker CP group (p0.05). Periodontal parameters were correlated with GCF LL-37 levels in non-smoker CP group (p0.05), but not in smoker CP group (p0.05).GCF LL-37 levels do not seem to be affected from smoking in periodontal health. However, smoking might have a suppressive effect on GCF LL-37 levels in CP. Non-surgical treatment is effective in decreasing GCF LL-37 levels in non-smoker CP patients but not in smokers with CP.

OBJECTIVE The aim of this study was to evaluate histologically the following treatment of bilateral localized gingival recessions with coronally advanced flap (CAF) combined with platelet-rich fibrin (PRF) or subepithelial connective tissue graft (SCTG). MATERIALS AND METHODS Tissue samples were harvested from 14 subjects either 1 or 6 months after the surgeries. The 2-mm punch biopsies were obtained from the mid-portion of the grafted sites. Neutral buffered formalin fixed, paraffin-embedded 5-μm thick tissue sections were stained with hematoxylin eosin and Masson's trichrome in order to analyze the collagen framework, epithelium thickness and rete-peg length. Multiple sequential sections were cut from paraffin-embedded blocks of tissue and immunohistochemically prepared for detection of vascular endothelial growth factor, CD31 and CD34, for the assessment of vascularization. RESULTS Rete peg formation was significantly increased in the sites treated with PRF compared to the SCTG group after 6 months (p

The objective of this study was to evaluate the levels of MMP-8, MMP-9, TIMP-1 and interleukin-1beta (IL-1β) in gingival crevicular fluid during the early and late stages of healing in gingival recession sites treated with coronally advanced flap plus platelet-rich fibrin (CAF+PRF) compared with CAF plus connective tissue graft (CAF+CTG).Twenty-four nonsmoking patients with Miller Class I or Class II localized gingival recession defects in bilateral sites received treatment with either CAF+PRF (PRF group) or CAF+CTG (CTG group). Gingival crevicular fluid samples were collected at baseline and at 10 d and 1 mo, 3 mo and 6 mo after surgery. The levels of MMP-8, MMP-9, TIMP-1 and IL-1β in gingival crevicular fluid were measured using a time-resolved immunofluorometric assay and ELISAs.Gingival crevicular fluid levels of IL-1β were significantly elevated in the CTG group at 10 d compared with baseline (p 0.05). At 10 d after surgery, the levels of TIMP-1 in gingival crevicular fluid showed a significant decrease in the CTG group compared with the PRF group (p 0.05). The levels of IL-1β and MMP-8 in gingival crevicular fluid were significantly lower in the PRF group than in the CTG group at 10 d (p 0.05). No significant differences were found in all clinical and biochemical parameters at 1, 3, and 6 mo between study groups (p 0.05).Root coverage with CAF+PRF has a significant effect on increasing gingival crevicular fluid TIMP-1 levels and suppressing gingival crevicular fluid MMP-8 and IL-β levels at 10 d. Gingival crevicular fluid levels of MMP-8, MMP-9, TIMP-1 and IL-1β did not seem to be affected by the technique at later phases of wound healing.

Interleukin (IL)-6 family of cytokines, including IL-6, oncostatin M (OSM), leukemia inhibitory factor (LIF), and IL-11, have fibrogenic features. The current study determines gingival crevicular fluid (GCF) levels of fibrosis-related IL-6-type cytokines in cyclosporine A (CsA)-induced gingival overgrowth (GO).Eighty non-smokers were included (40 CsA-medicated renal transplant patients with GO [GO+; n = 20] or without GO [GO-; n = 20], 20 individuals with gingivitis, and 20 healthy participants). Probing depth and plaque, papilla bleeding, and hyperplastic index scores were recorded. GCF samples were obtained from the mesio-buccal aspects of two teeth. GCF IL-6, IL-1β, OSM, LIF, and IL-11 levels were analyzed by enzyme-linked immunosorbent assay.The GO+ and GO- groups had higher IL-6 total amounts than the healthy group (P0.008). IL-1β total amounts in the GO+ group were significantly higher than in both the healthy and GO- groups (P0.008). OSM total amount was elevated in the GO+ and GO- groups compared with both the gingivitis and healthy groups (P0.008). All groups had similar LIF and IL-11 total amounts (P0.008). Moderate positive correlations were detected among IL-6, IL-1β, OSM, and IL-11 total amount in GCF and clinical parameters (P0.05).IL-6 and OSM increases in GCF as a result of CsA usage or an immunosuppressed state irrespective of the severity of inflammation and the presence of GO. The IL-6 family of cytokines might not be directly involved in biologic mechanisms associated with CsA-induced GO. Lack of an association between assessed IL-6 cytokines and CsA-induced GO might indicate distinct effects of these cytokines on fibrotic changes of different tissues.

The aim of the present study was to investigate the effect of cyclosporine-A (CsA) medication on gingival crevicular fluid (GCF) LL-37, human neutrophil peptide (HNP)1-3 and adrenomedullin (ADM) levels.CsA-treated renal transplant recipients with GO (CsA GO+) and without GO (CsA GO-), tacrolimus-medicated renal transplant recipients (n = 20/group), systemically healthy subjects with gingivitis (n = 21) and individuals free of periodontal and systemic diseases (n = 20) were included in the present study. Periodontal parameters were recorded and GCF samples were obtained from the study participants. GCF LL-37, HNP1-3 and ADM levels were analyzed by enzyme-linked immunosorbent assay.GCF LL-37 total amount was higher at GO+ sites than the other study sites (p0.05). Total amount of GCF HNP1-3 was higher in immunosuppressive treatment groups than healthy and gingivitis groups, regardless of GO presence (p0.05). GCF ADM total amount was similar in all study groups. GCF volume, papillary bleeding index and hyperplastic index (p0.05) were significantly correlated with GCF LL-37 total amounts (p0.05), but not with GCF HNP1-3 and ADM total amount at GO+ sites (p0.05).Neutrophil infiltration due to extended inflammation might have increased GCF LL-37 levels at GO+ sites and contributed to the pathogenesis of CsA-induced GO.

WOS: 000359064200018, PubMed ID: 26422857, Background/aim: The effect of smoking on inflammatory biomarkers in gingival crevicular fluid (GCF) is well established in the presence of periodontal inflammation. However, it is not clear if smoking has an influence on matrix metalloproteinase (MMP) and growth factor levels in the GCF of periodontally healthy subjects. The aim of this study was to investigate GCF levels of MMP-1, MMP-8, transforming growth factor (TGF)-beta 1, platelet-derived growth factor (PDGF)-AB, and vascular endothelial growth factor (VEGF) in smoking versus nonsmoking periodontally healthy subjects. Materials and methods: Thirty-two periodontally healthy subjects were included in this study. Probing depths, bleeding on probing, and plaque index was assessed. GCF levels of MMP-1, MMP-8, TGF- beta 1, PDGF-AB, and VEGF were analyzed by enzyme-linked immunosorbent assay. Results: No significant differences were observed in the distribution of demographic data between study groups. GCF total amount of PDGF-AB was significantly lower in smokers compared to nonsmokers (P = 0.014). Total amount of GCF MMP-1, MMP-8, TGF-beta 1, and VEGF levels were similar in both study groups (P = 0.022). Conclusion: Smoking has the effect of decreasing GCF PDGF-AB while it does not affect GCF MMP-1, MMP-8, TGF-beta 1, and VEGF in periodontally healthy subjects. Since increased levels of these molecules are involved in periodontal breakdown, our findings may emphasize the importance for maintenance of periodontal health in smokers.

The aim of the present study is to investigate matrix metalloproteinase (MMP)-8 and tissue inhibitor of MMP-1 (TIMP-1) gene polymorphisms in generalized aggressive periodontitis (GAgP) and to assess the effects of MMP-8 and TIMP-1 genotypes on the outcomes of non-surgical periodontal therapy.Genomic DNA was obtained from peripheral blood of 100 patients with GAgP and 167 periodontally healthy controls. MMP-8 +17 C/G, -799 C/T, -381 A/G and TIMP-1 372 T/C, *429 T/G polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism. Patients with GAgP received non-surgical periodontal therapy and were followed for 6 months. Clinical periodontal parameters and gingival crevicular fluid (GCF) samples were collected at baseline and at follow-up visits. GCF biomarkers were analyzed by immunofluorescence assay and enzyme-linked immunosorbent assay.Distribution of the MMP-8 -799 C/T genotypes was significantly different between the GAgP and control groups (P0.005). TIMP-1 372 T/C and *429 T/G genotypes in males were also significantly different between study groups (P0.004). GCF MMP-8 levels decreased until 3 months after non-surgical therapy compared with baseline in T and G alleles, as well as G and C allele carriers (P0.0125), whereas no significant decreased was observed in non-carriers (P0.0125).On the basis of the present findings, it can be suggested that MMP-8 -799 C/T and TIMP-1 372 T/C, *429 T/G gene polymorphisms in males may be associated with the susceptibility to GAgP in the Turkish population.

WOS: 000336597800001, PubMed ID: 24949444, Aim. Cathepsin C is the activator of the polymorphonuclear leukocyte-derived proteinase 3, which contributes to inflammatory processes. The aim of the present study was to investigate gingival crevicular fluid (GCF) proteinase 3 and cathepsin C levels in periodontal diseases. Design. Eighteen patients with chronic periodontitis (CP), 20 patients with generalized aggressive periodontitis (G-AgP), 20 patients with gingivitis, and 18 healthy subjects were included in the study. Periodontal parameters including probing depth, clinical attachment level, papilla bleeding index, and plaque index were assessed in all study subjects. GCF proteinase 3 and cathepsin C levels were analyzed by ELISA. Results. GCF proteinase 3 total amount was significantly higher in diseased groups compared to control group, after adjusting age(P0.05). Periodontal parameters of sampling sites were positively correlated with GCF proteinase 3 total amounts (P0.05). Conclusions. Elevated levels of GCF proteinase 3 in CP, G-AgP, and gingivitis might suggest that proteinase 3 plays a role during inflammatory periodontal events in host response. However, cathepsin C in GCF does not seem to have an effect on the pathogenesis of periodontal diseases., Ege University Research FoundationEge University, The authors greatly appreciate the statistical advice received from Professor Mehmet Orman, Department of Biostatistics and Medical Informatics, School of Medicine, Ege University. This study was supported by grant from Ege University Research Foundation.

The studyarea (Lakes Region) located in Western Anatolia part of Türkiye has a very richfloristic composition. However, these natural plants are used seldom forlandscape or economic purposes. In this context, ground cover plants are ofteninconspicuous elements of a landscape plantation design because of theirfunctional and aesthetics avantages. Although natural ground cover plants arevery important, knowledges on the subject are unsufficient. In this study,potantial natural ground cover plants were determined in survey studies fromthe Lakes Region for the first time. The study was carried out among 2000 and2004 years from different habitats in mountains of the region. By ourobservations in the study, concept of “ground cover plant” was defined newly.Collected and identified plants according to general flora books by the authorsare hold in Herbarium GUL of Süleyman Demirel University. According to ourfield observations, botanical and ecological characteristics of the plants weredetermined for ground covering. A list of these plants was listedalphabetically. According to the results,totally 175 taxa belonging to 79genera and 28 families were determined. 44of them are belonging to Fabaceae family. The familyhas more taxa than other families on the subject. Their distributions andration to life forms of the taxa are as follows: Chamaephytes 81 (46.2 %), Hemicryptophytes 71 (40.5 %), Terophytes 12(6.8 %), Geophytes 7 (4.0 %) and Phanerophytes 4 (2.2 %). 80 taxa of them are endemic to Türkiyeand endemism ratio is 46 %. On theother hand, distributions to phytogeographical regions of ground cover plantsgrowing in the study area Mediterraneanelements 48 (27.0 %), Irano-Turanian elements 44 (25.0 %), Euro-Siberianelements 13 (7.0 %), many regionous 10 (6.0 %) and unknown 60 (34.0.%).Some ground cover plants were proposed for different habitats and landscapeareas., Çalışma alanı(Göller Yöresi)Türkiye’nin Batı Anadolu kesiminde yer alır. Yöre, floristik açıdan çokzengindir. Bununla birlikte bu doğal bitkiler ekonomik amaçlı veya yer örtücü amaçlı olarak nadirenkullanılmaktadır. Bu manada, yer örtücü bitkiler yörede ekseriyetle peyzajdüzenlemelerde fazla dikkat çekmez. Ancak bu bitkilerin fonksiyonel ve estetikavantajları çok fazladır. Doğal yer örtücü bitkiler çok önemli olmasına rağmenbu konudaki bilgiler de oldukça yetersizdir. Bu çalışmada, floristikçalışmalarla yörenin potansiyel doğal yer örtücü bitkileri ilk kez belirlendi.Çalışma, 2000-2004 yıllarında yapıldı ve bitkilerin listesi yörenindağlarındaki farklı habitatlarda yapılan çalışmalarla belirlendi. Yerörtücü bitki anlamında çalışmaya dair gözlemlerimiz ilerleyen yıllarda yenidengözden geçirildi ve bazı tanımlamalar yapıldı. Tarafımızdan toplanan ve genelfloristik eserler yardımıyla tarafımızdan teşhisleri yapılan iletim demetli yer örtücübitki örnekleri Süleyman Demirel Üniversitesi’nin(Isparta) GUL Herbaryumu’unda muhafaza edilmektedir. Yer örtücü bitkiler içinGöller Yöresi’nin dağlarında yapılan arazi gözlemleri, bitkilerin botanik veekolojik özellikleri çalışmada belirlendi. Bu bitkiler alfabetik sırayadizilerek listelendi. Çalışmanınsonuçlarına göre; toplam olarak 79 cinsve 28 familyayaait 175 takson tayin edildi. Bunlardan 44takson Fabaceae familyasına aittir. Bu familya yer örtücü bitkiler açısından diğerfamilyalardan daha çok takson ihtiva eder. Yöreden tespit edilen yer örtücübitki taksonlarının hayat formları ve hayat formlarına dağılım oranlarıaşağıdaki gibidir: Kamafit 81 (% 46.2),Hemikriptofit 71 (% 40.5), Terofit 12 (% 6.8), Geofit 7 (% 4.0) ve Fanerofit 4(% 2.2). Bu taksonlardan 80’i Türkiye endemiğidir ve endemizm oranı %46’dır. Diğer taraftan yörenin yer örtücü bitkilerininbitki coğrafyası bölgelerine dağılımı ve oranları şöyledir: Mediterranean(Akdeniz) elementi 48 (% 27.0),Irano-Turanian(İran-Turan) elementi 44 (% 25.0), Euro-Siberian(Avrupa-Sibirya)elementi 13 (% 7.0), çok bölgeli 10 (% 6.0) ve coğfafi bölgesi bilinmeyen 60 (% 34.0).Bazı yer örtücü bitkiler peyzaj düzenlemeleri için ilk kez belirlendi ve farklıortamlarda uygulanması önerildi.

Transglutaminase (TGM)-2 has been shown to contribute to fibrosis by extracellular matrix accumulation in some organs and is activated by intracellular reactive oxygen species. The aim of this study is to investigate levels of gingival crevicular fluid (GCF) and plasma TGM-2 and oxidative stress markers (OSMs) in cyclosporin A (CsA)-induced gingival overgrowth (GO).The study enrolled 20 healthy (H) individuals; 20 patients with gingivitis (G); 20 CsA-medicated patients with GO (CsA GO+); and 20 CsA-medicated patients without GO (CsA GO-). GCF and plasma levels of TGM-2 were analyzed by enzyme-linked immunosorbent assay. Spectrofluorometry was used to analyze thiobarbituric acid reactive substance (TBARS); ferric-reducing antioxidant power (FRAP); total oxidant status (TOS); and total antioxidant capacity (TAC).GCF TGM-2 level was elevated in CsA GO+ compared with G (P = 0.048) and H (P = 0.001) groups. GCF TBARS level was elevated in CsA GO+ compared with other groups (CsA GO- group: P = 0.003; G group: P0.001; and H group: P0.001) and was higher in CsA GO- than in H (P = 0.048). GCF FRAP level was lower in CsA GO- than in H (P = 0.04). Both CsA GO+ and CsA GO- groups had lower GCF TOS levels than H (P0.001 and P = 0.002) and G (P = 0.003 and P = 0.04). GCF TAC was higher in CsA GO+ than in H (P = 0.02). Plasma TGM-2 level was elevated in CsA GO+ compared with G (P = 0.048) and H (P = 0.002). Plasma FRAP level was higher in H and CsA GO- than in CsA GO+ (P = 0.008 and P = 0.02).CsA use significantly alters GCF and plasma levels of TGM-2 and OSMs. TGM-2 may contribute to CsA-induced GO in CsA-treated patients by changing GCF and plasma levels of OSMs. Further studies are needed to prove causality and its direction.

This study was aimed to evaluate the gingival crevicular fluid (GCF) and plasma transglutaminase-2 (TGM-2), total antioxidant capacity (TAC), total oxidant status (TOS), ferric reducing antioxidant power (FRAP) and thiobarbituric acid reactive substances (TBARS) in patients with chronic periodontal disease.Twenty patients with chronic periodontitis (CP), 20 patients with gingivitis and 20 healthy subjects were enrolled in the study. Clinical periodontal parameters including probing depth, clinical attachment level, plaque index and papillary bleeding index were recorded. GCF and plasma levels of TGM-2, TAC, TOS, TBARS and FRAP were analyzed.GCF TGM-2 was significantly lower in CP group than in gingivitis patients (P=0.006). GCF FRAP in CP and gingivitis groups was significantly lower than in healthy subjects (P0.001). Plasma FRAP level was lower in gingivitis group when compared to healthy subjects (P=0.003). There was no significant difference in GCF and plasma TAC, TOS, TBARS and plasma TGM-2 levels among the study groups (P0.05). GCF TGM-2 level was positively correlated with GCF TAC and negatively correlated with CAL.Decreased FRAP in GCF and plasma indicating lower antioxidant status of CP patients might suggest the role of oxidative stress in periodontitis. GCF TGM-2 data might suggest that TGM2 is associated with stabilization of the extracellular matrix and wound healing in periodontium rather than gingival inflammation.

The aim of the present study is to investigate gingival crevicular fluid (GCF) and plasma acute-phase cytokines, interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-11 (IL-11), oncostatin M (OSM), and leukemia inhibitory factor (LIF) levels in patients with different periodontal diseases.Eighty individuals were included in this study; 20 with chronic periodontitis (CP), 20 with generalized aggressive periodontitis (GAgP), 20 with gingivitis, and 20 classified as healthy (H). Probing depth, clinical attachment level, plaque index, and papilla bleeding index were recorded. Plasma and GCF IL-1β, IL-6, IL-11, OSM, and LIF levels were analyzed by enzyme-linked immunosorbent assay.CP and GAgP groups had significantly higher GCF IL-1β, IL-6, and IL-11 levels when compared with the H group (P0.05). Conversely, GCF LIF levels of the CP and GAgP groups were lower than those of the H group (P0.05). GCF OSM levels did not differ significantly among study groups. Plasma levels of all the cytokines studied were not significantly different among the study groups.Based on the present data, elevated IL-1β, IL-6, and IL-11 GCF levels, but not plasma levels, are suggested as reliable inflammatory biomarkers in periodontal diseases. Decreased LIF levels in diseased groups might reflect the possible beneficial effects of LIF in the modulation of inflammatory response in gingiva.

Introduction: Subepithelial connective tissue graft (SCTG) + coronally advanced flap (CAF) has been suggested as the most predictable technique for treatment of recession defects. However, the SCTG technique necessitates a second surgical site and increases the risk of morbidity linked with harvesting the autogenous palatal donor mucosa. Platelet-rich fibrin (PRF) has been shown to accelerate soft-tissue healing. The use of PRF in treatment of gingival recessions eliminates the requirement of a donor site. The aim of this case report is to evaluate the clinical effectiveness of the CAF+PRF combined technique and CAF+SCTG in the treatment of bilateral gingival recessions.Case Presentation: The surgical treatment of a 23-year-old female patient with bilateral gingival recessions in maxillary cuspids is discussed in this case report. Sites were randomly assigned to CAF+PRF (test) or CAF+SCTG (control) sites. Clinical periodontal parameters were recorded and clinical photographs were taken at baseline; 1, 3, ...

Emingil G, Han B, Ozdemir G, Tervahartiala T, Vural C, Atilla G, Baylas H, Sorsa T. The effect of azithromycin, as an adjunct to nonsurgical periodontal treatment, on microbiological parameters and gingival crevicular fluid biomarkers in generalized aggressive periodontitis. J Periodont Res 2012; 47: 729–739. © 2012 John Wiley & Sons A/S Background and Objective: To study the effectiveness of azithromycin in combination with nonsurgical periodontal therapy on clinical and microbiological parameters, and on the MMP-8 and TIMP-1 levels in gingival crevicular fluid, over a 6-mo time-period in patients with generalized aggressive periodontitis. Material and Methods: Thirty-two patients with generalized aggressive periodontitis were included in this randomized, double-blind, placebo-controlled, parallel-arm study. They were randomly assigned to azithromycin or placebo groups (500 mg once daily for 3 d). Probing depth, clinical attachment levels, presence of bleeding on probing and plaque were recorded. Gingival crevicular fluid samples were obtained from one single-rooted tooth, while microbiological samples were obtained from two single-rooted teeth, all with a probing depth of ≥ 6 mm. Microbiological parameters were analyzed by quantitative real-time PCR for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Fusobacterium nucleatum, Prevotella intermedia and total bacteria. Gingival crevicular fluid biomarkers were determined by immunofluorometric assay and ELISA. Results: All clinical parameters improved, and microbiological parameters and gingival crevicular fluid MMP-8 levels significantly decreased, over the 6-mo period (p

Mantyla P, Buduneli E, Emingil G, Tervahartiala T, Pussinen PJ, Baris N, Akilli A, Atilla G, Sorsa T. Acute myocardial infarction elevates serine protease activity in saliva of patients with periodontitis. J Periodont Res 2012; 47: 345–353. © 2011 John Wiley & Sons A/S Background and Objective: There are indications that acute myocardial infarction (AMI) may have an effect on the oral environment, which is reflected in the expression of salivary and gingival proteinases. According to our knowledge, no studies have been carried out to investigate the effect of AMI on the activities of two major tissue-destructive serine protease and microbial effectors, elastase and cathepsin G, produced by oral fluid polymorphonuclear granulocytes (PMN). Therefore, we compared the activities of elastase and cathepsin G in saliva from patients with AMI and from systemically healthy subjects (non-AMI) with similar periodontal conditions. Material and Methods: A total of 92 patients (47 AMI and 28 non-AMI patients with gingivitis or periodontitis, and 17 systemically and periodontally healthy subjects as a control group) were recruited. Clinical periodontal measurements were recorded, and stimulated whole-saliva samples were collected. The patients with AMI were clinically examined within 3–4 d after admission to the coronary care unit. The activities of saliva neutrophil elastase and cathepsin G were measured after collection, at specific time-points during incubation (from baseline to 23 h) by specific synthetic peptide substrate assays. Results: The saliva of patients with AMI and periodontitis had a significant trend for the highest elastase activities among the study groups. Elastase and cathepsin G activities correlated significantly with each other in the AMI periodontitis group (r = 0.8, p

The aim of this cross-sectional study is to investigate gingival crevicular fluid (GCF) osteocalcin, cross-linked N-terminal telopeptide (NTx), and calprotectin levels in cyclosporin A (CsA)-induced gingival overgrowth (GO).Forty medicated patients with CsA including 20 with GO (CsA GO+), 10 without GO (CsA GO-), 10 with GO and chronic periodontitis (CsA CP) and 60 patients with CP alone, 20 patients with gingivitis, and 20 healthy patients were enrolled. Probing depth, clinical attachment level, plaque index, and papillary bleeding index were recorded. GCF calprotectin, osteocalcin, and NTx levels were analyzed by enzyme-linked immunosorbent assay. Parametric tests were used for statistical analysis.The CsA GO+ and CP groups had significantly lower GCF osteocalcin levels and osteocalcin/NTx ratio than the healthy group, whereas GCF osteocalcin levels and osteocalcin/NTx ratio in the gingivitis group were higher than the CsA GO+, CsA GO-, CsA CP, and CP groups (P0.05). The CP group had elevated GCF calprotectin levels compared to the other study groups (P0.05). The CsA GO+ and CsA GO- groups also had higher GCF calprotectin levels compared to the CsA CP, gingivitis, and healthy groups (P0.05).Increased GCF calprotectin and decreased GCF osteocalcin levels in the CsA GO+ and CsA GO- groups might suggest that CsA plays a role on the levels of these markers. The similarity of GCF osteocalcin, NTx, and calprotectin levels in the CsA GO+ and CsA GO- groups might suggest that these molecules are not involved in the pathogenesis of GO.

Objective Recent studies have showed that genetic factors involved in the host responses might determine the severity of periodontitis. hCAP-18/LL-37 is a part of the innate immune response in the oral cavity. The aim of the present study was to investigate the mutation of CAMP gene encoding hCAP-18/LL-37 in the patients with different periodontal diseases. Design Seventy-eight chronic periodontitis, 72 generalized aggressive periodontitis, and 149 controls were analysed for mutation of CAMP gene using direct DNA sequencing method. Frequencies of p.S34N mutation were compared by Pearson chi-square test. Logistic regression analysis was used to analyse the association between periodontitis and p.S34N mutation adjusting for bleeding on probing, age and gender. Results Twenty-five subjects had a novel missense mutation of CAMP gene. Single base substitution (c.101G>A) in exon 1 led to p.S34N mutation. All amino acid substitutions were heterozygous mutation. The patients with generalized aggressive periodontitis had significantly higher p.S34N mutation prevalence compared to the others, whilst there was no significant difference in prevalence of p.S34N mutation between the patients with chronic periodontitis and the control subjects. Logistic regression analysis adjusted for BOP, age and gender revealed that the patients with generalized aggressive periodontitis were 5.32 times more likely to have p.S34N mutation compared to the controls (OR = 5.32, 95% CI: 1.3–22.1). Conclusion We report a novel missense mutation of CAMP gene. p.S34N mutation in CAMP gene seems to be contributing factor for generalized aggressive periodontitis, but not for chronic periodontitis.

Objective Immune cell recognition of lipopolysaccharides via CD14 and Toll-like receptor 4 (TLR4) complexes plays a crucial role in linking innate and adaptive immune responses. This study was aimed to investigate the expression of TLR4 and membrane-bound CD14 (mCD14) in the gingival tissues of patients with gingivitis, periodontitis and CsA-induced gingival overgrowth. Design Gingival tissues were obtained from 10 renal transplant patients receiving cyclosporine-A (CsA) and having gingival overgrowth (GO), 10 patients with chronic periodontitis, 10 generalized aggressive periodontitis, 10 gingivitis and 10 healthy subjects. Immunohistochemistry was performed in order to determine the localization of TLR4 and mCD14 in tissue specimens. Results TLR4 and mCD14 expressions were detected in all tissues including healthy gingival biopsies. TLR4 and mCD14 positive cells were predominantly confined to the epithelium–connective tissue interface area, and were highly expressed in the basal cell layer of patients with CsA GO and chronic periodontitis, compared to healthy group ( P Conclusion The present study suggests that TLR4 and mCD14 protein expressions may be interrelated and appear to be associated with periodontal disease. CsA usage seemed not to affect TLR4 and mCD14 expressions in CsA induced GO tissues.

The aim of this study was to examine the effectiveness of chlorhexidine mouthrinse (CHX) in addition to daily plaque control on subgingival microbiota in patients with untreated gingivitis. Fifty gingivitis patients were randomized to CHX or placebo groups. CHX group rinsed with 0.2% CHX, while placebo group rinsed with placebo mouthrinse for 4 weeks. Subgingival plaque samples were collected and plaque index (PI), papilla bleeding index (PBI), calculus index, and probing pocket depth (PPD) were recorded at baseline and at 4 weeks. The amounts of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Fusobacterium nucleatum, and total bacteria were detected by quantitative real-time PCR method. In the CHX group the total bacteria count was significantly reduced in posterior teeth at 4 weeks (p 0.05). CHX mouthrinse as an adjunct to daily plaque control could be useful in the management of plaque-associated gingivitis and in reducing the subgingival total bacteria count especially in posterior teeth.

Bostanci N, Saygan B, Emingil G, Atilla G, Belibasakis GN. Effect of periodontal treatment on receptor activator of NF-κB ligand and osteoprotegerin levels and ratio in gingival crevicular fluid. J Clin Periodontol 2011; 38: 428–433. doi: 10.1111/j.1600-051X.2011.01701.x. Abstract Aim: Receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) have an established role in the pathogenesis of periodontitis, which is characterized by an increased RANKL/OPG ratio. The present study aims to investigate changes of RANKL, OPG and their relative ratio in gingival crevicular fluid (GCF) of periodontitis patients after non-surgical periodontal treatment. Materials and Methods: GCF was obtained from chronic periodontitis (n=14), generalized aggressive periodontitis (G-AgP; n=13) patients at baseline. The patients received scaling and root planing and were recalled after 2, 3 and 4 months for follow-up clinical examination and sampling. The total amounts and concentrations of RANKL and OPG in GCF were measured by enzyme-linked immunosorbent assay, and their relative ratio was calculated. Results: The RANKL/OPG ratio remained unchanged and did not correlate with clinical parameters throughout the monitoring period, despite the improved clinical outcome. This trend was similar in both chronic and G-AgP. Conclusions: Although the RANKL/OPG ratio has a potential diagnostic value for untreated periodontitis, it may not be a suitable predictor of clinically successful treatment outcome. As conventional therapy does not negatively modulate this ratio, the host could still be susceptible to further periodontal tissue destruction, warranting the consideration of adjunctive treatments.

Molecular Medicine Laboratory, Department of Pediatrics, School of Medicine, Ege University,Izmir, TurkeyOBJECTIVE: To investigate the levels of antimicrobialpeptide hCAP-18/LL-37 protein and mRNA expression ingingival tissues with different periodontal disease.MATERIALS AND METHODS: Ten patients with gen-eralized aggressive periodontitis, 10 patients with chronicperiodontitis, and 10 healthy controls were included inthis study. Periodontal parameters including probingdepth, clinical attachment level, plaque index, and papillableeding index were assessed in study subjects. hCAP-18/LL-37 mRNA analysis by RT-PCR and immunohisto-chemistry were performed in 19 samples providedenough RNA in terms of concentration and integrity.RESULTS: This study demonstrated that hCAP-18/LL-37was a product of neutrophils. Tissue samples of chronicperiodontitis patients had significantly higher immuno-staining of hCAP-18/LL-37 on neutrophils infiltrating inboth epithelium and connective tissue compared withcontrols. hCAP-18/LL-37 mRNA expression levels in tis-sue samples of chronic periodontitis patients seemed tobe upregulated compared with controls. While two gen-eralized aggressive periodontitis patients showed down-regulated hCAP-18/LL-37 mRNA expression levels, onegeneralized aggressive periodontitis patient showedslightly increased hCAP-18/LL-37 mRNA level comparedwith controls.CONCLUSIONS: hCAP-18/LL-37 has an important rolein innate response during periodontal inflammation.Local deficiency in hCAP-18/LL-37 might be a confound-ing effect in the pathogenesis of generalized aggressiveperiodontitis.Oral Diseases (2011) 17, 60–67Keywords: periodontitis; cathelicidin antimicrobial peptide; geneexpression; immunohistochemistry

Belibasakis GN, Emingil G, Saygan B, Turkoglu O, Atilla G, Bostanci N. Gene expression of transcription factor NFATc1 in periodontal diseases. APMIS 2011; 119: 167-172. Periodontitis is a disease of infectious aetiology that causes inflammatory destruction of the tooth-supporting tissues. Activated T cells are central to the pathogenesis of the disease, by producing receptor activator of nuclear factor κB (NF-κB) ligand (RANKL) that stimulates bone resorption. Antigenic activation of T cells is regulated by the induction of transcription factor nuclear factor of activated T cells, cytoplasmic 1 (NFATc1). There is as yet no information on the potential involvement of NFATc1 in periodontal diseases. This study aimed to investigate NFATc1 gene expression levels in periodontal diseases, and analyse the potential correlation with RANKL expression and clinical periodontal parameters. In this cross-sectional study, gingival tissue biopsies were obtained from healthy (n = 10) and periodontally diseased (n = 58) sites. NFATc1 and RANKL gene expression levels in these samples were analysed by quantitative real-time polymerase chain reaction. Compared with healthy subjects, patients with gingivitis, chronic and aggressive periodontitis, exhibited higher NFATc1 expression, which proved to be statistically significant in the periodontitis groups. NFATc1 and RANKL expression levels strongly correlated with each other, and with clinical periodontal parameters. The increased expression of NFATc1 in periodontitis denotes a role for this transcription factor in the pathogenesis of the disease.

Fluctuations in sex steroid hormones, which are also noticeable through the menstrual cycle of women, may impact periodontal health. The aim of this study is to evaluate the effect of hormonal changes occurring in the menstrual cycle on gingival inflammation and the gingival crevicular fluid (GCF) levels of interleukin 6 (IL-6), prostaglandin E(2) (PGE(2)), tissue plasminogen activator (t-PA), and plasminogen activator inhibitor-2 (PAI-2).Twenty-five gingivitis patients and 25 periodontally healthy subjects having regular menstrual cycles were seen at menstruation (ME) (1 to 2 days of menstruation), ovulation (OV) (12 to 14 days), and premenstrual phases (PM) (22 to 24 days). GCF and saliva samples were collected and clinical parameters including plaque index and bleeding on probing were recorded at each menstrual phase. Salivary estrogen and progesterone levels were analyzed to determine exact menstrual cycle days. GCF levels of IL-6, PGE(2), t-PA, and PAI-2 were measured by enzyme-linked immunosorbent assay.The percentages of sites with bleeding on probing were significantly higher in ME (60.85 +/- 18.36) and OV (58.92 +/- 25.04) than in the PM (40.12 +/- 20.10) phase in the gingivitis group (P0.001; repeated measures analysis of variance), whereas it was similar for all phases in the healthy group (P0.05; repeated measures analysis of variance). GCF levels of IL-6 were significantly elevated in gingivitis patients compared to healthy subjects in all phases (P = 0.004, P = 0.041, and P = 0.046 for ME, OV, and PM, respectively; Mann-Whitney U test). GCF levels of IL-6, PGE(2), t-PA, and PAI-2 were unchanged in different menstrual phases in both groups (P0.05; Friedman test).The present study suggests that changes in the sex steroid hormones during menstrual cycles might have a limited effect on the inflammatory status of gingiva, but GCF cytokine levels were not affected.

Antimicrobial peptides participating in the innate host response are important contributors for maintaining the balance between health and disease. The aim of the present study is to investigate the levels of gingival crevicular fluid (GCF) adrenomedullin and human neutrophil peptides 1 through 3 (HNP1-3) in patients with different periodontal diseases.A total of 77 subjects, including 20 patients with chronic periodontitis, 18 patients with generalized aggressive periodontitis, 20 patients with gingivitis, and 19 healthy subjects, were included in the present study. The probing depth, clinical attachment level, plaque index, and papilla bleeding index were assessed in all study subjects. GCF samples were analyzed for evaluating adrenomedullin and HNP1-3 levels by enzyme-linked immunosorbent assay.The present study demonstrated that the periodontitis groups had a significantly higher total amount of GCF adrenomedullin compared to the gingivitis and healthy control groups after adjusting for age and gender (P0.05). Additionally, GCF adrenomedullin levels were positively correlated with clinical periodontal parameters of sampling sites (P0.05). The total amount of GCF HNP1-3 was not different among the study groups, and there was no correlation between the total amount of GCF HNP1-3 and clinical periodontal parameters (P0.05).Our results suggest a defensive role for adrenomedullin during the host response in periodontal disease. Additionally, the lack of the HNP1-3 antimicrobial peptide might point to the deficiency of one of the protective mechanisms for periodontal tissues.

Aim: The aims of this study were to investigate (a) the matrix metalloproteinase-13 (MMP-13) promoter polymorphisms in severe, generalized chronic periodontitis (CP), (b) the relationship of periodontal therapy outcome with these genotypes and (c) gingival crevicular fluid (GCF) MMP-13 level–MMP-13 genotype correlation. Materials and Methods: Genomic DNA was obtained from peripheral blood of 102 patients with severe, generalized CP, and 98 periodontally healthy subjects. MMP-13 −77A/G and 11A/12A polymorphisms were determined by the polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing methods, respectively. Fifty-eight CP patients received non-surgical periodontal therapy and were followed for 6 months. Clinical periodontal parameters and GCF samples were collected at baseline and at 6 months. GCF MMP-13 levels were analysed by an enzyme-linked immunosorbent assay. Results: The distribution of MMP-13 −77AG genotypes and allele frequencies did not differ significantly between study groups (p>0.05). Study subjects, except 3, had the 11A/11A genotype. MMP-13 −77G allele carriers had similar GCF MMP-13 levels and clinical periodontal parameters compared with AA genotypes after non-surgical periodontal therapy (p

Host response to periodontopathic microorganisms can be modulated by genetic factors. Accumulated evidence highlighted the role of renin-angiotensin system (RAS) in inflammatory response thus potential implication of this molecular system in the pathogenesis of periodontitis can be suggested. The present study investigated common genetic variants of molecules within the RAS family namely angiotensin-converting enzyme (ACE), angiotensinogen (AGT) and angiotensin II type 1 receptor (AT1R) in relation to generalized aggressive periodontitis (G-AgP).DNA was obtained from peripheral blood of 103 G-AgP patients and 100 periodontally healthy subjects. ACE I/D, AGT M235T and AT1R A1166C polymorphisms were genotyped by polymerase chain reaction and restriction fragment length polymorphism method. Chi-square, ANOVA and logistic regression were used in statistical analyses.Both ACE I/D and AT1R polymorphisms were similar in G-AgP and healthy groups (p0.05). G-AgP subjects exhibited decreased AGT TT genotype and T allele frequency as compared to healthy subjects (p0.05). The same trend was also observed in the nonsmoker subgroup regarding investigated RAS polymorphisms.Present findings suggest that AGT M235T TT genotype and T allele might be associated with decreased risk for G-AgP in Turkish population.

Aim: Evidence suggests that the ultimate product of the renin-angiotensin system (RAS), angiotensin II, exerts inflammatory actions. The present study aimed to evaluate the inter-relation between gene polymorphisms of the RAS components; angiotensin converting enzyme (ACE), angiotensinogen (AGT) and angiotensin II type-I receptor (AT1R), and severe chronic periodontitis (CP). Material and Methods: DNA was obtained from peripheral blood of 90 CP patients and 126 periodontally healthy subjects, and the clinical parameters were recorded. ACE I/D, AGT M235T and AT1R A1166C polymorphisms were genotyped by the PCR-RFLP method. Chi-square, ANOVA and logistic regression methods were used in statistical analyses. Results: The frequency of the ACE D allele was significantly lower in the CP group than the healthy group (p corr = 0.015). CP subjects exhibited increased C allele carriage and C allele frequency of the AT1R gene (p corr = 0.03 and p corr = 0.03, respectively). All clinical parameters of CP patients were found to be similar in variant allele-carrying and non-carrying subjects (p>0.05). Conclusions: The present findings suggest that ACE I/D and AT1R polymorphisms might be associated with susceptibility to CP but not with disease severity. The D allele of ACE I/D might be associated with decreased, whereas the C variant of AT 1R A1166C might be associated with an elevated risk for CP in Turkish population.

Matrix metalloproteinases (MMPs) play important roles in tissue-destruction mechanisms-associated periodontitis. MMP-8 and -13 are the predominant collagenases that are important in the extracellular matrix degradation in periodontal tissues. MMP-14 is a membrane-type MMP, whereas laminin-5 indicates basal membrane modification and epithelial induction. The purpose of the present study was to evaluate the effects of celecoxib and omega-3 fatty acid administration on the gingival tissue expression of MMP-8, -13, and -14, tissue inhibitor of MMP (TIMP)-1, and laminin (Ln)-5gamma2-chain in rat experimental periodontitis induced by Escherichia coli endotoxin (lipopolysaccharide [LPS]).Experimental periodontitis was induced in rats by repeated LPS injection. Fifty-one adult male Sprague-Dawley rats were divided into six study groups: saline control, LPS, LPS + celecoxib, LPS + therapeutic omega-3 (TO3), prophylactic omega-3 + LPS + omega-3 (P+TO3), and LPS + celecoxib + omega-3 fatty acid. Celecoxib and omega-3 fatty acid were given as a single agent or as combination therapy for 14 days. On day 15, all rats were sacrificed, and gingival tissues were analyzed immunohistochemically for the expression of MMP-8, -13, and -14, TIMP-1, and Ln-5gamma2-chain. Alveolar bone loss was evaluated morphometrically under a stereomicroscope. Data were tested statistically by Kruskal-Wallis and Mann-Whitney tests and Spearman correlation analysis.Alveolar bone loss was significantly higher in all study groups compared to the saline control group (all P0.01). MMP-8 expression was significantly higher in the LPS group than in the saline group (P = 0.001). Very low expression of MMP-8 was found in the celecoxib, P+TO3, and combination groups. TO3 increased TIMP-1 expression significantly compared to the LPS group (P0.05). Individual celecoxib and P+TO3 administration increased MMP-14 significantly compared to saline control and LPS groups (P0.05). No significant differences were found among the study groups with regard to Ln-5gamma2-chain and MMP-13 expressions (P0.05).Selective cyclooxygenase-2 inhibitor, prophylactic omega-3 fatty acid, and a combination of these two agents can inhibit gingival tissue MMP-8 expression. Moreover, the individual administration of therapeutic omega-3 may increase gingival TIMP-1 expression in contrast to no effect on MMP-8, -13, and -14 expressions in experimental periodontitis. These experimental findings in a rat model of LPS-induced periodontitis need to be verified by clinical human studies.

Aim Matrix metalloproteinases (MMPs) are involved in periodontal tissue remodeling and degradation. MMP polymorphisms could alter transcription and function of these enzymes. The aim of this study was to investigate MMP-2 , MMP-9 and MMP-12 gene polymorphisms in relation to susceptibility to severe chronic periodontitis (CP). Methods Genomic DNA was obtained from peripheral blood of 87 severe CP patients and 107 periodontally healthy subjects. MMP-2 − 735C / T , MMP-9 − 1562C / T and MMP − 12 357Asn / Ser gene polymorphisms were genotyped by polymerase chain reaction and restriction fragment length polymorphism. Probing depth, clinical attachment loss, supragingival plaque accumulation and bleeding on probing were recorded. The data were analyzed by chi-square, logistic regression and Mann–Whitney- U -tests. Results The genotype distributions and allele frequencies of MMP-2 , MMP-9 and MMP-12 genes were similar in CP and healthy subjects ( p > 0.05). Differences between rare allele carriage rates of CP and healthy groups regarding MMP-2 , MMP-9 and MMP-12 gene polymorphisms were not significant ( p > 0.05). However, T allele carriers of MMP-9 − 1562 gene had less risk for CP (OR = 0.36; 95% CI = 0.16–0.81). Conclusion These data suggest that MMP-2 − 735C / T , MMP-9 − 1562C / T and MMP-12 357Asn / Ser polymorphisms are not associated with susceptibility to severe CP in Turkish population. However, T allele of MMP-9 − 1562 gene might be associated with decreased susceptibility to severe CP.

The aim of the present study was to investigate whether chronic periodontitis caused the elevated levels of anti-cardiolipin antibodies (anti-CL) and oxidized low-density lipoprotein (oxLDL) in subjects with essential hypertension.Seventy-two subjects were categorized as healthy controls, subjects with essential hypertension and periodontal health (healthy-hypertension group), subjects with essential hypertension and gingivitis (gingivitis-hypertension group), or subjects with essential hypertension and chronic periodontitis (periodontitis-hypertension group). Individuals with essential hypertension who had been taking antihypertensive medicationor =2 years were included in the present study. The presence of supragingival plaque, bleeding on probing (BOP), probing depth (PD), and clinical attachment level were recorded, and blood samples were collected. Serum immunoglobulin M (IgM) and immunoglobulin G (IgG) anti-CL and oxLDL levels were assessed by enzyme-linked immunosorbent assay. For IgM and IgG anti-CL assays, positive tests were defined asor =15 IgM phospholipid units andor =10 IgG phospholipid units, respectively.The mean IgM anti-CL level and the prevalence of subjects positive for IgM anti-CL were significantly higher in the periodontitis-hypertension group compared to the other groups (P = 0.001). No significant differences were observed in the mean IgG anti-CL and oxLDL levels or in the number of subjects positive for IgG anti-CL and positive for IgM or IgG anti-CL among the study groups. The Pearson correlation analysis revealed positive correlations between IgM anti-CL levels and supragingival plaque, BOP, and PD scores.Chronic periodontitis might play a causal role in the elevated serum levels of anti-CL antibodies in individuals with essential hypertension. These elevated anti-CL levels that are due to chronic periodontitis might contribute to an increased risk for atherosclerosis in individuals with essential hypertension.

The aim of the present study was to determine gingival crevicular fluid (GCF) levels of monocyte chemotactic protein-1 (MCP-1), regulated on activation, normal T-cell expressed and secreted protein (RANTES) and macrophage migration inhibitory factor (MIF) in metabolic syndrome patients with gingivitis.Twenty metabolic syndrome patients with gingivitis (MSG), 20 MetS patients with clinically healthy periodontium (MSH), 20 systemically healthy subjects with gingivitis and 20 subjects who were both systemically and periodontally healthy were included. Periodontal and systemical parameters were recorded. GCF MCP-1, RANTES and MIF levels were assayed by enzyme-linked immunosorbent assay method.MSG and MSH groups had elevated blood pressure, triglyceride, waist circumference and fasting glucose values in comparison to gingivitis and healthy groups (P0.0001). Clinical periodontal parameters were higher in MSG and gingivitis groups when compared to those of the MSH and healthy groups (P0.0001). MCP-1 and RANTES levels (ng/mg total protein) of MSG group were higher than those of the MSH groups (P=0.005, P=0.0001, respectively). Also gingivitis group had higher MCP-1, RANTES and MIF levels compared to the healthy group (P=0.011, P=0.0001, P=0.011 respectively). The RANTES level of MSG group was significantly higher than those of the gingivitis group (P=0.01), but MCP-1 and MIF levels were similar in the MSG and gingivitis groups (P0.05).Elevated levels of GCF RANTES in MetS patients with gingivitis might associate with the presence of increased gingival inflammation by MetS. Low-grade systemic inflammation associated with MetS and adipose tissue-derived RANTES might lead to altered GCF RANTES levels in the presence of gingival inflammation.

Aim: Receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) are a system of molecules that regulate bone resorption. This study aims to compare the levels of RANKL, OPG and their relative ratio in gingival crevicular fluid (GCF) of healthy and periodontal disease subjects. Material and Methods: GCF was obtained from healthy (n=21), gingivitis (n=22), chronic periodontitis (n=28), generalized aggressive periodontitis (n=25) and chronic periodontitis subjects under immunosuppressant therapy (n=11). RANKL and OPG concentrations in GCF were measured by enzyme-linked immunosorbent assays. Results: RANKL levels were low in health and gingivitis groups, but increased in all three forms of periodontitis. OPG levels were higher in health than all three periodontitis, or gingivitis groups. There were no differences in RANKL and OPG levels between chronic and generalized aggressive periodontitis groups, whereas these were lower in the immunosuppressed chronic periodontitis group. The RANKL/OPG ratio was significantly elevated in all three periodontitis forms, compared with health or gingivitis, and positively correlated to probing pocket depth and clinical attachment level. Conclusion: GCF RANKL and OPG levels were oppositely regulated in periodontitis, but not gingivitis, resulting in an enhanced RANKL/OPG ratio. This ratio was similar in all three periodontitis groups and may therefore predict disease occurrence.

Aim: Tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) have important roles in proteolytic events in periodontitis. The aim of this study was to investigate TPA and PAI-1 gene polymorphisms in relation to susceptibility to generalized aggressive periodontitis (G-AgP). Methods: Genomic DNA was obtained from peripheral blood of 90 G-AgP patients and 154 periodontally healthy subjects. 4G/5G polymorphism in the promoter region of the PAI-1 gene and Alu-repeat insertion/deletion (I/D) polymorphism in intron 8 of the TPA gene were genotyped by polymerase chain reaction and endonuclease digestion. Results: The genotype distributions of TPA and PAI-1 genes were similar between G-AgP and healthy subjects (p>0.05). The distribution of TPA genotypes in G-AgP patients was 33.4% D/D, 44.4% I/D, and 22.2% I/I and was 26.3% D/D, 40.4% I/D, and 33.3% I/I in healthy subjects. The D allele was 55.6% in G-AgP and 46.6% in healthy subjects. There was a significant difference among study groups in D allele frequencies (p=0.044). The PAI-1 genotype distribution in G-AgP was 29.1% 4G/4G, 43.0% 4G/5G, and 27.9% 5G/5G, while it was 35.7% 4G/4G, 43.8% 4G/5G, and 20.5% 5G/5G in healthy subjects. Conclusion: These data suggest that the D polymorphic allele of TPA gene polymorphism could be associated with susceptibility to G-AgP in Turkish subjects.

Matrix metalloproteinases (MMPs) play important roles in tissue destruction mechanisms of periodontitis. MMP-8 and -13 are the major collagenases that act in extracellular matrix degradation in periodontal tissues. MMP-14 is a membrane-type MMP, and laminin (Ln)-5 is a basal membrane component. The aim of the present study was to evaluate the effects of doxycycline and alendronate on gingival tissue expression of MMP-8, -13, and -14; tissue inhibitors of MMP (TIMP)-1; and Ln-5 gamma2 chain in experimental periodontitis induced by Escherichia coli endotoxin (LPS) in rats.Experimental periodontitis was induced by repeated injection of LPS. Forty-four adult male Sprague-Dawley rats were divided into five study groups: saline control, LPS, LPS + doxycycline, LPS + alendronate, and LPS + doxycycline + alendronate. Doxycycline and alendronate were given as a single agent or as combination therapy during the 7 days of the experimental study period. On day 7, the rats were sacrificed, and the gingival tissues were analyzed immunohistochemically for expression of MMP-8, -13, and -14, Ln-5 gamma2 chain, and TIMP-1. Alveolar bone loss was evaluated morphometrically under a stereomicroscope. Data were tested statistically by Kruskal-Wallis and Mann-Whitney tests and Spearman correlation analysis.Alveolar bone loss was significantly higher in the LPS, doxycycline, alendronate, and combination groups than in the saline control group (all P0.01). MMP-8 expression was significantly higher in the LPS group than in the saline control group (P = 0.001). Individual administration of doxycycline or alendronate significantly decreased the expression of MMP-8 compared to LPS (P = 0.01). Combined drug administration reduced MMP-14 significantly compared to doxycycline (P = 0.004). No significant differences in Ln-5 gamma2 chain expression were found between the study groups (P0.05). MMP-14 significantly correlated with the Ln-5 gamma2 chain in the LPS + alendronate group (P = 0.04) and with the amount of alveolar bone loss in the LPS + doxycycline + alendronate group (P = 0.03).Our findings suggest that alendronate and/or doxycycline may inhibit MMP-8 expression significantly; particularly, their combined administration may provide beneficial effects in periodontal treatment. Moreover, individual administration of alendronate and doxycycline results in significant increases in TIMP-1 expression in gingiva. However, these effects of combined low-dose doxycycline and alendronate on MMPs and TIMP should be verified by clinical human trials before these agents are used in dental practice.

Cyclophilin A (CypA) is able to regulate inflammatory responses and matrix metalloproteinase production via its interaction with extracellular matrix metalloproteinase inducer (EMMPRIN). EMMPRIN is the cell surface receptor of CypA. The aim of the present study was to evaluate the gingival crevicular fluid (GCF) CypA and EMMPRIN levels in patients with chronic periodontitis (CP), generalized aggressive periodontitis (G-AgP) and periodontally healthy controls.Twenty CP patients, 19 G-AgP patients and 20 healthy control subjects were included in the present study. All study participants were non-smokers. Full mouth clinical periodontal parameters including probing depth, clinical attachment level, plaque index, and papilla bleeding index were recorded. GCF CypA and EMMPRIN levels were analyzed by enzyme-linked immunosorbent assay. Data were analyzed statistically with parametric and non-parametric tests.GCF CypA total amount was higher in the G-AgP group compared to healthy controls (p0.05), whereas CypA total amounts were similar in CP and healthy controls (p0.05). No significant difference in GCF CypA total amount between CP and G-AgP was observed (p0.05). Also, there was no significant difference in GCF EMMPRIN total amounts among the study groups (p0.05).Higher levels of GCF CypA in patients with G-AgP might demonstrate that CypA is associated with the inflammatory infiltrate and alveolar bone destruction of G-AgP. However, GCF CypA level does not seem to be affected by CP. Similar GCF EMMPRIN levels in diseased and healthy groups might suggest that EMMPRIN has role in the turn over of connective tissues in physiological conditions as well as pathological state.