Effect of GARP on osteogenic differentiation of bone marrow mesenchymal stem cells via the regulation of TGFβ1 in vitro

Mesenchymal stem cells (MSCs), which have multipotential differentiation and self-renewal potential, are possible cells for tissue engineering. Transforming growth factor β1 (TGFβ1) can be produced by MSCs in an inactive form, and the activation of TGFβ1 functions as an important regulator of osteog...

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Published in:PeerJ (San Francisco, CA) CA), 2019-05, Vol.7, p.e6993-e6993, Article e6993
Main Authors: Li, Ruixue, Sun, Jian, Yang, Fei, Sun, Yang, Wu, Xingwen, Zhou, Qianrong, Yu, Youcheng, Bi, Wei
Format: Article
Language:English
Subjects:
Alkaline phosphatase
Bone marrow
Bone marrow mesenchymal stem cell
Bone regeneration
Cbfa-1 protein
Cell self-renewal
Dentistry
Extracellular matrix
Flow cytometry
GARP
Growth factors
Mesenchymal stem cells
Mesenchyme
mRNA
Osteoblastogenesis
Proteins
Secretion
Stem cells
TGFβ1
Tissue engineering
Transforming growth factor-b1
Translational Medicine
Transplants & implants
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Summary:Mesenchymal stem cells (MSCs), which have multipotential differentiation and self-renewal potential, are possible cells for tissue engineering. Transforming growth factor β1 (TGFβ1) can be produced by MSCs in an inactive form, and the activation of TGFβ1 functions as an important regulator of osteogenic differentiation in MSCs. Recently, studies showed that Glycoprotein A repetitions predominant (GARP) participated in the activation of latent TGFβ1, but the interaction between GARP and TGFβ1 is still undefined. In our study, we successfully isolated the MSCs from bone marrow of rats, and showed that GARP was detected in bone mesenchymal stem cells (BMSCs). During the osteogenic differentiation of BMSCs, GARP expression was increased over time. To elucidate the interaction between GARP and TGFβ1, we downregulated GARP expression in BMSCs to examine the level of active TGFβ1. We then verified that the downregulation of GARP decreased the secretion of active TGFβ1. Furthermore, osteogenic differentiation experiments, alkaline phosphatase (ALP) activity analyses and Alizarin Red S staining experiments were performed to evaluate the osteogenic capacity. After the downregulation of GARP, ALP activity and Alizarin Red S staining significantly declined and the osteogenic indicators, ALP, Runx2, and OPN, also decreased, both at the mRNA and protein levels. These results demonstrated that downregulated GARP expression resulted in the reduction of TGFβ1 and the attenuation of osteoblast differentiation of BMSCs .
ISSN:2167-8359
2167-8359
DOI:10.7717/peerj.6993