Abstract
Mesenchymal stem cells (MSCs) are important in regenerative medicine because of their potential for multi-differentiation. Bone marrow, chorion and placenta have all been suggested as potential sources for clinical application. However, the osteogenic differentiation potential of MSCs derived from chorion or placenta is not very efficient. Bone morphogenetic protein-2 (BMP-2) plays an important role in bone development. Its effect on osteogenic augmentation has been addressed in several studies. Recent studies have also shown a relationship between miRNAs and osteogenesis. We hypothesized that miRNAs targeted to Runt-related transcription factor 2 (Runx-2), a major transcription factor of osteogenesis, are responsible for regulating the differentiation of MSCs into osteoblasts. This study examines the effect of BMP-2 on the osteogenic differentiation of MSCs isolated from chorion and placenta in comparison to bone marrow-derived MSCs and investigates the role of miRNAs in the osteogenic differentiation of MSCs from these sources. MSCs were isolated from human bone marrow, chorion and placenta. The osteogenic differentiation potential after BMP-2 treatment was examined using ALP staining, ALP activity assay, and osteogenic gene expression. Candidate miRNAs were selected and their expression levels during osteoblastic differentiation were examined using real-time RT-PCR. The role of these miRNAs in osteogenesis was investigated by transfection with specific miRNA inhibitors. The level of osteogenic differentiation was monitored after anti-miRNA treatment. MSCs isolated from chorion and placenta exhibited self-renewal capacity and multi-lineage differentiation potential similar to MSCs isolated from bone marrow. BMP-2 treated MSCs showed higher ALP levels and osteogenic gene expression compared to untreated MSCs. All investigated miRNAs (miR-31, miR-106a and miR148) were consistently downregulated during the process of osteogenic differentiation. After treatment with miRNA inhibitors, ALP activity and osteogenic gene expression increased over the time of osteogenic differentiation. BMP-2 has a positive effect on osteogenic differentiation of chorion- and placenta-derived MSCs. The inhibition of specific miRNAs enhanced the osteogenic differentiation capacity of various MSCs in culture and this strategy might be used to promote bone regeneration. However, further in vivo experiments are required to assess the validity of this approach.
Highlights
Mesenchymal stem cells (MSCs) are important in regenerative medicine because of their potential for multi-differentiation
The down-regulation of these miRNA expression levels during the osteogenic differentiation of CHMSCs and PL-MSCs was lesser in extent compared with those of bone marrow-derived MSCs (BM-MSCs), resulting in significantly higher levels of miR-31, miR-106a, and miR-148a in CH-MSCs and PL-MSCs in comparison to those of BM-MSCs throughout the osteogenic differentiation process (Fig. 6). These results suggest that the higher expression levels of these miRNAs might be responsible for the lower osteogenic differentiation potential of CH-MSCs and PLMSCs compared with their bone marrow counterpart
The results revealed that inhibition of each miRNA, anti-miR-31, anti-miR-106a or anti-miR-148a, and inhibition using a combination of all three anti-miRNAs, enhanced the expression of ALP in both CH-MSCs and PL-MSCs compared to MSCs cultured in osteogenic differentiation medium without miRNA inhibitor and MSCs cultured in osteogenic differentiation medium with negative control (Fig. 8)
Summary
Mesenchymal stem cells (MSCs) are important in regenerative medicine because of their potential for multi-differentiation. This study examines the effect of BMP-2 on the osteogenic differentiation of MSCs isolated from chorion and placenta in comparison to bone marrow-derived MSCs and investigates the role of miRNAs in the osteogenic differentiation of MSCs from these sources. MSCs isolated from chorion and placenta exhibited self-renewal capacity and multi-lineage differentiation potential similar to MSCs isolated from bone marrow. BMP-2 has a positive effect on osteogenic differentiation of chorion- and placenta-derived MSCs. The inhibition of specific miRNAs enhanced the osteogenic differentiation capacity of various MSCs in culture and this strategy might be used to promote bone regeneration. Mesenchymal stem cells (MSCs) are multipotent stem cells that were first described in bone marrow as bone-forming progenitors by Friedenstein et al in 19763 These cells demonstrate self-renewal capacity and multi-lineage differentiation potential[4]. The precise mechanisms and reasons for this difference between bone marrow- and chorion- or placenta-derived MSCs are unclear
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