权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

A novel inhibitor in bone formation from periodontopathic bacteria

牙周病细菌骨形成的新型抑制剂

基本信息

批准号：
07557137
负责人：
TAKEHARA Tadamichi
金额：
$ 4.99万
依托单位：
Kyushu Dental College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

In order to study the effects of sonicated extracts from Prevotella intermedia. Actinobacillus actinomycetemcomitans, Porphyomonas gingivalis, and other oral-related bacteria, as well as Escherichia coli on bone formation, clone MC3T3-E_1 cells, which have retained osteoblastic activity, were cultured with various bacterial extracts.The addition of the sonicated extracts from Prevotella intermeedia and Actinobacillus actinomycetemcomitans decreased the alkaline phosphatase activity in a dosedependent fashion over the concentration range of 1-1000 ng ml^<-1> compared with the control. By contrast, the sonicated extracts from other oral bacteria including Prophyromonas gingivalis, Capnocytophaga ochracea, Streptococcus milleri and Streptococcus sanguis, and Escherichia coli did not decrease the alkaline phosphatase activity even in the presence of 100 ng ml^<-1> protein.The addition of prevotella intermedia and Actinobacillus actinomycetemcomitans extracts that had been treated with heat and trypsin to the cell cultures also inhibited alkaline phosphatase activity in the cells, suggesting that inhibitory factors are not proteinaceous. Polymyxin B did not change the alkaline phosphatase activity in the cells treated with the extracts from Prevotella intermedia and Actinobacillus actinomycetemcomitans, suggesting that the inhibitory activity of the extracts is not lipopolysaccharide.The inhibitory effect of both extracts was observed in the molecular mass over 290 kDa eluted from Sephadex G-200 column. The inhibitory substances of Prevotella intermedia were partially purified and showed broad band with estemated molecular weight of 170-190 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.These results indicate that Prevotella intermedia and Actinobacillus actinomycetemcomitans may play an important role in inhibiting bone formation as well as in stimulating bone resorption.

为了研究中间普雷沃氏菌超声提取物的效果。 Actinobacillus actinomycetemcomitans、牙龈卟啉单胞菌和其他口腔相关细菌，以及对骨形成的大肠杆菌，用各种细菌提取物培养保留成骨活性的克隆 MC3T3-E_1 细胞。添加来自 Prevotella intermeedia 和 Actinobacillus 的超声提取物与对照相比，伴放线菌在1-1000ng ml ^ -1 的浓度范围内以剂量依赖性方式降低碱性磷酸酶活性。相比之下，来自其他口腔细菌(包括牙龈原单胞菌、赭二氧化碳噬菌体、米勒链球菌和血链球菌以及大肠杆菌)的超声提取物即使在存在100ng ml^ -1 蛋白质的情况下也不会降低碱性磷酸酶活性。经过热和胰蛋白酶处理的细胞培养物放线杆菌提取物也抑制细胞中的碱性磷酸酶活性，表明抑制因子不是蛋白质。多粘菌素B没有改变用中间普雷沃氏菌和伴放线放线杆菌提取物处理的细胞中的碱性磷酸酶活性，表明提取物的抑制活性不是脂多糖。在Sephadex G-200柱洗脱的分子量超过290 kDa时观察到两种提取物的抑制作用。中间普雷沃氏菌的抑制物质得到部分纯化，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示出宽带，估计分子量为170-190 kDa。这些结果表明，中间普雷沃氏菌和伴放线放线杆菌可能在抑制骨形成和刺激骨吸收方面发挥重要作用。