Effects of theaflavin and transforming growth factor beta 1 on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells into chondrocytes

Xianfeng WU

ABSTRACT

BACKGROUND:

Recent studies have shown that transforming growth factor(TGF)-?1 can convert bone marrow mesenchymal stem cells(BMSCs) into chondrocyte effectively.Theaflavin exhibits an important role in cell transformation.

OBJECTIVE:

To observe the effects of theaflavin and TGF-?1 on the proliferation and chondrogenic differentiation of rabbit BMSCs compared with the effects of TGF-?1 alone.DESIGN,TIME AND

SETTING:

The biological trial on stem cells was performed at the Central Laboratory of Anhui Provincial Hospital from May to August 2008.MATERIALSHealthy 8-week New Zealand rabbits were selected for BMSC isolation and primary culture.

METHODS:

BMSCs from the third passage were divided into 3 groupscontrol group(complete medium + dexamethasone 10-8 mmol/L,vitamin C 10 mmol/L),TGF?1 group(complete medium + dexamethasone 10-8 mmol/L,TGF-?1 5 ?g/L,vitamin C 10 mmol/L),and theaflavin group(complete medium + dexamethasone 10-8 mmol/L,TGF-?1 5 ng/mL,vitamin C 10 mmol/L,theaflavin 30 mg/L).MAIN OUTCOME

MEASURES:

Inverted microscope was used to observe changes in cell growth and morphology.Toluidine blue staining and alcian blue staining were employed to glucose aminoglycan content in each well of each plate.Absorbance value was measured using immune detector.

RESULTS:

BMSCs isolated from rabbit bone marrow proliferated actively in vitro.BMSCs in monolayer cultures treated with TGF-?1 and theaflavin showed growth deceleration.Following treatment of induction,cell nodule was found in the TGF?1 and theaflavin groups.More cell nodule and some radial cell colonies were visible in the theaflavin group.No significant changes were detected in the control group.Positive reaction was found using immunohistochemistry in the TGF?1 group,and strong positive reaction was send in the theaflavin group.No positive cells were detected in the control group.Compared with the control group,absorbance values were increased in the TGF?1 and theaflavin groups(P