Expression dynamics of integrin α2, α3, and αV upon osteogenic differentiation of human mesenchymal stem cells

Abstract

Background: The differentiation of human mesenchymal stem cells (hMSCs) into osteoblasts (OBs) is a prerequisite for bone formation. However, little is known about the definitive surface markers for OBs during osteogenesis. Methods: To study the surface markers on OBs, we generated and used monoclonal antibodies (MAbs) against surface molecules on transforming growth factor-β1 (TGF-β1)-treated cancer cells. The generated MAbs were further selected toward expression changes on hMSCs cultured with TGF-β1/bone morphogenetic protein-2 (BMP-2) or osteogenic differentiation medium (ODM) by flow cytometry. Immunoprecipitation and mass spectrometry were performed to identify target antigens of selected MAbs. Expression changes of the target antigens were evaluated in hMSCs, human periodontal ligament cells (hPDLCs), and human dental pulp cells (hDPCs) during osteogenic and adipogenic differentiation by quantitative polymerase chain reaction (qPCR) and flow cytometry. hMSCs were also sorted by the MAbs using magnetic-activated cell sorting system, and osteogenic potential of sorted cells was evaluated via Alizarin Red S (ARS) staining and qPCR. Results: The binding reactivity of MR14-E5, one of the MAbs, was downregulated in hMSCs with ODM while the binding reactivity of ER7-A7, ER7-A8, and MR1-B1 MAbs was upregulated. Mass spectrometry and overexpression identified that MR14-E5, ER7-A7/ER7-A8, and MR1-B1 recognized integrin α2, α3, and αV, respectively. Upon osteogenic differentiation of hMSCs, the expression of integrin α2 was drastically downregulated, but the expression of integrin α3 and αV was upregulated in accordance with upregulation of osteogenic markers. Expression of integrin α3 and αV was also upregulated in hPDLCs and hDPCs during osteogenic differentiation. Cell sorting showed that integrin αV-high hMSCs have a greater osteogenic potential than integrin αV-low hMSCs upon the osteogenic differentiation of hMSCs. Cell sorting further revealed that the surface expression of integrin αV is more dramatically induced even in integrin αV-low hMSCs.

Simulated Microgravity Remodels Extracellular Matrix of Osteocommitted Mesenchymal Stromal Cells

“…The decreased expression of these molecules may be compensatory negative feedback to osteogenic stimulation. A similar effect has been previously demonstrated [ 45 ]. The important role of CD90 and CD73 in osteo-differentiation was demonstrated in vivo since relevant knockout mice exhibit signs of osteopenia phenotype [ 46 , 47 ].…”

Integrin α10 regulates adhesion, migration, and osteogenic differentiation of alveolar bone marrow mesenchymal stem cells in type 2 diabetic patients who underwent dental implant surgery

“…The protein group on which cell adhesion depends includes mainly ECM proteins, transmembrane proteins, and cytoskeleton proteins [ 39 ]. Among these proteins, integrins, which are an important class of transmembrane proteins, can not only influence cell adhesion, but also regulate the differentiation ability of stem cells by activating downstream signaling pathways [ 40–42 ]. This study found that ITGA10 expression was significantly lower in BMSCs from diabetic patients with implant failure than that in BMSCs from diabetic patients with implant success and normal patients without diabetes.…”

Performance of the Polydopamine-Graphene Oxide Composite Substrate in the Osteogenic Differentiation of Mouse Embryonic Stem Cells

“…Osteoblast adhesion on certain ECM proteins was achieved through binding to αvβ1 integrin [ 39 ]. Various studies have demonstrated that dynamic expression of different integrins is required for the osteogenic differentiation of MSCs [ 40 , 41 ]. The use of graphene material suggests the pivotal role of integrin β1 in ECM roughness recognition, which is involved in osteoblast maturation and MSC differentiation on graphitic carbon-coated surfaces [ 42 ].…”