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タイトル: | Preparation of antibody-immobilized gelatin nanospheres incorporating a molecular beacon to visualize the biological function of macrophages |
著者: | Yoshimoto, Yu Jo, Jun-ichiro Tabata, Yasuhiko |
著者名の別形: | 吉本, 雄 城, 潤一郎 田畑, 泰彦 |
キーワード: | Gelatin nanospheres Antibody immobilization Molecular beacon microRNA Macrophages Inflammatory response |
発行日: | Jun-2020 |
出版者: | Elsevier B.V. |
誌名: | Regenerative Therapy |
巻: | 14 |
開始ページ: | 11 |
終了ページ: | 18 |
抄録: | [Introduction]Inflammatory response plays an important role in the disease progress or therapeutic effect. In this context, it is highly required to develop a technology to visualize the inflammatory response. In this study, macrophages and their microRNA (miRNA) which are involved in the inflammatory response, were focused while a system of molecular beacon (MB) to detect the miRNA of macrophages was designed and prepared. [Methods]Gelatin nanospheres were prepared by the conventional coacervation method. An antibody with an affinity for the surface receptor of macrophages was immobilized onto the gelatin nanospheres by several methods. A nucleic acid-based MB for a pro-inflammatory miRNA 155–5p was designed and incorporated into the antibody-immobilized gelatin nanospheres (MB-gelatin NS). Macrophages before and after the polarization into pro-inflammatory or anti-inflammatory phenotypes were cultured with the MB-gelatin NS and change in the intracellular fluorescence was observed. [Results]The antibody-immobilized gelatin nanospheres prepared by a coupling between the amino groups of gelatin and the sugar chains of antibody with NaIO4 showed the highest affinity for cellular receptor. MB complexed with the cell-penetrating (CP) peptide was successfully incorporated into the antibody-immobilized gelatin nanospheres. When cultured with pro-inflammatory macrophages, MB-gelatin NS efficiently detected the miRNA 155–5p to emit fluorescence. [Conclusions]By the NaIO₄ method, the antibody was immobilized onto gelatin nanospheres with a high affinity remaining while the MB was incorporated into the antibody-immobilized gelatin nanospheres. The MB incorporated allowed mRNA to visualize the pro-inflammatory nature of macrophages. |
著作権等: | ©2020, The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license(http://creativecommons.org/licenses/by-nc-nd/4.0/). |
URI: | http://hdl.handle.net/2433/245476 |
DOI(出版社版): | 10.1016/j.reth.2019.12.009 |
PubMed ID: | 31970268 |
出現コレクション: | 学術雑誌掲載論文等 |
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