title: Regenerating cartilages by engineered ASCs: prolonged TGF-β3/BMP-6 expression improved articular cartilage formation and restored zonal structure.
abstract: Adipose-derived stem cells (ASCs) hold promise for cartilage regeneration but their chondrogenesis potential is inferior. Here, we used a baculovirus (BV) system that exploited FLPo/Frt-mediated transgene recombination and episomal minicircle formation to genetically engineer rabbit ASCs (rASCs). The BV system conferred prolonged and robust TGF-β3/BMP-6 expression in rASCs cultured in porous scaffolds, which critically augmented rASCs chondrogenesis and suppressed osteogenesis/hypertrophy, leading to the formation of cartilaginous constructs with improved maturity and mechanical properties in 2-week culture. Twelve weeks after implantation into full-thickness articular cartilage defects in rabbits, these engineered constructs regenerated neocartilages that resembled native hyaline cartilages in cell morphology, matrix composition and mechanical properties. The neocartilages also displayed cartilage-specific zonal structures without signs of hypertrophy and degeneration, and eventually integrated with host cartilages. In contrast, rASCs that transiently expressed TGF-β3/BMP-6 underwent osteogenesis/hypertrophy and resulted in the formation of inferior cartilaginous constructs, which after implantation regenerated fibrocartilages. These data underscored the crucial role of TGF-β3/BMP-6 expression level and duration in rASCs in the cell differentiation, constructs properties and in vivo repair. The BV-engineered rASCs that persistently express TGF-β3/BMP-6 improved the chondrogenesis, in vitro cartilaginous constructs production and in vivo hyaline cartilage regeneration, thus representing a remarkable advance in cartilage engineering.
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Chia-Hsin
authors.initials: CH
authors.lastname: Lu
authors.affiliation: 1] Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan [2] Division of Cardiovascular Surgery, Department of Surgery, Taipei Medical University Hospital, Taipei, Taiwan.
authors.forename: Tsung-Szu
authors.initials: TS
authors.lastname: Yeh
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Chia-Lin
authors.initials: CL
authors.lastname: Yeh
authors.affiliation: Molecular Imaging Center, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
authors.forename: Yu-Hua Dean
authors.initials: YH
authors.lastname: Fang
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Li-Yu
authors.initials: LY
authors.lastname: Sung
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Shih-Yeh
authors.initials: SY
authors.lastname: Lin
authors.affiliation: 1] Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taoyuan, Taiwan [2] College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Tzu-Chen
authors.initials: TC
authors.lastname: Yen
authors.affiliation: 1] College of Medicine, Chang Gung University, Taoyuan, Taiwan [2] Department of Orthopaedic, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
authors.forename: Yu-Han
authors.initials: YH
authors.lastname: Chang
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Yu-Chen
authors.initials: YC
authors.lastname: Hu
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D001696:Biomechanical Phenomena; D055418:Bone Morphogenetic Protein 6 / Q000235:genetics / Q000378:metabolism; D002356:Cartilage / Q000166:cytology / Q000378:metabolism; D020219:Chondrogenesis / Q000235:genetics; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D048091:Guided Tissue Regeneration; D008297:Male; D011817:Rabbits; D013234:Stem Cells / Q000378:metabolism*; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D014161:Transduction, Genetic; D053782:Transforming Growth Factor beta3 / Q000235:genetics / Q000378:metabolism
chemical_list: D055418:Bone Morphogenetic Protein 6; D053782:Transforming Growth Factor beta3