Journal
TISSUE ENGINEERING PART A
Volume 20, Issue 13-14, Pages 1858-1869Publisher
MARY ANN LIEBERT, INC
DOI: 10.1089/ten.tea.2013.0330
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Funding
- Deutsche Forschungsgemeinschaft (DFG) graduate school BioInterface 1035
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Objectives: A vascular supply network is essential in engineered tissues >100-200-mu m thickness. To control vascular network formation in vitro, we hypothesize that capillarization can be achieved locally by using fibers to position and guide vessel-forming endothelial cells within a three-dimensional (3D) matrix. Materials and Methods: Biofunctionalization of poly-(L-lactic acid) (PLLA) fibers was performed by amino-functionalization and covalent binding of RGD peptides. Human foreskin fibroblasts (HFFs) and human umbilical vein endothelial cells (HUVECs) were seeded on the fibers in a mould and subsequently embedded in fibrin gel. After 9-21 days of coculture, constructs were fixed and immunostained (PECAM-1). Capillary-like structures with lumen in the 3D fibrin matrix were verified and quantified using two-photon microscopy and image analysis software. Results: Capillary-like networks with lumen formed adjacent to the PLLA fibers. Increased cell numbers were observed to attach to RGD-functionalized fibers, resulting in enhanced formation of capillary-like structures. Cocultivation of HFFs sufficiently supported HUVECs in the formation of capillary-like structures, which persisted for at least 21 days of coculture. Conclusions: The guidance of vessel growth within tissue-engineered constructs can be achieved using bio-functionalized PLLA microfibers. Further methods are warranted to perform specified spatial positioning of fibers within 3D formative scaffolds to enhance the applicability of the concept.
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