RETRACTED ARTICLE: Schnurri-3 regulates BMP9-induced osteogenic differentiation and angiogenesis of human amniotic mesenchymal stem cells through Runx2 and VEGF

Abstract

Human amniotic mesenchymal stem cells (hAMSCs) are multiple potent progenitor cells (MPCs) that can differentiate into different lineages (osteogenic, chondrogenic, and adipogenic cells) and have a favorable capacity for angiogenesis. Schnurri-3 (Shn3) is a large zinc finger protein related to Drosophila Shn, which is a critical mediator of postnatal bone formation. Bone morphogenetic protein 9 (BMP9), one of the most potent osteogenic BMPs, can strongly upregulate various osteogenesis-and angiogenesis-related factors in MSCs. It remains unclear how Shn3 is involved in BMP9-induced osteogenic differentiation coupled with angiogenesis in hAMSCs. In this investigation, we conducted a comprehensive study to identify the effect of Shn3 on BMP9-induced osteogenic differentiation and angiogenesis in hAMSCs and analyze the responsible signaling pathway. The results from in vitro and in vivo experimentation show that Shn3 notably inhibits BMP9-induced early and late osteogenic differentiation of hAMSCs, expression of osteogenesis-related factors, and subcutaneous ectopic bone formation from hAMSCs in nude mice. Shn3 also inhibited BMP9-induced angiogenic differentiation, expression of angiogenesis-related factors, and subcutaneous vascular invasion in mice. Mechanistically, we found that Shn3 prominently inhibited the expression of BMP9 and activation of the BMP/Smad and BMP/MAPK signaling pathways. In addition, we further found activity on runt-related transcription factor 2 (Runx2), vascular endothelial growth factor (VEGF), and the target genes shared by BMP and Shn3 signaling pathways. Silencing Shn3 could dramatically enhance the expression of Runx2, which directly regulates the downstream target VEGF to couple osteogenic differentiation with angiogenesis. To summarize, our findings suggested that Shn3 significantly inhibited the BMP9-induced osteogenic differentiation and angiogenesis in hAMSCs. The effect of Shn3 was primarily seen through inhibition of the BMP/Smad signaling pathway and depressed expression of Runx2, which directly regulates VEGF, which couples BMP9-induced osteogenic differentiation with angiogenesis.

BMP9 exhibits dual and coupled roles in inducing osteogenic and angiogenic differentiation of mesenchymal stem cells

“…At the same time, we identified that Notch1 signaling potentiated BMP9-induced osteogenic differentiation of MSCs by potentiating the osteogenesis-angiogenesis coupling [16]. In addition, we clarified that Schnurri-3 simultaneously regulates BMP9-induced osteogenic differentiation and blood vessel formation in human amniotic MSCs [31]. The research indicated that angiogenic signaling participated in BMP9-induced bone formation of MSCs.…”

“…Meanwhile, BMP9 was proved to regulate postnatal angiogenesis, which indicated the angiogenic differentiation potential of BMP9 [45,46]. In previous studies, we have identified that angiogenic signaling is essential for BMP9-mediated osteogenic differentiation of MSCs [16,30,31], however, BMP9-induced angiogenic differentiation of MSCs and the relationship between BMP9-mediated osteogenic and angiogenic differentiation of MSCs were not well established. In the present study, we found that BMP9 exhibited dual roles in inducing MSCs osteogenic and angiogenic differentiation in vitro, and further in vivo assays confirmed that BMP9-mediated bone formation are coupled with angiogenesis.…”

Silencing Smad7 Potentiates BMP2-induced Chondrogenic Differentiation and Inhibits Endochondral Ossification in Human Synovial Derived Mesenchymal Stem Cells

“…1D) The hSMSCs express most of the MSCs markers and capable of multidirectional differentiation potential According to the criteria for mesenchymal stem cell identi cation,we identify the surface marker of hSMSCs using ow cytometry. [29,32] Flow cytometric results showed that P1 hSMSCs were positive for MSC markers CD73, CD90, and CD105 and weakly expressed hematopoietic markers CD34, CD14, CD45, which indicated that P1 hSMSCs express most of the consensus MSC markers, suggesting that these cells may possess MSC-like characteristics. (Fig.…”

Silencing Smad7 potentiates BMP2-induced chondrogenic differentiation and inhibits endochondral ossification in human synovial-derived mesenchymal stromal cells

“…Briefly, to identify the multiple differentiation potential [32,33], cells were seeded in a 6-well plate, and the differentiation medium was replaced when the cell density reached 60%. Osteogenic medium was composed of DMEM with 10% FBS, 10 mM bglycerophosphate (Sigma-Aldrich), 50 mg/mL ascorbate (Sigma-Aldrich), and 100 nM dexamethasone (Sigma-Aldrich).…”