Abstract:

BACKGROUND: LIM mineralization protein 1 (LMP-1), an intracellular non-secreted protein, can recruit a large number of osteogenic factors to participate in osteoblast differentiation. The role of LMP-1 in the upper levels of osteogenic differentiation suggested the powerful roles in bone reconstruction.
OBJECTIVE: To observe the effect of recombinant adenoviruson Ad-LMP-1 infection on bone marrow mesenchymal stem cells (BMSCs) and the effect on its differentiation.
METHODS: Rat BMSCs were isolated and cultured. The expression of specific marker of BMSCs, CD44, was successfully detected. Mineralized solution was added into the study group medium to induce osteogenic differentiation and the calcified nodules were detected by alizarin red staining. After infecting with recombinant adnovirus Ad-LMP-1, the infection efficiency was evaluated by observing the green fluorescent protein (GFP) signals. The upregulated expression of LMP-1 was confirmed by RT-PCR and Western blot, while the effects of Ad-LMP-1 on morphological and proliferative changes were observed at 2, 7 and 14 days.
RESULTS AND CONCLUSION: The morphology of subcultured BMSCs was consistent and arranged closely. After induction by mineralized solution, osteogenous changes were observed in cell morphology. Alizarin red staining results showed that the calcified nodules were produced after induction. Observation of GFP signal showed that the infection efficiency of recombinant adenovirus Ad-LMP-1 was about 85%. After infection with Ad-LMP-1, the morphological changes appeared at day 7, and calcified nodules were observed at day 14, but not accompanied with any change in proliferative ability. Results indicated that recombinant adenovirus Ad-LMP-1 can effectively infect primary cultured BMSCs, and induce osteogenic differentiation.