Chondrogenic Differentiation of Adipose-Derived Stromal Cells Induced by Decellularized Cartilage Matrix/Silk Fibroin Secondary Crosslinking Hydrogel Scaffolds with a Three-Dimensional Microstructure

Abstract

Finding an ideal scaffold is always an important issue in the field of cartilage tissue engineering. Both decellularized extracellular matrix and silk fibroin have been used as natural biomaterials for tissue regeneration. In this study, a secondary crosslinking method of γ irradiation and ethanol induction was used to prepare decellularized cartilage extracellular matrix and silk fibroin (dECM-SF) hydrogels with biological activity. Furthermore, the dECM-SF hydrogels were cast in custom-designed molds to produce a three-dimensional multi-channeled structure to improve internal connectivity. The adipose-derived stromal cells (ADSC) were seeded on the scaffolds, cultured in vitro for 2 weeks, and implanted in vivo for another 4 and 12 weeks. The double crosslinked dECM-SF hydrogels exhibited an excellent pore structure after lyophilization. The multi-channeled hydrogel scaffold presents higher water absorption ability, surface wettability, and no cytotoxicity. The addition of dECM and a channeled structure could promote chondrogenic differentiation of ADSC and engineered cartilage formation, confirmed by H&E, safranin O staining, type II collagen immunostaining, and qPCR assay. In conclusion, the hydrogel scaffold fabricated by the secondary crosslinking method has good plasticity and can be used as a scaffold for cartilage tissue engineering. The multi-channeled dECM-SF hydrogel scaffolds possess a chondrogenic induction activity that promotes engineered cartilage regeneration of ADSC in vivo.

Silk Fibroin Materials: Biomedical Applications and Perspectives

“…(d,e) 3D sca [155], (f,g) patches [156], and (h) hydrogels [157] are some concrete applications of these devic the regeneration of cardiovascular, neural, skin, cartilage, bone, abdominal, pancreas, and tissues, respectively (figure adapted from the mentioned papers). Created using Biorender.co [152], (c) films [153,154], (d,e) 3D scaffolds [155], (f,g) patches [156], and (h) hydrogels [157] are some concrete applications of these devices for the regeneration of cardiovascular, neural, skin, cartilage, bone, abdominal, pancreas, and breast tissues, respectively (figure adapted from the mentioned papers). Created using Biorender.com.…”

“…Figure 4. SF-based materials prepared from amorphous aqueous solution and related device applications: (a) electrospun silk fibroin tubes [151], (b) fibers[152], (c) films[153,154], (d,e) 3D scaffolds[155], (f,g) patches[156], and (h) hydrogels[157] are some concrete applications of these devices for the regeneration of cardiovascular, neural, skin, cartilage, bone, abdominal, pancreas, and breast tissues, respectively (figure adapted from the mentioned papers). Created using Biorender.com.…”