Response of human gingival microvascular endothelial cells to mechanical stress

1.人牙龈微血管内皮细胞的分离：经胶原酶处理后，将人牙龈标本压缩以释放内皮细胞。将细胞悬浮液均匀地引入96孔板的每一孔中。3~4周后，取融合处形态均一的鹅卵石样细胞，经鉴定：融合处鹅卵石样形态，有von Willebrand因子存在，摄取乙酰化低密度脂蛋白(Ac-LDL)，Matrigel上形成毛细血管样结构，鉴定为HGEC。供者10-20岁时，16份标本中有5份(31.3%)分离到HGEC。另一方面，在30岁以上的供者中，35例中有6例(17.1%)分离到HGECs。3.HGECs的生长介质：HGECs在含有20%胎牛血清(FBS)、75mug/ml内皮细胞生长补充剂(ECGs)和30mug/ml肝素的MCDB 107培养液中生长良好。4.HGECs的特征：与人脐静脉内皮细胞(HUECs)不同，HGECs对ac-LDL的摄取较弱，对FBS浓度的依赖性较大。前列腺素E_2是人脐静脉内皮细胞的主要蛛丝蛋白代谢产物。5.机械应力：用Flexcell应变装置对人脐静脉内皮细胞施加机械应力。间歇力促进内皮细胞的增殖。6.研究机械应力作用下内皮细胞的代谢产物。

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.