PURPOSE: To assess the effects of co-delivering osteoinductive DNA and/or small interfering RNA in directing the osteogenic differentiation of human adipose-derived stem cells (hADSCs) using a combinatorial, non-viral gene delivery approach. METHODS: hADSCs were transfected using combinations of the following genes: BMP2, siGNAS and siNoggin using poly(β-amino esters) or lipid-like molecules. A total of 15 groups were evaluated by varying DNA doses, timing of treatment, and combinations of signals. All groups were cultured in osteogenic medium for up to 37 days, and outcomes were measured using gene expression, biochemical assays, and histology. RESULTS: Biomaterials-mediated gene delivery led to a dose-dependent up-regulation of BMP2 and significant gene silencing of GNAS and Noggin in hADSCs. BMP2 alone slightly up-regulates osteogenic marker expression in hADSCs. In contrast, co-delivery of BMP2 and siGNAS or siNoggin significantly accelerates the hADSC differentiation towards osteogenic differentiation, with marked increase in bone marker expression and mineralization. CONCLUSIONS: We report a combinatorial platform for identifying synergistic interactions among multiple genetic signals associated with osteogenic differentiation of hADSCs. Our results suggest that inductive or suppressive genetic switches interact in a complex manner, and highlight the promise of combinatorial approaches towards rapidly identifying optimal signals for promoting desired stem cell differentiation.
PURPOSE: To assess the effects of co-delivering osteoinductive DNA and/or small interfering RNA in directing the osteogenic differentiation of human adipose-derived stem cells (hADSCs) using a combinatorial, non-viral gene delivery approach. METHODS: hADSCs were transfected using combinations of the following genes: BMP2, siGNAS and siNoggin using poly(β-amino esters) or lipid-like molecules. A total of 15 groups were evaluated by varying DNA doses, timing of treatment, and combinations of signals. All groups were cultured in osteogenic medium for up to 37 days, and outcomes were measured using gene expression, biochemical assays, and histology. RESULTS: Biomaterials-mediated gene delivery led to a dose-dependent up-regulation of BMP2 and significant gene silencing of GNAS and Noggin in hADSCs. BMP2 alone slightly up-regulates osteogenic marker expression in hADSCs. In contrast, co-delivery of BMP2 and siGNAS or siNoggin significantly accelerates the hADSC differentiation towards osteogenic differentiation, with marked increase in bone marker expression and mineralization. CONCLUSIONS: We report a combinatorial platform for identifying synergistic interactions among multiple genetic signals associated with osteogenic differentiation of hADSCs. Our results suggest that inductive or suppressive genetic switches interact in a complex manner, and highlight the promise of combinatorial approaches towards rapidly identifying optimal signals for promoting desired stem cell differentiation.
Authors: Osamu Sugiyama; Dong Sung An; Sam P K Kung; Brian T Feeley; Seth Gamradt; Nancy Q Liu; Irvin S Y Chen; Jay R Lieberman Journal: Mol Ther Date: 2005-03 Impact factor: 11.454
Authors: Oju Jeon; Jong Won Rhie; Il-Kuen Kwon; Jae-Hwan Kim; Byung-Soo Kim; Soo-Hong Lee Journal: Tissue Eng Part A Date: 2008-08 Impact factor: 3.845
Authors: Akin Akinc; Andreas Zumbuehl; Michael Goldberg; Elizaveta S Leshchiner; Valentina Busini; Naushad Hossain; Sergio A Bacallado; David N Nguyen; Jason Fuller; Rene Alvarez; Anna Borodovsky; Todd Borland; Rainer Constien; Antonin de Fougerolles; J Robert Dorkin; K Narayanannair Jayaprakash; Muthusamy Jayaraman; Matthias John; Victor Koteliansky; Muthiah Manoharan; Lubomir Nechev; June Qin; Timothy Racie; Denitza Raitcheva; Kallanthottathil G Rajeev; Dinah W Y Sah; Jürgen Soutschek; Ivanka Toudjarska; Hans-Peter Vornlocher; Tracy S Zimmermann; Robert Langer; Daniel G Anderson Journal: Nat Biotechnol Date: 2008-04-27 Impact factor: 54.908
Authors: Seung-Woo Cho; Michael Goldberg; Sun Mi Son; Qiaobing Xu; Fan Yang; Ying Mei; Said Bogatyrev; Robert Langer; Daniel G Anderson Journal: Adv Funct Mater Date: 2009-08-25 Impact factor: 18.808
Authors: Fan Yang; Seung-Woo Cho; Sun Mi Son; Said R Bogatyrev; Deepika Singh; Jordan J Green; Ying Mei; Sohyun Park; Suk Ho Bhang; Byung-Soo Kim; Robert Langer; Daniel G Anderson Journal: Proc Natl Acad Sci U S A Date: 2009-10-05 Impact factor: 11.205
Authors: Minh Khanh Nguyen; Alexandra McMillan; Cong Truc Huynh; Daniel S Schapira; Eben Alsberg Journal: J Mater Chem B Date: 2016-12-14 Impact factor: 6.331
Authors: Jiabing Fan; Hyejin Park; Matthew K Lee; Olga Bezouglaia; Armita Fartash; Jinku Kim; Tara Aghaloo; Min Lee Journal: Tissue Eng Part A Date: 2014-05-09 Impact factor: 3.845