| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1987: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1986: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Research Abstract |
The important subject are as the basic research for the study of mechanisms of adherence of oral microorganisms to the cell by an electron microscope. Gin-1(gingival fibroblast,apparent normal;human)ATCC CRL 1292 was used as the host cell, and cariogenic Streptococci were chosen as control cells of adherence. However, we have a vary great difficuity to stabilize tissue culture of gingival cells of animals, then in the 1986, the present study was performed to establish culture and inoculation of fibroblasts Gin-1. So that we can inoculate fibroblasts Gin-1 at cell concentration of 25,000 cells/cm^2 under 37゜C, 5%CO_2-95%air and a saturated steam. And Streptococcus mutans ATCC 25175, S.oralis ATCC 35037, S.sanguis ATCC 10556, S.salivarius ATCC 25975 and S.mitis ATCC 33399 were confirmed biochemical characteristics by Facklam, and examined adherence of these bacteria to cells except the host cell.
In the 1987, furthermore the adherence of estimate the interaction between oral micro- organi
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sms and tissue culture cells was studied for photograph through an electron micro- scope( SEM and TEM ). Bacterial strains used were S.mutans ATCC 25175, S.oralis ATCC 35037, S.sanguis ATCC 10556, S.salivarius ATCC 25975 and S.mitis ATCC 33399. Dell strain used was Gin-1.
Samples for SEM were fixed in a containing 2% glutaraldehyde and 2% osmic acid, dehydrated in ethanol solutions, coated by Au and observed by SEM(X-560,Hitachi), acceleration voltage 20kV. Samples for TEM were fixed in a containing 2% glutaraldehyde ,postfixed for 16 h at 4゜C in 1% osmic acid-0.1M cacodylic acid-HCl buffer( pH 7.2 ), dehydrated in ethanol solutions, embedded in Epon for 72 h at 60 ゜C, stained by 2% uranyl acetate and lead citrate and observed by TEM(H-800,Hitachi), acceleration voltage 100kV.
In conclusion, according to ultrastructure of SEM and TEM, the order of adhesive activity is S.mitis, S.salivarius, S.sanguis, S.oralis, S.mutans. S.oralis and the microvillus of the host cell appear to related. S.mitis had the strongest adhesive activity to Gin-1 cells ,and S.mutans had the most weak activity. Further experiment will be in the future. Less
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