title: Cartilage tissue formation from dedifferentiated chondrocytes by codelivery of BMP-2 and SOX-9 genes encoding bicistronic vector.
abstract: 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.
authors: Department of Biomedical Science, CHA University, Seoul, Republic of Korea.
authors: Byung-Hyun
authors: BH
authors: Cha
authors: Jae-Hwan
authors: JH
authors: Kim
authors: Sun-Woong
authors: SW
authors: Kang
authors: Hyun-Jin
authors: HJ
authors: Do
authors: Ju-Woong
authors: JW
authors: Jang
authors: Yon Rak
authors: YR
authors: Choi
authors: Hansoo
authors: H
authors: Park
authors: Byung-Soo
authors: BS
authors: Kim
authors: Soo-Hong
authors: SH
authors: Lee
mesh_terms: D000328:Adult; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D002358:Cartilage, Articular / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*; D054337:Cell Dedifferentiation / Q000235:genetics; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D005260:Female; D018014:Gene Transfer Techniques; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D055758:SOX9 Transcription Factor / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human