title: Programmed Application of Transforming Growth Factor β3 and Rac1 Inhibitor NSC23766 Committed Hyaline Cartilage Differentiation of Adipose-Derived Stem Cells for Osteochondral Defect Repair.
abstract: Hyaline cartilage differentiation is always the challenge with application of stem cells for joint repair. Transforming growth factors (TGFs) and bone morphogenetic proteins can initiate cartilage differentiation but often lead to hypertrophy and calcification, related to abnormal Rac1 activity. In this study, we developed a strategy of programmed application of TGFβ3 and Rac1 inhibitor NSC23766 to commit the hyaline cartilage differentiation of adipose-derived stem cells (ADSCs) for joint cartilage repair. ADSCs were isolated and cultured in a micromass and pellet culture model to evaluate chondrogenic and hypertrophic differentiation. The function of Rac1 was investigated with constitutively active Rac1 mutant and dominant negative Rac1 mutant. The efficacy of ADSCs with programmed application of TGFβ3 and Rac1 inhibitor for cartilage repair was studied in a rat model of osteochondral defects. The results showed that TGFβ3 promoted ADSCs chondro-lineage differentiation and that NSC23766 prevented ADSC-derived chondrocytes from hypertrophy in vitro. The combination of ADSCs, TGFβ3, and NSC23766 promoted quality osteochondral defect repair in rats with much less chondrocytes hypertrophy and significantly higher International Cartilage Repair Society macroscopic and microscopic scores. The findings have illustrated that programmed application of TGFβ3 and Rac1 inhibitor NSC23766 can commit ADSCs to chondro-lineage differentiation and improve the efficacy of ADSCs for cartilage defect repair. These findings suggest a promising stem cell-based strategy for articular cartilage repair.
authors.affiliation: Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China; Zhejiang Provincial Key Laboratory for Tissue Engineering and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China.
authors.forename: Shouan
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China; Zhejiang Provincial Key Laboratory for Tissue Engineering and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Chen
authors.affiliation: Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China; Zhejiang Provincial Key Laboratory for Tissue Engineering and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China; Zhejiang Provincial Key Laboratory for Tissue Engineering and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China.
authors.forename: Si
authors.initials: S
authors.lastname: Xiong
authors.affiliation: Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China; Zhejiang Provincial Key Laboratory for Tissue Engineering and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China.
authors.forename: Heng
authors.initials: H
authors.lastname: Sun
authors.affiliation: Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China; Zhejiang Provincial Key Laboratory for Tissue Engineering and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China.
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Xia
authors.affiliation: Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China; Zhejiang Provincial Key Laboratory for Tissue Engineering and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China.
authors.forename: Libing
authors.initials: L
authors.lastname: Shi
authors.affiliation: Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China; Zhejiang Provincial Key Laboratory for Tissue Engineering and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China hwoy@zju.edu.cn hhliu012@gmail.com.
authors.forename: Huanhuan
authors.initials: H
authors.lastname: Liu
authors.affiliation: Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China; Zhejiang Provincial Key Laboratory for Tissue Engineering and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China hwoy@zju.edu.cn hhliu012@gmail.com.
authors.forename: Hong Wei
authors.initials: HW
authors.lastname: Ouyang
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D000634:Aminoquinolines / Q000008:administration & dosage*; D000818:Animals; D015153:Blotting, Western; D002357:Cartilage Diseases / Q000628:therapy*; D002454:Cell Differentiation / Q000502:physiology; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects; D004195:Disease Models, Animal; D005434:Flow Cytometry; D051457:Hyaline Cartilage / Q000166:cytology*; D008297:Male; D008863:Microspheres; D011743:Pyrimidines / Q000008:administration & dosage*; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D033581:Stem Cell Transplantation / Q000379:methods*; D013234:Stem Cells / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods; D053782:Transforming Growth Factor beta3 / Q000037:antagonists & inhibitors; D020830:rac1 GTP-Binding Protein / Q000037:antagonists & inhibitors
chemical_list: D000634:Aminoquinolines; C487513:NSC 23766; D011743:Pyrimidines; D053782:Transforming Growth Factor beta3; D020830:rac1 GTP-Binding Protein
keywords: Adipose-derive stem cells; Hypertrophy; Osteochondral defect; Rac1