Cartilage Particles can Promote Chondrogenesis of Adipose-Derived Stromal Cells on Poly(ε-Caprolactone)/Fibrin Hybrid Constructs Prepared via Sandwich Model

Abstract

Electrospun fibers have demonstrated a remarkable potential as a framework structure in the fabrication of cartilage tissue engineering (CTE) scaffolds. Various extracellular matrices have been incorporated into electrospun scaffolds to mimic and simulate the extracellular environment. The objective of this study was to fabricate hybrid constructs using composite electrospun scaffolds based on poly (ε-caprolactone) (PCL) and cartilage-derived matrix (CDM) and fibrin hydrogel to improve the viability and differentiation of human adipose-derived stromal cells (ADSCs) for CTE applications.Initially, PCL and PCL-CDM electrospun mats were fabricated. Fibrin/ ADSCs hydrogel were seeded on PCL- CDM mats and arranged layer-by-layer using sandwich technique. This method has been employed to increase cell seeding and infiltration efficiency through the entire mass of the scaffold. Real-time reverse-transcription polymerase chain reaction (RT- PCR), were performed to examine the expression of collagen types II and X, SOX9 and aggrecan. The production of glycosaminoglycan (GAG) was also tested in vitro by Toluidine blue stain and biochemical assay in the cultured scaffolds.The findings demonstrated that incorporation of CDM in PCL fibers results in improved cell viability. Hematoxylin and eosin staining showed that the sandwich method resulted in homogenous cell seeding within the scaffold. Overall, the RT- PCR, biochemical and histological results, showed that incorporation of the CDM into PCL/fibrin sandwich scaffolds stimulated ADSCs chondrogenesis and produced the products which increased expression of chondrogenic genes. It also, enhanced GAG synthesis compared to PCL/fibrin scaffolds.These findings suggest PCL-CDM/fibrin can be considered as an appropriate hybrid scaffold for CTE applications.