title: Engineered N-cadherin and L1 biomimetic substrates concertedly promote neuronal differentiation, neurite extension and neuroprotection of human neural stem cells.
abstract: We investigated the design of neurotrophic biomaterial constructs for human neural stem cells, guided by neural developmental cues of N-cadherin and L1 adhesion molecules. Polymer substrates fabricated either as two-dimensional (2-D) films or three-dimensional (3-D) microfibrous scaffolds were functionalized with fusion chimeras of N-cadherin-Fc alone and in combination with L1-Fc, and the effects on differentiation, neurite extension and survival of H9 human-embryonic-stem-cell-derived neural stem cells (H9-NSCs) were quantified. Combinations of N-cadherin and L1-Fc co-operatively enhanced neuronal differentiation profiles, indicating the critical nature of the two complementary developmental cues. Notably, substrates presenting low levels of N-cadherin-Fc concentrations, combined with proportionately higher L1-Fc concentration, most enhanced neurite outgrowth and the degree of MAP2+ and neurofilament-M+ H9-NSCs. Low N-cadherin-Fc alone promoted improved cell survival following oxidative stress, compared to higher concentrations of N-cadherin-Fc alone or combinations with L1-Fc. Pharmacological and antibody blockage studies revealed that substrates presenting low levels of N-cadherin are functionally competent so long as they elicit a threshold signal mediated by homophilic N-cadherin and fibroblast growth factor signaling. Overall, these studies highlight the ability of optimal combinations of N-cadherin and L1 to recapitulate a "neurotrophic" microenvironment that enhances human neural stem cell differentiation and neurite outgrowth. Additionally, 3-D fibrous scaffolds presenting low N-cadherin-Fc further enhanced the survival of H9-NSCs compared to equivalent 2-D films. This indicates that similar biofunctionalization approaches based on N-cadherin and L1 can be translated to 3-D "transplantable" scaffolds with enhanced neurotrophic behaviors. Thus, the insights from this study have fundamental and translational impacts for neural-stem-cell-based regenerative medicine.
authors.affiliation: Department of Biomedical Engineering, Rutgers University, 599 Taylor Road, Piscataway, NJ 08854, USA.
authors.forename: Jocie F
authors.initials: JF
authors.lastname: Cherry
authors.affiliation: Department of Biomedical Engineering, Rutgers University, 599 Taylor Road, Piscataway, NJ 08854, USA.
authors.forename: Neal K
authors.initials: NK
authors.lastname: Bennett
authors.affiliation: W.M. Keck Center for Collaborative Neuroscience, Department of Cell Biology and Neuroscience, Rutgers University, 599 Taylor Road, Piscataway, NJ 08854, USA; Center for Neuroscience, Shantou University Medical College, 22 Xin Ling Road, Shantou 515041, People's Republic of China.
authors.forename: Melitta
authors.initials: M
authors.lastname: Schachner
authors.affiliation: Department of Biomedical Engineering, Rutgers University, 599 Taylor Road, Piscataway, NJ 08854, USA; Department of Chemical and Biochemical Engineering, Rutgers University, 599 Taylor Road, Piscataway, NJ 08854, USA. Electronic address: moghe@rutgers.edu.
authors.forename: Prabhas V
authors.initials: PV
authors.lastname: Moghe
mesh_terms: D015703:Antigens, CD / Q000494:pharmacology*; D040761:Biomimetic Materials / Q000494:pharmacology*; D015820:Cadherins / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000494:pharmacology*; D039842:Neural Cell Adhesion Molecule L1 / Q000494:pharmacology*; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism*; D016501:Neurites / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D044968:Regenerative Medicine; D055153:Stem Cell Niche; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D015703:Antigens, CD; C050361:CDH2 protein, human; D015820:Cadherins; D007141:Immunoglobulin Fc Fragments; D039842:Neural Cell Adhesion Molecule L1; D011993:Recombinant Fusion Proteins
keywords: Biomimetic surface; Electrospun scaffolds; Human embryonic stem cells; L1; N-cadherin; Neural stem cells