title: Calcification of primary human osteoblast cultures under flow conditions using polycaprolactone scaffolds for intravascular applications.
abstract: Total atherosclerotic occlusion is a leading cause of death. Recent animal models of this disease are devoid of cell-mediated calcification and arteries are often not occluded gradually. This study is part of a project with the objective of developing a new model featuring the above two characteristics, using a tissue-engineering scaffold. The amount and distribution of calcium deposits in primary human osteoblast (HOB) cultures on polycaprolactone (PCL) scaffolds under flow conditions were investigated. HOBs were cultured on PCL scaffolds with TGF-β1 loadings of 0 (control), 5 and 50 ng. HOB-PCL constructs were cultured in spinner flasks. Under flow conditions, cell numbers present in HOB cultures on PCL scaffolds increased from day 7 to day 14, and most calcification was induced at day 21. TGF-β1 loadings of 5 and 50 ng did not show a significant difference in ALP activity, cell numbers and amount of calcium deposited in HOB cultures, but calcium staining showed that 50 ng TGF-β1 had higher calcium deposited on both days 21 and 28 under flow conditions compared with 5 ng of loading. Amount of calcium deposited by HOBs on day 28 showed a decrease from their levels on day 21. PCL degradation may be a factor contributing to this loss. The results indicate that cell-induced calcification can be achieved on PCL scaffolds under flow conditions. In conclusion, TGFβ1-HOB loaded PCL can be applied to create a model for total atherosclerotic occlusion with cell-deposited calcium in animal arteries.
authors.affiliation: Department of Biomedical Engineering, College of Engineering, University of Texas at San Antonio, TX, USA.
authors.forename: Beili
authors.initials: B
authors.lastname: Zhu
authors.affiliation: Department of Biomedical Engineering, College of Engineering, University of Texas at San Antonio, TX, USA.|Janey Briscoe Center for Cardiovascular Research, Janey and Dolph Briscoe Division of Cardiology, Department of Medicine, University of Texas Health Science Center at San Antonio, TX, USA.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Bailey
authors.affiliation: Department of Biomedical Engineering, College of Engineering, University of Texas at San Antonio, TX, USA.
authors.forename: C Mauli
authors.initials: CM
authors.lastname: Agrawal
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D050197:Atherosclerosis / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D019149:Bioreactors; D002114:Calcinosis; D002118:Calcium / Q000378:metabolism; D002452:Cell Count; D002454:Cell Differentiation; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D006801:Humans; D008422:Materials Testing; D008955:Models, Cardiovascular; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D011091:Polyesters / Q000737:chemistry*; D012212:Rheology; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry*; D053773:Transforming Growth Factor beta1 / Q000008:administration & dosage / Q000737:chemistry
chemical_list: D020099:Coated Materials, Biocompatible; D011091:Polyesters; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; C016240:polycaprolactone; D000469:Alkaline Phosphatase; D002118:Calcium