Performance of epiphyseal cartilage tissue engineering scaffold constructed by chitosan and extracellular matrix of cartilage

Qiang LU; Jiang PENG; Jingxiang HUANG; Li ZHANG; Quanyi GUO; Shibi LU

ABSTRACT

BACKGROUND:

Choose an ideal vector for amplified chondrocytes arouses more and more attention during the construction of epiphyseal cartilage tissue engineering.

OBJECTIVE:

To explore the feasibility and performance of epiphyseal cartilage tissue engineering scaffold constructed by chitosan and extracellular matrix of cartilage. DESIGN,TIME AND

SETTING:

An in vitro study was performed at Department of Orthopedics,General Hospital of Chinese PLA from December 2007 to March 2003. MATERIALSChitosan(deacetylation90%;Mr10?105) was provided by Haihui Bioengineering Company,Qingdao;articular cartilage of swine was collected from market.

METHODS:

Fresh porcine articular cartilages were obtained and shattered in the iso-osmia liquid. After pulverization and gradient centrifugation,3% artilage microfilament suspension was equally mixed with 2% chitosan. Three-dimensional porous scaffolds were fabricated using a simple freeze-drying method. MAIN OUTCOME

MEASURES:

After the second gradient ethanol treatment,the scaffolds were investigated by histological staining and scanning electron microscopy to measure aperture,porosity,and water absorption rate. MTT test was also done to assess cytotoxicity of the scaffolds. After induced by transforming growth factor-?1(TGF-?1) ,bone marrow mesenchymal stem cells(BMSCs) of rabbits were incubated onto the scaffolds. Cell proliferation and differentiation were analyzed using inverted microscopy and scanning electron microscopy.

RESULTS:

The three-dimensional porous scaffold had good pore interconnectivity with pore diameter(161?31) ?m,(90.1?1.6) % porosity and(2 361?132) % water absorption rate. The histological staining showed that toluidine blue,safranin O and anti-collagen II immunohistochemistry staining were positive. The intrinsic cytotoxicity assessment of the scaffolds using MTT test showed that the scaffolds had no cytotoxic effect on BMSCs. Most of the BMSCs attached and covered the surface of the scaffolds with matrix secretion.

CONCLUSION:

The three-dimensional porous scaffold constructed by extracellular matrix of cartilage and chitosan has good pore diameter and porosity,non-toxicity and good biocompatibility,so it is a suitable scaffold for epiphyseal cartilage tissue engineering.