Fabrication and properties of a composite chitosan/type II collagen scaffold for tissue engineering cartilage

Objective: To develop a novel porous three-dimensional scaffold and to investigate its physico-chemical properties for tissue engineering cartilage.

Methods: Refined 88% deacetylation degree chitosan was prepared and dissolved in 0.2 mol/L acetate acid and fully mixed with highly purified porcine type II collagen in 0.5 mol/L acetate acid solution in a ratio of 4 to 1 (wt/wt). Freeze-drying process was employed to fabricate the composite scaffold. The construct was cross-linked by use of 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). A mechanical tester was utilized to determine the tensile strength change before and after cross-linking. The microstructure was observed via scanning electron microscopy (SEM). The lysozyme degradation was performed to evaluate the degradability of the scaffold in vitro.

Results: A bulk scaffold with desired configuration was obtained. The mechanical test showed that the cross-linking treatment could enhance the mechanical strength of the scaffold. The SEM results revealed that the two constituents evenly distributed in the scaffold and that the matrix was porous, sponge-like with interconnected pore sizing 100-250 microm. In vitro lysozyme degradation indicated that cross-linked or uncross-linked composite scaffolds had faster degradation rate than the chitosan matrix.

Conclusions: Chitosan and type II collagen can be developed into a porous three-dimensional scaffold. The related physico-chemical tests suggest that the composite scaffold meets requirements for tissue engineered scaffold and may serve as an alternative cell-carrier for tissue engineering cartilage.