In vitro proliferation of mesenchymal stem cells on three-dimensional macroporous scaffolds of chitosan-gelatin-basic fibroblast growth factor composite

Hui LI; Dongfeng CHEN; Jinyuan LIU; Jianhong ZHOU; Shaohui DU; Yiwei LI; Rudong DENG; Saixia ZHANG; Heping ZENG

ABSTRACT

BACKGROUND:

Stem cell differentiation potential is strongly correlated with culture condition. The alteration in scaffold material surface function, three dimensional (3D) structure, and addition of growth factors can control stem cell proliferation and differentiation.

OBJECTIVE:

To develop 3D macroporous scaffolds with optimal porosity and porous structure to provide a microenvironment that promotes the growth of multi-potent stem cells.

DESIGN:

Repetitive measurement.

SETTING:

Department of Anatomy, College of Basic Medicine, Guangzhou University of Chinese Medicine.MATERIALS Healthy adult SD rats were provided by the Experimental Animal Center in Guangzhou University of Chinese Medicine. Chitosan and basic fibroblast growth factor (bFGF) were purchased from Sigma Corporation (St. Louis,MO).

METHODS:

The experiment was performed at the Department of Anatomy, College of Basic Medicine, Guangzhou University of Chinese Medicine from March 2003 to December 2006. Using a freeze-drying method, 3D macroporous scaffolds made of different ratios of chitosan-gelatin with bFGF were fabricated that could release bFGF with controlled porosity and porous structure. Bone marrow was obtained from the femur and tibia of SD rats, and mesenchymal stem cells (MSCs) were isolated, cultured and seeded on the scaffolds with bFGF. MSCs seeded on scaffolds with no bFGF served as control. The procedure during experiment was accorded with animal ethical requirements.MAIN OUTCOME

MEASURES:

3D structure and release performance of the scaffolds were observed by ELISA and scanning electron microscope; the effect of 3D macroporous scaffolds that released bFGF on MSC growth and viability were observed by HE staining, MTT, cell counting and SEM.

RESULTS:

There was no significant difference in pore size between scaffolds with and without bFGF (P ＞ 0.05). Scaffolds with bFGF significantly improved MSC survival rate, promoted cell adhesion, proliferation, and viability compared with scaffolds without bFGF (P ＜ 0.05).

CONCLUSION:

The results suggest that 3D macroporous scaffolds with bFGF release improve MSC survival on scaffolds,and lay a foundation for its application in tissue engineering.