Mural cell composition and functional analysis in the healing process of human gingiva from periodontal intrabony defects.

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.

Association between Smokeless Tobacco Use and Risk of Periodontitis in Asian Countries: A Systematic Review and Meta-Analysis.

BACKGROUND: Individual studies conducted in Asian countries have reported higher risk of periodontitis among smokeless tobacco (SLT) users in comparison to non-users. Therefore, a systematic review was conducted to summarize the available evidence on this topic. METHODS: Prominent electronic databases were searched using pre-decided MeSH terms and keywords. Screening of titles and abstracts, full text reading, quality assessment and data extraction was done by two investigators independently. The Newcastle-Ottawa scale was used for risk of bias assessment of eligible studies. Meta-analysis was performed for four periodontal outcomes (periodontal pocket depth, loss of attachment, clinical attachment level and gingival recession). A sensitivity analysis was also performed. RESULTS: Of the 546 citations, 367 were screened for eligibility. Finally, 89 studies were shortlisted for full text reading, of which, 36 were found eligible for qualitative analysis. Most of the studies were conducted in India (n=22), were of cross-sectional design (n=33), utilized purposive sampling and 24 studies were included for meta-analysis (n=28) and done on hospital-based population (n=26). Only 13 (37.1%) studies achieved a score of more than 50% (5/10 stars) on quality assessment scale. SLT users had higher odds of greater periodontal pocket depth greater than 4 mm (OR=3.64), gingival recession (OR=1.71) and loss of attachment 4-5 mm (OR=2.83) and mean difference of 1.7 mm for Clinical Attachment Level compared to non-users. CONCLUSION: The studies included in this review suggests that SLT users have poorer periodontal health in comparison to non-users. But most of this evidence comes from cross-sectional studies. Longitudinal studies with rigorous methodology are required to support this elucidation. Registration: This systematic review protocol has been registered in PROSPERO (CRD42019122964).

Epstein-Barr virus-infected plasma cells in periodontitis lesions.

Growing evidence supports that the Epstein-Barr virus (EBV) is a putative periodontal pathogen, but little is known regarding EBV behavior in periodontitis. Here, EBV infection was monitored in saliva and periodontal pocket (PP), at baseline and 3 months after periodontal non-surgical therapy (p-NST) in 20 patients diagnosed with periodontitis. After the treatment, the patients with the improved periodontal condition (good responders) showed a significant decrease in salivary EBV load. In contrast, in poor responders, EBV load was slightly increased. Moreover, after the therapy, most patients showed clear signs of EBV infection in a deep PP (≥5 mm) selected as a study site. To investigate how EBV can persist in a PP, we further investigate cellular sites of viral replication in PP. We identified large amounts of infiltrated EBV-infected cells, mostly overlapping with CD138+ plasma cells (PC). EBV-infected PCs formed high-density clusters within the infiltrate and along the periodontal epithelium which were commonly associated with CD3+ T-cells and CD20+ B-cells to evoke diffuse ectopic lymphoid-like structures. Taking together, this study provides new insights to support a model where the periodontal condition may play a major role in oral EBV shedding. Since PC harbors the late productive phases of EBV replication, the periodontal condition may favor B-cell differentiation with possible amplification of periodontal EBV infection and viral spreading. PCs have long been recognized as pathogenic markers in inflammatory lesions. Our finding sheds new light on the role of EBV infection and PC in periodontitis.

Quantification of TNF-α in Patients with Periodontitis and Type 2 Diabetes.

OBJECTIVE: The present study aimed to compare variations in quantified tumor necrosis factor-alpha (TNF-α) levels in patients with periodontitis stage 2 grade B (POD2B) and/or type 2 diabetes (T2D) and to identify any relationships between this cytokine and these diseases. METHODS: Levels of the cytokine TNF-α in gingival crevicular fluid in patients with POD2B and/or T2D were evaluated. A total of 160 subjects were distributed into four groups: those with POD2B (n=44); those with T2D (n=37); those with POD2B/T2D (n=40); and healthy subjects (n=39). Glycosylated hemoglobin (HbA1c) and blood glucose (BG) levels were quantified in each subject. Data were collected on body mass index (BMI), loss of insertion (LI), and probe depth (PD). Gingival crevicular fluid samples were collected from the most acutely affected periodontal pocket and gingival sulcus in each subject, and TNF-α was quantified by multiplex analysis. RESULTS: Kruskal Wallis tests was used to identify differences in TNF-α levels, LI, PD, BMI, BG, and HbA1c by group. Differences (p<0.001) were found for LI, PD, BG, and HbA1c. A Spearman test was used to calculate possible correlations between TNF-α levels and LI or PD identified a weak but significant negative correlation of TNF-α with LI (Rho=-0199; p=0.012), and a moderately positive correlation of LI with PD (Rho=0.509; p < 0.001). CONCLUSIONS: No variation was found between TNF-α levels and the presence of POD2B, POD2B/T2D, or T2D, suggesting the absence of any direct relationship between progression of these diseases and TNF-α levels. However, a correlation was present between low TNF-α concentrations and greater LI.

Association of initial mucogingival status with clinical outcome of non-surgical periodontal therapy: A retrospective analysis of 204 patients.

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.

Scaling and Root Planing Effect to mRNA Expression of Matrix Metalloproteinase-9 and Periodontal Clinical Parameters on Chronic Periodontitis

Abstract Objective: To evaluate the relationship between the mRNA transcription level of Matrix Metalloproteinase-9 (MMP-9) and the selected clinical periodontal healing at one month of scaling and root planing. Material and Methods: A total of six chronic periodontitis patients and one periodontally healthy subject were recruited. The gingival crevicular fluid was collected from all subjects, and the expression level of MMP-9 mRNA was measured by quantitative real-time PCR. Pocket depth, papilla bleeding index, and clinical attachment loss were measured on day 1 at baseline and day 30. Scaling and root planing was performed on day 1. Data were analyzed using SPSS 22.0 software Results: In comparison to the control, periodontal clinical parameters in the treatment group were significantly reduced after scaling and root planing. MMP-9 mRNA expression did not show a significant change after the 30th day. A weak correlation was noted between the MMP-9 mRNA transcription level and the changed PBI measurement Conclusion: Scaling and root planing is clinically effective for chronic periodontitis with a 4-6 mm pocket, whereas the expression of MMP-9 mRNA was not altered. Further studies with a more extended observation period are needed to confirm or reject the present findings.

Clinical and microbiological evaluation of the effect of Lactobacillus reuteri in the treatment of mucositis and peri-implantitis: A triple-blind randomized clinical trial.

BACKGROUND AND OBJECTIVE: Oral probiotics appear to improve the treatment of periodontal diseases but there is limited evidence on their efficacy in the treatment of peri-implant diseases. The objective of the present study was to evaluate, clinically and microbiologically, the effect of the oral probiotic, Lactobacillus reuteri Prodentis, as adjuvant to non-surgical mechanical therapy in implants with mucositis or peri-implantitis, placed in patients with a history of periodontal disease. MATERIAL AND METHODS: A randomized, controlled, parallel-design, triple-blind prospective clinical study was designed. Patients included in the study were partially edentulous and had implants with mucositis or peri-implantitis. Implants with radiographic bone loss of ≥5 mm and/or ≥50% of the implant length were excluded, and only one implant per patient was included. After non-surgical mechanical therapy, subjects were randomly assigned to take either 1 probiotic lozenge or 1 placebo lozenge every day for 30 days. Clinical measurements were taken in the whole mouth (general plaque index and general bleeding on probing) and at the implant site (probing pocket depth, plaque index and bleeding on probing) at baseline and 30 and 90 days Microbiological examination (to identify Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Peptostreptococcus micros, Fusobacterium nucleatum, Campylobacter rectus and Eikenella corrodens) was performed at the same study time points that clinical measurements were made. RESULTS: A total of 44 patients - 22 with mucositis and 22 with peri-implantitis - randomly received treatment with either probiotic or placebo. The probiotic L. reuteri, together with mechanical therapy, produced an additional improvement over treatment with mechanical therapy alone, both in the general clinical parameters of patients with mucositis (bleeding on probing) and at the level of implants with mucositis (probing pocket depth) or peri-implantitis (bleeding on probing and probing pocket depth). However, L. reuteri had a very limited effect on the peri-implant microbiota because the only parameter in which a significant decrease was found was the bacterial load of P. gingivalis in implants with mucositis (P = .031). CONCLUSION: The administration of a daily lozenge of L. reuteri for 30 days, together with mechanical debridement of the whole mouth, improved the clinical parameters of implants with mucositis or peri-implantitis over a period of at least 90 days, but the microbiological effect was much more limited. Probiotics provide an alternative therapeutic approach to consider in the prevention and treatment of peri-implant diseases, but further long-term prospective studies with standardized variables are needed.

Clinical and radiographic periodontal parameters and release of collagen degradation biomarkers in naswar dippers.

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.

Relationship between clinical periodontal parameters and changes in liver enzymes levels over an 8-year period in an elderly Japanese population.

AIMS: An association between periodontal disease and liver enzymes has been reported previously. This study examined the relationship between clinical periodontal parameters and changes in liver enzymes levels in 265 non-institutionalized Japanese elderly aged 72 years over 8 years. MATERIALS AND METHODS: The number of sites with probing pocket depth (PPD) ≥ 6 mm and clinical attachment level (CAL) ≥ 6 mm were measured. Changes in liver enzymes (alanine transaminase [ALT] and aspartate transaminase [AST]) was grouped based on any elevation of concentrations. The relationship was determined by logistic regression with adjustment for confounders. Interaction test and stratified analysis were then performed according to smoking status and alcohol drinking habits, separately. RESULTS: Elevation in ALT, but not AST, was significantly associated with PPD (odds ratio [OR] = 1.10) and CAL (OR = 1.03). A significant interaction of periodontal parameters on ALT was observed with smoking status, but not with alcohol drinking habit. The stratified analysis suggested that ALT was found to be significantly correlated with PPD (OR = 1.20) and CAL (OR = 1.04) to those who were smoker. CONCLUSION: The elevation in ALT levels might be associated with clinical periodontal parameters among non-institutionalized Japanese elderly, and this association was modified by smoking status.

Periodontal inflamed surface area and periodontal case definition classification.

OBJECTIVE: To assess whether a higher periodontal inflamed surface area (PISA) may reflect more severe periodontitis, and if so, to calculate which cut-off point should be used to classify a patient as suffering from periodontitis. MATERIAL AND METHODS: Eighty subjects were selected based on their periodontal status and were divided into 4 groups of 20 each; periodontally healthy, mild periodontitis, moderate periodontitis and severe periodontitis. In addition, demographic data, smoking status and socioeconomic status were also recorded. RESULTS: The highest mean PISA value was obtained for the severe periodontitis group (2309.42 ± 587.69 mm2) and the least for the periodontally healthy (34.30 ± 16.48 mm2). The PISA values corresponding to the three categories of periodontitis severity were significantly higher than the periodontally healthy group. When receiver operating characteristic (ROC) analysis was performed, a PISA value ≥130.33 mm2 predicted the presence of periodontitis with a sensitivity of 98% and a specificity of 100%. CONCLUSIONS: PISA is a periodontal parameter that may be used in conjunction with the Centers for Disease Control and Prevention - American Academy of Periodontology case definition classification in periodontal medicine research.

The periodontal pocket: pathogenesis, histopathology and consequences.

The conversion of junctional epithelium to pocket epithelium is regarded as a hallmark in the development of periodontitis. Knowledge of factors contributing to the initiation and progression of pocket formation is important and may result in the development of better preventive measures and improve healing outcomes after therapeutic interventions. The periodontal pocket is a pathologically deepened gingival sulcus. In healthy periodontal conditions, the defense mechanisms are generally sufficient to control the constant microbiological challenge through a normally functioning junctional epithelium and the concentrated powerful mass of inflammatory and immune cells and macromolecules transmigrating through this epithelium. In contrast, destruction of the structural integrity of the junctional epithelium, which includes disruption of cell-to-cell contacts and detachment from the tooth surface, consequently leading to pocket formation, disequilibrates this delicate defense system. Deepening of the pocket apically, and also horizontal expansion of the biofilm on the tooth root, puts this system to a grueling test. There is no more this powerful concentration of defense cells and macromolecules that are discharged at the sulcus bottom and that face a relatively small biofilm surface in the gingival sulcus. In a pocket situation, the defense cells and the macromolecules are directly discharged into the periodontal pocket and the majority of epithelial cells directly face the biofilm. The thinning of the epithelium and its ulceration increase the chance for invasion of microorganisms and their products into the soft connective tissue and this aggravates the situation. Depending on the severity and duration of disease, a vicious circle may develop in the pocket environment, which is difficult or impossible to break without therapeutic intervention.

The use of hyaluronic acid as an adjuvant in the management of peri-implantitis.

It is well known in dentistry that there are numerous chronic conditions that require ongoing and constant management over time, the most noteworthy being periodontal disease, gingivitis and periodontitis. Yet, in recent years, with the increase in the number of implants being placed, mucositis and peri-implantitis have become more and more prevalent pathologies. The results of the statistical analysis demonstrate that there was a slight difference between the pocket depth as measured in the treated sites at time 0 (pre-treatment) and time 1 (15 days post-treatment), although the difference was so small as to render it statistically irrelevant. Bleeding on probing as measured at time 0 and time 1 indicated an improvement on both sides, but with no greater improvement noted on the side treated with HA.

[The relevance of Candida spp. in chronic periodontal disease]. / Vyiavlenie drozhzhepodobnykh gribov roda u patsientov s khronicheskim generalizovannym parodontitom.

The aim of the study was to assess the correlation of Candida spp. incidence in periodontal tissues with various clinical manifestations of chronic periodontal disease (CPD). Ninety patients with CPD were included in the study in which Candida spp. was evaluated in periodontal pockets content and gingival biopsy material. In severe CPD more Candida spp. were seen in gingival biopsy than in periodontal pockets (p=0.0006). Candida spp. incidence and quantity correlated directly with the disease grade showing incidence increase from 40 to 73.3% and quantity increase from 0.8±0.18 до 3.6±0.49 lg CFU/ml in light and severe CPD, correspondingly Candida spp. had statistically significant association with cyanotic gingival color (p=0.0018), tongue plaque and swelling (р=0.0042), lip exfoliation (р=0.0030), periodontal pockets depth >5 mm (р=0.0030), oral mucosa hyperemia (р=0.0157), alveolar bone destruction >1/2 of root length (р=0.0157). These data prove the relevance of Candida spp. and mycological assessment of gingival biopsy in CPD patients.

Steatosis caused by experimental periodontitis is reversible after removal of ligature in rats.

BACKGROUND AND OBJECTIVE: Periodontitis may promote harmful systemic effects such as changes in hepatic tissues. The purpose of this study was to investigate whether the steatosis and oxidative stress caused by experimental periodontitis are reversible in the liver. MATERIAL AND METHODS: Twenty-four rats were divided into three groups: control, periodontitis and P20-20 (20 days with experimental periodontitis and 20 days without experimental periodontitis, to verify the reversibility of hepatic injuries). The following parameters were assessed: gingival bleeding index, probing pocket depth, myeloperoxidase activity, alveolar bone loss for periodontal tissues; liver weights, histopathological scores for steatosis, inflammation and necrosis in liver; glutathione, malondialdehyde, total cholesterol and triglyceride concentrations in hepatic tissues; and blood levels of aspartate aminotransferase, alanine aminotransferase, albumin, gamma-glutaryl transferase, total cholesterol and random glucose. RESULTS: Gingival bleeding index, probing pocket depth, myeloperoxidase and alveolar bone loss parameters demonstrated the development of periodontitis. There was a significant reduction in the steatosis score of animals from the P20-20 group when compared with the periodontitis group. P20-20 group presented significantly higher glutathione (11 times) and lower malondialdehyde (nearly 23%), total cholesterol (both in blood and hepatic tissue) and triglyceride concentrations compared with the periodontitis group. For levels of aspartate aminotransferase, alanine aminotransferase, albumin, gamma-glutaryl transferase and random glucose, a significant difference between the groups was not observed. CONCLUSION: Our results demonstrate that the microvesicular steatosis caused by periodontitis in rats is reversible after removal of the ligature, which is associated with the increase in oxidative stress and lipid peroxidation in the liver.

Changes of Periodontal Parameters after Apical Surgery: Correlation of Clinical and Cone-beam Computed Tomographic Data.

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.

Is there a relationship between periodontal conditions and number of medications among the elderly?

OBJECTIVE: To investigate possible correlations of clinical attachment level and pocket depth with number of medications in elderly individuals. METHODS: Intra-oral examinations for 139 patients visiting Tufts dental clinic were done. Periodontal assessments were performed with a manual UNC-15 periodontal probe to measure probing depth (PD) and clinical attachment level (CAL) at 6 sites. Complete lists of patients' medications were obtained during the examinations. Statistical analysis involved Kruskal-Wallis, chi square and multivariate logistic regression analyses. RESULTS: Age and health status attained statistical significance (p< 0.05), in contingency table analysis with number of medications. Number of medications had an effect on CAL: increased attachment loss was observed when 4 or more medications were being taken by the patient. Number of medications did not have any effect on periodontal PD. In multivariate logistic regression analysis, 6 or more medications had a higher risk of attachment loss (>3mm) when compared to the no-medication group, in crude OR (1.20, 95% CI:0.22-6.64), and age adjusted (OR=1.16, 95% CI:0.21-6.45), but not with the multivariate model (OR=0.71, 95% CI:0.11-4.39). CONCLUSION: CAL seems to be more sensitive to the number of medications taken, when compared to PD. However, it is not possible to discriminate at exactly what number of drug combinations the breakdown in CAL will happen. We need to do further analysis, including more subjects, to understand the possible synergistic mechanisms for different drug and periodontal responses.

Migration and Differentiation of GFP-transplanted Bone Marrow-derived Cells into Experimentally Induced Periodontal Polyp in Mice.

Perforation of floor of the dental pulp is often encountered during root canal treatment in routine clinical practice of dental caries. If perforation were large, granulation tissue would grow to form periodontal polyp. Granulation tissue consists of proliferating cells however their origin is not clear. It was shown that the cells in granulation tissue are mainly from migration of undifferentiated mesenchymal cells of the bone marrow. Hence, this study utilized GFP bone marrow transplantation mouse model. The floor of the pulp chamber in maxillary first molar was perforated using ½ dental round bur. Morphological assessment was carried out by micro CT and microscopy and GFP cell mechanism was further assessed by immunohistochemistry using double fluorescent staining with GFP-S100A4; GFP-Runx2 and GFP-CD31. Results of micro CT revealed alveolar bone resorption and widening of periodontal ligament. Histopathological examination showed proliferation of fibroblasts with some round cells and blood vessels in the granulation tissue. At 2 weeks, the outermost layer of the granulation tissue was lined by squamous cells with distinct intercellular bridges. At 4 weeks, the granulation tissue became larger than the perforation and the outermost layer was lined by relatively typical stratified squamous epithelium. Double immunofluorescent staining of GFP and Runx2 revealed that both proteins were expressed in spindle-shaped cells. Double immunofluorescent staining of GFP and CD31 revealed that both proteins were expressed in vascular endothelial cells in morphologically distinct vessels. The results suggest that fibroblasts, periodontal ligament fibroblasts and blood vessels in granulation tissue were derived from transplanted-bone marrow cells. Thus, essential growth of granulation tissue in periodontal polyp was caused by the migration of undifferentiated mesenchymal cells derived from bone marrow, which differentiated into fibroblasts and later on differentiated into other cells in response to injury.

Gingival crevicular fluid vascular endothelial cell growth factor and platelet-derived growth factor-BB release profile following the use of perforated barrier membranes during treatment of intrabony defects: a randomized clinical trial.

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.

Enhancing gingival biotype through chorion membrane with innovative step in periodontal pocket therapy.

A thick gingival biotype is a requisite for good periodontal health. It has important role in resisting trauma and subsequent gingival recession. The gingival thickness is a significant predictor of clinical outcome of periodontal surgeries. Various surgical procedures are used to increase the gingival thickness. The present study incorporated the innovative step of placement of chorion membrane to objectively evaluate the increase in thickness of gingival biotype during periodontal pocket therapy. The patients in age group between 25 and 45 years with chronic periodontitis, indicated for flap surgery were selected for the study. The sites with pocket depth of 6-8 mm in the mandibular anterior teeth were divided into test and control sites. Periodontal flap surgery was carried at both the sites and chorion membrane was placed at the test sites. The gingival thickness measurement was assessed using a markings marked on injection needle, these markings were read using digital vernier caliper, pre and post operatively. The baseline values of gingival thickness at test site (1.04 ± 0.19 at mid buccal region, 1.24 ± 0.20 at mid papillary) and control site (0.94 ± 0.11 at mid buccal region, 1.14 ± 0.11 at mid papillary region) showed no statistically significant difference. At test sites, 6 weeks post treatment (1.36 ± 0.16 at mid buccal region and 1.48 ± 0.17 at mid papillary region) as compared to control sites (1.06 ± 0.11 at mid buccal region, 1.24 ± 0.11 at mid papillary) showed statistically significant increase in gingival thickness (p ≤ 0.05*). The innovative step of placement of chorion membrane during periodontal pocket therapy facilitated increase in the gingival thickness in the areas with thin gingival biotype.

Subgingival microbiome in patients with healthy and ailing dental implants.

Dental implants are commonly used to replace missing teeth. However, the dysbiotic polymicrobial communities of peri-implant sites are responsible for peri-implant diseases, such as peri-implant mucositis and peri-implantitis. In this study, we analyzed the microbial characteristics of oral plaque from peri-implant pockets or sulci of healthy implants (n = 10), peri-implant mucositis (n = 8) and peri-implantitis (n = 6) sites using pyrosequencing of the 16S rRNA gene. An increase in microbial diversity was observed in subgingival sites of ailing implants, compared with healthy implants. Microbial co-occurrence analysis revealed that periodontal pathogens, such as Porphyromonas gingivalis, Tannerella forsythia, and Prevotella intermedia, were clustered into modules in the peri-implant mucositis network. Putative pathogens associated with peri-implantitis were present at a moderate relative abundance in peri-implant mucositis, suggesting that peri-implant mucositis an important early transitional phase during the development of peri-implantitis. Furthermore, the relative abundance of Eubacterium was increased at peri-implantitis locations, and co-occurrence analysis revealed that Eubacterium minutum was correlated with Prevotella intermedia in peri-implantitis sites, which suggests the association of Eubacterium with peri-implantitis. This study indicates that periodontal pathogens may play important roles in the shifting of healthy implant status to peri-implant disease.