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.

3D brain angiogenesis model to reconstitute functional human blood–brain barrier in vitro

“…A similar result was obtained when fibrin and HA were mixed at a ratio of 2.5:1.5 (final concentrations of fibrin and HA were 2.5 and 1.5 mg/ml, respectively), and most astrocytes had protrusions. This result agreed well with previous reports (Arulmoli et al, ; Levy et al, ; Placone et al, ). Vascular morphology, such as vessel width and lumen structure, was not significantly affected up to the addition of 1.5 mg/ml HA, whereas the effect of HA on astrocyte morphology was saturated at this ratio.…”

“…Brain tissue is composed of complex ECM components, including HA, proteoglycans, and tenascins (Zimmermann & Dours‐Zimmermann, ). Among cells in the human BBB microenvironment, astrocyte phenotype and morphology are known to be influenced greatly by the local microenvironment (Arulmoli et al, ; Levy et al, ). This was clearly demonstrated by Placone et al (), who manipulated the type and ratio of the ECM including HA and then analyzed astrocyte morphology and active states under each ECM condition.…”

Platelet-functionalized three-dimensional poly-ε-caprolactone fibrous scaffold prepared using centrifugal spinning for delivery of growth factors

“…[42][43][44] Bardsley et al 43 showed in his study high initial in vitro degradation of fibrin gel between 0 and 5 days. It was shown that the degradation rate can be slowed down by combining with hyaluronic acid and laminin 42 or polyethylenglycol. 45 While fibrin-only gel totally degrades after 14 days of cultivation, PEGylated fibrin stayed intact.…”

Proteomic Profiling of Neuroblastoma Cells Adhesion on Hyaluronic Acid-Based Surface for Neural Tissue Engineering

“…Previous study indicated that combination scaffolds consisting of fibrin with HA and laminin provide biomaterial properties to enable polymerization with cells. This mimics the native tissue of the brain and supports differentiation of human neural stem/progenitor cell (hNSPC) function [27]. Currently, HA-based biomaterials are used to regulate the cell differentiation and studied for tissue engineering purposes in combination with growth factors or ECM components for tissue repair [28–33].…”