Integrated Trilayered Silk Fibroin Scaffold for Osteochondral Differentiation of Adipose-Derived Stem Cells

Abstract

Repairing osteochondral defects (OCD) remains a formidable challenge due to the high complexity of native osteochondral tissue and the limited self-repair capability of cartilage. Osteochondral tissue engineering is a promising strategy for the treatment of OCD. In this study, we fabricated a novel integrated trilayered scaffold using silk fibroin and hydroxyapatite by combining paraffin-sphere leaching with a modified temperature gradient-guided thermal-induced phase separation (TIPS) technique. This biomimetic scaffold is characterized by three layers: a chondral layer with a longitudinally oriented microtubular structure, a bony layer with a 3D porous structure and an intermediate layer with a dense structure. Live/dead and CCK-8 tests indicated that this scaffold possesses good biocompatibility for supporting the growth, proliferation, and infiltration of adipose-derived stem cells (ADSCs). Histological and immunohistochemical stainings and real-time polymerase chain reaction (RT-PCR) confirmed that the ADSCs could be induced to differentiate toward chondrocytes or osteoblasts in vitro at chondral and bony layers in the presence of chondrogenic- or osteogenic-induced culture medium, respectively. Moreover, the intermediate layer could play an isolating role for preventing the cells within the chondral and bony layers from mixing with each other. In conclusion, the trilayered and integrated osteochondral scaffolds can effectively support cartilage and bone tissue generation in vitro and are potentially applicable for OC tissue engineering in vivo.

Silk fibroin/cartilage extracellular matrix scaffolds with sequential delivery of TGF-β3 for chondrogenic differentiation of adipose-derived stem cells

“…SF solution was prepared as described. 27 Other materials, including biochemical and chemical reagents, are listed in supporting information.…”

“…27 All experimental protocols were approved by the Animal Experimental Ethics Committee of Tianjin Hospital. All animals were handled in accordance with the animal care guidelines of Animal Experimental Ethics Committee of Tianjin Hospital.…”

“…The leaching fluids of the scaffolds were obtained as described. 27 ADSCs (3×10 3 in 200 μL suspension) were seeded into 96-well plates and …”

Extracellular Calcium Modulates Chondrogenic and Osteogenic Differentiation of Human Adipose-Derived Stem Cells: A Novel Approach for Osteochondral Tissue Engineering Using a Single Stem Cell Source

“…Although previous studies have used a single stem cell source to develop osteochondral tissue, [18][19][20][21][22][23][24][25][26] the approaches usually involve local delivery of growth factors through microspheres to the different layers, 19 or complex fabrication of multiphasic scaffolds using different materials or composites to resemble the different architectures of the native osteochondral tissue, 18,22,23 or used a combination of osteogenic (beta-glycerophosphate) and chondrogenic (insulin) factors within the scaffold, 24 complex double-chamber bioreactors, 25,27 or complex integrated scaffolds. 26 Graded TCP electrospun scaffolds have also been used with mouse preosteoblast cells (MC3T3-E1) to induce continuous mineralized tissue representative of the bone-cartilage interface, but not cartilage tissue.…”

“…26 Graded TCP electrospun scaffolds have also been used with mouse preosteoblast cells (MC3T3-E1) to induce continuous mineralized tissue representative of the bone-cartilage interface, but not cartilage tissue. 28 Our study uses a simple calcium gradient scaffold approach to simultaneously modulate both osteogenesis and chondrogenesis of human adiposederived stem cells without the need of bioreactors or different types of media.…”