Human EPO adenoviral particles

Regeneration of large bone defects is a common clinical problem. Recent studies have shown that mesenchymal stem cells (MSCs) have become a promising alternative to traditional surgical techniques. However, how to enhance the osteogenic potential of MSCs for possible clinical trials remains a critical issue. Here, researchers investigated the effects of adenovirus-mediated erythropoietin (Ad-EPO) transfer on BMSCs. Flow cytometry analysis and MTT results showed that EPO could promote BMSC proliferation. QPCR data showed that EPO increased the expression of Runx2, Sp7, and Col1 in osteoblasts at different time points, and also increased alkaline phosphatase activity and calcium deposition. These results suggest that EPO can increase osteoblast differentiation. Importantly, in vivo experiments demonstrated that EPO can effectively induce new bone formation in a bone defect model. These findings strongly suggest that EPO can affect osteoblast differentiation and play an important role in bone regeneration, thereby increasing bone formation.

Here, MOI of 400 was a better value for efficient transduction of BMSCs (Figure 1a). The transfection efficiency of EPO to BMSCs cells was 80%. PCR data of the Ad-EPO experimental group showed an increase in EPO expression (Figure 1b). Cell cycle analysis showed that the proportion of S phase cells increased compared with the Ad-EPO experimental group (Figure 1c). The proportion of G2 and S phase cells in Ad-EPO was higher than that in the Advehicle negative control group and the control group. Therefore, when DNA replication begins, the S phase begins, and the amount of DNA in the cell has actually doubled, although the ploidy of the cell remains unchanged. In addition, MTT analysis showed an increase in the number of cells (Figure 1d).

Figure 1. The transfection efficiency determined by fluorescence inverted microscope, 72 h after transfection (X100). (Li C, et al., 2014)