|Budget Amount *help
¥2,100,000 (Direct Cost : ¥2,100,000)
Fiscal Year 1997 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 1996 : ¥1,300,000 (Direct Cost : ¥1,300,000)
Many patientens who are suffering from pain and disorder due to the internalderangement on temporomandibular join (TMJ) visited our clinic but the it's etiology still unclear.
Especially the fuction of TMJ synoviocytes in the case of internalderangement is not investigated at all. We tried to establish the culture synovial celllines from the internalderangement of TMJ.At first, we tried just the same method as the synovial cell culture method on knee joint by Dayr JC.With this method, in the end of the screening the synovial tissue materials, the number of rest of synoviocytes was very little and it's activity was not so hight as synoviocytes of rheumatism arthritis on knee joints. With this method, we could not get the cultured synovial cellline from our TMJ materials. Therefore we added some arrangement to this method ; after that, we found that DMEM medium with 10% fetal bovine serum (BSA) was best to culture TMJ synovial cell. In this condition, one of cellline could grow after 38 h
ours later, another of cellline takes more than 70 hours. What was interesting for us the hormer cellline derived from the patient who had much more pain in TMJ than the latter cellline.
We also studied the cellline with anti-cytokeratin, anti-vimentin, anti-leukocyte common antigen, and anti-macrophage antigen monoclonal antibodies immunohistochemically. As the result of these study only anti-vimentin monoclonal antibodies reacted strongly a lot of numbers of culture cells, with another antibodies, no positive reaction could be seen.
Farther more, with the culture supernatants from synoviocytes, the consentration of trans forming growth facto beta, interleukin-1 beta and interleukin-6 were analized with ELISA method. As the result of these study, only the consentration of interleukin-6 was hight level in every celllines but there ware no capable difference between them. The other consentration of another cytokines was not detectable with ELISA.
The other hand, on the hyperplastic synovial tissues in these disease m-RNA production could be recognized with in-situ-hybridization. Less