Bio-inspired polymer surfaces for reverse transfection of siRNA to enhance osteogenic differentiation and bone formation of human adipose-derived stem cells

Abstract

Event Abstract Back to Event Bio-inspired polymer surfaces for reverse transfection of siRNA to enhance osteogenic differentiation and bone formation of human adipose-derived stem cells Jisoo Shin1, Jung-Ho Cho1, Yoonhee Jin1, Hyun-Ji Park1, Jong Seung Lee1, Kisuk Yang1 and Seung-Woo Cho1 1 Yonsei University, Department of Biotechnology, Korea Introduction: Controlling stem cell fate by regulating gene expression using small interfering RNA (siRNA) has been recently studied. However, conventional siRNA transfection methods are not efficient to induce guided stem cell differentiation due to low intracellular transfer of siRNA and short target silencing. To overcome these limitations, we developed bio-inspired polymer surfaces for reverse transfection of siRNA. We efficiently immobilized siRNA onto the catechol-modified surface of biodegradable polymer scaffolds to transfect siRNA into human stem cells. We tested whether this system could offer enhanced, long-term siRNA silencing to stem cells for promoting osteogenic differentiation and bone formation of stem cells. Materials and Methods: To prepare bio-inspired scaffolds for siRNA reverse transfection, biodegradable poly(lactide-co-glycolide) (PLGA) film was coated with catechol group, known as a main adhesive component of mussel pads. Then siRNA complexed with lipidoid nanoparticles was immobilized onto the polymer surfaces for non-viral gene delivery. Human adipose-derived stem cells (hADSCs) were seeded onto this scaffold system to reverse transfect siRNA targeting an osteogenic differentiation inhibitor to enhance osteogenic differentiation of hADSCs. Finally, hADSCs differentiated onto the siRNA-immobilized scaffolds were transplanted into critical-sized calvarial bone defects for in vivo bone formation. Results and Discussion: Immobilization efficiency of siRNA-lipidoid complexes on the catechol-modified surfaces was more than 90%. Target knockdown efficacy in hADSCs cultured on the surfaces modified with siRNA-lipidoid was around 90% with no significant cytotoxicity. The expression of osteogenic markers (OPN and ColI) was significantly upregulated in hADSCs differentiated on the surfaces with siRNA targeting osteogenic differentiation suppressor in vitro. Finally, we confirmed transplantation of hADSCs with siRNA-lipidoid immobilized PLGA scaffolds significantly enhanced in vivo bone formation in critical-sized calvarial bone defects confirmed by micro-CT and immunohistochemistry analysis. Conclusion: In summary, we developed bio-inspired polymer surfaces for siRNA reverse transfection platform exhibiting highly effective intracellular siRNA transfer and long-lasting target silencing. The polymer surfaces developed in this study could provide efficient genetic manipulation for improving stem cell engineering and tissue regeneration. This work was supported by a grant (HI14C1588) from the Korea Health Technology R&D Project funded by the Ministry of Health and Welfare, Republic of Korea; This work was supported by the Brain Korea 21 plus (BK21PLUS) program. J. Shin is a fellowship awardee by BK21 plus program Keywords: Bone Regeneration, stem cell, Surface modification, siRNA delivery Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Biomaterials in constructing tissue substitutes Citation: Shin J, Cho J, Jin Y, Park H, Lee J, Yang K and Cho S (2016). Bio-inspired polymer surfaces for reverse transfection of siRNA to enhance osteogenic differentiation and bone formation of human adipose-derived stem cells. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.00670 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Jisoo Shin Jung-Ho Cho Yoonhee Jin Hyun-Ji Park Jong Seung Lee Kisuk Yang Seung-Woo Cho Google Jisoo Shin Jung-Ho Cho Yoonhee Jin Hyun-Ji Park Jong Seung Lee Kisuk Yang Seung-Woo Cho Google Scholar Jisoo Shin Jung-Ho Cho Yoonhee Jin Hyun-Ji Park Jong Seung Lee Kisuk Yang Seung-Woo Cho PubMed Jisoo Shin Jung-Ho Cho Yoonhee Jin Hyun-Ji Park Jong Seung Lee Kisuk Yang Seung-Woo Cho Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.