siRNA Nanoparticle Functionalization of Nanostructured Scaffolds Enables Controlled Multilineage Differentiation of Stem Cells

• Published in: Molecular therapy 2010-11, Vol.18 (11), p.2018-2027
• Main Authors: Andersen, Morten Ø, Nygaard, Jens V, Burns, Jorge S, Raarup, Merete K, Nyengaard, Jens R, Bünger, Cody, Besenbacher, Flemming, Howard, Kenneth A, Kassem, Moustapha, Kjems, Jørgen
• Format: Article
• Language: English
• Subjects: Adipocytes - cytology; Adipocytes - metabolism; Animals; Apoptosis; Apoptosis Regulatory Proteins - antagonists & inhibitors; Apoptosis Regulatory Proteins - genetics; Apoptosis Regulatory Proteins - metabolism; Blotting, Western; Calcium-Calmodulin-Dependent Protein Kinases; Cell Differentiation - genetics; Cell Proliferation; Cells, Cultured; Chondrogenesis; DNA-Activated Protein Kinase - physiology; DNA-Binding Proteins - physiology; Female; Flow Cytometry; Gene Silencing; Genes; Green Fluorescent Proteins - genetics; Humans; Immunoenzyme Techniques; Intracellular Signaling Peptides and Proteins - antagonists & inhibitors; Intracellular Signaling Peptides and Proteins - genetics; Intracellular Signaling Peptides and Proteins - metabolism; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; Mice; Mice, Inbred NOD; Mice, SCID; Nanoparticles; Nanoparticles - ultrastructure; Nanoscience; Nanostructures - chemistry; Nanostructures - ultrastructure; Nuclear Proteins - physiology; Original; Osteoblasts - cytology; Osteoblasts - metabolism; Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; RNA, Small Interfering - genetics; Standard deviation; Stem cells; Tissue Engineering; Tissue Scaffolds
• ISSN: 1525-0016; 1525-0024
• DOI: 10.1038/mt.2010.166

The creation of complex tissues and organs is the ultimate goal in tissue engineering. Engineered morphogenesis necessitates spatially controlled development of multiple cell types within a scaffold implant. We present a novel method to achieve this by adhering nanoparticles containing different small-interfering RNAs (siRNAs) into nanostructured scaffolds. This allows spatial retention of the RNAs within nanopores until their cellular delivery. The released siRNAs were capable of gene silencing BCL2L2 and TRIB2, in mesenchymal stem cells (MSCs), enhancing osteogenic and adipogenic differentiation, respectively. This approach for enhancing a single type of differentiation is immediately applicable to all areas of tissue engineering. Different nanoparticles localized to spatially distinct locations within a single implant allowed two different tissue types to develop in controllable areas of an implant. As a consequence of this, we predict that complex tissues and organs can be engineered by the in situ development of multiple cell types guided by spatially restricted nanoparticles.