Hmox1 promotes osteogenic differentiation at the expense of reduced adipogenic differentiation induced by BMP9 in C3H10T1/2 cells

Abstract

Mesenchymal stem cells (MSCs) are multipotent progenitors that can differentiate into a variety of cell types under proper stimuli. Bone morphogenetic protein 9 (BMP9) is able to simultaneously induce both adipogenic and osteogenic differentiation of MSCs although the regulatory molecules involved remain to be fully identified and characterized. Heme oxygenase 1 (Hmox1) plays an essential role not only in fat metabolism, but also in bone development. In the present study, we investigated the functional role of Hmox1 in BMP9-induced osteogenic/adipogenic differentiation in MSCs line C3H10T1/2 and probed the possible mechanism involved. We found that BMP9 promoted the endogenous expression of Hmox1 in C3H10T1/2 cells. Overexpression of Hmox1 or cobalt protoporphyrin (CoPP), an inducer of Hmox1, increased BMP9-induced osteogenic differentiation in vitro. Subcutaneous stem cell implantation in nude mice further confirmed that Hmox1 potentiated BMP9-induced ectopic bone formation in vivo. In contrast, Hmox1 reduced BMP9-induced adipogenic differentiation in C3H10T1/2 cells. Although had no obvious effect on BMP9-induced Smad1/5/8 phosphorylation, Hmox1 enhanced phosphorylation of p38, and AKT, while decreased phosphorylation of ERK1/2. Furthermore, Hmox1 increased total β-catenin protein level, and promoted the nuclear translocation of β-catenin in C3H10T1/2 cells. Taken together, our study strongly suggests that Hmox1 is likely to potentiate osteogenic differentiation and yet decrease adipogenic differentiation induced by BMP9 possibly through regulation of multiple signaling pathways.

Nrf2 Activation Is Involved in Cyclic Mechanical Stress-Stimulated Osteogenic Differentiation in Periodontal Ligament Stem Cells via PI3K/Akt Signaling and HO1-SOD2 Interaction

“…HO1 possesses powerfully cytoprotective and antioxidant properties through the enzymatic degradation of heme (Consoli et al, 2021;Nitti et al, 2021). Some studies showed improve the osteogenic differentiation in mesenchymal stem cells (MSCs) line C3H10T1/2 and promote ectopic bone formation in nude mice (Liu et al, 2018).…”

“…Panax ginseng fruit extract (PGFE) induced the expression levels of Nrf2 and HO1, and recovered the osteoblast differentiation restrained by Porphyromonas gingivalis lipopolysaccharide (PG-LPS) in the human periodontal ligament cells, while tin protoporphyrin IX (SnPP), the inhibitor of HO1, suppressed the osteoblast differentiation ( Kim et al, 2020 ). Up-regulation of HO1 expression could improve the osteogenic differentiation in mesenchymal stem cells (MSCs) line C3H10T1/2 and promote ectopic bone formation in nude mice ( Liu et al, 2018 ).…”

Engineering elastic bioactive composite hydrogels for promoting osteogenic differentiation of embryonic mesenchymal stem cells

“…Among these MSCs, EMSCs have the advantages of strong self-renewal ability, strong proliferation ability, multidirectional differentiation capacity, strong immunosuppressive effect, and convenient acquisition with large amounts ( Saleh et al, 2020 ). Notably, C3H10T1/2 cells, as mouse EMSCs, can differentiate into osteoblasts under the stimulation of various growth factors and active substances, which exhibit potential therapeutic value in bone tissue regeneration ( Lin et al, 2009 ; Liu et al, 2018 ; Jiang et al, 2019 ). Therefore, it is a key factor in effectively promoting the osteogenic differentiation of C3H10T1/2 cells in bone tissue engineering.…”

TAZ promotes osteogenic differentiation of mesenchymal stem cells line C3H10T1/2, murine multi-lineage cells lines C2C12, and MEFs induced by BMP9

“…Therefore, it is necessary to understand the regulatory mechanisms behind the MSCs differentiation, which was regulated by BMP9 to improve the clinical application of BMP9. It has been reported that other cytokines combined with BMP9 can promote osteogenic differentiation of MSCs induced by BMP9, which will further optimize the bone-promoting effect of BMP9 [ 34 , 35 ]. Nevertheless, the exact biological molecular mechanism involved is still under investigation.…”