成果報告書詳細
管理番号20110000001135
タイトル*平成22年度中間年報 ナノテク・先端部材実用化研究開発/虚血下肢の切断回避を実現する細胞移植用ナノスキャフォールドの開発
公開日2011/9/9
報告書年度2010 - 2010
委託先名学校法人常翔学園大阪工業大学 公立大学法人大阪市立大学 グンゼ株式会社 株式会社ソフセラ 学校法人近畿大学
プロジェクト番号P05023
部署名電子・材料・ナノテクノロジー部
和文要約和文要約等以下本編抜粋:1. 研究開発の内容及び成果等研究開発項目(1)「ナノスキャフォールドの開発」《近畿大学担当》 本研究開発項目では、高分子微粒子の製造時に一般的に用いられる有機界面活性剤の代わりにハイドロキシアパタイト(HAp)ナノ単結晶を用いることで、生体吸収性高分子を基体粒子とし、HApナノ単結晶を外殻とするナノスキャフォールドの開発を目的とする。 平成22年度においては、これまでの研究開発によって得られた実験室レベルでの製造法に関する知見をもとにナノスキャフォールド量産化を図った。まず、溶液粘度の観点から決定した濃度に調製した生体吸収性高分子溶液と、分散性の観点から決定した濃度に調製したHApナノ単結晶水分散体とを混合した。この際、スケールアップに伴う混合時の撹拌速度を最適化することでナノスキャフォールドの粒子径を制御した。さらに、ポットミル架台を用いて同エマルションからの溶媒の蒸発挙動を制御することで、5g/バッチのナノスキャフォールド製造を達成した。
英文要約In this NEDO project, we focused on the development of nano-composite scaffolds with biosafety and cell affinity, and aimed at establishing therapeutic angiogenesis treatment with cell transplantation to avoid extremity amputation. Kinki University conducted the development of nano-composite scaffolds via Pickering emulsion method using nano-sized hydroxyapatite (HAp) as an emulsifier and dichloromethane as a solvent for bioabsorbable polymer. Kinki University conducted scale-up tests for producing nano-composite scaffolds, and successfully produced more than 5 g of nano-composite scaffolds by controlling the conditions. Osaka Institute of Technology (OIT) conducted characterization of nano-composite scaffolds in terms of molecular weight and mass during the incubation in phosphate buffered saline for one year. OIT successfully found suitable nano-composite scaffolds whose molecular weight decreased down to below 40,000 g/mol after 24 weeks. The mass started to decrease after 16 weeks and 20 wt% mass loss was detected after 1 year. Osaka City University (OCU) examined the usefulness of the nano-composite scaffolds (NS) with ischemic-limb models. OCU successfully revealed much higher induction of angiogenesis in ischemic tissues by NS+BMC than BMC alone or PLLA microspheres (LA)+BMNC groups. Immunohistochemical analysis also showed that proangiogenic cytokines, vascular endothelial growth factor (VEGF) and fibroblast growth factor-2 (FGF2), were mostly co-localized with implanted GFP-positive BMCs around NS, with some diffusing out of the clusters of BMCs and NS. NS-enhanced therapeutic angiogenesis and collateral blood flow confirmed by 3D-CT angiography showed good correlations with increased intramuscular levels of VEGF and FGF2. OCU also successfully revealed that optimum ratio of NSs to BMCs is 103-105 to 1.0X106 cells to anchor cells in ischemic tissues. GUNZE Ltd. conducted the establishment of the manufacturing process of the suitable bioabsorbable polymers for nano-composite scaffolds and provided three kinds of polymer over 1000g for Kinki University. Bioabsorbable polymer and monomer characterizations were conducted for deciding product and raw material specifications, respectively. SofSera Corporation conducted the establishment of the manufacturing process of high-dispersibility HAp nanocrystals. HAp nanocrystals were successfully fabricated in 1kg yield by calcinations using anti-sintering agent surrounding the particles, followed by removal of the agent. The crystal size and the coefficient of variance (CV) of the HAp nanocrystals were 45 nm and 27%.
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