Cell-based therapy in the management of Class III Miller's recession - A case report with 45-month follow-up.

Miller's Class III gingival recessions (GRs) have always posed a challenge to the clinicians in terms of achieving complete root coverage (CRC). In the present case, a cell-based therapy with autologous fibroblasts seeded onto a Type 1 collagen membrane, through an in-vitro culturing method was utilized. The fibroblasts-seeded membrane was surgically placed under a laterally repositioned flap. The patient presented with a CRC, which was stable even at the postoperative period of 45 months. In addition, a 3-mm substantial gain in the width of keratinized tissue was achieved and maintained throughout the postoperative period. Hence, the results of the cell-seeded therapy emphasize that it can serve as an effective alternative method for the management of Miller's Class III GRs.

Comparative evaluation of subgingival microbiome in healthy periodontium and gingivitis using next-generation sequencing technology: A case-control study.

Background: Human dental plaque is a complex microbial community containing millions of species. Gingivitis is a dysregulated immune-inflammatory response induced by dysbiotic plaque biofilm that interrupts symbiosis. The emergence of next-generation sequencing with 16S rRNA gene has greatly contributed in understanding the complexity of microbiota. However, studies focusing on microbiome in gingivitis are limited. The whole bacterial community is important in causing periodontal disease than a small number of periodontal pathogens. In this study, we attempted to profile the subgingival microbiome from individuals with healthy gingiva and in patients with gingivitis using next-generation sequencing technology. Materials and Methods: Subgingival plaque samples from 15 healthy periodontium (Group I) and 15 gingivitis (Group II) were collected and 16s rRNA sequencing was done in Illumina Solexa Sequencer. Data analysis using 16s metagenomics tool from BaseSpace onsite operational taxonomic units was assigned to each sequence using HOMD database. Individual variation in the microbiome of the subgingival samples between the two groups was also evaluated. Results: The comparison of top 20 species between Group I and Group II revealed no significant species group between them. Synergistetes was absent in Group I samples but found in Group II. At the genus level, HACEK group species were found in both the groups, while Dialister and Aneroglobus were found abundantly in the Group II. Conclusion: The presence of unique genera and species seen in Group II samples could point toward a dysbiotic shift that could be taking place in the subgingival environment leading to gingivitis.

Serum and salivary adipokines in type 2 diabetes - Results of a pilot study in India.

BACKGROUND AND AIMS: Association of serum and salivary adiponectin, apelin, visfatin and vaspin were studied in type 2 diabetes (T2DM) among Asian Indians. Their concentrations in periodontitis were also studied. METHODS: In this cross-sectional analysis, men and women aged ≥35 years, with no history of diabetes, were screened for ≥3 risk factors for T2DM (n = 615). Eligible persons underwent a 75 gm oral glucose tolerance test and were categorized as Group A (Normal and Impaired Glucose Tolerant, n = 65) and Group B (Incident T2DM, n = 25). Screening for periodontitis was done. Saliva samples were collected in the morning. Participants refrained from food intake for about 2 hours prior to collection . Serum and saliva were stored for analysis. RESULTS: Serum adiponectin was low (p = 0.006) in T2DM and correlated with its salivary levels (r = 0.46, p < 0.001). Serum apelin levels were similar, but salivary concentrations were higher (p = 0.014) in T2DM. Higher serum (p = 0.016) and salivary (p = 0.03) visfatin levels were seen in T2DM. Vaspin levels showed no significant difference in the two groups, either in blood or saliva. Serum adipokines did not differ in the presence of periodontitis. In saliva, higher vaspin (p = 0.034) and lower visfatin (p = 0.018) concentrations were observed. CONCLUSIONS: The selected adipokines were measurable in saliva, in lower concentrations. Salivary adiponectin and visfatin measurements may be useful in studies on T2DM.

Evaluation of estrogen receptor and circulating estradiol levels in pre- and postmenopausal women with periodontal disease.

Although the presence of estrogen receptors (ERs) in the gingival tissues has been confirmed, there is as yet insufficient literature regarding its expression in periodontal health and disease. Gingival samples were collected from 40 subjects who were divided into four groups. (Group A, premenopausal health; Group B, premenopausal periodontitis; Group C, postmenopausal health; Group D, postmenopausal periodontitis). ERs were identified with an anti-ER monoclonal antibody (Bio Genex). Circulating estradiol (E2) levels were estimated using the Genix E2 EIA commercial kit. An inverse relationship between E2 and ER levels in gingiva was observed in all the samples. There was significantly reduced expression of ERs in the gingiva of subjects with chronic periodontitis when compared to those with healthy periodontium in the postmenopausal group.

Free gingival graft in the treatment of class III gingival recession.

AIM: The purpose of this study was to assess the success and predictability of root coverage and esthetics obtained with free gingival grafts (FGGs) in the treatment of early class III gingival recessions for a period of 12 months. MATERIALS AND METHODS: Ten patients contributed to 12 sites, each with early class III recession with interdental bone loss < or = 4 mm from cemento enamel junction(CEJ). Clinical parameters recorded at baseline and at 1, 6, and 12 months were probing depth (PD), recession depth (RD), recession width (RW), and clinical attachment level (CAL). RESULTS: Reduction of recession resulted in a significant gain in CAL and PD at the end of 12 months. A statistically significant mean root coverage of 41.25 +/- 21.07% was obtained at the end of 12 months. A statistically significant improvement in Visual Analog Scale score was seen after a 12-month follow-up period. CONCLUSION: In a south Indian population, early class III gingival recessions treated with FGG procedures resulted in 40-50% root coverage with fairly acceptable esthetics.

Differential expression of E-cadherin and cytokeratin 19 and net proliferative rate of gingival keratinocytes in oral epithelium in periodontal health and disease.

BACKGROUND: The effects of periodontal disease on the oral gingival epithelium (OGE) have not been documented fully because they may not be as dramatic as those seen on the junctional epithelium. The aim of this study was to estimate the changes occurring in the OGE with respect to its proliferation and E-cadherin and cytokeratin 19 (K19) expression during pocket formation. METHODS: Gingival samples were collected from 17 periodontally healthy subjects and 18 subjects with chronic periodontitis. K19 and E-cadherin levels were analyzed immunohistochemically. The net proliferative rate was calculated as the difference between the proliferative rate and the apoptotic rate as determined by immunohistochemical analysis of Ki-67 and p53, respectively. RESULTS: There was a significant increase in the net proliferative rate of the OGE during pocket formation (periodontitis group, 220.90+/-46.85; healthy group, 107.60+/-25.86; P<0.001). There was a significant reduction in E-cadherin expression (periodontitis group, 0.837+/-0.428; healthy group, 1.846+/-0.555) and a significant increase in K19 expression during pocket formation (periodontitis group, 1.45+/-0.686; healthy group, 0.533+/-0.410). CONCLUSION: OGE appears to undergo significant changes in proliferation and differentiation during pocket formation that do not seem to be restricted to proteolytic destruction by the invading microorganisms.

Isolated lesions of gingiva: A case series and review.

Isolated lesions of gingiva arise in succession to the hyperinflammatory reactions in response to the underlying local irritants. Despite their overlapping clinical and histological features, these lesions are distinctive regarding their biological behavior. Recurrence has been reported after surgical excision because of the incomplete removal of underlying local irritants. This article describes the clinical and histological features of four localized gingival lesions, adding a note on their molecular pathogenesis and surgical management.

Evaluation of transcription factor that regulates T helper 17 and regulatory T cells function in periodontal health and disease.

BACKGROUND: The differentiation of naοve T helper (Th) cells towards Th17 and regulatory T cells (Treg) is regulated by the transcription factors retinoic acid related orphan receptor gamma transcription (RORYt) and Forkhead box p3 (Foxp3), respectively. An imbalance in the activity of these transcription factors could result in the dysregulation of Th17/Treg response. MATERIALS AND METHODS: Total RNA was isolated from gingival tissue obtained from 10 patients, each from periodontally healthy and diseased groups. The gene expression of RORYt and Foxp3 was measured by real-time reverse transcription polymerization chain reaction using total RNA isolates from gingival tissues group when compared to the healthy group, while Foxp3 demonstrated a 6.68 ± 0.03 fold decrease of expression in diseased group when compared to healthy group. CONCLUSION: Our results indicate a functional imbalance in the Th17/Treg response in periodontal disease group when compared to the periodontally healthy group.

Evaluation of Salivary Levels of Pyridinoline Cross Linked Carboxyterminal Telopeptide of Type I Collagen (ICTP) in Periodontal Health and Disease.

BACKGROUND: Traditional parameters (Pocket depth, bleeding on probing, clinical attachment loss, radiographic findings) have been used for a long time for the assessment of periodontal disease conditions. However, these parameters only indicate towards the periodontal damage that has already taken place but do not give any idea regarding the current status of the periodontal health or disease. Hence, the present study is aimed at evaluating the concentration of the bone biomarker ICTP in saliva, which can give a better real time assessment of periodontal health and disease. MATERIALS AND METHODS: Forty three patients were selected and divided into three groups based on the recorded clinical parameters of probing pocket depth, attachment loss and bleeding on probing. Group I (Healthy, n = 11), Group II (Gingivitis, n = 17), Group III (Periodontitis. n = 15). Salivary samples were collected before scaling and root planning to avoid contamination by blood. ICTP levels were evaluated in the salivary samples by using enzyme linked immunosorbent assay (ELISA). STATISTICAL ANALYSIS USED: Kruskal Wallis test was used to compare the mean ICTP level of the three groups. RESULTS: ICTP was detected in all the samples. Highest mean ICTP concentrations in saliva were obtained for group III (periodontitis group) and the lowest mean ICTP concentrations were seen in group I (healthy group). This suggests that the level of ICTP in saliva increases proportionally from periodontal health to diseased conditions (gingivitis & periodontitis). CONCLUSION: There is a substantial increase in the salivary concentration of ICTP in chronic periodontitis patients than in gingivitis and healthy patients. Salivary ICTP levels were the maximum in chronic periodontitis patients followed by gingivitis patients and the least in healthy individuals. ICTP may be considered as a biomarker in periodontal disease progression.

Evaluation of circulatory and salivary levels of heat shock protein 60 in periodontal health and disease.

BACKGROUND: Self-antigens such as heat shock protein 60 (HSP 60) have recently been implicated in the periodontal disease pathogenesis. There is scant evidence regarding HSP 60 levels in circulation and saliva following periodontal disease and its possible relation to systemic inflammation. AIM OF THE STUDY: The aim was to evaluate the circulatory and salivary levels of HSP 60 in periodontal health and disease and to correlate it with high sensitivity C-reactive protein (hs-CRP). MATERIALS AND METHODS: Forty-five peripheral blood samples were collected from two groups of patients (periodontally healthy - Group A [22 patients] and periodontal disease - Group B [23 patients]). Serum, cell lysates, and saliva samples were used to detect HSP 60 levels in both groups by enzyme linked immunosorbent assay technique. Measurement of hs-CRP was performed using an immunoturbidimetric assay. Statistical analysis was done using the student t-test and Pearson's correlation. RESULTS: Circulatory HSP 60 was significantly increased in periodontal disease compared to health (P - 0.038). There was a significant correlation between the totals circulating HSP 60 and hs-CRP (P - 0.052), but there was no significant correlation between the salivary HSP 60 and hs-CRP levels in periodontal disease. CONCLUSION: Circulating HSP 60 levels may play a role in the systemic inflammatory state produced by periodontal disease. Salivary HSP 60 may not be used as a surrogate to determine systemic inflammation.

Plasticity of T helper cell subsets: Implications in periodontal disease.

T helper (Th) cells have an important role in host defence as well in the pathogenesis of periodontal disease. Th cells differentiate from naive cells into various subsets, each of which is associated with a set of inducing and effector cytokines. Previously, it was thought that this differentiation was an irreversible event. Recent evidence suggest that even differentiated Th cells, retain the flexibility to transform from one lineage to another, a phenomenon referred to as plasticity. This plasticity is thought to be brought about by epigenetic modifications that are regulated by external and internal signals in the micro-environment of these cells. The factors and mechanisms which affect the plasticity of these cells and their potential role in the etio-pathogenesis of periodontal disease has been described in this article.

Mathematical analysis of furcation angle in extracted mandibular molars.

BACKGROUND: Multi-rooted teeth with furcation involvement exhibit a poorer prognosis when compared to single rooted teeth. The furcation angle (formed by the divergent roots and the roof) may exert a considerable influence on the accessibility for both home care maintenance and instrumentation during periodontal therapy. As there are few anatomy based reports, the furcation angle has not yet been delineated. MATERIALS AND METHODS: Furcation angle (FA) was mathematically evaluated in extracted mandibular first and second molar teeth, using the Computer-aided design - computer-aided manufacturing technology. RESULTS: THE FURCATIONS WERE DIVIDED INTO THREE GROUPS (GROUP I: <30°, Group II: 30°-60°, Group III: >60°) based on the furcation angle and their prevalence. The first molar showed greater prevalence of group II FA, while second molar showed a greater prevalence of group III FA. CONCLUSION: Linear, two dimensional measurements may not accurately reflect the complexities of the furcation area which exhibits considerable intermolar and intramolar (buccal and lingual furcations of second molar) variation.

Collagen with simvastatin promotes cell metabolism in osteoblast-like SaOS-2 cells.

BACKGROUND: Simvastatin (SMV) is one of the cholesterol-lowering pharmacological drugs. Recent studies demonstrate that it has a bone stimulatory effect. The present study was designed to investigate the effect of SMV along with collagen membrane on osteoblast-like SaOS-2 cells and also to standardize the dosage of SMV to be incorporated into the collagen membrane to achieve regeneration. MATERIALS AND METHODS: SMV at doses of 0.5, 1, 1.5, and 2 mg was incorporated into the collagen membrane and cell metabolism was assessed by (3-[4,5-dimethylthiazolyl-2]-2,5-diphenyltetrazolium bromide) (MTT) assay for 24 h. RESULTS: SMV enhanced cell metabolism dose dependently at 24-h time and the maximum effect was obtained at a concentration of 1.5 mg of SMV. CONCLUSION: These results indicate that collagen with 1.5 mg SMV exhibits positive effect on cell metabolism of human osteoblast-like SaOS-2 cells.

T-helper cells in the etiopathogenesis of periodontal disease: A mini review.

Our traditional understanding of the T-helper (Th)1/Th2 paradigm in periodontal disease has undergone considerable changes in recent years. This review focuses on the Th subsets, including the recently identified cells of the CD4 lineage, their activation pathways and effector function in periodontal disease. The roles of Th17 and regulatory T (Treg) cells in disease pathogenesis have been explored. Newer Th subsets such as Th9 and Th22 cells and their potential role in periodontal disease have also been outlined.

Clinical evaluation of the efficacy of a GTR membrane (HEALIGUIDE) and demineralised bone matrix (OSSEOGRAFT) as a space maintainer in the treatment of Miller's Class I gingival recession.

BACKGROUND: Periodontal plastic surgical procedures aimed at coverage of exposed root surface have evolved into routine treatment modalities. The present study was designed to evaluate the effectiveness and predictability of using a collagen barrier along with a demineralized bone matrix in the treatment of recession defects in a single surgical procedure. MATERIALS AND METHODS: Seventeen patients with Miller's class I recession were treated with a combination of a collagen barrier used along with a bone graft and coronally advanced flap technique. Clinical parameters were recorded at baseline, 3 months, 6 months, and 9 months. RESULTS: The study showed a highly significant reduction in the recession depth (70.29 ± 21.96%) at the end of the study. This study showed that the use of this technique for recession coverage is highly predictable and highly esthetic root coverage can be obtained.

E-cadherin and CD1a expression in gingival epithelium in periodontal health, disease and post-treatment.

BACKGROUND: Epithelial integrity is important for maintenance of periodontal health. It is not fully known if non-surgical periodontal therapy is capable of recreating the epithelial barrier in its functional state. PATIENTS AND METHODS: Sixty-five patients (31 males and 34 females) were included in the study. They were divided into group A (healthy gingiva 16 patients), group B (gingivitis 17 patients), group C (periodontitis 17 patients), and group D (post-treatment 15 patients). Gingival samples were collected and immunohistochemical study was done using E-cadherin and CD1a antibody. Statistical analysis was done using analysis of variance (ANOVA), followed by Tukey-Kramer multiple comparison test for CD1a and Tukey's highly significant difference (HSD) test for E-cadherin. RESULT: There was a statistically significant difference (P < 0.001) in the expression of E-cadherin between healthy (1.846±0.555), gingivitis (1.100±0.994), and periodontitis group (0.700±0.483). Similarly, there was a statistically significant difference (P < 0.001) in the expression of CD1a between healthy (75.70±3.09), gingivitis (42.53±3.09), and periodontitis group (29.07±3.08). However, the expression of E-cadherin (1.242±0.653) and CD1a in post-treatment samples (52.18±2.90) was lower with no statistically significant difference when compared to health. DISCUSSION: The significant reduction in E-cadherin and CD1a levels in periodontal disease when compared to health could possibly be a result of invasion by the periodontopathogens and its subsequent sequel. Although, the post-treatment samples showed significant improvement when compared to disease, the reduction in E-cadherin and CD1a levels when compared to gingival health suggests that the epithelial barrier was not yet fully established in its functional state.

Osteoblast response to commercially available demineralized bone matrices--an in-vitro study.

OBJECTIVE: Reconstruction of lost attachment apparatus is a major goal of periodontal therapy. Although various osteoinductive bone replacement grafts (BRGs) have been used with apparent clinical success, unequivocal evidence of osteoinductivity may be obtained only through the demonstration of increased osteoblastic/osteoclastic differentiation following exposure to these materials. MATERIALS AND METHODS: Bone marrow stem cells (BMSCs) obtained from rat femur were cultured in Dulbecco's Modified Eagles Medium (DMEM) and 10% fetal bovine serum (FBS). They were then exposed to two demineralized bone matrices (DBM's)--Grafton and Osseograft, and divided into three groups, comprising of a negative control (BMSC + DMEM + 10% FBS), Grafton, Osseograft. An osteogenic medium (OM) (10 hm dexamethasone, 10 hm b-glycerophosphate, and 50 microg/ml ascorbic acid) was added to create three subgroups comprising of a positive control (OM), Grafton with OM, Osseograft with OM. RESULTS: After an initial phase (up to day 5), both Grafton and Osseograft induced an increased proliferative activity in the BMSCs, which reached a plateau after day 10. These grafts also induced increased alkaline phosphatase activity when compared to the control groups and to BMSCs with an OM. CONCLUSION: Both Osseograft and Grafton are capable of inducing osteoblastic proliferation and differentiation.

Retrograde peri-implantitis.

Retrograde peri-implantitis constitutes an important cause for implant failure. Retrograde peri-implantitis may sometimes prove difficult to identify and hence institution of early treatment may not be possible. This paper presents a report of four cases of (the implant placed developing to) retrograde peri-implantitis. Three of these implants were successfully restored to their fully functional state while one was lost due to extensive damage. The paper highlights the importance of recognizing the etiopathogenic mechanisms, preoperative assessment, and a strong postoperative maintenance protocol to avoid retrograde peri-implant inflammation.

Clinical and histological evaluation of two dressing materials in the healing of palatal wounds.

BACKGROUND: Free gingival grafts have been used extensively for gingival augmentation procedures, but are associated with postoperative morbidity because of the open palatal wound. This study compares the clinical efficiency of two dressing materials, a non-eugenol-based dressing (Coe-Pak™) and a collagen dressing (Colla Cote(®)) on palatal wound healing. MATERIALS AND METHODS: Thirty-two patients in the age group of 25-50 years, who required gingival augmentation, were selected. Free gingival graft was harvested from the palatal mucosa and the wound was then protected using Coe-pak(®) in control group and Colla Cote(®) in test group. The subjective parameters pain and burning sensation were recorded on the 2(nd) and 7(th) day and the objective parameters colour and consistency were recorded on the 7(th) and 42(nd) day, using a visual analog scale. Thickness of the mucosa was measured using K file at baseline and 42(nd) day. Histological examination was done on 42(nd) day. RESULTS: The subjective and objective parameters showed significant improvement in the test group when compared to control group. Histologically, there was a greater evidence of collagen formation and turn over in the test group than control group. CONCLUSIONS: Collagen-based dressing may thus offer significantly greater advantages over the traditional non-eugenol dressings.

Osteoblast response (initial adhesion and alkaline phosphatase activity) following exposure to a barrier membrane/enamel matrix derivative combination.

BACKGROUND AND OBJECTIVE: The enamel matrix derivative (EMD) has been used in combination with barrier membranes to optimize regeneration in vertical osseous defects. However, the osteoblast response when exposed to the EMD/barrier membrane combination has not yet been evaluated. The osteoblast behavior when exposed to a combination of regenerative materials must be evaluated to fully understand their effect on bone regeneration. Therefore, the present study was undertaken to estimate the initial adhesion and alkaline phosphatase (ALP) activity of an osteoblast cell line (SaOS-2) when exposed to four commercially available resorbable membranes and determine if the addition of EMD had any modulatory effect on osteoblast behavior. MATERIALS AND METHODS: 5 x 104 SaOS-2 cells between passages 7-10 were cultured in two 24-well culture plates. Plate A was used for the adhesion assay and Plate B was used for the ALP assay. A MTT (3-[4, 5-dimethylthiazolyl-2]-2, 5-diphenyltetrazolium bromide) assay was done after 24 hours to determine the adhesion of the osteoblastic cells to four barrier membranes: 1) a non cross-linked porcine Type I and III collagen membrane (BG), 2) a weakly cross-linked Type I collagen membrane (HG), 3) a glutaraldehyde cross-linked bovine Type I collagen (BM), and 4) a resorbable polymer membrane (CP). Osteoblast differentiation was studied using an ALP assay with p-nitro phenyl phosphate as the substrate at 24 hours, 72 hours, and 1 week. A total of 50 microg/ml of EMD dissolved in 10 mM acetic acid was added into each well and the entire experimental protocol outlined above was repeated. RESULTS: The osteoblast adhesion to collagen barriers showed a statistically insignificant reduction following the addition of EMD. Adhesion to the polymer barrier, although significantly lower when compared with collagen barriers, was unaffected by the addition of EMD. ALP activity after 1 week among the various groups was as follows: EMD alone (75.59+/-2.5)>EMD/BG(64.78+/-3.04)>EMD/HG(55.40+/-3.89) approximately EMD/BM(54.75+/-4.17)>BG (51.32+/-2.76)>HG(49.92+/-2.4)>BM(48.14+/-1.4)>Control(46.29+/-1.39)>EMD/CP (37.46+/-3.54)>CP(32.12+/-1.49) CONCLUSION: There was no additive effect on osteoblast adhesion/ALP activity following exposure to an EMD/polymer combination. EMD/collagen positively influences osteoblast differentiation in a time dependent manner.