title: Combination scaffolds of salmon fibrin, hyaluronic acid, and laminin for human neural stem cell and vascular tissue engineering.
abstract: Human neural stem/progenitor cells (hNSPCs) are good candidates for treating central nervous system (CNS) trauma since they secrete beneficial trophic factors and differentiate into mature CNS cells; however, many cells die after transplantation. This cell death can be ameliorated by inclusion of a biomaterial scaffold, making identification of optimal scaffolds for hNSPCs a critical research focus. We investigated the properties of fibrin-based scaffolds and their effects on hNSPCs and found that fibrin generated from salmon fibrinogen and thrombin stimulates greater hNSPC proliferation than mammalian fibrin. Fibrin scaffolds degrade over the course of a few days in vivo, so we sought to develop a novel scaffold that would retain the beneficial properties of fibrin but degrade more slowly to provide longer support for hNSPCs. We found combination scaffolds of salmon fibrin with interpenetrating networks (IPNs) of hyaluronic acid (HA) with and without laminin polymerize more effectively than fibrin alone and generate compliant hydrogels matching the physical properties of brain tissue. Furthermore, combination scaffolds support hNSPC proliferation and differentiation while significantly attenuating the cell-mediated degradation seen with fibrin alone. HNSPCs express two fibrinogen-binding integrins, αVβ1 and α5β1, and several laminin binding integrins (α7β1, α6β1, α3β1) that can mediate interaction with the scaffold. Lastly, to test the ability of scaffolds to support vascularization, we analyzed human cord blood-derived endothelial cells alone and in co-culture with hNSPCs and found enhanced vessel formation and complexity in co-cultures within combination scaffolds. Overall, combination scaffolds of fibrin, HA, and laminin are excellent biomaterials for hNSPCs.
  
  
  
  Interest has increased recently in the development of biomaterials as neural stem cell transplantation scaffolds to treat central nervous system (CNS) injury since scaffolds improve survival and integration of transplanted cells. We report here on a novel combination scaffold composed of fibrin, hyaluronic acid, and laminin to support human neural stem/progenitor cell (hNSPC) function. This combined biomaterial scaffold has appropriate physical properties for hNSPCs and the CNS, supports hNSPC proliferation and differentiation, and attenuates rapid cell-mediated scaffold degradation. The hNSPCs and scaffold components synergistically encourage new vessel formation from human endothelial cells. This work marks the first report of a combination scaffold supporting human neural and vascular cells to encourage vasculogenesis, and sets a benchmark for biomaterials to treat CNS injury.
authors.affiliation: Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA; Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Janahan
authors.initials: J
authors.lastname: Arulmoli
authors.affiliation: Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA; Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Heather J
authors.initials: HJ
authors.lastname: Wright
authors.affiliation: Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Duc T T
authors.initials: DTT
authors.lastname: Phan
authors.affiliation: Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Urmi
authors.initials: U
authors.lastname: Sheth
authors.affiliation: Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Que
authors.affiliation: Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA.
authors.forename: Giovanni A
authors.initials: GA
authors.lastname: Botten
authors.affiliation: Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Keating
authors.affiliation: Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA; The Edwards Lifesciences Center for Advanced Cardiovascular Technology, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Elliot L
authors.initials: EL
authors.lastname: Botvinick
authors.affiliation: Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA; Department of Physiology & Biophysics, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Medha M
authors.initials: MM
authors.lastname: Pathak
authors.affiliation: BioTime, Inc., 1301 Harbor Parkway, Alameda, CA 94502, USA.
authors.forename: Thomas I
authors.initials: TI
authors.lastname: Zarembinski
authors.affiliation: Disc Comfort, Inc., 351 Hospital Road, Suite 202, Newport Beach, CA 92663, USA.
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Yanni
authors.affiliation: Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA; Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Olga V
authors.initials: OV
authors.lastname: Razorenova
authors.affiliation: Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA; Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA 92697, USA; The Edwards Lifesciences Center for Advanced Cardiovascular Technology, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Christopher C W
authors.initials: CCW
authors.lastname: Hughes
authors.affiliation: Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA; Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA; Department of Neurology, University of California, Irvine, Irvine, CA 92697, USA. Electronic address: lisa.flanagan@uci.edu.
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Flanagan
mesh_terms: D000818:Animals; D001808:Blood Vessels / Q000187:drug effects / Q000502:physiology*; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D005337:Fibrin / Q000494:pharmacology*; D006801:Humans; D006820:Hyaluronic Acid / Q000494:pharmacology*; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology; D016023:Integrins / Q000378:metabolism; D007797:Laminin / Q000494:pharmacology*; D018919:Neovascularization, Physiologic / Q000187:drug effects; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects; D058105:Polymerization / Q000187:drug effects; D012474:Salmon; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D016023:Integrins; D007797:Laminin; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D005337:Fibrin; D006820:Hyaluronic Acid
keywords: Biomaterial scaffold; Co-culture; Hyaluronic acid; Hydrogel; Integrin; Laminin; Matrix mechanics; Neural stem cell; Neural tissue engineering; Neurovascular niche; Salmon fibrin