Grant-in-Aid for Scientific Research (B)
|Allocation Type||Single-year Grants |
|Research Institution||OKAYAMA UNIVERSITY |
KANYAMA Manabu Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Assistant Professor, 医学部・歯学部附属病院, 講師 (90294420)
KUBOKI Takuo Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (00225195)
TAKIGAWA Masaharu Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (20112063)
ARAKAWA Hikaru Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Research Associate, 大学院・医歯薬学総合研究科, 助手 (30304314)
SUZUKI Kouji Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Research Associate, 医学部・歯学部附属病院, 助手 (30304322)
FUJISAWA Takuo Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Research Associate, 大学院・医歯薬学総合研究科, 助手 (20325096)
中西 徹 岡山大学, 大学院・医歯学総合研究科, 助教授 (30243463)
|Project Period (FY)
2003 – 2005
Completed (Fiscal Year 2005)
|Budget Amount *help
¥14,800,000 (Direct Cost: ¥14,800,000)
Fiscal Year 2005: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2003: ¥10,800,000 (Direct Cost: ¥10,800,000)
|Keywords||bone marrow cells / Osteoblasts / Titanium / Osseointegration / xab-2 / sod-1 / galection-1 / デンタルインプラント / チタン特異的遺伝子 / 骨芽細胞 / 骨組織再生 / 幹葉系幹細胞|
The aim of this study is to clarify the cell behavior induced by titanium and to isolate specific genes that promote the titanium implant-bone integration. Especially, to identify the master key genes related osseointegration to titanium, the osteoblastic cell line, MC3T3-E1 cells were cultured on titanium coated glasses and the gene expression were analyzed.
1)Titanium and chrome coated glasses
Ti and Cr were coated to polished glass surfaces (33 mm in diameter by 1.5 mm in thickness) in a high vacuum condition. Ti and Cr coated glass disks were placed in a 6 well plate and fixed by silicone rings. Non-coated glass disks were served as controls.
2)Cell attachment, proliferation and differentiation assessments.
Initial cell attachment and proliferation of MC3T3-E1 cells were estimated by MTS-assay (CellTiter 96【○!R】 AQueous One Solution, Promega, USA), and cell differentiation was evaluated by alkaline phosphatase activity assay. The mean relative amount of the attached cells on the Ti-coa
ted glass was significantly higher than those on non-coated and Cr-coated glass at 3 hours after seeding. On the same way the mean cell number on Ti-coated glass at 3 days after seeding was significantly higher than those of other conditions. The mean alkaline phosphatase(ALP) activity of the seeded osteoblasts also increased with time in these three conditions, while the ALP activity level on the Ti-coated glass at 14 days after seeding was significantly higher than those of other conditions. These results suggest that, Ti-coated glass plate accelerated the cell attachment, proliferation and differentiation of the MC3T3-E1 cells, compared to the non- and Cr-coated glass plates
3)Effect of gene expression of osteoblast on titanium
Osteoblasts were cultured on Ti coated, Cr coated and non-coated glass plates. And then differentially expressed genes were identified by cDNA subtractive hybridization. Twenty independent clones were isolated and by nucleotide sequencing of these clones, seven clones were identified including EST genes ; xab-2,sod-1,galectin-1,actin related protein 2/3 mRNA, RIKEN cDNA 2210013021 gene, EST 601086505F1, and EST 01439. And galectin-1,xab-2,sod-1 gene expression on Ti-coated glass were higher than non-coated, Cr-coated glass.
These results suggest that Ti-coating is more advantageous for osteoblastic cell attachment, proliferation and differentiation than Cr or non-coating and galectin-1,xab-2 and sod-1 up-regulation could be related to the Ti-bone integration. Less