Stimulation of osteoblasts with Emdogain increases the expression of specific mineralization markers.

OBJECTIVE: The purpose of this study was to determine the effects of enamel matrix derivative on mRNA expression of markers related to periodontal healing. STUDY DESIGN: Murine osteoprogenitor cells (MC3T3-E1) were grown for 12 and 16 days in mineralization media and stimulated with 100 microg/mL Emdogain (EMD). Cell cultures treated with 2% and 10% fetal calf serum (FCS) served as control. The mRNA expression of bone sialoprotein (BSP), osteopontin (OPN), and runt-related protein 2 (Runx2) was analyzed by real-time polymerase chain reaction. One-way analysis of variance was used for statistical analysis. RESULTS: Stimulation with EMD significantly (P < .01) enhanced mRNA expression of BSP up to 13.9-fold and of OPN up to 3.2-fold at day 16 compared with the 2% FCS control. The expression of mRNA for transcription factor Runx2 was not significantly changed. CONCLUSION: The beneficial effects seen in periodontal regeneration after treatment with EMD may be related to an increase of the mineralization markers BSP and OPN at mRNA level.

Influence of periodontal therapy on the regulation of soluble cell adhesion molecule expression in aggressive periodontitis patients.

BACKGROUND: Inflammatory periodontal disease is associated with an increased risk of cardiovascular disease. Circulating cell adhesion molecules (CAM) (intercellular adhesion molecule-1 [ICAM-1], vascular cell adhesion molecule-1 [VCAM-1], and E-selectin) have been suggested as potential candidate markers of endothelial dysfunction, which contribute to the pathogenesis of cardiovascular diseases. The regulation of CAM in subjects with severe periodontitis and the influence of periodontal intervention on systemic CAM levels are not clear. The aim of this study was to determine whether intensive periodontal therapy reduces serum levels of CAM in patients with generalized aggressive periodontitis. METHODS: Blood samples were collected at six treatment time points from 21 patients with previously untreated generalized aggressive periodontitis (mean age: 34.6 +/- 4.3 years). Patients received subgingival scaling and root planing and antibiotic therapy and were monitored over a 6-month recall period. Serum levels of soluble ICAM-1 (sICAM-1), VCAM-1 (sVCAM-1), and E-selectin (sE-selectin) were measured by enzyme-linked immunosorbent assay. RESULTS: sE-selectin plasma levels decreased significantly (P <0.01) during periodontal therapy. Mean plasma levels were 65.95 ng/ml before treatment and 44.71 ng/ml 6 months after antibiotic therapy. sICAM-1 and sVCAM-2 serum levels were unaffected by therapeutic intervention. CONCLUSIONS: Periodontal therapy reduces plasma sE-selectin levels. Whether this leads to a reduction in risk of future cardiovascular events in patients with aggressive periodontal disease warrants further studies.

Effects of an enamel matrix derivative on human osteoblasts and PDL cells grown in organoid cultures.

OBJECTIVE: The objective of this study was to investigate cellular effects of enamel matrix derivative (EMD) in human derived, primary osteoblasts and periodontal ligament (PDL) cells grown in organoid cultures. STUDY DESIGN: Cell replication was assessed by BrdU-incorporation. [(3)H]-proline incorporation was measured to determine the synthesis of proline-containing proteins, such as collagen. In addition, calcium accumulation and alkaline-phosphatase-activity were quantified. Electron microscopy for morphological analysis was performed. RESULTS: Our results showed that EMD enhances BrdU-incorporation in PDL cells and osteoblasts. Also, in osteoblast organoid cultures [3H]-proline incorporation was 3-fold increased (P < .01). Extensive matrix deposition was noted in osteoblast cultures by electron microscopy. In osteoblasts, high levels of calcium accumulation and alkaline-phosphatase-activity were found. However, EMD did not promote mineralization. CONCLUSION: Our results indicate that under organoid culture conditions EMD is able to promote the synthesis of proline-containing proteins such as collagen but not matrix mineralization of primary human osteoblastic cells.

Moderate effect of enamel matrix derivative (Emdogain Gel) on Porphyromonas gingivalis growth in vitro.

OBJECTIVE: The aim of this study was to analyse the antibacterial effects of Emdogain Gel or its constituents on the growth of the suspected periodontopathogen Porphyromonas gingivalis. STUDY DESIGN: The effects of the proteins of enamel matrix derivative (EMD), the commercial product Emdogain Gel or its vehicle propylene glycol alginate (PGA) (Straumann, Switzerland) on P. gingivalis growth were determined by two methods: broth dilution assay (BDA) and agar diffusion assay (ADA). RESULTS: BDA-Emdogain Gel inhibited moderately the growth of P. gingivalis, whereas EMD showed no effect. The PGA vehicle inhibited the growth completely. ADA-Emdogain Gel resulted in some inhibition in growth but was not significantly different from control. EMD revealed no zone of inhibition. PGA demonstrated statistically significant zones of inhibition. CONCLUSION: Emdogain Gel shows moderate antibacterial activities against P. gingivalis. These properties seem to be due to the PGA component of the gel preparation.

Effects of enamel matrix derivative on proliferation and differentiation of primary osteoblasts.

OBJECTIVE: The aim of this study was to investigate the effects of enamel matrix derivative (EMD) on proliferation, protein synthesis, and mineralization in primary mouse osteoblasts. STUDY DESIGN: Osteoblasts were obtained from mouse calvaria by enzymatic digestion and grown in monolayer together with EMD (2-100 microg/ml). Metabolic activity and cell proliferation were determined by tetrazolium salt assay (MTT) and by 5-bromo-2'-deoxyuridine (BrdU) incorporation. For differentiation studies, a 3-dimensional organoid culture system was used. Osteoblastic differentiation was estimated by alkaline phosphatase (ALP) activity and calcium content. Collagen synthesis was assessed by [(3)H]-proline incorporation. Morphologic observations were made by electron microscopy. RESULTS: EMD treatments increased metabolic cell activity and BrdU incorporation. In the organoid cultures, ALP activity and calcium accumulation were enhanced by EMD treatment, but [(3)H]-proline incorporation was not. Morphologically, an increased deposition of mineralized nodules was found. CONCLUSIONS: EMD treatment enhanced cellular activities of primary osteoblasts and might support the regeneration of periodontal bony defects.

Salivary IgA in response to periodontal treatment.

There is evidence that the quantity of antigen load is crucial for the activation of IgA immune responses. In order to investigate the relevance of these findings in aggressive periodontitis, salivary antibody responses were measured during non-surgical and antibiotic treatment. Twenty-one patients with generalized aggressive periodontitis were monitored for total salivary IgA and IgA reactive to Porphyromonas gingivalis in resting and stimulated whole saliva. Non-surgical treatment included full-mouth professional tooth cleaning and subgingival scaling and root planing (SRP) under local anesthesia. Patients were recalled at 3 months and 6 months following systemic antibiotic treatment. Non-parametric statistics showed significant improvements in the clinical parameters in all patients. Between baseline and 4 wk following SRP, median concentrations of total IgA decreased both in resting (-46%) and in stimulated (-33%) saliva. The P. gingivalis-specific IgA activity showed a twofold increase at 4 wk after SRP. In addition to these changes, periodontal treatment of aggressive periodontitis did not appear to affect salivary IgA, and there were no significant correlations of IgA to the clinical parameters. In conclusion, salivary IgA responses during periodontal treatment were not found to have a diagnostic or prognostic significance.

White blood cell count in generalized aggressive periodontitis after non-surgical therapy.

BACKGROUND: Periodontal bacteria are known to invade the systemic circulation. Chronic low-level bacteremia and a systemic inflammatory response have been suggested as a pathogenetic link between periodontal disease and atherosclerosis. The purpose of this study was to examine the systemic effect of a non-surgical therapy on white blood cell count (WBC count) and differential blood count in smoking and non-smoking generalized aggressive periodontitis (GAP) patients. METHODS: 27 adult periodontitis patients (13 smokers and 14 non-smokers) with previously untreated GAP were subjected to 3 sessions of oral hygiene procedure. Afterwards, the patients were treated by scaling and root planing under local anaesthesia. Periodontal examinations were performed after supragingival pretreatment and three months after subgingival therapy. Pocket probing depth (PPD) and relative attachment level (RAL) were measured with Florida probe and disc probe. Accompanying clinical evaluation venous blood samples were taken to analyse the WBC counts and differential blood counts. For statistical analysis non-parametric tests were utilized. RESULTS: No clinical or demographic differences were found between smokers (n=13) and non-smokers (n=14). PPD, bleeding on probing (BoP) and suppuration improved significantly after therapy both in smokers and non-smokers. Following periodontal treatment WBC counts, neutrophil and platelet counts decreased significantly in non-smokers (p< or =0.004), while in smokers only platelet counts were significantly reduced (p=0.006). Non-smokers showed a significantly higher reduction of WBC counts (p=0.005) and neutrophils (p=0.001) compared to smokers. CONCLUSION: The results indicate that a therapeutical intervention may have a systemic effect on the blood count in GAP patients. This effect seems to differ between smokers and non-smokers.

Comparative study of Emdogain and coronally advanced flap technique in the treatment of human gingival recessions. A prospective controlled clinical study.

OBJECTIVES: Various surgical techniques have been proposed for coverage of denuded root surfaces. The aim of this study was to evaluate a comparison of coronally repositioned flap procedure with or without the use of enamel matrix proteins in the treatment of recession defects. MATERIAL AND METHODS: This study was an intra-individual longitudinal test of 12 months duration conducted as a blinded, split-mouth, placebo-controlled and randomised design. It was performed in 2 dental schools. 36 patients, aged 22-62 years, with 2 paired buccal recession defects of at least 3 mm participated. Surgical recession coverage was performed as coronally-advanced flap technique at both sites in the same session. One site was additionally treated with commercially-available enamel matrix proteins (Emdogain) and the other site with placebo (propylene glycol alginate) in accordance with the randomisation list. A blinded examiner assessed pre- and post-surgical measurements. Clinical measurements and photographs were taken pre-surgically and after 1 week, 3 weeks, 3 months, 6 months and 12 months, postoperatively. Measurements comprised height and width of the gingival recession, height of keratinized tissue, probing attachment level, probing pocket depth and alveolar bone level by periodontal probe, Florida Probe or caliper to the nearest 0.5 mm. RESULTS: 12 months after therapy, both treatment modalities showed significant root coverage and probing attachment gain. Gingival recession decreased from 3.7 mm to 0.8 mm for the Emdogain treated sites and from 3.9 mm to 1.0 mm for the control sites, corresponding to mean root coverages of 80% and 79%, respectively. This difference was not significant. With the exception of keratinized tissue gain, which was significantly higher (p=0.003) in the Emdogain group, all other clinical variables were not different in the between-group comparison. CONCLUSIONS: As the additional use of Emdogain together with coronally advanced flap technique for recession coverage showed no difference in the overall clinical outcome, there is no clear benefit to combine Emdogain with this surgical technique.