Fabrication and characterization of a novel cartilage acellular matrix/chitosan hybrid scaffold for cartilage tissue engineering / 中华骨科杂志

ABSTRACT

Objective To explore the feasibility of fabricating a novel cartilage acellular matrix/chitosan hybrid scaffold for cartilage tissue engineering. Methods Human cartilage microfilaments about 100 nm-5 μm were prepared after pulverization and made into 1% suspension after decellularization. The suspension was mixed with 2% chitosan acetic acid solution, and then hybrid scaffolds were fabricated using a simple freeze-drying method. The scaffolds were cross-linked and were investigated by histological staining,SEM observation, porosity measurement, water absorption rate, biomechanical properties, and biocompatibility analysis. MTT test was also done to assess the cytotoxicity of scaffold leaching liquor. Canine chondrocytes were isolated and seeded into the scaffold. Cell proliferation and differentiation were analyzed using inverted microscope and SEM. Results The histological staining showed no chondrocyte fragments remained in the scaffolds, and anti-col Ⅱ immunohistochemistry staining were positive. SEM observation show the scaffold has good pore interconnectivity with pore diameter (136.2±34.9) μm, 81.4%±3.5% porosity and 1525.7%±129.3% water absorption rate. The longitudinal elastic modulus of the scaffold was (1.940±0.335) MPa. MTT test showed that the scaffold leaching liquor did not exert any cytotoxic effect on BMSCs. Inverted microscope and SEM micrographs indicatod that cells covered the scaffolds uniformly, and majority of the cells showed the round or elliptic morphology with much matrix secretion. Conclusion Novel cartilage acellular matrix/chitosan hybrid scaffold had similar extracellular matrix as cartilage, good pore diameter and porosity,appropriate biomechanical character, non-toxicity and good biocompatibility, which make it a suitable candidate as an alternative cell-carrier for cartilage tissue engineering.