Chronic High Glucose Concentration Induces Inflammatory and Remodeling Changes in Valvular Endothelial Cells and Valvular Interstitial Cells in a Gelatin Methacrylate 3D Model of the Human Aortic Valve
Letitia Ciortan,
Razvan Daniel Macarie,
Sergiu Cecoltan,
Mihaela Vadana,
Monica Madalina Tucureanu,
Andreea Cristina Mihaila,
Ionel Droc,
Elena Butoi,
Ileana Manduteanu
Affiliations
Letitia Ciortan
Institute of Cellular Biology and Pathology “Nicolae Simionescu”, Biopathology and Therapy of Inflammation, 8, B.P. Hasdeu Street, P.O. Box 35-14, 050568 Bucharest, Romania
Razvan Daniel Macarie
Institute of Cellular Biology and Pathology “Nicolae Simionescu”, Biopathology and Therapy of Inflammation, 8, B.P. Hasdeu Street, P.O. Box 35-14, 050568 Bucharest, Romania
Sergiu Cecoltan
Institute of Cellular Biology and Pathology “Nicolae Simionescu”, Biopathology and Therapy of Inflammation, 8, B.P. Hasdeu Street, P.O. Box 35-14, 050568 Bucharest, Romania
Mihaela Vadana
Institute of Cellular Biology and Pathology “Nicolae Simionescu”, Biopathology and Therapy of Inflammation, 8, B.P. Hasdeu Street, P.O. Box 35-14, 050568 Bucharest, Romania
Monica Madalina Tucureanu
Institute of Cellular Biology and Pathology “Nicolae Simionescu”, Biopathology and Therapy of Inflammation, 8, B.P. Hasdeu Street, P.O. Box 35-14, 050568 Bucharest, Romania
Andreea Cristina Mihaila
Institute of Cellular Biology and Pathology “Nicolae Simionescu”, Biopathology and Therapy of Inflammation, 8, B.P. Hasdeu Street, P.O. Box 35-14, 050568 Bucharest, Romania
Ionel Droc
Cardiovascular Surgery Department, Central Military Hospital, 010825 Bucharest, Romania
Elena Butoi
Institute of Cellular Biology and Pathology “Nicolae Simionescu”, Biopathology and Therapy of Inflammation, 8, B.P. Hasdeu Street, P.O. Box 35-14, 050568 Bucharest, Romania
Ileana Manduteanu
Institute of Cellular Biology and Pathology “Nicolae Simionescu”, Biopathology and Therapy of Inflammation, 8, B.P. Hasdeu Street, P.O. Box 35-14, 050568 Bucharest, Romania
Calcific aortic valve disease (CAVD), a degenerative disease characterized by inflammation, fibrosis and calcification, is accelerated in diabetes. Hyperglycemia contributes to this process by mechanisms that still need to be uncovered. We have recently developed a 3D model of the human aortic valve based on gelatin methacrylate and revealed that high glucose (HG) induced osteogenic molecules and increased calcium deposits in a pro-osteogenic environment. To further understand the events leading to calcification in diabetic conditions in CAVD, we analyzed here the inflammatory and remodeling mechanisms induced by HG in our 3D model. We exposed valvular endothelial cells (VEC) and interstitial cells (VIC) to normal glucose (NG) or HG for 7 and 14 days, then we isolated and separated the cells by anti-CD31 immunomagnetic beads. The changes induced by HG in the 3D model were investigated by real-time polymerase chain reaction (RT-PCR), Western blot, enzyme-linked immunosorbent assay (ELISA) and immunofluorescence. Our results showed that HG induced expression of different cytokines, cell adhesion molecules and matrix metalloproteinases in VEC and VIC. In addition, protein kinase C was increased in VEC and VIC, indicating molecular mechanisms associated with HG induced inflammation and remodeling in both valvular cells. These findings may indicate new biomarkers and targets for therapy in diabetes associated with CAVD.
