牙龈卟啉菌表面相关物质的提取、纯化及骨吸收和免疫特性研究

目的 :通过对牙龈卟啉菌Pg表面相关物质 (SAM)的提取、纯化 ,研究其潜在的骨吸收活性 ,以揭示pg的SAM在牙周病过程中的致病作用。方法 :1)细菌培养。 2 )SAM粗品的制备。Sephacryl-S2 0 0凝胶过滤、冻干、纯化。3)DEAE -Sephacel阴离子交换层析。 4 )SAM介导的小鼠颅骨吸收试验。结果 :1)细菌培养得到冻干的细菌 4 .1g。2 )分离SAM得到粗品 0 .16g。Sephacryl -S2 0 0凝胶过滤纯化SAM粗品 ,冻干后得到较纯的SAM约为 0 .0 2 2 1g。DEAE -Sephacel阴离子交换层析进一步纯化SAM获得SAM纯品 3mg。 3)纯化的SAM骨吸收活性鉴定。骨吸收作用可通过体外小鼠颅骨培养基中Ca2 + 浓度的增加而间接证实。SAM的骨吸收活性随着浓度的增加而增加 ,与对照组相比 ,P <0 .0 5。SAM具有免疫原性。结论 :SAM在骨吸收方面具有极大的潜能。提示SAM在牙周病骨损害方面起到重要的作用

Isolation and Purification of Surface-associated Material from Porphyromonas Gingivalis to Study its Bone Resorbing Activity and Antigenicity

Objective: Study the effect of bone resorption of Surface-associated material(SAM) of Porphyromonas gingivalis in the Periodontal disease by abstracting and purification of SAM and reveal the cause diseases of SAM of Porphyromonas gingivalis during periodontal diseases. Methods: 1) bacterial culture. 2) extraction of SAM. 3) Purification of SAM(4)Bone resorption: Bone resorption is measured by the release of calcium of mouse calvaria. Results: 1) we obained lyophilized bacterial 4.1g from bacterial culture. 2)crude SAM from bacterial wall was 0.16g. Gel filtration of the crude SAM on Sephacryl-S200 and then purification of SAM by DEAE-sephacel ,purified SAM was obtained about 3mg. 3) Bone resorption tests indicated that activity of Bone resorption increased with concentration of SAM,compare with control group P<0.05.SAM has antigenicity. Conclusions: SAM has grest potential in bone resoption indicaed that SAM play important role in bone lesion respect in periodontal diseases.