Three-year clinical evaluation of a polyacid-modified resin composite in minimally invasive occlusal cavities

OBJECTIVE: The aim of this study was to evaluate the 3-year clinical performance of one polyacid-modified resin composite material (PMRC). Dyract, in minimally invasive occlusal cavities and its neighbouring fissures. METHODS: One hundred and sixteen restorations of the material investigated were placed by a single operator in a group of selected children under controlled conditions. Isolation of the restorations was accomplished with the use of cotton rolls and aspiration. Using modified US Public Health Service (USPHS) codes and criteria, the restorations were reviewed clinically within 1 week of placement (baseline), and thereafter at 6 months, 1, 2 and 3 years. RESULTS: After 3 years, marginal discolouration was present in 8.6% of the restorations. The marginal adaptation was rated as partly sealed (Oscar-Alpha) in 107 (92.2) of the restorations. Five restorations had lost their sealant components, while four restorations were partly sealed with explorer-catch after 3 years. Although wear of the restorations was considerable, restorations rated as 'partly sealed' had at least two-thirds of their sealant components fully retained. Recurrent caries was associated with four (3.4%) restorations. CONCLUSION: In this clinical study, the retention rate of the tested PMRC material was good, although a marked occlusal wear was evident. The marginal adaptation of the PMRC at the enamel site would probably have been better by the use of enamel-etching. Provided the marginal adaptation and wear resistance of the material is further improved, clinical use of PMRCs in minimally invasive occlusal cavities can be advocated.     

Porcine fetal enamel matrix derivative enhances bone formation induced by demineralized freeze dried bone allograft in vivo

BACKGROUND: Embryonic enamel matrix proteins are involved in the formation of acellular cementum during development of the periodontal attachment apparatus, suggesting that these proteins might be used clinically to promote periodontal regeneration. At present, it is unknown if these proteins are osteoinductive, osteoconductive, or osteopromotive. To address this question, we examined the ability of a commercially prepared embryonic porcine enamel matrix derivative to induce new bone formation in nude mouse calf muscle, or to enhance the bone induction ability of a demineralized freeze-dried bone allograft (DFDBA). METHODS: Porcine fetal enamel matrix derivative (EMD) was implanted bilaterally in the calf muscle of 4 male Nu/Nu mice per treatment group (N = 8 implants): 2 mg EMD alone; 4 mg EMD alone; inactive human DFDBA alone; inactive DFDBA + 2 mg EMD; inactive DFDBA + 4 mg EMD; active DFDBA alone; active DFDBA + 2 mg EMD; and active DFDBA + 4 mg EMD. Implants were harvested after 56 days and examined histologically for bone induction using a semi-quantitative score and histomorphometrically for area of new bone, cortical bone, bone marrow, and residual DFDBA. RESULTS: Implants containing inactive DFDBA, 2 mg EMD, 4 mg EMD, and inactive DFDBA + 2 or 4 mg EMD did not induce new bone. Active DFDBA and active DFDBA + 2 mg EMD induced new bone to a similar extent. In contrast, active DFDBA + 4 mg EMD resulted in enhanced bone induction, area of new bone, and cortical bone. Residual DFDBA was also increased in this group. CONCLUSIONS: EMD is not osteoinductive. However, it is osteopromotive, due in part to its osteoconductive properties, but a threshold concentration is required.     

细菌素免疫蛋白调控变形链球菌抗菌敏感性及生物膜形成的实验研究

目的 观察细菌素免疫蛋白相关基因对变形链球菌抗菌敏感性及生物膜形成的影响,探讨细菌素免疫蛋白与细菌抗菌剂耐受性的关系,为生物膜抗菌敏感性的研究提供基础数据.方法筛选培养细菌素免疫蛋白基因突变株,绘制生长曲线.酶标仪检测不同质量浓度氨苄青霉素(0.04、0.05、0.06、0.07及0.08 mg/L)、氟化钠(50、100、150、200及250 mg/L)及不同质量分数的次氯酸钠(0.078%、0.156%、0.313%、0.625%及1.250%)作用下变形链球菌标准株、△immA-和△immB-突变株菌液的吸光度值.应用最小生物膜清除浓度(minimal biofilm eradicatin concentration,MBEC)桩钉96孔板以连续稀释法检测醋酸氯己定对3种菌株生物膜的MBEC.应用激光共聚焦扫描显微镜(confocal laser scanning microscope,CLSM)定量分析标准株和突变株生物膜结构.结果 △immA-和△immB-突变株的迟缓期和稳定生长期均比标准株延时1 h.氨苄青霉素为0.06 mg/L时,标准株、△immA-突变株和△immB-突变株菌液吸光度值分别为0.334±0.016、0.027±0.016及0.047±0.018;氟化钠质量浓度为150 mg/L时,3种菌株菌液吸光度值分别为0.254±0.018、0.129±0.011及0.167±0.01;当次氯酸钠质量分数为0.313%时,3种菌株菌液吸光度值分别为0.467±0.008、0.017±0.006及0.050±0.006,以上各组抗菌剂中,标准株与突变株吸光度值差异均有统计学意义(P<0.01).醋酸氯己定对3种菌株的MBEC分别为6.25、1.57及3.13 mg/L.标准株生物膜厚度显著高于△immA-和△immB-突变株(P<0.01);标准株各层活菌比例均高于△immA-突变株(P<0.05);标准株中、外层活菌比例高于△immB-突变株(P<0.01),但内层活菌比例差异无统计学意义(P=0.191).结论细菌素免疫蛋白参与调控浮游细菌生长,尤其在生长初期;细菌素免疫蛋白相关基因缺陷使浮游态变形链球菌抗菌敏感性提高、抗菌剂MBEC降低及生物膜结构不成熟.     

Measurement of proteases in human subgingival dental plaque by fluorescence polarization

Fluorescence polarization (FP) was examined as a rapid quantitative method to assay the proteases in subgingival plaque. Protease activity was measured by a decrease in FP at 0.5-min intervals over 5 min, using BODIPY®--casein, a protein substrate. To quantitate activity, the least absolute deviation (LAD) slope for each assay was determined. Protease activity increased with the quantity of plaque (r=0.416, P<0.001). Of the 208 subgingival plaque samples, 87 contained detectable protease activity, with a mean of about 4 μg trypsin equivalents above a general background of 1 μg per site. The mean plaque protease activity of 89 paired samples from 15 individuals had decreased by 1.1 μg trypsin equivalents per site when measured at 8 months after tooth scaling and root planing (P<0.01). Most isolates of Porphyromonas gingivalis, Treponema denticola, Prevotella nigrescens, and Prevotella intermedia implicated in the pathogenesis of adult periodontitis exhibited high activity in the FP assay. The assay is rapid, quantitative and requires only one-tenth of the plaque sampled using a single pass with a Gracey curette at a single tooth site.     

Healing of onlay mandibular bone grafts covered with collagen membrane or bovine bone substitutes: a microscopical and immunohistochemical study in the sheep

The objective of this study was to evaluate the role of collagen membrane and Bio-Oss coverage in healing of an onlay graft to the mandible. Twelve adult sheep each received an onlay bone graft (experiment 1), bone graft+Bio-Gide (experiment 2), and bone graft+Bio-Oss/Bio-Gide (experiment 3) on the lateral surface of the mandible. The animals were euthanized at 4, 8, 12 or 16 weeks after surgery, and findings were analysed by routine microscopy and immunohistochemistry for proliferation (Ki67) and apoptotic (Caspase-3) markers. Grafts were fully incorporated in all specimens. Pronounced resorption was observed in experiment 1. Minimal loss of graft volume was seen in experiment 2 specimens without membrane displacement. A remarkable increase in the augmented region of the mandible was observed in experiment 3. A high number of osteoclasts were expressed within the grafts during the early healing period, and thereafter declined markedly. Osteoblasts within the grafts expressed a moderate level of Ki67 at 8 weeks, which thereafter declined markedly. The strongest expression of Caspase-3 on the bone surface was observed after 16 weeks. In conclusion, the effect of collagen membrane coverage on bone graft volume maintenance was dependent on membrane stability during healing. An autogenous bone graft covered with Bio-Oss particles resulted in a remarkable increase in augmented lateral surface of the mandible. The late stage of bone graft healing was associated with a high apoptotic induction pathway of osteoblasts lining the surfaces of the new bone, demonstrated by strong positive Caspase-3 immunoreactivity.     

Protease-active extracellular protein preparations from Porphyromonas gingivalis W83 induce N-cadherin proteolysis, loss of cell adhesion, and apoptosis in human epithelial cells

BACKGROUND: The protease-induced cytotoxicity of P. gingivalis may partly result from alteration of the extracellular matrix and/or surface receptors that mediate interaction between the host cells and their matrix. While P. gingivalis-induced degradation of E-cadherin has been documented, there is no information on the effects of P. gingivalis proteases on other members of this family of cell adhesion proteins. METHODS: Human epithelial KB cells were exposed to protease-active extracellular protein preparations from isogenic mutants of P. gingivalis. Quantification of apoptosis was performed by visualization of nuclei stained with 4,6'-diamidino-2-phenylindole. Alteration of cell adhesion proteins was examined by immunoblotting of cell lysates using monoclonal antibodies to those proteins. RESULTS: Treated cells exhibited loss of cell adhesion properties with apoptotic cell death subsequently observed. These effects correlated with the different levels of cysteine-dependent proteolytic activities of the isogenic mutants tested. Cleavage of N-cadherin was observed in immunoblots of lysates from detached cells. There was a direct correlation between the kinetics of N-cadherin cleavage and loss of cell adhesion properties. Loss of cell adhesion, as well as N-cadherin cleavage, could be inhibited by preincubation of P. gingivalis protease active extracellular protein preparations with the cysteine protease inhibitor TLCK. In control experiments, the cleavage of N-cadherin was detected after treatment of KB cells with trypsin but not after cell dissociation by a non-enzymatic method. CONCLUSIONS: These results suggest that extracellular proteases from P. gingivalis can induce degradation of N-cadherin, which could have implications for the pathogenicity of this bacterium.