Surface-Functionalized Silk Fibroin Films as a Platform To Guide Neuron-like Differentiation of Human Mesenchymal Stem Cells

Abstract

Surface interactions at the biomaterial-cellular interface determine the proliferation and differentiation of stem cells. Manipulating such interactions through the surface chemistry of scaffolds renders control over directed stem cell differentiation into the cell lineage of interest. This approach is of central importance for stem cell-based tissue engineering and regenerative therapy applications. In the present study, silk fibroin films (SFFs) decorated with integrin-binding laminin peptide motifs (YIGSR and GYIGSR) were prepared and employed for in vitro adult stem cell-based neural tissue engineering applications. Functionalization of SFFs with short peptides showcased the peptide sequence and nature of functionalization-dependent differentiation of bone marrow-derived human mesenchymal stem cells (hMSCs). Intriguingly, covalently functionalized SFFs with GYIGSR hexapeptide (CL2-SFF) supported hMSC proliferation and maintenance in an undifferentiated pluripotent state and directed the differentiation of hMSCs into neuron-like cells in the presence of a biochemical cue, on-demand. The observed morphological changes were further corroborated by the up-regulation of neuronal-specific marker gene expression (MAP2, TUBB3, NEFL), confirmed through semiquantitative reverse-transcription polymerase chain reaction (RT-PCR) analysis. The enhanced proliferation and on-demand directed differentiation of adult stem cells (hMSCs) by the use of an economically viable short recognition peptide (GYIGSR), as opposed to the integrin recognition protein laminin, establishes the potential of SFFs for neural tissue engineering and regenerative therapy applications.

Controlling Cell Behavior through the Design of Biomaterial Surfaces: A Focus on Surface Modification Techniques

“…The authors mentioned that NHS/EDC linker contributed to covalent attachment of COOH groups from sulfated glycosaminoglycan (GAG) with NH 2 groups of collagen in sulfated GAG–collagen matrices and provided considerable resistance of tissue scaffolds to enzymatic degradation . In another study, Manchineella et al showed that covalent modifications of silk fibroin (SF) films led to the increased proliferation of human multipotent mesenchymal stromal cells (hMSCs) relative to physical modification of SF films . These findings suggest that covalent modification would be appropriate approach for the long‐term culture experiments .…”

Differentiation of Bone Marrow Stem Cells into Schwann Cells for the Promotion of Neurite Outgrowth on Electrospun Fibers

“…26,39–41 The presence of proteins on their surface can also modulate the interactions between the fibers and the cells, thereby affecting the differentiation of stem cells. 42,43 …”

Electroactive 3D Scaffolds Based on Silk Fibroin and Water‐Borne Polyaniline for Skeletal Muscle Tissue Engineering

“…Hence, designing 3D tissue engineering scaffolds with excellent biocompatibility and controllable biodegradability which can mimic the extracellular matrix and guide 3D tissue regeneration is highly required. Owing to the remarkable biocompatibility, silk fibroin (SF) has been widely used in biomaterials with a variety of forms, such as electrospun nanofibers, films, sponges, hydrogels, and scaffolds . Researchers recently used salt‐leaching method or organic solvent to prepare the water‐stable SF scaffolds for tissue engineering.…”