Isolation and characterization of human gingival microvascular endothelial cells.
OBJECTIVE
Endothelial cells have a substantial role in maintaining vascular homeostasis, and their dysregulation can contribute to the development of pathology. The plasminogen activators and their inhibitors may, arguably, be the single most important proteolytic system of the endothelium for vascular maintenance by controlling plasminogen activation and other proteolytic cascades that impact on clotting, hemodynamics, angiogenesis and the character of the vascular wall. In chronic periodontal disease, significant changes to the microvasculature occur in association with the severity of the disease. Investigation of the role played by endothelial cells in periodontal health and disease has been limited to in situ immunolocalization or to the use of endothelial cells of nongingival origin, such as human umbilical vein endothelial cells. The objective of this research was to establish a replicable protocol for isolating microvascular endothelial cells from the gingiva.


METHODS
From inflamed gingiva, isolated cells were characterized by morphology, the expression of factor VIII-related antigen, the expression of UEA-1 ligand, the uptake of acetylated low-density lipoprotein, network formation on Matrigel, and by the expression levels of urokinase plasminogen activator, tissue plasminogen activator, plasminogen activator inhibitor-1 and collagen IV.


CONCLUSIONS
Gingival endothelial cells were most readily obtained from inflamed gingival tissues, and these endothelial cells, when isolated by the protocol established herein, demonstrated endothelial characteristics and constitutively secreted plasminogen activators and plasminogen activator inhibitor-1 in culture.
DeCarlo
A A
AA
Nova South-eastern University College of Dental Medicine, Department of Periodontology, Fort Lauderdale, FL, USA. DeCarlo@agentabio.com
Cohen
J A
JA
Aguado
A
A
Glenn
B
B
D018929:Cell Culture Techniques; D002469:Cell Separation; D002478:Cells, Cultured; D003094:Collagen; D024141:Collagen Type IV / Q000096:biosynthesis; D004338:Drug Combinations; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005881:Gingiva / Q000098:blood supply*; D005891:Gingivitis / Q000473:pathology; D006801:Humans; D007797:Laminin; D008077:Lipoproteins, LDL / Q000378:metabolism; D008833:Microcirculation / Q000166:cytology / Q000378:metabolism*; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011509:Proteoglycans; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis; D054457:Tissue Scaffolds; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis; D014841:von Willebrand Factor / Q000302:isolation & purification
D024141:Collagen Type IV; D004338:Drug Combinations; D007797:Laminin; D008077:Lipoproteins, LDL; D017395:Plasminogen Activator Inhibitor 1; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011509:Proteoglycans; C032629:acetyl-LDL; D014841:von Willebrand Factor; C059485:matrigel; D003094:Collagen; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator