Laser-layered microfabrication of spatially patterned functionalized tissue-engineering scaffolds.

Understanding cell behavior inside complex, three-dimensional (3D) microenvironments with controlled spatiotemporal patterning of physical and biochemical factors would provide significant insights into the basic biology of organ development and tissue functions. One of the fundamental limitations in studying such behavior has been the inability to create patterned microenvironments within 3D scaffold structures. Here a simple, layer-by-layer stereolithography (SL) method that can precisely pattern ligands, extracellular-matrix (ECM) components, and growth factors, as well as controlled release particles inside a single scaffold, has been developed. The process also allows fabrication of predesigned internal architectures and porosities. Photocrosslinkable poly(ethylene glycol) dimethacrylate (PEGDMA) was used as the basic structural component of these microfabricated scaffolds. PEG acrylates, covalently modified with the cell adhesive peptide arginine-glycine-aspartic acid (RGD) or the ECM component heparan sulfate, was incorporated within the scaffolds to facilitate cell attachment and to allow spatial sequestration of heparan-binding growth factors. Fluorescently labeled polymer microparticles and basic fibroblast growth factor (FGF-2) were chosen to illustrate the capability of SL to spatiotemporally pattern scaffolds. The results demonstrate that a precise, predesigned distribution of single or multiple factors within a single 3D structure can be created, and specific internal architectures can be fabricated. Functionalization of these scaffolds with RGD is demonstrated, and heparan sulfate allows efficient cell attachment and spatial localization of growth factors. Such patterned scaffolds might provide effective systems to study cell behavior in complex microenvironments and could eventually lead to engineering of complex, hybrid tissue structures through predesigned, multilineage differentiation of a single stem-cell population.

[1]  S. Hollister,et al.  Optimal design and fabrication of scaffolds to mimic tissue properties and satisfy biological constraints. , 2002, Biomaterials.

[2]  Won-Gun Koh,et al.  Poly(ethylene glycol) hydrogel microstructures encapsulating living cells. , 2002, Langmuir : the ACS journal of surfaces and colloids.

[3]  Linda G Griffith,et al.  Emerging Design Principles in Biomaterials and Scaffolds for Tissue Engineering , 2002, Annals of the New York Academy of Sciences.

[4]  E. Sachlos,et al.  Making tissue engineering scaffolds work. Review: the application of solid freeform fabrication technology to the production of tissue engineering scaffolds. , 2003, European cells & materials.

[5]  Milan Mrksich,et al.  Micropatterned Surfaces for Control of Cell Shape, Position, and Function , 1998, Biotechnology progress.

[6]  Christopher G Williams,et al.  In vitro chondrogenesis of bone marrow-derived mesenchymal stem cells in a photopolymerizing hydrogel. , 2003, Tissue engineering.

[7]  P. Gleizes,et al.  Basic fibroblast growth factor (FGF-2) internalization through the heparan sulfate proteoglycans-mediated pathway: an ultrastructural approach. , 1995, European journal of cell biology.

[8]  Adel Alhadlaq,et al.  Adult Stem Cell Driven Genesis of Human-Shaped Articular Condyle , 2004, Annals of Biomedical Engineering.

[9]  S. Bryant,et al.  Hydrogel properties influence ECM production by chondrocytes photoencapsulated in poly(ethylene glycol) hydrogels. , 2002, Journal of biomedical materials research.

[10]  Kristi S Anseth,et al.  In vitro osteogenic differentiation of human mesenchymal stem cells photoencapsulated in PEG hydrogels. , 2004, Journal of biomedical materials research. Part A.

[11]  N. Kikuchi,et al.  A novel method for biomaterial scaffold internal architecture design to match bone elastic properties with desired porosity. , 2004, Journal of biomechanics.

[12]  P. Rudland,et al.  Interaction of Heparan Sulfate from Mammary Cells with Acidic Fibroblast Growth Factor (FGF) and Basic FGF , 1998, The Journal of Biological Chemistry.

[13]  K. Anseth,et al.  A review of photocrosslinked polyanhydrides: in situ forming degradable networks. , 2000, Biomaterials.

[14]  Kristi S Anseth,et al.  Controlled release from crosslinked degradable networks. , 2002, Critical reviews in therapeutic drug carrier systems.

[15]  P H Krebsbach,et al.  Indirect solid free form fabrication of local and global porous, biomimetic and composite 3D polymer-ceramic scaffolds. , 2003, Biomaterials.

[16]  K. Marra,et al.  Composition options for tissue-engineered bone. , 2002, Tissue engineering.

[17]  J. Elisseeff,et al.  Experimental model for cartilage tissue engineering to regenerate the zonal organization of articular cartilage. , 2003, Osteoarthritis and cartilage.

[18]  D. Mooney,et al.  Polymeric system for dual growth factor delivery , 2001, Nature Biotechnology.

[19]  N. Quarto,et al.  Heparan sulfate proteoglycans as transducers of FGF-2 signalling. , 1994, Journal of cell science.

[20]  M. Filla,et al.  Role of heparan sulfate as a tissue-specific regulator of FGF-4 and FGF receptor recognition , 2001, The Journal of cell biology.

[21]  Stephen E. Feinberg,et al.  Image-Based Biomimetic Approach to Reconstruction of the Temporomandibular Joint , 2001, Cells Tissues Organs.

[22]  Alyssa Panitch,et al.  Biologically engineered protein-graft-poly(ethylene glycol) hydrogels: a cell adhesive and plasmin-degradable biosynthetic material for tissue repair. , 2002, Biomacromolecules.

[23]  Christopher S. Chen,et al.  Simple approach to micropattern cells on common culture substrates by tuning substrate wettability. , 2004, Tissue engineering.

[24]  R. Vivès,et al.  Regulation of FGF-1 mitogenic activity by heparan sulfate oligosaccharides is dependent on specific structural features: differential requirements for the modulation of FGF-1 and FGF-2. , 2000, Glycobiology.

[25]  Robert Langer,et al.  Photopolymerizable degradable polyanhydrides with osteocompatibility , 1999, Nature Biotechnology.

[26]  Jussi Taipale,et al.  Growth factors in the extracellular matrix , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[27]  A. Mikos,et al.  Controlled release of an osteogenic peptide from injectable biodegradable polymeric composites. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[28]  D. Moscatelli,et al.  Fibroblast growth factor (FGF)-2 mediates cell attachment through interactions with two FGF receptor-1 isoforms and extracellular matrix or cell-associated heparan sulfate proteoglycans. , 2000, Biochemical and biophysical research communications.

[29]  G. Whitesides,et al.  Cell shape provides global control of focal adhesion assembly. , 2003, Biochemical and biophysical research communications.

[30]  L G Griffith,et al.  Integration of surface modification and 3D fabrication techniques to prepare patterned poly(L-lactide) substrates allowing regionally selective cell adhesion. , 1998, Journal of biomaterials science. Polymer edition.

[31]  Stephen E. Feinberg,et al.  An image-based approach for designing and manufacturing craniofacial scaffolds. , 2000, International journal of oral and maxillofacial surgery.

[32]  S J Bryant,et al.  Cytocompatibility of UV and visible light photoinitiating systems on cultured NIH/3T3 fibroblasts in vitro , 2000, Journal of biomaterials science. Polymer edition.

[33]  A. Metters,et al.  Fundamental studies of biodegradable hydrogels as cartilage replacement materials. , 1999, Biomedical sciences instrumentation.

[34]  Robert Langer,et al.  Controlled‐release of IGF‐I and TGF‐β1 in a photopolymerizing hydrogel for cartilage tissue engineering , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[35]  N. Perrimon,et al.  Heparan sulfate proteoglycans are essential for FGF receptor signaling during Drosophila embryonic development. , 1999, Development.

[36]  Jason A Burdick,et al.  Photoencapsulation of osteoblasts in injectable RGD-modified PEG hydrogels for bone tissue engineering. , 2002, Biomaterials.

[37]  J. Elisseeff,et al.  Photoencapsulation of chondrocytes in poly(ethylene oxide)-based semi-interpenetrating networks. , 2000, Journal of biomedical materials research.