Development of Functional Biomaterials Using RNA Aptamer

Event Abstract Back to Event Development of functional biomaterials using RNA aptamer Yusuke Nomura1, Chie Fukui1, Hitomi Toida1, Yuki Morishita1, Shingo Niimi1, Shin Miyakawa2, Ling Jin2, Yoshikazu Nakamura2, 3 and Yuji Haishima1 1 National Institute of Health Sciences, Division of Medical Devices, Japan 2 RIBOMIC Inc., Japan 3 University of Tokyo, Institute of Medical Science, Japan Introduction: Biomaterials used for implant interact with cells and tissues through the protein layer present at the biointerface. These proteins could adversely affect the function and biocompatibility of the materials. Their adsorption behavior is basically dependent on the physicochemical property of the materials, but it is possible to provide favorable functions to the materials by artificially controlling the protein adsorption behavior on the material surface. In this study, we developed a novel and functional biomaterial immobilized the RNA aptamer (RNA-AP) which specifically captures active FGF2. Materials and Methods: Human FGF2, FGF2 receptor and Lysozyme were purchased from Peprotec, R&D systems and SIGMA, respectively. RNA-APs were prepared by in vitro transcription using the DuraScribe T7 Transcription kit. RNA-AP-SH, poly dT-SH (pdT-SH) and PEG (PEG2k-SH and PEG5k-SH) were immobilized on the Au-coated glass plate. The inhibition of FGF2-mediated phosphorylation in NIH 3T3 cells was analyzed by western blotting using phospho-specific antibody (P-FRS2, P-ERK). RNA-AP/protein interaction and cell proliferation were analyzed using BIAcore2000 and BZ-9000, respectively. Results and Discussion: Candidate of RNA-APs against the human FGF2 were selected by surface plasmon resonance (SPR) method, that did not inhibit the binding of FGF2 to FGF2 receptor in SPR analysis and the phosphorylation of FRS2 and ERK in the NIH 3T3 cells. The several SPR sensor chips were prepared and used for evaluating the protein adsorption. The bare gold and pdT-immobilized surfaces did not show a difference between Lysozyme and FGF2 adsorption, indicating that only nonspecific adsorption of the proteins was observed. Although the specific adsorption of FGF2 was observed on the RNA-AP-immobilized surface, the nonspecific adsorption of Lysozyme was still high. On the other hand, the RNA-AP/PEG2k/PEG5k-coimmobilized surfaces showed a remarkable reduction of the nonspecific adsorption due to the excluded volume effect of the PEG chains[1]. In addition, cell proliferation on the RNA-AP/PEG-coated surfaces was significantly increased in comparison with the pdT/PEG-coated surfaces, indicating that RNA-AP/PEG-immobilized surfaces effectively capture and concentrate the active FGF2. Conclusion: In this study, we succeeded in selectively capturing and concentrating the target protein on the biomaterial surface by immobilizing the functional RNA-AP. It is expected that biomaterials combinationally immobilized several RNA-APs may exhibit the potency to control different function such as proliferation and differentiation of various stem cells. Thus, the innovative biomaterials using RNA aptamer are useful for not only the application to regenerative medicine product, but also other medical devices. References:[1] Uchida K., Otsuka H., Kaneko M., Kataoka K. and Nagasaki Y., Anal.Chem., 77, 1075. 2005 Keywords: Regenerative Medicine, growth factor, Implant, Novel material Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Regenerative medicine: biomaterials for control of tissue induction Citation: Nomura Y, Fukui C, Toida H, Morishita Y, Niimi S, Miyakawa S, Jin L, Nakamura Y and Haishima Y (2016). Development of functional biomaterials using RNA aptamer. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02676 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 Yusuke Nomura Chie Fukui Hitomi Toida Yuki Morishita Shingo Niimi Shin Miyakawa Ling Jin Yoshikazu Nakamura Yuji Haishima Google Yusuke Nomura Chie Fukui Hitomi Toida Yuki Morishita Shingo Niimi Shin Miyakawa Ling Jin Yoshikazu Nakamura Yuji Haishima Google Scholar Yusuke Nomura Chie Fukui Hitomi Toida Yuki Morishita Shingo Niimi Shin Miyakawa Ling Jin Yoshikazu Nakamura Yuji Haishima PubMed Yusuke Nomura Chie Fukui Hitomi Toida Yuki Morishita Shingo Niimi Shin Miyakawa Ling Jin Yoshikazu Nakamura Yuji Haishima 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.