Osteogenic differentiation potential of adipose-derived stem cells from ovariectomized mice

Osteoporosis (OP) is a systemic disease caused by imbalance between bone resorption and bone formation, commonly resulting from post-menopausal oestrogen deficiency. Although osteogenic differentiation potential of adipose-derived stem cells (ASCs) has been demonstrated, the effect of OP on osteogenic differentiation of ASCs remains unclear. Here, our work has been designed to compare proliferative capacity and osteogenic differentiation ability of ASCs obtained from osteoporotic mice and normal control mice.

Twenty 14-week-old female C57BL/6 mice were randomly divided into two groups: one, the ovariectomy (OVX) group (n=10), the other being the sham operated (Sham) group (n=10). ASCs and OP-ASCs were obtained from subcutaneous fat of female inguinal sites. Cells were passaged three times prior to subsequent experimentation. The xCELLigence system was used to monitor cell adhesion and proliferation. Mineralized nodules of differentiated ASCs and OP-ASCs were analysed using Alizarin red staining after osteogenic induction. Expressions of osteogenic-specific genes including osteopontin (Opn) and runt-related transcription factor 2 (Runx2) were assessed by real-time PCR and expression of bone-related proteins was detected by Western blotting.

Numbers of cells in all groups increased steadily for 6 days; rate of cell proliferation in the Sham group was found to be higher than in the OVX group after 48 hours. Mineralized bone nodular structures were significantly more concentrated in the Sham group than in the OVX group by day 21, and mRNA levels of Runx2 in the OVX group were significantly lower than in the Sham group. Transcript levels of genes coding for Opn showed a similar pattern to those of Runx2. Western blot results indicated that protein expression levels of OPN and RUNX2 in the OVX group were lower than those in the Sham group, at each time point.

These results indicated that the proliferative capacity and osteogenic potential of ASCs were significantly impaired in osteoporotic mice compared to normal controls. However, use of autologous transplantation of modified OP-ASCs for treatment of OP, or combination of composite scaffolds and modified OP-ASCs for repair of osteoporotic bone defects, can overcome shortcomings of other methods.