An investigation of the effect of heat stimulation for cartilage metabolism in mRNA

2005 Fiscal Year Final Research Report Summary

• Project/Area Number: 16591506
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: HOJO Tatsuya Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Assistant, 医学研究科, 講師 (40298740)
• Co-Investigator(Kenkyū-buntansha): KUBO Toshikazu Kyoto Prefectural University of Medicine, Graduate School of Medical Science, professor, 医学研究科, 教授 (20178031) ; TAKAHASHI Kenji Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Assistant, 医学研究科, 助手 (30347447)
• Project Period (FY): 2004 – 2005
• Keywords: chondrocyte / heat / cell viability / aggrecan / type II collagen / mRNA

Research Abstract

Chondrocyte like cell (HCS-2/8) established from human chondrosarcoma were cultured in flat plate. They were stimulated by heat within a water bath which was setting to 37, 39, 41, 43 degrees Celsius with each temperature for 15 minutes and 30 minutes. We extracted mRNA by guanidine acid phenol chloroform method after having continued culture for 4 hours at 37 degrees Celsius after heat stimulation and analyzed a manifestation of mRNA of aggrecan and type II collagen to be related to cartilage metabolism by Real-time PCR method. In addition, we measured cell viability by Tetrazolium salt-based assay after having continued culture after heat stimulation for 72 hours. We used 37 degrees Celsius group as control. As for the manifestation of mRNA of aggrecan, there were not most changes by 39 degrees Celsius stimulation of 15 minutes, but it was enhanced by 39 degrees Celsius stimulation for 30 minutes. There were slight decrease by 41 degrees Celsius stimulation both for 15 minutes and 30 minutes. and obvious decrease by 43 degrees Celsius stimulation especially for 30 minutes. As for the manifestation of mRNA of type II collagen, there were slight increase by 39 degrees Celsius stimulation of 15 minutes and 30 minutes, and by 41 degrees Celsius stimulation for 15 minutes and decrease by 41 degrees Celsius for 30 minutes and 43 degrees Celsius. The cell viability did not have a dominant difference in 39 and 41 degrees Celsius, but there were a dominant decrease at 43 degrees Celsius. For these results, it was necessary for heat stimulation for chondrocyte to take into consideration in the quantity of that was able to put stimulation temperature and stimulation time. An adequate heat stimulation promoted cartilage metabolism and cell viability and an excessive heat stimulation decreased them.