caa a22 4500 445311827 CHVBK 20180317142632.0 cr unu---uuuuu 170323e20110601xx s 000 0 eng 10.1007/s11095-010-0305-5 doi (NATIONALLICENCE)springer-10.1007/s11095-010-0305-5 Optimal Construction and Delivery of Dual-Functioning Lentiviral Vectors for Type I Collagen-Suppressed Chondrogenesis in Synovium-Derived Mesenchymal Stem Cells [Elektronische Daten] [Feng Zhang, Yongchang Yao, Ruijie Zhou, Kai Su, Fudiman Citra, Dong-An Wang] ABSTRACT: Purpose: This study aims to deliver both transforming growth factor β3 (TGF-β3) and shRNA targeting type I collagen (Col I) by optimal construction and application of various dual-functioning lentiviral vectors to induce Col I-suppressed chondrogenesis in synovium-derived mesenchymal stem cells (SMSCs). Methods: We constructed four lentiviral vectors (LV-1, LV-2, LV-3 and LV-4) with various arrangements of the two expression cassettes in different positions and orientations. Col I inhibition efficiency and chondrogenic markers were assessed with qPCR, ELISA and staining techniques. Among the four vectors, LV-1 has two distant and reversely oriented cassettes, LV-2 has two distant and same-oriented cassettes, LV-3 has two proximal and reversely oriented cassettes, and LV-4 has two proximal and same-oriented cassettes. Col I and chondrogenic markers, including type II collagen (Col II), aggrecan and glycosaminoglycan (GAG), were examined in SMSCs cultured in 3-D alginate hydrogel. Results: All of the four vectors showed distinct effects in Col I level as well as diverse inductive efficiencies in upregulation of the cartilaginous markers. Based on real-time PCR results, LV-1 was optimal towards Col I-suppressed chondrogenesis. Conclusion: LV-1 vector is competent to promote Col I-suppressed chondrogenesis in SMSCs. Springer Science+Business Media, LLC, 2010 dual-functioning lentiviral vector nationallicence synovial mesenchymal stem cells (SMSCs) nationallicence TGF-β3 nationallicence type I collagen nationallicence chondrogenesis nationallicence Col I : type I collagen nationallicence Col II : type II collagen nationallicence LV-dual : recombinant lentiviral vector that expresses both shRNA targeting type I collagen and TGF- β3 nationallicence LV-T : recombinant lentiviral vector that specifically expresses TGF-β3 nationallicence shRNA : short hairpin RNA nationallicence SMSC : synovium-derived mesenchymal stem cell nationallicence TGF-β3 : transforming growth factor-β3 nationallicence Zhang Feng Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut Yao Yongchang Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut Zhou Ruijie Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut Su Kai Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut Citra Fudiman Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut Wang Dong-An Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/6(2011-06-01), 1338-1348 0724-8741 28:6<1338 2011 28 11095 https://doi.org/10.1007/s11095-010-0305-5 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-010-0305-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Feng Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut NATIONALLICENCE 700 1- Yao Yongchang Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut NATIONALLICENCE 700 1- Zhou Ruijie Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut NATIONALLICENCE 700 1- Su Kai Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut NATIONALLICENCE 700 1- Citra Fudiman Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut NATIONALLICENCE 700 1- Wang Dong-An Division of Bioengineering School of Chemical and Biomedical Engineering, Nanyang Technological University, 70 Nanyang Drive, N1.3-B2-13, 637457, Singapore, Republic of Singapore aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/6(2011-06-01), 1338-1348 0724-8741 28:6<1338 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer