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Objective: To evaluate the composition and function of mural cell populations in human gingival tissues DESIGN: A cross-sectional study was conducted on seven periodontitis (stage Ⅲ) patients. Gingival tissues were collected two months after scaling and root planing and divided into 3 groups: 1, h_h group (horizontal bone resorption, residual pocket depth ≤3 mm); 2, v_h group (vertical bone resorption >4 mm, residual pocket depth ≤3 mm); 3, v_i group (vertical bone resorption >4 mm, residual pocket depth ≥6 mm). Single-cell RNA sequencing (10X genomics) and subsequent bioinformatics analysis were performed. Protein expression of selected genes was confirmed by histological staining., Results: Two mural cell clusters, RGS5 + THY1 + and ACTA2 + MYH11 + subpopulations, were identified and confirmed by histological staining and cross-validation with three different single-cell RNA sequencing datasets in the GEO database. RGS5 + THY1 + cluster in perivascular areas possessed cellular protrusions and exhibited immunomodulatory and synthetic phenotypes. In contrast, the ACTA2 + MYH11 + cluster strictly distributed around vessel walls was characterized by a contractile phenotype. Mural cells closely interacted with endothelial cells through PDGF and NOTCH3 signaling. Mural cell loss was detected in the v_i group and in hopeless periodontal teeth, which might be caused by tumor necrosis factor-alpha induced apoptosis., Conclusions: Gingival mural cells can be classified into two distinct clusters according to their gene signatures and cell morphology. The loss of mural cells may indicate periodontitis progression., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The aim of this cross-sectional study was to investigate associations between salivary active matrix-metalloproteinase 8 (aMMP-8) and periodontitis severity, potentially periodontal pathogenic bacteria as well as blood parameters in generally healthy participants. Therefore, 188 participants with a mean age of 48.9 ± 8 years were examined. The periodontitis severity was assessed based on periodontal probing depth and clinical attachment loss. Both, aMMP-8 and microbiological analysis were performed using a validated, commercially available test system. Blood values were utilized from regular differential blood count. The aMMP-8 findings were associated with the periodontitis severity (P < 0.01), as well as with the prevalence of Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia, Parvimonas micra, Camphylobacter rectus and Eubacterium nodatum (P i  < 0.05). No associations between aMMP-8 and the examined blood parameters were found (P i  > 0.05). In conclusion, salivary aMMP-8 findings seem to reflect periodontal disease severity as a result of an immunoreaction, especially against bacteria with high periodontal pathogenic potential., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Objective: To monitor early periodontal disease progression and to investigate clinical and molecular profile of inflamed sites by means of crevicular fluid and gingival biopsy analysis., Methodology: Eighty-one samples of twenty-seven periodontitis subjects and periodontally healthy individuals were collected for the study. Measurements of clinical parameters were recorded at day -15, baseline and 2 months after basic periodontal treatment aiming at monitoring early variations ofthe clinical attachment level. Saliva, crevicular fluid and gingival biopsies were harvested from clinically inflamed and non-inflamed sites from periodontal patients and from control sites of healthy patients for the assessment of IL-10, MMP-8, VEGF, RANKL, OPG and TGF-β1 protein and gene expression levels., Results: Baseline IL-10 protein levels from inflamed sites were higher in comparison to both non-inflamed and control sites (p<0.05). Higher expression of mRNA for IL-10, RANK-L, OPG, e TGF-β1 were also observed in inflamed sites at day -15 prior treatment (p<0.05). After the periodontal treatment and the resolution of inflammation, seventeen percent of evaluated sites still showed clinically detectable attachment loss without significant differences in the molecular profile., Conclusions: Clinical attachment loss is a negative event that may occur even after successful basic periodontal therapy, but it is small and limited to a small percentage of sites. Elevated inflammation markers of inflamed sites from disease patients reduced to the mean levels of those observed in healthy subjects after successful basic periodontal therapy. Significantly elevated both gene and protein levels of IL-10 in inflamed sites prior treatment confirms its modulatory role in the disease status.

Serum hepcidin levels may increase in response to infection and inflammation. The present study investigated the effect of nonsurgical periodontal therapy (NSPT) on levels of serum hepcidin, inflammatory markers, and iron markers. An interventional study was conducted on 67 patients (age 30-65 years) without other diseases, except for chronic periodontitis (CP). Patients were allocated to either CP or control groups. The CP group received supragingival and subgingival scaling and root planing procedures, whereas the control group received supragingival scaling. Probing depth (PD), bleeding on probing, clinical attachment level (CAL), visible plaque index (VPI), serum hepcidin and interleukin-6 (IL-6) levels, high-sensitivity C-reactive protein (hs-CRP), hematological markers, and iron markers were measured at baseline and at 90 days after NSPT. The CP group had statistically significant lower mean values for mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) (p ≤ 0.05). The control group had statistically significant reductions in hemoglobin, hematocrit, MCV, and MCH (p ≤ 0.05). Serum hepcidin, IL-6, and erythrocyte sedimentation rate (ESR) levels were significantly decreased in both groups after NSPT. Periodontal markers were more markedly reduced in the CP group compared with the control group (p ≤ 0.05). These findings suggest that NSPT may reduce the serum levels of IL-6, hepcidin, and periodontal parameters.

Background/purpose: This study was conducted to evaluate the influence of mucogingival parameters, including keratinized mucosa (KM) and attached gingiva (AG), on the outcome of non-surgical periodontal therapy (NSPT)., Methods: A total of 204 non-smoking patients with generalized chronic periodontitis who received NSPT between 2012 and 2014 were included. The Mantel-Haenszel chi-square test was used to assess the associations between initial mucogingival parameters and initial clinical parameters on the buccal aspect, and the associations between initial mucogingival parameters and outcome clinical parameters on the buccal aspect of the sites with severe periodontal destruction. The generalized liner model was used to evaluate the contribution of initial clinical parameters to the outcome of NSPT., Results: KM ≥ 3 mm was associated with greater probing pocket depth (PD), less gingival recession (REC), and less clinical attachment level (CAL), and AG < 1 mm was associated with greater PD, REC, and CAL before NSPT. At the sites with severe periodontal destruction, KM ≥ 3 mm was associated with greater PD reduction (0.25 ± 0.08 mm) and CAL gain (0.25 ± 0.09 mm), and AG < 1 mm was associated with greater CAL gain (0.15 ± 0.08 mm) after NSPT. Initial PD ≥ 7 mm and non-molar teeth showed greater contribution to the outcome of NSPT., Conclusion: Less AG (<1 mm) was associated with greater periodontal destruction at baseline. At the sites with severe periodontal destruction, greater KM (≥3 mm) and less AG (<1 mm) resulted in better outcomes of NSPT., (Copyright © 2018. Published by Elsevier B.V.)

It has previously been shown that the presence of Aggregatibacter actinomycetemcomitans in subgingival plaque is significantly associated with increased risk for clinical attachment loss. The highly leukotoxic JP2 genotype of this bacterium is frequently detected in adolescents with aggressive forms of periodontitis. The aims of the study were to quantify the levels of JP2 and non-JP2 genotypes of A. actinomycetemcomitans in saliva of Moroccan adolescents with the JP2 genotype earlier detected in the subgingival plaque. The salivary concentrations of inflammatory proteins were quantified and linked to the clinical parameters and microbial findings. Finally, a mouth rinse with leukotoxin-neutralizing effect was administrated and its effect on the levels the biomarkers and A. actinomycetemcomitans examined. The study population consisted of 22 adolescents that previously were found to be positive for the JP2 genotype in subgingival plaque. Periodontal registration and sampling of stimulated saliva was performed at baseline. A mouth rinse (active/placebo) was administrated, and saliva sampling repeated after 2 and 4 weeks rinse. The salivary levels of JP2 and non-JP2 were analyzed by quantitative PCR and inflammatory proteins by ELISA. Both the JP2 and the non-JP2 genotype were detected in all individuals with significantly higher levels of the non-JP2. Enhanced levels of the JP2 genotype of A. actinomycetemcomitans was significantly correlated to the presence of attachment loss (≥3 mm). Salivary concentrations of inflammatory biomarkers did not correlate to periodontal condition or levels of A. actinomycetemcomitans . The use of active or placebo leukotoxin-neutralizing mouth rinse did not significantly interfered with the levels of these biomarkers. Saliva is an excellent source for detection of A. actinomycetemcomitans on individual basis, and high levels of the JP2 genotype were significantly associated with the presence of clinical attachment loss., Competing Interests: The authors declare that there is no conflict of interest.

Purpose: The grey-bluish discoloration of gingiva (known as "amalgam tattoo") does not appear only in the presence of amalgam restorations. It may also be seen in cases of teeth restored with cast dowels and porcelain-fused-to-metal (PFM) restorations. The aim of this article was to determine the clinical characteristics of abutment teeth with gingival discoloration., Materials and Methods: This research was conducted on 25 patients referred for cast dowel and PFM restorations. These restorations were manufactured from Ni-Cr alloys. Ninety days after cementing the fixed prosthodontic restorations, the abutment teeth (n = 61) were divided into a group with gingival discoloration (GD) (n = 25) and without gingival discoloration (NGD) (n = 36). The control group (CG) comprised the contralateral teeth (n = 61). Plaque index, gingival index, clinical attachment level, and probing depth were assessed before fabrication and also 90 days after cementation of the PFM restorations., Results: The gingival index, clinical attachment level, and probing depths of the abutment teeth that had GD were statistically higher before restoration, in comparison with the abutment teeth in the NGD and control groups. Ninety days after cementation, the abutment teeth with GD had significantly lower gingival indexes and probing depths, compared to the abutment teeth in the NGD group. Both abutment teeth groups (GD and NGD) had significantly higher values of clinical attachment levels when compared to the control group. There were no statistically significant differences in plaque index values between the study groups., Conclusions: The results of this study indicated that impairment of periodontal status of abutment teeth seemed to be related to the presence of gingival discolorations. Therefore, fabrication of fixed prosthodontic restorations requires careful planning and abutment teeth preparation to minimize the occurrence of gingival discolorations., Clinical Relevance: With careful preparation of abutment teeth for cast dowels and crown restorations it may be possible to decrease the frequency of gingival discolorations adjacent to abutment teeth., (© 2017 by the American College of Prosthodontists.)

Purpose: The aim of this study was to assess if there is an association between buccal mucosa thickness and periimplant attachment loss after 1 year of function., Materials and Methods: A total of 28 patients (14 periimplantitis implants and 14 healthy implants) were included. The buccal mucosal thickness was assessed using K-files at 3 mm apical to the soft tissue margin of the implant. Probing depth, recession (REC), clinical attachment level (CAL), bleeding on probing, and radiographic bone loss on mesial and distal sites of the implant were recorded., Results: The data showed that there was a statistically significant difference in midfacial REC between thin and thick buccal mucosa groups. However, the CAL was not statistically significant different between both groups. In addition, there was no statistically significant difference in mesial and distal bone loss between implants with thin and thick mucosa., Conclusion: When the midfacial soft tissue thickness was thin, the midfacial REC was greater and the CAL also tended to be higher. There was no association between buccal mucosa thickness and periimplant bone loss on mesial and distal sites of the implant after 1 year of function.

Background and Objective: Attachment loss of the junctional epithelium and alveolar bone destruction are signs of periodontitis, which is mainly caused by an inflammatory response to dental plaque. Glycyrrhetinic acid (GA), a component of the licorice herb, has been shown to have important anti-inflammatory activities; however, there are no previous reports on the ability of its inhibitory effects to prevent periodontal diseases. Hence, in this study, using our experimental periodontitis model, we attempted to evaluate whether GA had an effect on the prevention of attachment loss and alveolar bone loss., Material and Methods: Rats were intraperitoneally immunized with Escherichia coli lipopolysaccharide (LPS). The LPS group (n = 5) received 3 topical applications of 50 μg/μL of LPS followed by one application of the vehicle (propylene glycol:ethyl alcohol:phosphate-buffered saline [PBS] = 8:1:1) into the gingival sulcus. This protocol was repeated twice per day for 10 days. The low (n = 5) and high (n = 5) groups received topical application of LPS and 0.03% or 0.3% GA, respectively. The control group received topical application of PBS and vehicle. The rats were killed on the 10th day. Attachment loss, alveolar bone level and inflammatory cell infiltration were investigated histometrically. The formation of immune complexes and infiltration of LPS were evaluated immunohistologically., Results: Attachment loss, formation of immune complexes and infiltration of inflammatory cells were increased in the LPS group compared with the control group, and were completely inhibited in the low and high groups compared with the LPS group. The LPS group showed greater alveolar bone destruction compared with the control group and GA-treated groups. In addition, invasion of LPS was detected in the LPS group, was absent in the control group and was weaker in the GA-treated groups than in the LPS group., Conclusion: In the present study, we showed that GA inhibits periodontal destruction in the rat experimental periodontitis model., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background and Objective: The aim of the present study was to compare clinical periodontal parameters and to assess the release of C-telopeptides pyridinoline cross-links (ICTP) and C-terminal crosslinked telopeptide (CTX) from gingival collagen of naswar (NW) and non-naswar (control) dippers., Material and Methods: Eighty-seven individuals (42 individuals consuming NW and 45 controls) were included. Clinical (plaque index, bleeding on probing, probing depth and clinical attachment loss) and radiographic (marginal bone loss) periodontal parameters were compared among NW and control groups. Gingival specimens were taken from subjects in NW and control groups, assessed for ICTP and CTX levels (using ELISA) and analyzed using micro-Raman spectroscopy. The significance of differences in periodontal parameters between the groups was determined using Kruskal-Wallis and Mann-Whitney U tests. The percent loss of dry mass over exposure time and the rate of release of ICTP and CTX from all groups were compared using the paired t-test to examine the effects of exposure time., Results: Clinical and radiographic periodontal parameters were significantly higher in the NW group than the control group (P < .01). In the Raman spectrum, the strongest and sharpest band occurred at 1260 cm -1 amongst NW users. A Raman band at Amide I was observed with slight shifts in wave numbers. The rate of ICTP and CTX release was significantly higher in subjects from the NW group compared with those from the control group (P < .05). Both factors, the type of groups and time, had a significant effect on release of ICTP and CTX (P < .05)., Conclusion: Within the limits of the present study, it may be concluded that clinical and radiographic periodontal parameters were worse among subjects in the NW group than in those of the control group. There is a higher degree of collagen breakdown in the connective tissue of subjects in the NW group as a result of naswar usage., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Introduction: The purpose of this study was to evaluate the changes of the marginal periodontium 1 year after apical surgery., Methods: Clinical and radiographic (cone-beam computed tomographic) examinations of 54 teeth treated with buccal access flaps for apical surgery were performed at baseline and after 1 year. Clinical assessment included measurements of probing pocket depth, the level of gingival margin (GM), and the width of keratinized tissue. Subsequently, the clinical attachment level (CAL) and the width of the attached gingiva were calculated. On bucco-oral cone-beam computed tomographic sections, the height and thickness of the crestal bone and the thickness of the alveolar bone were measured at different levels., Results: In general, the calculated mean changes of periodontal tissue and crestal/alveolar bone were only minimal. Significant mean changes included only GM and CAL on midoral aspects and the distance from the cementoenamel junction or restoration margin on midbuccal sites. CAL was further correlated with the thickness of the alveolar bone at 3 mm below the cementoenamel junction or restoration margin. None of the clinically and radiographically calculated mean changes were correlated with sex, biotype, or incision techniques. With regard to age, older patients showed significantly more gingival recession on the buccal aspect compared with younger individuals. Furthermore, mean changes of the midbuccal width of the attached gingiva were positively correlated with the healing outcome, whereas mean changes of the midoral GM and CAL were negatively correlated with the healing outcome., Conclusions: Within an observation period of 1 year, the marginal periodontium and its underlying bone structures did not suffer from significant changes after apical surgery., (Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: The periodontal morphotype is a leading feature to be born in mind during surgical, implant, prosthetic, restorative as well as orthodontic treatment. A fragile morphotype will rapidly trigger severe and worrying clinical repercussions for the patient., Materials and Methods: In order to minimize the risks involved, sound knowledge of the different morphotypes and of the techniques for clinically evaluating them is a major requirement. These factors need to be regularly reassessed since morphotypes can vary throughout the course of treatment., Results: Detecting and controlling mucogingival risk factors will enable an appropriate clinical approach designed to avoid the onset of secondary recessions or other gingival pathologies., (© EDP Sciences, SFODF, 2017.)

Helicobacter pylori (H. pylori), a pathogen inducing peptic disease, is recently found to be binding to the progress of periodontitis. Most previous studies are case-controlled, and they investigate the risk of H. pylori infection in disease the development of while few studies evaluate the correlation between H. pylori and periodontal pathogens. Therefore, we investigated the correlation between H. pylori infection with periodontal parameters, periodontal pathogens and inflammation. The results indicated that patients with H. pylori showed significantly higher probing depth and attachment loss than those without (p < 0.05). Among 28 subgingival plaque samples from 14 patients, the frequencies of Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum and Treponema denticola were significantly higher with H. pylori infection than those without H. pylori infection (p < 0.05). However, the frequency of Aggregatibacter actinomycetemcomitans was lower (p < 0.05). Furthermore, after human acute monocytic leukemia cell line (THP-1) was stimulated with cagA-positive standard strains (cagA+ H. pylori 26695), the expression of periodontitis-related molecules Wnt5a, interleukin 8 (IL-8), interleukin 6 (IL-6) and interferon gamma (IFN-γ) significantly increased (p < 0.05). Conversely, the expression of tumor necrosis factor alpha (TNF-α) was almost stable. Meanwhile, cagA+ H. pylori promoted significantly higher expression of IL-8 and Wnt5a than isogenic cagA mutants strains (cagA- H. pylori 26695) did. Taken together, our data suggested that H. pylori might promote the growth of some periodontal pathogens and aggravate the progress of chronic periodontitis.

Purpose: In recent years, dental implants made from zirconia have been further developed and are considered a reliable treatment method for replacing missing teeth. The aim of this study was to analyze dental implants made from zirconia regarding their clinical performance compared with natural teeth (control)., Materials and Methods: One hundred six zirconia implants in 38 adults were analyzed in a clinical study after 1 year of loading. The plaque index (PI), bleeding on probing (BOP), probing pocket depth (PPD), probing attachment level (PAL), and creeping or recession (CR/REC) of the gingiva were detected and compared with natural control teeth (CT). Furthermore, the papilla index (PAP), Periotest values (PTV), microbial colonization of the implant/dental sulcus fluid, and patient satisfaction were assessed., Results: The survival rate was 100%. No statistical significance was observed between implants and teeth regarding BOP, PPD, and PAL. A statistical significance was detected regarding PI and CR/REC with significantly less plaque accumulation and recession in the study group. Mean PAP was 1.76 ± 0.55, whereas the mean PTV was -1.31 ± 2.24 (range from -5 to +6). A non-statistically significant higher colonization of periodontitis/peri-implantitis bacteria was observed in the implant group. The questionnaire showed that the majority of the patients were satisfied with the overall treatment., Conclusion: One-piece zirconia dental implants exhibited similar clinical results (BOP, PPD, and PAL) compared with natural teeth in regard to adhesion of plaque (PI) and creeping attachment (CR/REC); zirconia implants performed even better. The favorable results for PAL and CR/REC reflect the comparable low affinity of zirconia for plaque adhesion. Patient satisfaction indicated a high level of acceptance for zirconia implants. However, a long-term follow-up is needed to support these findings.

Background and Objective: Perforated barrier membranes open channels between the suprabony and intrabony compartments of the defect, which could allow for more physiologic cellular interactions between different components of the periodontium during guided tissue regeneration surgery. To test this assumption, this study was designed to evaluate levels of vascular endothelial cell growth factor (VEGF) and platelet-derived growth factor (PDGF)-BB in gingival crevicular fluid during the early stages of healing of localized intrabony defects treated with perforated membranes (PMs) or non-PMs, as compared with open flap debridement., Material and Methods: Thirty non-smoking patients with severe chronic periodontitis participated in this prospective, randomized and single blinded trial. Each patient contributed one interproximal defect that was randomly assigned to the PM group (n = 10), occlusive membrane (OM) group (n = 10) or open flap debridement (OFD) group (n = 10). Plaque index, gingival index, probing depth, clinical attachment level and the intrabony depth of the defect were measured at baseline and reassessed at 6 and 9 mo after therapy. Gingival crevicular fluid samples were collected on days 1, 3, 7, 14, 21 and 30 d after therapy for the changes in VEGF and PDGF-BB levels., Results: During the early stages of healing (1, 3 and 7 d), the mean VEGF and PDGF-BB concentrations at sites treated with PMs and OFD peaked with a statistically significant difference as compared with the OM-treated group. VEGF and PDGF-BB levels at sites treated with PMs and OFD were not statistically different. Growth factor levels decreased sharply in the samples obtained at days 21 and 30 with non-significant differences between the three groups. Nine months after therapy, the PM-treated group showed a statistically significant improvement in probing depth, clinical attachment level and intrabony defect compared to the OM and OFD groups., Conclusions: Within the limits of the present study, one can conclude that PM coverage of periodontal defects is associated with initial gingival crevicular fluid growth factor upregulation that could improve the clinical outcomes of guided tissue regeneration surgery., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Purpose: The aim of this cohort study was to evaluate the width of keratinized mucosa at implant sites of partially edentulous patients who were treated for generalized aggressive periodontitis., Materials and Methods: Dental implants were placed in 35 patients who were treated for generalized aggressive periodontitis and 18 periodontally healthy individuals (controls). At baseline, the keratinized mucosa of all implants was ≥ 2 mm. Follow-up examinations were conducted every 3 months over a 4-year period., Results: The implant survival rate was 97.3% in patients with generalized aggressive periodontitis and 100% in the control group. Four years after implant insertion, patients with generalized aggressive periodontitis had significantly higher clinical attachment levels at the teeth and implants compared with the controls. At all time points, in both groups the mean probing depth at the implants was significantly larger than at the teeth. The mean widths of keratinized mucosa and keratinized gingiva were not significantly different between the two groups. In both groups, the widths of keratinized mucosa and keratinized gingiva were significantly higher at the maxilla than at the mandible. Four years after baseline, the implants in the mandible showed the smallest keratinized mucosa (mean: ≤ 1 mm)., Conclusion: During the first 4 years after implant placement, no significant changes in the keratinized mucosa at implants could be shown, either in periodontally healthy patients or in patients treated for generalized aggressive periodontitis. The keratinized gingiva at the teeth was generally significantly wider than the keratinized mucosa at the implants.

Background and Objective: Insufficient information on the cellular composition of long-standing gingivitis lesions without signs of attachment loss makes an understanding of differences in cellular composition between "destructive" and "nondestructive" periodontal lesions difficult. The aim of the current study was to analyze differences in cell characteristics between lesions representing long-standing gingivitis and severe periodontitis., Material and Methods: Two groups of patients were recruited. One group consisted of 36 patients, 33-67 years of age, with severe generalized periodontitis (periodontitis group). The second group consisted of 28 patients, 41-70 years of age, with overt signs of gingival inflammation but no attachment loss (gingivitis group). From each patient a gingival biopsy was obtained from one selected diseased site and prepared for immunohistochemical analysis., Results: Periodontitis lesions were twice as large and contained significantly larger proportions, numbers and densities of cells positive for CD138 (plasma cells) and CD68 (macrophages) than did gingivitis lesions. The proportion of B cells that expressed the additional CD5 marker (B-1a cells) was significantly larger in periodontitis lesions than in gingivitis lesions. The densities of T cells and B cells did not differ between periodontitis lesions and gingivitis lesions. T cells were not the dominating cell type in gingivitis lesions, as B cells together with their subset plasma cells comprised a larger number and proportion than T cells., Conclusion: Periodontitis lesions at teeth with advanced attachment and bone loss exhibit quantitative and qualitative differences in relation to gingivitis lesions at teeth with no attachment and bone loss. It is suggested that the large number and high density of plasma cells are the hallmarks of advanced periodontitis lesions and the most conspicuous difference in relation to long-standing gingivitis lesions., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: In a secondary data analysis, this pilot study evaluated the relationship between subgingival biofilm morphotypes and chronic periodontitis progression in treated adults., Methods: Periodontal parameters in 47 adults with chronic periodontitis were assessed by a calibrated examiner at baseline and a mean 4.5 years after a non-surgical periodontal therapy regimen. Microbial and inflammatory cell morphotypes in subgingival biofilm specimens from each patient were evaluated with phase-contrast microscopy at baseline, and at post-treatment intervals. Chronic periodontitis progression in patients was defined as ≥ 2 teeth exhibiting ≥ 3 mm interproximal clinical periodontal attachment loss from baseline evaluations. Bivariate and odds ratio analysis assessed baseline and post-treatment variables relative to chronic periodontitis progression., Results: Eight (17%) patients had chronic periodontitis progression. No baseline clinical, radiographic or microbiological variables, and no post-treatment clinical variables demonstrated statistically significant relationships with chronic periodontitis progression. Elevated post-treatment counts of subgingival spirochetes, medium to large-sized motile rods, and crevicular leukocytes, both alone and concurrently, appeared more frequently in patients experiencing chronic periodontitis progression. A post-treatment occurrence of high concurrent counts of subgingival spirochetes and crevicular leukocytes exhibited the strongest association with chronic periodontitis progression (odds ratio = 10.1; 95% Cl = 2.2, 45.4; p = 0.004), which was greater than with either morphotype alone., Conclusions: Joint morphotype analysis of subgingival spirochetes and crevicular leukocytes, as simplified biomarkers of pathogenic biofilm infection and host inflammatory responses in periodontal pockets, may be diagnostically useful in assessing risk of progressive disease in treated chronic periodontitis patients.

Objective: To study non-osteoclastic sources of cathepsin K in periodontitis., Materials and Methods: Tissue samples were obtained from 10 otherwise healthy periodontitis pati-ents during routine periodontal flap operations and 10 systemically and periodontally healthy individuals who underwent extraction operations for retained third molars. Methods used were immunohistochemistry, image analysis, immunofluorescence double-staining, gingival fibroblast culture, tumour necrosis factor-α (TNF-α) stimulation and Western blotting., Results: Macrophage-like cells, fibroblast-like cells, vascular endothelial cells and gingival epithelial cells were more intensively stained for cathepsin K and also more frequent in periodontitis than in controls (665 ± 104 vs 258 ± 40 cells mm(-2) , P < 0.01). Some cathepsin K(+) cells in periodontal tissues were CD68(+) , but some were CD68(-) and probably fibroblasts. Indeed, in gingival fibroblast culture, resting fibroblasts released cathepsin K, more 43 kD procathepsin K than 29 kD active cathepsin K. TNF-α increased the release of the activated cathepsin K 4- to 5-fold., Conclusions: Results suggest that GCF-cathepsin K is not only osteoclast-derived, but in periodontitis, also other cells contribute to it. GCF-cathepsin K, perhaps together with intracellular, lysosomal collagenolytically active cathepsin K in fibroblasts, macrophages and gingival epithelial cells, can contribute to the loss of attachment and destruction of the periodontal ligament., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Smoking is one of the strongest predictors of attachment and bone loss. Smokers demonstrate reduced inflammatory clinical signs, which could be due to local vasoconstriction and increased gingival epithelial thickness. The byproducts originating from tobacco oxidation modify the clinical characteristics and progression of periodontal disease. The aim of this study was to investigate the relationship between the thickness of marginal gingival oral epithelium, sulcular bleeding, and vascular caliber and density of the microvessels in smokers and nonsmokers with and without periodontitis and to better understand the role of smoking in relation to periodontal disease. One hundred twenty individuals were enrolled in this study and divided into four groups comprising 30 participants each. The clinical measurements carried out included probing depth, clinical attachment loss, and bleeding index, along with gingival biopsy specimens, which were subjected to immunohistochemical and histomorphometric analysis. Correlation of the clinical and histologic features revealed that smokers presented with fewer inflammatory signs, had fewer vascular elements in the subepithelial connective tissue layer, and showed a resultant increase in epithelial thickness irrespective of the presence of periodontitis. There was a mean increase of epithelial thickness of 181.3 μm (suprapapillary epithelial thickness [SET]) to 380.2 μm (maximal epithelial thickness [MET]) in smokers with periodontitis as compared to 157.4 μm (SET) to 325.3 μm (MET) in nonsmokers with periodontitis. The mean microvascular density in smokers with periodontitis was 325.4 per mm, which was found to be statistically significantly less than that of nonsmokers with periodontitis, who had a mean value of 412.13 per mm. The vessel caliber also was reduced in smokers, with a mean value ranging from 4.7 to 6.1 μm compared with a mean of 6.2 to 9.2 μm in nonsmokers, irrespective of the presence of periodontitis. Statistically significant differences were found in vascular density and thickness of gingival epithelium between smokers and nonsmokers with and without periodontitis. These differences may impact the progression of periodontal disease.

Background and Objective: Green tea extract exerts a variety of biological effects, including anti-inflammatory activities. However, there has been no report on the effect of green tea extract on loss of attachment, which is an important characteristic of periodontitis. Here, we examined the inhibitory effects of green tea extract on the onset of periodontitis in a rat model., Material and Methods: Rats were immunized intraperitoneally with Escherichia coli lipopolysaccharide (LPS). The LPS group (n = 12) received a topical application of LPS onto the palatal gingival sulcus every 24 h. The green tea extract group (n = 12) received a topical application of LPS mixed with green tea extract, sunphenon BG, every 24 h. The phosphate-buffered saline (PBS) group (n = 6) received a topical application of PBS every 24 h. The levels of anti-LPS immunoglobulin G (IgG) in serum were determined using ELISA. Rats in the LPS and green tea extract groups were killed after the 10th and 20th applications. Rats in the PBS group were killed after the 20th application. Loss of attachment, level of alveolar bone and inflammatory cell infiltration were investigated histopathologically and histometrically. RANKL-positive cells and the formation of immune complexes were evaluated immunohistologically., Results: There was no significant difference in the serum levels of anti-LPS IgG between the LPS group and the green tea extract group. In contrast, loss of attachment, level of alveolar bone, inflammatory cell infiltration and RANKL expression in the green tea extract group were significantly decreased compared with those in the LPS group., Conclusion: These findings demonstrate that green tea extract suppresses the onset of loss of attachment and alveolar bone resorption in a rat model of experimental periodontitis.

Objective: The aim of this study was to evaluate the effect of the depth of curette sample collection from periodontal lesions on the recovery of putative periodontal pathogens using real-time polymerase chain reaction (PCR)., Methods: Twenty-two periodontal pockets 6 to 8 mm deep with bleeding on probing at a single-rooted tooth were sampled, yielding 66 separate samples. Curette samples were obtained at three different levels of the periodontal lesion (orifice, shallow--2 mm into the pocket; or base of lesion), and processed using PCR to identify 10 periodontal pathogens. The chi-square procedure was used to determine whether probe depth affected the distribution of bacterial counts observed. A repeated measures analysis of variance tested the hypotheses related to level of probe and microorganism on mean rank of bacterial counts., Results: The effect of probe level on mean bacterial counts depends on the type of microorganism. Likewise, the effect of microorganism type on mean bacterial counts significantly depends on probe level, where sampling from 2 mm into the periodontal pocket was found to yield significantly higher results than sampling from the orifice. Overall mean counts of pathogenic microorganisms were found to differ significantly across the three probe depths. The microorganisms differed in their observed levels over all three probe levels. Further analysis found several significant differences that characterize the nature of the interaction between probe level and microorganism type., Conclusion: There is significant difference in the amount of putative periodontal pathogens at varying depths of the pocket when sampled with a periodontal curette.

Background: Monocyte subsets with low CD14 expression that coexpress CD16 (CD14+CD16+) are called non-classic or hyperinflammatory monocytes. Previous studies have reported an increase in the percentage of CD14+CD16+ monocytes in the peripheral blood of patients with chronic periodontitis (CP). To our knowledge, there are no reports demonstrating the presence of CD14+CD16+ monocyte-derived macrophages (MDMs) in the gingival tissue. The objective of this study is to identify the proportion of non-classic (CD14+CD16+) monocytes/macrophages in peripheral blood and gingiva of healthy individuals and patients with CP., Methods: A total of 60 individuals (n = 30 per group) were recruited for the study. Group 1 included 30 individuals with healthy gingiva, and group 2 included 30 patients with CP. Direct immunofluorescent staining was done in 200 μL whole-blood and single-cell suspensions obtained from gingival tissue, with fluorochrome-conjugated monoclonal antibodies against CD14, CD16, and human leukocyte antigen-DR (HLA-DR), and subjected to flow cytometric analysis., Results: The mean percentage of CD14+CD16+ monocytes in the peripheral blood of healthy individuals was 9.10% ± 1.39%, and for patients with CP it was 14.18% ± 2.69% (P <0.05). The mean percentage of CD14+CD16+ MDMs in the gingival tissue of healthy individuals was found to be 0.93% ± 0.33%, whereas in patients with CP, it was 1.92% ± 0.78% (P <0.01). Non-classic monocytes/macrophages showed a high median fluorescent intensity for HLA-DR (DR++)., Conclusion: This study demonstrates an increased proportion of CD14+CD16+HLA-DR++ monocytes/macrophages in the peripheral blood and gingiva of patients with CP.

Background: Restoring papillae in the gingival embrasures of the esthetic zone is a key consideration in periodontal, restorative, and orthodontic treatment. This study analyzes the existence of interdental papillae with regard to the vertical dimensions between the contact point and the alveolar crest and horizontal dimensions between adjacent roots., Methods: A total of 259 papillae, in 60 patients, were examined. All patients were indicated for full-thickness periodontal flap surgery. A mucoperiosteal flap was raised and, after thorough debridement, horizontal and vertical dimensions between adjacent roots were recorded. The incidence of papillae was analyzed using χ(2) for linear trend analysis., Results: Interdental papillae were present in 85.7% of the cases when the vertical dimensions were 4 mm (P <0.05). Also, papillae were present in 78.5% of cases when the horizontal dimensions were between 0.5 and 1 mm (P <0.05)., Conclusion: The vertical and horizontal dimensions of interproximal areas had a substantial, independent, and combined effect on the existence of interproximal papillae.

Background and Objective: Occlusal trauma is an important factor that influences the progression of periodontitis, but it is unclear whether occlusal trauma influences periodontal destruction at the onset of periodontitis. We established an experimental periodontitis model with both site-specific loss of attachment and alveolar bone resorption. The purpose of the present study was to investigate the effects of occlusal trauma on periodontal destruction, particularly loss of attachment, at the onset of experimental periodontitis., Material and Methods: Sixty rats were used in the present study. Forty-eight rats immunized with lipopolysaccharide (LPS) intraperitoneally were divided into four groups. In the trauma (T) group, occlusal trauma was induced by placing an excessively high metal wire in the occlusal surface of the mandibular right first molar. In the inflammation (I) group, periodontal inflammation was induced by topical application of LPS into the palatal gingival sulcus of maxillary right first molars. In the trauma + inflammation (T+I) group, both trauma and periodontal inflammation were simultaneously induced. The PBS group was administered phosphate-buffered saline only. Another 12 nonimmunized rats (the n-(T+I) group) were treated as described for the T+I group. All rats were killed after 5 or 10 d, and their maxillary first molars with surrounding tissues were observed histopathologically. Loss of attachment and osteoclasts on the alveolar bone crest were investigated histopathologically. To detect immune complexes, immunohistological staining for C1qB was performed. Collagen fibers were also observed using the picrosirius red-polarization method., Results: There were significant increases in loss of attachment and in the number of osteoclasts in the T+I group compared with the other groups. Moreover, widespread distribution of immune complexes was observed in the T + I group, and collagen fibers oriented from the root surface to the alveolar bone crest had partially disappeared in the T, T+I and n-(T+I) groups., Conclusion: When inflammation was combined with occlusal trauma, immune complexes were confirmed in more expanding areas than in the area of the I group without occlusal trauma, and loss of attachment at the onset of experimental periodontitis was increased. Damage of collagen fibers by occlusal trauma may elevate the permeability of the antigen through the tissue and result in expansion of the area of immune-complex formation and accelerating inflammatory reaction. The periodontal tissue destruction was thus greater in the T+I group than in the I group., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: To date, flow cytometric immunophenotyping has not been used to investigate immune patterns in saliva samples from individuals with inflammatory processes in the oral cavity, such as chronic periodontitis (CP). Saliva analysis could be a non-invasive method for evaluating oral health. The objective of this study is to determine the phenotype of leukocytes and total immunoglobulin A (IgA), IgG, and IgM titers in the saliva of individuals with CP., Methods: Saliva samples were obtained from patients with CP (n = 12) and from a control group (n = 27) without oral diseases. Flow cytometry was performed to determine the frequency of T cells (CD4(+) and CD8(+)), B cells, and natural killer (NK) cells as well as the total leukocyte population. Immunoglobulin titers were determined by dot enzyme-linked immunosorbent assay., Results: Cell immunophenotyping revealed that patients with CP had a higher frequency of total leukocytes (47.94% ± 5.1%; P < 0.001), B cells (43.93% ± 6.2%; P = 0.006), NK cells (0.16% ± 0.04%; P = 0.03), and CD4(+) T cells (38.99% ± 4.4%; P = 0.002) than individuals without oral pathologies (24.75% ± 2.2%, 20.60% ± 2.7%, 0.09% ± 0.03%, and 16.82% ± 3.5%, respectively). No significant differences in salivary total IgA, IgG, and IgM titers were found between the two cohorts studied. Nevertheless, higher total IgG levels were observed in patients with CP, which could indicate a possible correlation between clinical attachment level and salivary IgG (P = 0.07; r(2) = 0.08)., Conclusion: These results show that cell phenotyping by flow cytometry could be an effective tool for determining leukocyte profiles in saliva samples from patients with CP and healthy individuals.

Background: Gingival recession is both unpleasant and unesthetic. Meeting the esthetic and functional demands of patients with multiple gingival recessions remains a major therapeutic challenge. We compared the clinical effectiveness of Zucchelli's technique and tunnel technique with subepithelial connective tissue graft (SECTG) for multiple gingival recessions., Methods: Twenty systemically and periodontally healthy subjects having 75 recession defects (Miller's class I or II, 39 test and 36 control sites) were included. After initial nonsurgical therapy, test sites were treated with Zucchelli's technique and control sites with tunnel technique with SECTG. Plaque index, bleeding index, pocket depth, recession depth, clinical attachment level, and keratinized gingiva height were evaluated at baseline, 3 and 6 months post-surgery., Results: The mean root coverage was 89.33% +/- 14.47% and 80.00% +/- 15.39% in the test and control groups respectively, with no significant difference between groups. Statistically significant root coverage was obtained for 82.50% +/- 23.72% and 71.40% +/- 20.93% of defects in the test and control groups, respectively., Conclusion: Zucchelli's technique is effective for the treatment of multiple adjacent recessions in terms of both root coverage and keratinized tissue gain, irrespective of the number of defects. Moreover, this technique does not require an additional surgical site as required in the gold standard SECTG.

Background: In guided tissue regenerative surgery, membrane perforations may serve as a mechanism for the passage of cells and biologic mediators from the periosteum and overlying gingival connective tissue into the periodontal defects. To test this assumption, this study was designed to evaluate levels of bone morphogenetic protein-2 (BMP-2) in gingival crevicular fluid (GCF) during the early stages of healing for sites treated with modified perforated membranes (MPMs) as compared with occlusive membranes (OMs)., Methods: Fifteen non-smoking patients with severe chronic periodontitis participated in this prospective, randomized and single-blinded clinical trial. Each patient contributed two interproximal contralateral defects that were randomly assigned to either an experimental modified perforated membrane group (15 sites) or a control occlusive membrane group (15 sites). Plaque index, gingival index, probing depth(PD), clinical attachment level (CAL) and the relative intrabony depth of the defect (rIBD) were measured at baseline and reassessed at three, six and nine months after therapy. Gingival crevicular fluid samples were collected on day 1 and 3, 7, 14, 21, and 30 days after therapy., Results: The MPM-treated group showed a statistically significant improvement in PD reduction and clinical attachment gain compared to the OM control group. Similarly, rIBD was significantly reduced in MPM-treated sites as compared with those of the OM group. BMP-2 concentrations peaked in the MPM samples obtained during the early postoperative period (days 1, 3 and 7) with a statistically significant difference compared with OM-treated groups. BMP-2 levels decreased sharply in the samples obtained at days 14, 21 and 30 with non-significant higher levels in MPM samples as compared with those of OM sites., Conclusion: Within the limits of the present study, one can conclude that MPM coverage of periodontal defects is associated with a significant initial increase in GCF levels of BMP-2, a factor that could improve the clinical outcomes of guided tissue regenerative surgery.

Background: Subantibiotic doses of doxycycline (low-dose doxycycline [LDD]) have been widely used in periodontal treatment for enzymatic inhibition and related anti-inflammatory properties. The aim of the present study is to verify the possible effects of LDD on oxidative stress in relation to periodontal attachment loss associated with ligature-induced experimental periodontal disease in rats., Methods: Thirty female Wistar albino rats were divided into three study groups as follows: 1) control (C) rats; 2) rats with experimental periodontitis (PED); and 3) rats with PED that were treated with doxycycline (PED + LDD). PED was induced by placing ligatures around the cervix of the maxillary second molars for 21 days. The PED + LDD group was treated orally with doxycycline (6 mg/kg) for 21 days after the ligature was placed. After 21 days, the rats were euthanized, and samples of the right maxilla were defleshed and used for histologic and morphometric analyses. The gingival tissue of the left maxilla was used for the analysis of lipid peroxidation (malondialdehyde [MDA]) and antioxidant enzymes (catalase, glutathione peroxidase, and superoxide dismutase). Levels of serum total antioxidant status (TAS)/total oxidant status (TOS) and oxidative stress index (OSI) were also analyzed., Results: Alveolar bone loss was significantly higher in the PED group compared with the PED + LDD and C groups (P <0.05). Doxycycline exhibited the most prominent inhibition on gingival tissue levels of MDA and antioxidant enzymes (P <0.05). Doxycycline also significantly reduced TOS and OSI levels (P <0.05) but increased the TAS level., Conclusion: Doxycycline helps to prevent periodontal tissue breakdown by inhibiting local and systemic oxidative stress.

Background: Although dental dilaceration disinclusion is an accepted treatment modality, few studies have evaluated the prognosis for dilacerated maxillary incisors and changes in clinical periodontal parameters still need to be demonstrated. The objectives of this study were to evaluate the prognosis and changes in clinical attachment level (CAL), probing depth (PD), and soft tissue recession (REC) for disincluded dilacerated maxillary incisors., Methods: Ten impacted dilacerated teeth were scheduled for disinclusion. Five of them were disincluded with the apically repositioned flap technique and the other five with the closed eruption technique and brought into alignment with light orthodontic forces., Results: All the dilacerated teeth were disincluded as planned. CAL, PD, and REC were the same as natural teeth. Among the two surgical techniques, no statistically significant differences have been found., Conclusion: The choice to disinclude dilacerated central maxillary incisors reached the goals planned. Periodontal parameters obtained in a short- and long-term follow-up allow to affirm that the disinclusion of dilacerated teeth has a good survival rate.

The 2 major forms of periodontitis, chronic (CP) and aggressive (AgP), do not display sufficiently distinct histopathological characteristics or microbiological/immunological features. We used molecular profiling to explore biological differences between CP and AgP and subsequently carried out supervised classification using machine-learning algorithms including an internal validation. We used whole-genome gene expression profiles from 310 'healthy' or 'diseased' gingival tissue biopsies from 120 systemically healthy non-smokers, 65 with CP and 55 with AgP, each contributing with ≥ 2 'diseased' gingival papillae (n = 241; with bleeding-on-probing, probing depth ≥ 4 mm, and clinical attachment loss ≥ 3 mm), and, when available, a 'healthy' papilla (n = 69; no bleeding-on-probing, probing depth ≤ 4 mm, and clinical attachment loss ≤ 4 mm). Our analyses revealed limited differences between the gingival tissue transcriptional profiles of AgP and CP, with genes related to immune responses, apoptosis, and signal transduction overexpressed in AgP, and genes related to epithelial integrity and metabolism overexpressed in CP. Different classifying algorithms discriminated CP from AgP with an area under the curve ranging from 0.63 to 0.99. The small differences in gene expression and the highly variable classifier performance suggest limited dissimilarities between established AgP and CP lesions. Future analyses may facilitate the development of a novel, 'intrinsic' classification of periodontitis based on molecular profiling.

Background: With increase in the incidence of resistance to antibiotics, probiotics are emerging as a promising adjunctive periodontal therapy. The authors of this study evaluate the influence of probiotic (PROB) supplementation on ligature-induced periodontitis (LIP) and intestinal morphology in rats., Methods: Thirty-two rats were randomly divided into four groups: control (C), LIP, PROB, and LIP/PROB. In groups PROB and LIP/PROB, the PROB was administered orally by addition to the drinking water of the animals for 44 days. In groups LIP and LIP/PROB, the mandibular right first molar of the animals received a cotton ligature that was left in the same position for 14 days. All animals were euthanized 44 days after the start of the PROB supplementation. The jaws were resected and histomorphometric analyses were performed. The measurements included evaluation of attachment loss (AL) and alveolar bone level (ABL) on the distal root of the mandibular first molar. Samples of the duodenum, jejunum, and ileum were also dissected from each animal to evaluate the villous height (VH) and crypt depth (CD). The data obtained were subjected to statistical analyses (analysis of variance, Tukey; P <0.05)., Results: Mean values of AL and ABL were significantly higher in group LIP compared with group LIP/PROB (AL: 3.05 ± 0.57 mm and 1.78 ± 0.63 mm, respectively; ABL: 4.21 ± 0.42 mm and 3.38 ± 0.17 mm, respectively). In group LIP/PROB, the mean values of VH and CD of the jejunum were significantly higher than the ones from group LIP (VH: 672.1 ± 83.3 µm and 528.0 ± 51.7 µm, respectively; CD: 463.8 ± 100.9 µm and 269.0 ± 48.4 µm, respectively)., Conclusions: It can be concluded that PROB supplementation 1) reduces AL and alveolar bone loss in rats with LIP and 2) can protect the small intestine from reactive changes induced by LIP.

Background and Objective: The effects of tobacco smoking on the prevalence and severity of periodontal disease have been well documented. However, very few studies have assessed the effects of oral smokeless tobacco (ST) on the periodontium. Considering the widespread use of ST products globally, the effects of such products on the periodontal tissues may be important. The present study was performed to compare retrospectively the patterns of periodontal destruction among oral ST users and never-users with periodontitis., Material and Methods: Data from 149 patients with periodontitis (60 ST users and 89 never-users) were compared for mean scores of probing depth, recession (REC) and clinical attachment loss (CAL) and the mean percentage of sites with different ranges of probing depth, REC and CAL., Results: For full-mouth scores, mean REC and CAL were significantly higher in ST users than in never-users (p < 0.001 and p = 0.008, respectively). For different regions of the dentition, mean scores of REC were significantly higher among ST users than among never-users (p < 0.001 for all regions), and mean scores of CAL were significantly higher for ST users in mandibular (p < 0.001), buccal (p = 0.008), lingual (p = 0.022), anterior (p = 0.012) and molar (p = 0.009) sites. Generally, there were higher percentages of sites with shallow pockets (0-3 mm), REC of ≥ 1 mm and CAL of ≥ 3 mm in ST users than in never-users. However, only differences for REC categories were significant for all regions (p < 0.001). ST users had a significantly higher proportion of sites with CAL of ≥8 mm for full-mouth sites (p = 0.003), mandibular teeth (p < 0.001), buccal sites (p = 0.002), anterior teeth (p = 0.040) and molars (p = 0.007)., Conclusion: ST users tend to have more severe REC and CAL and a greater proportion of sites with higher values of REC and CAL compared with never-users. The greatest increase in severity of CAL was found to be localized to sites on mandibular teeth, buccal surfaces, anteriors and molars, which may be a result of the retention of the ST product in the oral cavity., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Aims: To investigate the influence of subject-, tooth- and site-level characteristics on intra-examiner reproducibility of direct and indirect clinical attachment level (CALDIR and CALIND ) recordings, and to quantify the impact of less-than-perfect reliability on our ability to assess periodontitis progression., Materials and Methods: Within a 1-week interval, a single examiner performed duplicate probing depth (PD), CALDIR and gingival recession (GR) recordings in six sites of all teeth present in 148 periodontitis patients. CALIND was calculated on the basis of the PD and GR., Results: Agreement was observed in 65%, 62%, 69% and 84% of the duplicate CALDIR , CALIND , PD and GR recordings, respectively, and >95% of the differences were within ±1 mm. This study identified multiple predictors for CAL measurement errors ≥1 mm, including tooth and site location, presence of supra- and subgingival calculus, bleeding on probing and suppuration. Measurement errors were more likely in patients diagnosed with "extensive" rather than "less extensive" periodontitis. In over half of the patients, measurement error frequencies were too high to allow for detection of possible CAL changes ≥2 mm with a false-positive rate ≤5%. Detection of CAL changes with low false-positives was more likely using recordings obtained by the direct method., Conclusions: From a measurement error point of view, CALDIR recordings are preferable over CALIND ., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: It is known that tumour necrosis factor (TNF)-α converting enzyme (TACE) plays a crucial role in fibrotic inflammatory diseases, and is specifically inhibited by tissue inhibitor of metalloproteinase (TIMP)-3. Fibrotic drug-induced gingival overgrowth (GO) is often combined with periodontitis. However, neither TACE nor TIMP-3 has been thoroughly examined in periodontal tissues to date. The aim of the present study was to analyse mRNA expression of TACE and TIMP-3, and protein localisation of TACE in gingival tissues removed from drug-(calcium-channel blocker) induced GO and periodontitis., Methods: A total of 30 gingival tissue samples were taken from 15 GO and 15 periodontitis patients. The mRNA expression levels were analysed by quantitative reverse transcription polymerase chain-reaction (qRT-PCR) and the protein localisation was investigated by immunohistochemistry. Statistical analysis was performed using the Mann-Whitney U-test., Results: TACE and TIMP-3 mRNA levels were significantly higher in GO compared to the periodontitis groups, as revealed by qRT-PCR (p<0.05). TACE-producing cells were immunohistochemically detected among monocytes/macrophages, plasma cells and some epithelial cells. TACE immunoreactivity was shown to be more intense in GO than in periodontitis-gingival tissue., Conclusions: We have demonstrated TACE expression in cells such as macrophages, plasma cells and epithelial cells, and its predominant expression in GO tissues. This data suggests that TACE expression in GO-gingiva could be involved in the pathogenesis of disease., (Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.)

Background: Mast cells appear to be associated with human periodontal disease. However, to the best of our knowledge, no studies have evaluated the presence of mast cells in individuals with human immunodeficiency virus (HIV) undergoing highly active antiretroviral therapy (HAART)., Methods: Gingival samples were obtained from 50 individuals with and without HIV who presented with chronic gingivitis and periodontitis. Immunohistochemical staining was performed to identify c-kit and tryptase mast cells. Inflammatory infiltrate was evaluated and quantified in the specimens of gingival tissue. In the inflammatory infiltrate subjacent to the pocket epithelium, the densities of the mast cells were calculated. These results were compared between the groups with and without HIV. Correlations could be drawn between the densities of c-kit and tryptase mast cells and the density of inflammatory infiltrate, cluster of differentiation 4 (CD4)-positive and CD8(+) T-lymphocyte levels, and viral loads., Results: Although the individuals with HIV had higher densities of c-kit or tryptase mast cells than those without HIV, both groups presented with chronic gingivitis or periodontitis, and no statistically significant differences could be observed. Both strong and negative correlations could be observed among the inflammatory infiltrate, c-kit, and tryptase in individuals both with and without HIV. Concerning the chronic gingivitis of the individuals with HIV, both strong and positive correlations could be observed between the density of c-kit and CD8(+) T-lymphocyte levels., Conclusions: Individuals with HIV undergoing HAART, compared with individuals without HIV, had no statistically significant differences in mast cell densities in gingival tissue.

Orthodontics contributes to improving self-esteem and function. However, a common refrain in ortho-perio relationships states that orthodontics has no deleterious effects on the healthy or reduced and treated periodontium. Though supported for decades, this idea has now been heavily challenged by a recent systematic review of the literature. The conclusion of this article is unfortunately very clear. There is a lack of reliable evidence showing the beneficial effects of orthodontic treatment on periodontal health with, at best, mild adverse effects. How can we reduce the periodontal cost of orthodontic treatment to a minimum? How can we ensure that this "at best" does not turn into "at worst"? To minimize the adverse effects of orthodontic treatment on periodontal tissues, the orthodontist must be able to determine to which patients he/she can consider providing orthodontic treatment and those for whom prior periodontal treatment is mandatory. In addition to the items collected for an orthodontic diagnosis, the orthodontist must be able to recognize the eight signs of attachment loss., (Copyright © 2013. Published by Elsevier Masson SAS.)

Background and Objective: Gingival crevicular fluid has been suggested as a possible source of biomarkers for periodontal disease progression. This paper describes a technique for the analysis of gingival crevicular fluid from individual sites using mass spectrometry. It explores the novel use of mass spectrometry to examine the relationship between the relative amounts of proteins and peptides in gingival crevicular fluid and their relationship with clinical indices and periodontal attachment loss in periodontal maintenance patients. The aim of this paper was to assess whether the mass spectrometric analysis of gingival crevicular fluid may allow for the site-specific prediction of periodontal disease progression., Material and Methods: Forty-one periodontal maintenance subjects were followed over 12 mo, with clinical measurements taken at baseline and every 3 mo thereafter. Gingival crevicular fluid was collected from subjects at each visit and was analysed using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Samples were classified based upon pocket depth, modified gingival index (MGI), plaque index and attachment loss, and were analysed within these groups. A genetic algorithm was used to create a model based on pattern analysis to predict sites undergoing attachment loss., Results: Three hundred and eighty-five gingival crevicular fluid samples were analysed. Twenty-five sites under observation in 14 patients exhibited attachment loss of > 2 mm over the 12-mo period. The clinical indices pocket depth, MGI, plaque levels and bleeding on probing served as poor discriminators of gingival crevicular fluid mass spectra. Models generated from the gingival crevicular fluid mass spectra could predict attachment loss at a site with a high specificity (97% recognition capability and 67% cross-validation)., Conclusions: Gingival crevicular fluid mass spectra could be used to predict sites with attachment loss. The use of algorithm-generated models based on gingival crevicular fluid mass spectra may provide utility in the diagnosis of periodontal disease., (© 2012 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background and Objective: Dentin sialophosphoprotein (DSPP) and its cleaved products, dentin phosphoprotein (DPP) and dentin sialoprotein (DSP), play important roles in biomineralization. Recently, we observed that DSPP is highly expressed in the alveolar bone and cementum, indicating that this molecule may play an important role in the formation and maintenance of a healthy periodontium, and its deletion may cause increased susceptibility to periodontal diseases. The objective of this investigation was to study the effects of Dspp ablation on periodontal tissues by analyzing Dspp null mice., Material and Methods: Newborn to 6-mo-old Dspp null mice were examined, and the 3- and 6-mo-old Dspp null mice were characterized in detail using X-ray radiography, histology and scanning electron microscopy (backscattered as well as resin-infiltrating). Wild-type mice of the same age groups served as the normal controls., Results: The Dspp null mice showed significant loss of alveolar bone and cementum, particularly in the furcation and interproximal regions of the molars. The alveolar bone appeared porous while the quantity of cementum was reduced in the apical region. The canalicular systems and osteocytes in the alveolar bone were abnormal, with reduced numbers of canaliculi and altered osteocyte morphology. The loss of alveolar bone and cementum along with the detachment of the periodontal ligaments (PDL) led to the apical migration of the epithelial attachment and formation of periodontal pockets., Conclusion: Inactivation of DSPP leads to the loss of alveolar bone and cementum and increased susceptibility to bacterial infections in PDL of Dspp null mice. The fact that the loss of DSPP results in periodontal diseases indicates that this molecule plays a vital role in maintaining the health of the periodontium., (© 2012 John Wiley & Sons A/S.)

Background: Bone loss is a feature of both periodontitis and osteoporosis, and several studies have analyzed whether the periodontal destruction could have been influenced by systemic bone loss. The aim of this study is to assess the association between clinical attachment level (CAL) and bone mineral density (BMD) at the lumbar spine and hip, lifestyle, smoking, sociodemographic factors, and dental clinical variables in postmenopausal women., Methods: One hundred forty-eight women were interviewed using a structured written questionnaire and clinically examined. The periodontal examination, which was performed by calibrated investigators, included CAL, probing depth, gingival recession, bleeding on probing (BOP), visible plaque, supragingival calculus, and mean tooth loss. The sample was stratified into two groups: moderate and severe CAL. The moderate group had all sites with CAL ≤5 mm. The severe group had ≥1 site with CAL >5 mm. BMD, measured using dual-energy x-ray absorptiometry, was assessed at the lumbar spine, femoral neck, and total femur (grams per square centimeters)., Results: Severe CAL was identified in 86 women (58.1%). The multiple linear regression analysis using CAL (dependent variable), adjusted by menopause, education, and family income, demonstrated an inverse relationship of severe CAL with the BMD of the femoral neck (P = 0.015), as well as a positive association of severe CAL with tooth loss (P = 0.000), BOP (P = 0.004), and heavy smokers (P = 0.001)., Conclusions: Our study demonstrated that severe CAL was associated with low BMD of the femoral neck and deleterious clinical dental parameters and smoking. Our findings suggest that, in addition to appropriate oral care, individuals with severe CAL may also require additional attention to their systemic bone health.

Objectives: The aim of the study was to clinically and histologically evaluate the healing of human intrabony defects treated with open flap surgery (OFD) and application of a new, resorbable, fully synthetic, unsintered, nanocrystalline, phase-pure hydroxyapatite (nano-HA)., Materials and Methods: Six patients, each of them displaying very advanced intrabony defects around teeth scheduled for extraction due to advanced chronic periodontitis and further prosthodontic considerations, were included in the study. Following local anaesthesia, mucoperiosteal flaps were reflected; the granulation tissue was removed, and the roots were meticulously debrided by hand and ultrasonic instruments. A notch was placed at the most apical extent of the calculus present on the root surface or at the most apical part of the defect (if no calculus was present) in order to serve as a reference for the histological evaluation. Following defect fill with nano-HA, the flaps were sutured by means of mattress sutures to allow primary intention healing. At 7 months after regenerative surgery, the teeth were extracted together with some of their surrounding soft and hard tissues and processed for histological analysis., Results: The postoperative healing was uneventful in all cases. At 7 months following surgery, mean PPD reduction and mean CAL gain measured 4.0 ± 0.8 and 2.5 ± 0.8 mm, respectively. The histological analysis revealed a healing predominantly characterized by epithelial downgrowth. Limited formation of new cementum with inserting connective tissue fibers and bone regeneration occurred in three out of the six biopsies (i.e. 0-0.86 and 0-1.33 mm, respectively). Complete resorption of the nano-HA was found in four out of the six biopsies. A few remnants of the graft particles (either surrounded by newly formed mineralized tissue or encapsulated in connective tissue) were found in two out of the six biopsies., Conclusion: Within their limits, the present results indicate that nano-HA has limited potential to promote periodontal regeneration in human intrabony defects., Clinical Relevance: The clinical outcomes obtained following surgery with OFD + nano-HA may not reflect true periodontal regeneration.

Aim: The aim of this randomized controlled clinical trial was to evaluate the clinical effects of chlorhexidine (CHX) application in a full-mouth disinfection (FMD) protocol in poorly controlled type-2 diabetic subjects with generalized chronic periodontitis., Material and Methods: Thirty-eight subjects were randomly assigned into FMD group (n=19): full-mouth scaling and root planing (FMSRP) within 24 h + local application of CHX gel + CHX rinses for 60 days or Control group (n = 19): FMSRP within 24 h + local application of placebo gel + placebo rinses for 60 days. Clinical parameters, glycated haemoglobin and fasting plasma glucose were assessed at baseline, 3, 6 and 12 months post-therapies., Results: All clinical parameters improved significantly at 3, 6 and 12 months post-therapies for both groups (p < 0.05). There were no significant differences between groups for any clinical parameters, and glycemic condition at any time-point (p > 0.05)., Conclusions: The treatments did not differ with respect to clinical parameters, including the primary outcome variable (i.e. changes in clinical attachment level in deep pockets), for up to 12 months post-treatments., (© 2012 John Wiley & Sons A/S.)

Background: Mast cells are tissue-resident immune cells that participate in a variety of allergic and inflammatory conditions. Limited attention has been given to the role of mast cells in periodontal diseases, and the effects of mast cell degranulation on the chronic stages of non-allergic inflammation, particularly in periodontitis, are not known. The present study analyzes the relationship between the mast cell degranulation and human periodontal disease progression., Methods: A total of 50 clinical specimens including moderate periodontitis (n = 17), advanced periodontitis (n = 18), and healthy control tissues (n = 15) were used in this study. All specimens were fixed in 10% buffered formalin and stained with hematoxylin and eosin for histopathology, with toluidine blue for identifying mast cells, and by immunohistochemistry for the expressions of mast cell tryptase in periodontal tissues. The total and degranulated mast cell densities (per high-power field) were quantified in the specimens., Results: Compared with healthy controls, there were significantly increased both total and degranulated mast cell densities in human moderate (P <0.01) and advanced (P <0.01) periodontitis groups by toluidine blue staining, and there were significantly higher densities of both total and degranulated tryptase-positive mast cell subpopulation in the moderate periodontitis group (P <0.01) and even significantly higher subpopulation densities in the advanced periodontitis group by immunohistochemical staining, in which both total and degranulated mast cell densities were significantly higher in the advanced periodontitis group than those in the moderate periodontitis group (P <0.01) by both toluidine blue staining and immunohistochemical staining. There was significantly more severe periodontal inflammatory pathology in the advanced periodontitis group than in the moderate periodontitis group (P <0.01)., Conclusion: These findings indicate a significant correlation among tryptase-positive mast cell density, the degree of their degranulation, and the human periodontitis severity, and the results of this study further indicate that mast cell degranulation appears to be associated with human periodontal disease.

Bone sialoprotein (BSP) is an extracellular matrix protein found in mineralized tissues of the skeleton and dentition. BSP is multifunctional, affecting cell attachment and signaling through an RGD integrin-binding region, and acting as a positive regulator for mineral precipitation by nucleating hydroxyapatite crystals. BSP is present in cementum, the hard tissue covering the tooth root that anchors periodontal ligament (PDL) attachment. To test our hypothesis that BSP plays an important role in cementogenesis, we analyzed tooth development in a Bsp null ((-/-)) mouse model. Developmental analysis by histology, histochemistry, and SEM revealed a significant reduction in acellular cementum formation on Bsp (-/-) mouse molar and incisor roots, and the cementum deposited appeared hypomineralized. Structural defects in cementum-PDL interfaces in Bsp (-/-) mice caused PDL detachment, likely contributing to the high incidence of incisor malocclusion. Loss of BSP caused progressively disorganized PDL and significantly increased epithelial down-growth with aging. Bsp (-/-) mice displayed extensive root and alveolar bone resorption, mediated by increased RANKL and the presence of osteoclasts. Results collected here suggest that BSP plays a non-redundant role in acellular cementum formation, likely involved in initiating mineralization on the root surface. Through its importance to cementum integrity, BSP is essential for periodontal function.

The Triggering Receptor Expressed on Myeloid cells 1 (TREM-1) is a cell-surface receptor of the immunoglobulin superfamily, involved in the propagation of the inflammatory response to bacterial challenge. Soluble (s)TREM-1 is released from the cell surface during the course of infection and is a useful inflammatory biomarker in the early diagnosis of systemic sepsis. The hypothesis of this study was that oral and systemic levels of sTREM-1 are elevated in periodontitis. Therefore, the aim was to investigate, by ELISA, the sTREM-1 concentrations in saliva and serum of individuals without periodontitis (control) and persons with chronic or generalized aggressive periodontitis. In saliva, sTREM-1 concentrations were higher in chronic and aggressive periodontitis than in the control group, by 3.3-fold and 5.6-fold, respectively. In serum, these differences were 1.7-fold and 2-fold, respectively. However, there were no significant differences between the two forms of periodontitis, neither in saliva nor in serum. Salivary and serum sTREM-1 levels positively correlated with full-mouth clinical periodontal parameters. In conclusion, the increased oral and systemic levels of sTREM-1 in periodontitis denote a value for this molecule as a biomarker for the disease and may also have implications in the association between periodontal infections and systemic inflammatory response.

Background and Objective: The clinical efficacy of EMDs for the treatment of periodontal infrabony defects has been reported. However, recent publications have questioned the validity of results from early findings. Hence, the purpose of this study was to compare the results obtained from early and late studies when EMD was used as an adjunct in treating human intrabony defects during flap surgery. The aim of this meta-analysis was to evaluate the validity of results published from early studies compared with those published from later studies., Material and Methods: PubMed and MEDLINE searches were performed. The evaluation period was 1997-2010 and it was divided into two groups of equal periods of time: early studies (1997-2003) and late studies (2004-2010). The clinical parameters assessed were clinical attachment level (CAL), probing pocket depth and bone gain (BG; measured as a percentage or in mm)., Results: No statistically significant difference was found between the results obtained from early studies (1997-2003) and late studies (2004-2010) with regards to CAL gain, probing pocket depth reduction and BG. Nonetheless, both study periods showed a benefit for using EMD to treat periodontal infrabony defects when compared with the groups without EMD during open flap surgery., Conclusions: The results obtained from this study failed to show any potential differences between the results published from early studies and late studies with regards to the clinical effectiveness of EMD in treating periodontal infrabony defects., (© 2012 John Wiley & Sons A/S.)

Objectives: The aim of this study was to compare the long-term clinical effects produced by subepithelial connective tissue graft (SCTG) and guided tissue regeneration combined with demineralized freeze-dried bone allograft (GTR-DFDBA) in the treatment of gingival recessions in a 30-month follow-up clinical trial., Methods: Twenty-four defects were treated in 12 patients who presented canine or pre-molar Miller class I and/or II bilateral gingival recessions. GTR-DFDBA and SCTG treatments were performed in a randomized selection in a split-mouth design. The clinical measurements included root coverage (RC), gingival recession (GR), probing depth (PD), clinical attachment level (CAL) and keratinized tissue width (KTW). These clinical parameters were evaluated at baseline and after 6, 18 and 30months post-surgery., Results: The changes in RC, GR, PD and CAL did not show significant differences between groups (p>0.05). Both procedures promoted similar RC (GTR-DFDBA: 87% and SCTG: 95.5%) and similar reduction in GR (GTR-DFDBA: 3.25mm and SCTG: 3.9mm), PD (GTR-DFDBA: 1.6mm and SCTG: 1.2mm) and CAL (GTR-DFDBA: 4.9mm and SCTG: 5.0mm). The increase in KTW was significantly higher (p=0.02) in the SCTG group (3.5mm) than in the GTR-DFDBA group (2.4mm)., Conclusions: Both techniques for treatment of gingival recession (SCTG and GTR-DFDBA) lead to favourable and long-term stable results, but SCTG promoted a more favourable increase in keratinized tissue., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background and Objective:   There are few data concerning the effect of scaling and root planing on the levels of immune and inflammatory mediators in gingival crevicular fluid from patients with chronic periodontitis. Therefore, in this study the influence of scaling and root planing was determined on amounts of interleukin (IL)-1β, IL-8 and MMP-8 in gingival crevicular fluid from patients with chronic periodontitis, in relation to clinical parameters., Material and Methods:   A total of 51 patients were enrolled in this study. The study population consisted of 30 patients with generalized advanced chronic periodontitis, while 21 periodontally healthy subjects were recruited for the control group. The clinical parameters included approximal plaque index, gingival index, pocket depth and clinical attachment loss. The amounts of IL-1β, IL-8 and MMP-8 in gingival crevicular fluid were measured by ELISA. Periodontal parameters as well as gingival crevicular fluid humoral factor amounts were evaluated in the control group and in chronic periodontitis patients at baseline and at 1 and 4 wk after scaling and root planing treatment., Results:   At baseline, there were significant differences between control subjects and chronic periodontitis patients in terms of clinical attachment loss, pocket depth, gingival index (p < 0.001) and approximal plaque index (p < 0.01). The amounts of IL-1β, MMP-8 (p < 0.001) and IL-8 (p < 0.01) in gingival crevicular fluid were significantly lower in healthy subjects than in chronic periodontitis patients. Scaling and root planing led to improvement in all examined clinical parameters, apart from clinical attachment loss. Periodontal treatment also resulted in a significant decrease in the amounts of IL-1β, IL-8 and MMP-8 in comparison to baseline, especially 4 wk after scaling and root planing (p < 0.001); however, the amounts of these humoral factors were still higher than those in control group., Conclusion: Our observations indicated that short-term nonsurgical therapy resulted in a significant improvement in periodontal indices and in a marked decrease of IL-1β, IL-8 and MMP-8 gingival crevicular fluid levels. Nevertheless, no significant correlations were found between clinical parameters and amounts of humoral factors after therapy., (© 2012 John Wiley & Sons A/S.)

Objective: The aim of this study was to evaluate the presence and distribution of CD1a and S100 protein markers in states of gingival health and chronic periodontitis in human subjects., Materials and Methods: Gingival tissue samples were derived from 10 healthy and 10 chronic periodontitis-affected human subjects. The presence and distribution of CD1a and S100 protein was assessed using immunohistochemistry, and the cell types involved in their expression was determined., Results: The presence and distribution of CD1a was confined only to the gingival epithelium, whereas S100 was seen in the epithelium and connective tissue. However, increased expression of both CD1a and S100 protein was seen in periodontitis-affected gingival tissues compared with healthy gingiva. Immunohistochemistry demonstrated that CD1a- and S100-positive cells in the epithelium are Langerhans cells (LCs) and S100 positive cells in the connective tissue are dendritic cells (DCs)., Conclusion: Our findings suggest the transition of CD1a-positive LCs to S100-positive DCs from epithelium to connective tissue in response to an antigenic challenge. Demonstration of increased number of S100-positive DCs in the gingival connective tissue in chronic periodontitis possibly suggests their involvement in bone resorption in addition to their antigen presentation property., (© 2012 John Wiley & Sons A/S.)

Background and Objective: The causes of periodontitis are bacteria and the host immune system, but the role of the immune system in the onset and progression of periodontal disease is still unclear. Our previous report showed that the formation of an immune complex in the gingival sulcus induces periodontal destruction. This study was carried out to investigate how the immune system, particularly immunization, is involved in periodontal destruction., Material and Methods: Animals immunized intraperitoneally with lipopolysaccharide (LPS) were used as the immunized group. The nonimmunized group received only phosphate-buffered saline. LPS was applied daily onto the palatal gingival sulcus in both groups 1 d after the booster injection. Serum levels of anti-LPS IgG were determined. Loss of attachment and the level of alveolar bone were histopathologically and histometrically investigated. RANKL-bearing cells and the expression of C1qB were immunohistologically evaluated., Results: The serum levels of anti-LPS IgG were elevated in the early experimental period in the immunized group. There were significant increases in loss of attachment, level of alveolar bone and the number of RANKL-bearing cells in the immunized group. C1qB was observed in the junctional epithelium and adjacent connective tissue. The nonimmunized group showed similar findings at and after the time when the serum level of anti-LPS IgG was elevated., Conclusion: Topical application of LPS as an antigen induced periodontal destruction when the serum level of anti-LPS IgG was elevated in rats immunized with LPS. The presence of C1qB suggests that the formation of immune complexes is involved in this destruction., (© 2012 John Wiley & Sons A/S.)