caa a22 4500 44531186X CHVBK 20180317142633.0 cr unu---uuuuu 170323e20110601xx s 000 0 eng 10.1007/s11095-011-0406-9 doi (NATIONALLICENCE)springer-10.1007/s11095-011-0406-9 Non-viral Delivery of Inductive and Suppressive Genes to Adipose-Derived Stem Cells for Osteogenic Differentiation [Elektronische Daten] [Anusuya Ramasubramanian, Stacey Shiigi, Gordon Lee, Fan Yang] ABSTRACT: 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 37days, 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. Springer Science+Business Media, LLC, 2011 BMP2 nationallicence combinatorial nationallicence gene delivery nationallicence GNAS nationallicence Noggin nationallicence ADSCs : adipose-derived stem cells nationallicence ALP : alkaline phosphatase nationallicence BMP : bone morphogenetic protein nationallicence Cbfa1 : core binding factor alpha-1 nationallicence ELISA : enzyme-linked immunosorbent assay nationallicence GNAS : guanine nucleotide binding protein alpha stimulating activity polypeptide nationallicence OCN : osteocalcin nationallicence PBAE : poly(β-amino ester) nationallicence qRT-PCR : quantitative real-time polymerase chain reaction nationallicence Ramasubramanian Anusuya Program of Biomechanical Engineering, Stanford University, 94305, Stanford, California, USA aut Shiigi Stacey Department of Bioengineering, Stanford University, 94305, Stanford, California, USA aut Lee Gordon Division of Plastic and Reconstructive Surgery, Stanford University Hospital and Clinics, 94305, Stanford, California, USA aut Yang Fan Department of Bioengineering, Stanford University, 94305, Stanford, California, USA aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/6(2011-06-01), 1328-1337 0724-8741 28:6<1328 2011 28 11095 https://doi.org/10.1007/s11095-011-0406-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0406-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ramasubramanian Anusuya Program of Biomechanical Engineering, Stanford University, 94305, Stanford, California, USA aut NATIONALLICENCE 700 1- Shiigi Stacey Department of Bioengineering, Stanford University, 94305, Stanford, California, USA aut NATIONALLICENCE 700 1- Lee Gordon Division of Plastic and Reconstructive Surgery, Stanford University Hospital and Clinics, 94305, Stanford, California, USA aut NATIONALLICENCE 700 1- Yang Fan Department of Bioengineering, Stanford University, 94305, Stanford, California, USA aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/6(2011-06-01), 1328-1337 0724-8741 28:6<1328 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer