Genipin‐cross‐linked type II collagen scaffold promotes the differentiation of adipose‐derived stem cells into nucleus pulposus‐like cells

Abstract

Adipose-derived stem cells (ADSCs)-based tissue engineering was a promising method to treat intervertebral disc degeneration. Type II collagen is a native component in the nucleus pulposus (NP), and has the ability to promote ADSCs to differentiate into NP-like cells. In this article, we aimed to establish a genipin-cross-linked three-dimensional (3D) type II collagen scaffold, and determine the biological effects of the scaffold on ADSCs differentiating into a NP-like phenotype. Different concentrations of genipin were used to cross-link the 3D type II collagen scaffold. Microstructure, surface topography, mechanical strength, porosity, swelling property, and biological stability of the scaffolds were detected to evaluate the scaffold properties. Cell proliferation, gene and protein expression were measured to access the biological effects of the scaffolds on ADSCs, and the related molecular mechanism was investigated. Cross-linking by genipin increased the stability of the type II collagen scaffolds, but deformed the configuration of scaffolds and changed the intrinsic properties of type II collagen. scaffold cross-linked with 0.1% genipin improved the biostability on the basis of maintaining the configuration of scaffold. In addition, the 0.1% genipin-cross-linked scaffold promoted ADSCs proliferation and differentiation into NP-like cells, along with the increasing gene and protein expressions of Sonic Hedgehog (Shh). All these results suggested that 0.1% genipin was the optimal concentration to establish a bio-stable 3D type II collagen scaffold, which inducing ADSC proliferation and differentiation toward a NP-like phenotype through the activation of Shh signaling pathway. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1258-1268, 2018.

Biomaterials-Induced Stem Cells Specific Differentiation Into Intervertebral Disc Lineage Cells

“…Similarly, Shh protein expression level has also been shown an increase when crosslinked scaffolds promoted ADSC proliferation and differentiation into NP-like cells, indicating that the activation of the Shh signaling pathway might play a regulatory role in directional differentiation (Zhou et al, 2018b). In addition, NP-like cell differentiation induced by Collagen type II is related to FoxA2 overexpression and can be rescued by a Shh signaling pathway inhibitor (Zhou et al, 2018a).…”

“…Thus, to mimic the natural structure of IVD tissues, collagen I/II are widely used materials for IVD regeneration. However, biomaterials consisting of collagen alone are less efficient in promoting stem cell differentiation toward IVD cells, and supplementation with crosslinking agents of appropriate concentration is more likely to increase adipose-derived stem cells (ADSCs) gene and protein expression of Collagen type II, Sox-9 and aggrecan, suggesting a NP-like phenotype (Zhou et al, 2018b). Gelatine, which is a denatured form of collagen, has been used for NP-like differentiation of ADSCs and IVD repair.…”

“…Collagen type II alone has been shown to induce ADSC differentiation into NP-like cells (Zhou et al, 2018b), which was related to FoxA2 overexpression (Zhou et al, 2018a). Moreover, Collagen type II combined with other natural components and crosslinkers, such as chondroitin sulfate (Zhou et al, 2018c), hyaluronan (Calderon et al, 2010), genipin (Zhou et al, 2018b), N,N-(3-dimethylaminopropyl)-N -ethylcarbodiimide and N-hydroxysuccinimide (EDAC/NHS) , to modify its porosity and other micromechanical properties has also been shown to exert an improved effect on stem cell proliferation and directional differentiation.…”

Tissue Engineering Strategies for Intervertebral Disc Treatment Using Functional Polymers

“…In many cases, gelatin and collagen were used as an additional composite with other materials to improve biomechanical and metabolic properties [85,86,87]. Human AF cells have good adhesion and increased anabolic activity in fibronectin, collagen, and gelatin substrate.…”

FoxA2 regulates the type II collagen–induced nucleus pulposus–like differentiation of adipose‐derived stem cells by activation of the Shh signaling pathway

“…This evidence indicates that FoxA2 is important in generating NP cells. We have demonstrated that type II collagen increases the expression of FoxA2 in ADSCs during NP-like differentiation (22). An investigation of the function of FoxA2 and the relationship between FoxA2 and Shh signaling is needed to clarify the mechanism of type II collagen-induced NP-like differentiation of ADSCs.…”