Human endothelial cell growth and phenotypic expression on three dimensional poly(lactide-co-glycolide) sintered microsphere scaffolds for bone tissue engineering.
Bone tissue engineering offers promising alternatives to repair and restore tissues. Our laboratory has employed poly(lactide-co-glycolide) PLAGA microspheres to develop a three dimensional (3-D) porous bioresorbable scaffold with a biomimetic pore structure. Osseous healing and integration with the surrounding tissue depends in part on new blood vessel formation within the porous structure. Since endothelial cells play a key role in angiogenesis (formation of new blood vessels from pre-existing vasculature), the purpose of this study was to better understand human endothelial cell attachment, viability, growth, and phenotypic expression on sintered PLAGA microsphere scaffold. Scanning electron microscopy (SEM) examination showed cells attaching to the surface of microspheres and bridging the pores between the microspheres. Cell proliferation studies indicated that cell number increased during early stages and reached a plateau between days 10 and 14. Immunofluorescent staining for actin showed that cells were proliferating three dimensionally through the scaffolds while staining for PECAM-1 (platelet endothelial cell adhesion molecule) displayed typical localization at cell-cell contacts. Gene expression analysis showed that endothelial cells grown on PLAGA scaffolds maintained their normal characteristic phenotype. The cell proliferation and phenotypic expression were independent of scaffold pore architecture. These results demonstrate that PLAGA sintered microsphere scaffolds can support the growth and biological functions of human endothelial cells. The insights from this study should aid future studies aimed at enhancing angiogenesis in three dimensional tissue engineered scaffolds.
Jabbarzadeh
Ehsan
E
Department of Orthopaedic Surgery, University of Virginia, 400 Ray C. Hunt Drive, Suite 330, Charlottesville, Virginia 22903, USA.
Jiang
Tao
T
Deng
Meng
M
Nair
Lakshmi S
LS
Khan
Yusuf M
YM
Laurencin
Cato T
CT
D000199:Actins / Q000032:analysis; D001842:Bone and Bones / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D002448:Cell Adhesion; D049109:Cell Proliferation; D002470:Cell Survival; D019040:E-Selectin / Q000235:genetics; D042783:Endothelial Cells / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D011098:Polyglactin 910 / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D014471:Umbilical Veins / Q000166:cytology; D014841:von Willebrand Factor / Q000235:genetics
D000199:Actins; D019040:E-Selectin; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011137:Polystyrenes; D014841:von Willebrand Factor; D018799:Intercellular Adhesion Molecule-1; D011098:Polyglactin 910