Enhancement of bone regeneration by gene delivery of BMP2/Runx2 bicistronic vector into adipose-derived stromal cells.
Adipose tissue contains multipotent mesenchymal stem cells (MSCs) that are able to differentiate into various tissues. Bone morphogenetic protein 2 (BMP2) is known as one of the key osteogenesis induction factors in MSCs. Recently, several new transcription factors that contribute to osteogenic differentiation have been reported, among them Runx2, Osterix, and Dlx5. We hypothesized that adipose-derived stromal cells (ASCs) could be induced to efficiently differentiate into osteocytes by the co-expression of the BMP2 and Runx2 genes. To prove this hypothesis, we constructed a bicistronic vector encoding the BMP2 and Runx2 genes linked to the 'self-cleaving' 2A peptide sequence. BMP2/Runx2-ASCs showed a gradual increase in alkaline phosphatase activity for two weeks. RT-PCR analysis and alizarin red staining revealed a high expression of osteogenesis-related markers (osteopontin, osteocalcin and collagen type I) and increased mineralization in BMP2/Runx2-ASCs compared to BMP2-ASCs. Six weeks after in vivo transplantation, BMP2/Runx2-ASCs also showed a significant increase in bone formation compared to ASCs and BMP2-ASCs. These findings demonstrate that the co-transfection of two osteogenic lineage-determining genes can enhance osteogenic differentiation of ASCs.
Lee
Suk-Jun
SJ
Department of Medical Bioscience, CHA University, 606-16 Yeoksam 1-dong, Gangnam-gu, Seoul, Republic of Korea.
Kang
Sun-Woong
SW
Do
Hyun-Jin
HJ
Han
Inbo
I
Shin
Dong Ah
DA
Kim
Jae-Hwan
JH
Lee
Soo-Hong
SH
D017667:Adipocytes / Q000166:cytology / Q000502:physiology*; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism*; D001861:Bone Regeneration / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D010012:Osteogenesis / Q000502:physiology; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology*
D055396:Bone Morphogenetic Protein 2; D050659:Core Binding Factor Alpha 1 Subunit; C493709:RUNX2 protein, human