Cellular behavior in micropatterned hydrogels by bioprinting system depended on the cell types and cellular interaction.

The fabrication of patterned microstructures within three-dimensional (3D) matrices is a challenging subject in tissue engineering and regenerative medicine. A 3D, free-moving bioprinting system was developed and hydrogels were patterned by varying the process parameters of z-axis moving velocity and ejection velocity. The patterning of hydrogel based microfibers in a 3D matrigel was achieved with dimensions of 4.5 mm length and widths from 79 to 200 μm. Hyaluronan-based hydrogels mixed with fibroblasts (L929), mouse endothelial cells (MS1), or human mesenchymal stem cells (hMSCs) were patterned using a 3D moving axis bioprinter and cell behavior was monitored in culture for up to 16 days. L929 and MS1 cells and hMSCs in patterned hydrogel revealed cell-cell interactions and a morphological dependency on cell types. HMSCs formed spheres through cell aggregation, while L929 cells increased in cellular mass without cell aggregation and MS1 dispersed into the matrix instead of aggregating. The aggregation of hMSCs was attenuated by treatment with Rho kinase (ROCK) inhibitor and cadherin antibody. This reflected the close relationship between cell aggregation and migration with RhoA and cell-cell adhesion molecules. Angiogenic-specific gene expression profiles showed that expression of CD105 decreased to 22% in the ROCK inhibitor group compared to control group. These results showed that cell-based patterns in a 3D matrix are highly dependent on both cell aggregation and migration over time.

[1]  Yumiko Saga,et al.  Cell‐Cell Interaction Mediated by Cadherin‐11 Directly Regulates the Differentiation of Mesenchymal Cells Into the Cells of the Osteo‐Lineage and the Chondro‐Lineage , 2004, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[2]  Anthony P. Napolitano,et al.  Dynamics of the self-assembly of complex cellular aggregates on micromolded nonadhesive hydrogels. , 2007, Tissue engineering.

[3]  J. Wikswo,et al.  Microfabricated scaffold-guided endothelial morphogenesis in three-dimensional culture , 2011, Biomedical microdevices.

[4]  Vladimir Mironov,et al.  Bioprinting living structures , 2007 .

[5]  Glenn D Prestwich,et al.  Bioprinting vessel-like constructs using hyaluronan hydrogels crosslinked with tetrahedral polyethylene glycol tetracrylates. , 2010, Biomaterials.

[6]  P. Friedl,et al.  Migration of coordinated cell clusters in mesenchymal and epithelial cancer explants in vitro. , 1995, Cancer research.

[7]  W. Weis,et al.  Structure and mechanism of cadherins and catenins in cell-cell contacts. , 2007, Annual review of cell and developmental biology.

[8]  Jörg Fiedler,et al.  BMP‐2, BMP‐4, and PDGF‐bb stimulate chemotactic migration of primary human mesenchymal progenitor cells , 2002, Journal of cellular biochemistry.

[9]  T. Couffinhal,et al.  Secreted Frizzled‐Related Protein‐1 Enhances Mesenchymal Stem Cell Function in Angiogenesis and Contributes to Neovessel Maturation , 2008, Stem cells.

[10]  Jason A Burdick,et al.  Patterning network structure to spatially control cellular remodeling and stem cell fate within 3-dimensional hydrogels. , 2010, Biomaterials.

[11]  Brian Derby,et al.  Bioprinting: Inkjet printing proteins and hybrid cell-containing materials and structures , 2008 .

[12]  V. Barron,et al.  Behavior of Human Mesenchymal Stem Cells in Fibrin-Based Vascular Tissue Engineering Constructs , 2010, Annals of Biomedical Engineering.

[13]  Chrysanthi Williams,et al.  Small-diameter artificial arteries engineered in vitro. , 2005, Circulation research.

[14]  Kenneth M. Yamada,et al.  Cell-matrix adhesions in 3D. , 2011, Matrix biology : journal of the International Society for Matrix Biology.

[15]  F. Guillemot,et al.  Laser assisted bioprinting of engineered tissue with high cell density and microscale organization. , 2010, Biomaterials.

[16]  Stuart K Williams,et al.  Direct-write bioprinting three-dimensional biohybrid systems for future regenerative therapies. , 2011, Journal of biomedical materials research. Part B, Applied biomaterials.

[17]  P. Friedl.  Prespecification and plasticity: shifting mechanisms of cell migration. , 2004, Current opinion in cell biology.

[18]  Sami Alom Ruiz,et al.  Mechanical tugging force regulates the size of cell–cell junctions , 2010, Proceedings of the National Academy of Sciences.

[19]  J. Liao,et al.  Rho Kinase (ROCK) Inhibitors , 2007, Journal of cardiovascular pharmacology.

[20]  P. Friedl,et al.  Collective cell migration in morphogenesis, regeneration and cancer , 2009, Nature Reviews Molecular Cell Biology.

[21]  G. Forgacs,et al.  Surface tensions of embryonic tissues predict their mutual envelopment behavior. , 1996, Development.

[22]  M. S. Steinberg,et al.  The differential adhesion hypothesis: a direct evaluation. , 2005, Developmental biology.

[23]  Ding-Yuan Chen,et al.  Core-shell cell bodies composed of human cbMSCs and HUVECs for functional vasculogenesis. , 2011, Biomaterials.

[24]  Karl R Edminster,et al.  Multi-layered culture of human skin fibroblasts and keratinocytes through three-dimensional freeform fabrication. , 2009, Biomaterials.

[25]  L. Romer,et al.  Endothelial cell adhesion, signaling, and morphogenesis in fibroblast-derived matrix. , 2009, Matrix biology : journal of the International Society for Matrix Biology.

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

[27]  Frederick Grinnell,et al.  Cell motility and mechanics in three-dimensional collagen matrices. , 2010, Annual review of cell and developmental biology.

[28]  Clare M Waterman,et al.  Mechanical integration of actin and adhesion dynamics in cell migration. , 2010, Annual review of cell and developmental biology.

[29]  Lei Tian,et al.  Biomaterials to Prevascularize Engineered Tissues , 2011, Journal of cardiovascular translational research.

[30]  Gordana Vunjak-Novakovic,et al.  Micropatterned three‐dimensional hydrogel system to study human endothelial–mesenchymal stem cell interactions , 2010, Journal of tissue engineering and regenerative medicine.

[31]  W. Sun,et al.  Bioprinting cell-laden matrigel for radioprotection study of liver by pro-drug conversion in a dual-tissue microfluidic chip , 2011, Biofabrication.

[32]  Ali Khademhosseini,et al.  SAM-based cell transfer to photopatterned hydrogels for microengineering vascular-like structures. , 2011, Biomaterials.

[33]  Vladimir Mironov,et al.  Towards organ printing: engineering an intra-organ branched vascular tree , 2010, Expert opinion on biological therapy.

[34]  Vivian H. Fan,et al.  Epidermal Growth Factor as a Candidate for Ex Vivo Expansion of Bone Marrow–Derived Mesenchymal Stem Cells , 2006, Stem cells.

[35]  M. Ratajczak,et al.  Migration of Bone Marrow and Cord Blood Mesenchymal Stem Cells In Vitro Is Regulated by Stromal‐Derived Factor‐1‐CXCR4 and Hepatocyte Growth Factor‐c‐met Axes and Involves Matrix Metalloproteinases , 2006, Stem cells.

[36]  Lisa Mohamet,et al.  Abrogation of E-Cadherin-Mediated Cellular Aggregation Allows Proliferation of Pluripotent Mouse Embryonic Stem Cells in Shake Flask Bioreactors , 2010, PloS one.

[37]  Sang Hoon Lee,et al.  Bone regeneration using hyaluronic acid-based hydrogel with bone morphogenic protein-2 and human mesenchymal stem cells. , 2007, Biomaterials.

[38]  E. Seifried,et al.  Rho Inhibition Induces Migration of Mesenchymal Stromal Cells , 2007, Stem cells.

[39]  Jung-Joo Lee,et al.  A three-dimensional bioprinting system for use with a hydrogel-based biomaterial and printing parameter characterization. , 2010, Artificial organs.

[40]  Matthias P Lutolf,et al.  The effect of matrix characteristics on fibroblast proliferation in 3D gels. , 2010, Biomaterials.