Response of human gingival microvascular endothelial cells to mechanical stress

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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.