Response of human gingival microvascular endothelial cells to mechanical stress

Research Project

Project/Area Number	05404071
Research Category	Grant-in-Aid for General Scientific Research (A)
Allocation Type	Single-year Grants
Research Field	矯正・小児・社会系歯学
Research Institution	Okayama University
Principal Investigator	WATANABE Tatsuo Department of Preventive Dentistry, Okayama University Dental School, Professor, 歯学部, 教授 (20034176)
Co-Investigator(Kenkyū-buntansha)	ISHIKAWA Akira Department of Preventive Dentistry, Okayama University Dental Hospital, Lecturer, 歯学部・付属病院, 講師 (70222958) MORITA Manabu Department of Preventive Dentistry, Okayama University Dental Hospital, Lecturer, 歯学部・付属病院, 講師 (40157904) ICHIKAWA Hiroyuki 2nd Department of Oral Anatomy, Okayama University Dental School, Assistant Prof, 歯学部, 助教授 (20193435) SUGIMOTO Tomosada 2nd Department of Oral Anatomy, Okayama University Dental School, Professor, 歯学部, 教授 (50135729) 梶浦靖二岡山大学, 歯学部, 助手 (60243482) 山本龍生岡山大学, 歯学部, 助手 (20252984)
Project Period (FY)	1993 – 1995
Project Status	Completed (Fiscal Year 1995)
Budget Amount *help	¥18,000,000 (Direct Cost: ¥18,000,000) Fiscal Year 1995: ¥2,000,000 (Direct Cost: ¥2,000,000) Fiscal Year 1994: ¥5,300,000 (Direct Cost: ¥5,300,000) Fiscal Year 1993: ¥10,700,000 (Direct Cost: ¥10,700,000)
Keywords	Endothelial cells / Human gingivae / Periodontal disease / Mechanical stress / 細胞培養 / マッサージ / 歯肉血管内皮細胞 / 敷石状形態 / von Willebrand factor / 蛍光標識 acetylated LDL / 分離 / 同定 / 96穴プレート
Research Abstract	1.Isolation of human gingival microvascular endothelial cells (HGECs) : After collagenase treatment, human gingival samples were compressed to release endothelial cells. The cell suspension was introduced into every well of a 96-well plate equally. After 3-4 weeks later, the morphologically homogeneous cells with cobblestone appearance at confluence were harvested. 2.Identification of HGECs : The cells were identified as HGECs by the cobblestone morphology at confluence, presence of von Willebrand factor, uptake of acetylated low density lipoprotein (Ac-LDL) and formation of capillary-like structures on Matrigel. HGECs were isolated from 5 out of 16 samples (31.3%) at the donor's age of 10-20s. On the other hand, at the donors of more than 30 years old, HGECs were isolated from 6 out of 35 samples (17.1%). 3.Growth medium for HGECs : HGECs were well grown in MCDB 107 medium supplemented with 20% fetal bovine serum (FBS), 75mug/ml endothelial cell growth supplement (ECGS) and 30mug/ml heparin. 4.Characterization of HGECs : Unlike human umbilical vein endothelial cells (HUECs), HGECs exhibited the weaker uptake of Ac-LDL and the greater dependence upon the concentrations of FBS.Moreover, prostaglandin E_2 was the major arachdonic metabolite in HGECs. 5.Mehanical stress : Mechanical stress was applied to HUECs with the Flexercell Strain Unit. Intermittent force accelerated the proliferation of HUECs. 6.The metabolic products of endothelial cells with mechanical stress will be investigated.