Biofunctionalized microfiber-assisted formation of intrinsic three-dimensional capillary-like structures.

OBJECTIVES A vascular supply network is essential in engineered tissues >100-200-μm 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 biofunctionalized PLLA microfibers. Further methods are warranted to perform specified spatial positioning of fibers within 3D formative scaffolds to enhance the applicability of the concept.

[1]  Thomas Schmitz-Rode,et al.  Fibrin-based tissue engineering: comparison of different methods of autologous fibrinogen isolation. , 2013, Tissue engineering. Part C, Methods.

[2]  D. Mantovani,et al.  The biological response of poly(L-lactide) films modified by different biomolecules: role of the coating strategy. , 2012, Journal of biomedical materials research. Part A.

[3]  Y. Dor,et al.  Engineered Vascular Beds Provide Key Signals to Pancreatic Hormone-Producing Cells , 2012, PloS one.

[4]  Brendon M. Baker,et al.  Rapid casting of patterned vascular networks for perfusable engineered 3D tissues , 2012, Nature materials.

[5]  Ali Khademhosseini,et al.  Microfabrication of complex porous tissue engineering scaffolds using 3D projection stereolithography. , 2012, Biomaterials.

[6]  Z. Kam,et al.  Engineering vessel-like networks within multicellular fibrin-based constructs. , 2011, Biomaterials.

[7]  M. N. Nakatsu,et al.  The requirement for fibroblasts in angiogenesis: fibroblast-derived matrix proteins are essential for endothelial cell lumen formation , 2011, Molecular biology of the cell.

[8]  Didier Y. R. Stainier,et al.  Molecular control of endothelial cell behaviour during blood vessel morphogenesis , 2011, Nature Reviews Molecular Cell Biology.

[9]  C James Kirkpatrick,et al.  Co-culture systems for vascularization--learning from nature. , 2011, Advanced drug delivery reviews.

[10]  C. Doillon,et al.  Directional migration of endothelial cells towards angiogenesis using polymer fibres in a 3D co‐culture system , 2010, Journal of tissue engineering and regenerative medicine.

[11]  P. Vermette,et al.  The effects of co-culture with fibroblasts and angiogenic growth factors on microvascular maturation and multi-cellular lumen formation in HUVEC-oriented polymer fibre constructs. , 2010, Biomaterials.

[12]  D. Slaaf,et al.  Evaluation of magnetic resonance vessel size imaging by two‐photon laser scanning microscopy , 2010, Magnetic resonance in medicine.

[13]  Srivatsan Raghavan,et al.  Geometrically controlled endothelial tubulogenesis in micropatterned gels. , 2010, Tissue engineering. Part A.

[14]  Tadashi Sasagawa,et al.  Design of prevascularized three-dimensional cell-dense tissues using a cell sheet stacking manipulation technology. , 2010, Biomaterials.

[15]  Steven C George,et al.  Rapid anastomosis of endothelial progenitor cell-derived vessels with host vasculature is promoted by a high density of cotransplanted fibroblasts. , 2010, Tissue engineering. Part A.

[16]  Tal Dvir,et al.  Prevascularization of cardiac patch on the omentum improves its therapeutic outcome , 2009, Proceedings of the National Academy of Sciences.

[17]  A. J. Putnam,et al.  Endothelial cell traction and ECM density influence both capillary morphogenesis and maintenance in 3-D. , 2009, American journal of physiology. Cell physiology.

[18]  Steven C George,et al.  Prevascularization of a fibrin-based tissue construct accelerates the formation of functional anastomosis with host vasculature. , 2009, Tissue engineering. Part A.

[19]  P. Vermette,et al.  Polymer fibers as contact guidance to orient microvascularization in a 3D environment. , 2009, Journal of biomedical materials research. Part A.

[20]  S. Levenberg,et al.  Vascularization--the conduit to viable engineered tissues. , 2009, Tissue engineering. Part B, Reviews.

[21]  Guo-qiang Chen,et al.  Synthesis, Characterization and Biocompatibility of Biodegradable Elastomeric Poly(ether-ester urethane)s Based on Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) and Poly(ethylene glycol) via Melting Polymerization , 2009, Journal of biomaterials science. Polymer edition.

[22]  Taiji Sohmura,et al.  Three-Dimensional Cell and Tissue Patterning in a Strained Fibrin Gel System , 2007, PloS one.

[23]  L. Claes,et al.  Human mesenchymal progenitor cell responses to a novel textured poly(L-lactide) scaffold for ligament tissue engineering. , 2007, Journal of biomedical materials research. Part B, Applied biomaterials.

[24]  Jeroen Rouwkema,et al.  Endothelial cells assemble into a 3-dimensional prevascular network in a bone tissue engineering construct. , 2006, Tissue engineering.

[25]  Fen Chen,et al.  Biomimetic approach to cardiac tissue engineering: oxygen carriers and channeled scaffolds. , 2006, Tissue engineering.

[26]  Denys N Wheatley,et al.  Potential of fibroblasts to regulate the formation of three-dimensional vessel-like structures from endothelial cells in vitro. , 2006, American journal of physiology. Cell physiology.

[27]  Lucie Germain,et al.  Extracellular matrix deposition by fibroblasts is necessary to promote capillary‐like tube formation in vitro , 2006, Journal of cellular physiology.

[28]  Thorsten Walles,et al.  Engineering of a vascularized scaffold for artificial tissue and organ generation. , 2005, Biomaterials.

[29]  Dai Fukumura,et al.  Engineering vascularized tissue , 2005, Nature Biotechnology.

[30]  D. Kohane,et al.  Engineering vascularized skeletal muscle tissue , 2005, Nature Biotechnology.

[31]  Lucie Germain,et al.  Inosculation of Tissue‐Engineered Capillaries with the Host's Vasculature in a Reconstructed Skin Transplanted on Mice , 2005, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[32]  D. Klee,et al.  Oberflächenmodifizierung von Titan zur Verbesserung der Grenzflächenverträglichkeit , 2004 .

[33]  R. Sainson,et al.  Angiogenic sprouting and capillary lumen formation modeled by human umbilical vein endothelial cells (HUVEC) in fibrin gels: the role of fibroblasts and Angiopoietin-1. , 2003, Microvascular research.

[34]  B J Messmer,et al.  Tissue engineering: complete autologous valve conduit--a new moulding technique. , 2001, The Thoracic and cardiovascular surgeon.

[35]  J. Lahann,et al.  Bioactive immobilization of r-hirudin on CVD-coated metallic implant devices. , 2001, Biomaterials.

[36]  K. Shakesheff,et al.  Poly(L-lysine)-GRGDS as a biomimetic surface modifier for poly(lactic acid). , 2001, Biomaterials.

[37]  J. Hubbell,et al.  Incorporation of adhesion peptides into nonadhesive hydrogels useful for tissue resurfacing. , 1998, Journal of biomedical materials research.

[38]  E. Kastenbauer,et al.  Resorbable polyesters in cartilage engineering: affinity and biocompatibility of polymer fiber structures to chondrocytes. , 1996, Journal of biomedical materials research.

[39]  J. Hubbell,et al.  Polymer networks with grafted cell adhesion peptides for highly biospecific cell adhesive substrates. , 1994, Analytical biochemistry.

[40]  A. Logan,et al.  Angiogenesis , 1993, The Lancet.

[41]  A. M. Reed,et al.  Biodegradable polymers for use in surgery—poly(ethylene oxide) poly(ethylene terephthalate) (PEO/PET) copolymers: 1 , 1979 .

[42]  C A van Blitterswijk,et al.  Cell-seeding and in vitro biocompatibility evaluation of polymeric matrices of PEO/PBT copolymers and PLLA. , 1993, Biomaterials.

[43]  D. Klee,et al.  Surface modification of a biocompatible polymer based on polyurethane for artificial blood vessels , 1992 .