Abstract
Adipose-derived mesenchymal stem cells (ADSCs) are a class of pluripotent stem cells isolated from the adipose tissue; they can differentiate into osteoblasts after induction and play an important role in bone repair. EGFL6 protein is secreted by adipocytes and osteoblasts and can promote endothelial cell migration and angiogenesis. This study aimed to explore the effect of recombinant EGFL6 protein on the osteogenic differentiation of ADSCs. The cells were incubated with fluorescein isothiocyanate-conjugated antibodies and analyzed by flow cytometry. Alizarin red staining and alkaline phosphatase staining were used to detect the osteogenic differentiation ability. mRNA expression was analyzed by real-time quantitative polymerase chain reaction (RT-qPCR). Protein expression was determined using Western blotting. The osteogenic differentiation ability of ADSCs isolated from the adipose tissue was significantly weakened after EGFL6 knockdown; this ability was restored upon the addition of EGFL6 recombinant protein. BMP2 knockdown inhibited the effect of EGFL6 recombinant protein on osteogenic differentiation. EGFL6 recombinant protein promoted osteogenic differentiation of ADSCs through the BMP2/SMAD4 signaling pathway. This may provide a potential target for the osteogenic differentiation of ADSCs.
Keywords:
ADSCs; EGFL6 recombinant protein; osteogenic differentiation.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Adipocytes / drug effects
-
Bone Morphogenetic Protein 2 / genetics*
-
Bone Morphogenetic Protein 2 / metabolism
-
Calcium-Binding Proteins* / genetics
-
Calcium-Binding Proteins* / metabolism
-
Calcium-Binding Proteins* / pharmacology
-
Cell Adhesion Molecules* / genetics
-
Cell Adhesion Molecules* / metabolism
-
Cell Adhesion Molecules* / pharmacology
-
Cell Differentiation / drug effects
-
Cells, Cultured
-
Humans
-
Osteogenesis / drug effects*
-
Recombinant Proteins / genetics
-
Recombinant Proteins / metabolism
-
Recombinant Proteins / pharmacology
-
Signal Transduction / drug effects
-
Smad4 Protein / genetics*
-
Smad4 Protein / metabolism
-
Stem Cells / drug effects*
-
Transforming Growth Factor beta / genetics*
-
Transforming Growth Factor beta / metabolism
Substances
-
Bone Morphogenetic Protein 2
-
Calcium-Binding Proteins
-
Cell Adhesion Molecules
-
EGFL6 protein, human
-
Recombinant Proteins
-
SMAD4 protein, human
-
Smad4 Protein
-
Transforming Growth Factor beta
-
recombinant human bone morphogenetic protein-2
Grants and funding
The work was supported by grants from the Key Research & Development Project of Liaoning Province (No. 2018225082), the 2018 Scientist Partners of China Medical University (CMU) and Shenyang Branch of the Chinese Academy of Sciences (CAS) (No. <#AWARD-ID;>HZHB2018013) and the National Natural Science Foundation of China (NSFC) (Nos. 81571832).