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.

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 μg/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.

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.

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.