Cartilage Tissue Formation From Dedifferentiated Chondrocytes by Code livery of BMP-2 and SOX-9 Genes Encoding Bicistronic Vector

被引:41
作者
Cha, Byung-Hyun [1 ]
Kim, Jae-Hwan [1 ]
Kang, Sun-Woong [1 ]
Do, Hyun-Jin [1 ]
Jang, Ju-Woong [2 ]
Choi, Yon Rak [2 ]
Park, Hansoo [3 ]
Kim, Byung-Soo [4 ]
Lee, Soo-Hong [1 ]
机构
[1] CHA Univ, Dept Biomed Sci, Seoul, South Korea
[2] Korea Bone Bank, Seoul, South Korea
[3] Chung Ang Univ, Dept Integrat Engn, Seoul 156756, South Korea
[4] Seoul Natl Univ, Sch Chem & Biol Engn, Seoul, South Korea
基金
新加坡国家研究基金会;
关键词
Bone morphogenetic protein 2 (BMP-2); Sex-determining region Y box gene 9 (SOX-9); Codelivery; Dedifferentiation; Cartilage tissue formation; BONE MORPHOGENETIC PROTEIN-2; HUMAN ADIPOSE-TISSUE; ARTICULAR CHONDROCYTES; TRANSCRIPTION FACTOR; GROWTH-FACTOR; REDIFFERENTIATION; CELLS; DIFFERENTIATION; CHONDROGENESIS; RESPONSIVENESS;
D O I
10.3727/096368912X647261
中图分类号
Q813 [细胞工程];
学科分类号
100113 [医学细胞生物学];
摘要
Articular cartilage, when damaged by degenerative disease or trauma, has limited ability for self-repair. Recently, many trials have demonstrated that gene therapy combined with tissue engineering techniques would be a promising approach for cartilage regeneration. Bone morphogenetic protein 2 (BMP-2) is an important signal for upregulation of osteogenesis and chondrogenesis of stem cells. Sex-determining region Y box gene 9 (SOX-9) has also been reported as one of the key transcription factors for chondrogenesis. We hypothesized that codelivery of BMP-2 and SOX-9 genes would result in improved efficiency of recovery of normal chondrogenic properties in dedifferentiated chondrocytes. To this aim, we constructed a bicistronic vector encoding the BMP-2 and SOX-9 genes linked to the "self-cleaving" 2A peptide sequence. After gene delivery to dedifferentiated chondrocytes using a microporator transfection system, we confirmed over 65% delivery efficiency of the BMP-2 and SOX-9 genes. According to RT-PCR analysis and Alcian blue staining, simultaneous delivery of BMP-2/SOX-9 resulted in significantly increased expression of chondrogenesis-related markers (type II collagen and aggrecan) and GAG matrix formation compared with individual delivery of the BMP-2 or SOX-9 gene. Six weeks after in vivo transplantation, BMP-2/SOX-9 genes also showed a significant increase in cartilage formation compared with the BMP-2 or SOX-9 gene. These results demonstrate that codelivery of two chondrogenic lineage-determining genes can enhance normal chondrogenic properties of dedifferentiated chondrocytes followed by improved cartilage formation.
引用
收藏
页码:1519 / 1528
页数:10
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