成果報告書詳細
管理番号20140000000400
タイトル*平成25年度中間年報 「次世代機能代替技術の研究開発 次世代再生医療技術の研究開発 少量の細胞により生体内で自己組織の再生を促す自律成熟型再生デバイスの開発(生体内で自律的に成熟する臓器再生デバイスのための基盤研究開発)」
公開日2014/8/20
報告書年度2013 - 2013
委託先名国立大学法人東京大学 国立大学法人大阪大学 学校法人東京理科大学 野村ユニソン株式会社 国立大学法人神戸大学 学校法人福田学園
プロジェクト番号P10004
部署名バイオテクノロジー・医療技術部
和文要約
英文要約Title: Fundamental Research Project for Autonomous Regeneration Device (FY2010-FY2014) FY2013 Annual Report

Reconstruction of joints, based on autonomous regeneration of cartilages, would enable to promote generalization and industrialization of cartilage tissue engineering. In this project, we will establish an industrialization technology to apply the autonomous regeneration for reconstruction of joints. To develop of the cell culture method for autonomous regeneration devices for cartilage, the controlled release using the hydrogel with growth factors and micro-particles examined the optimal density of system, exquisite combination. It was clear that the sustained release system using the hydrogel with growth factors and micro-particles promotes cellular proliferation during 2 weeks. In vitro, we established that sustained release system for cell culture. To develop the orientation of the cell for autonomous regeneration devices for bone, we examined the mesh size, dynamic strength, tolerance and safety by the load using PLLA. To prevent cartilage and bone device and the slack between the host bone and devise, we narrowed down the most suitable condition. Regarding the hydrogel, we describe the one-pot fabrication of injectable hybrid hydrogel, which consists of Chitosan/poly(ethylene glycol) (PEG) and RAD16 self-assembling peptide. A primary network was formed based on a self-assembling of RAD16, and then a chemical cross-linking of Chitosan/PEG was proceeded as the independent secondary network. The interpenetrating polymer network (IPN) like structure was induced by the difference of the cross-linking mechanism and the gelation time between two networks. Actually, the beta-sheet structure attributed to the self-assembling of RAD16 molecules in hybrid hydrogel was observed obviously by circular dichroism spectrum measurement. Regarding the hollow fiber membrane, Poly(lactic acid) was used as a biodegradable polymer and poly(lactic acid) hollow fiber membranes were prepared by nonsolvent induced phase separation. The pure water permeabilities of the obtained membranes were from 39 L/m2 h atm to 210 L/m2 h atm (OD:1.3 mm-1.6 mm). As the development of a hollow fiber module becoming the reproduction organization lifeline, in the cell culture module which used a hollow fiber together with beta-TCP, we carried out the module design and fabrication of the shape that we put together in 1/4 of the joint shape of the pig which had the space that found beta-TCP and the class of cell culture gel of 3mm. We made a hole in beta-TCP and buried an hollow fiber and examined structure as an examination factor in consideration of the interval between the hollow fiber. In addition, we devised the structure that could be cut easily and that prevents a foreign body from invading during cell culture. As evaluation of autonomous cartilage regeneration device for animal experiments, we manufactured the cartilage bone defect model of the pig, and MSC and MSC derived from synovium were implanted into the defect site. After transplantation, we analyzed noninvasively by soft Xray, µCT and performed the quantitative and qualitative evaluation of the regeneration cartilage by histologic analysis. As evaluation of autonomous bone regeneration device for animal experiments, using gel and β-TCP mixture material making technique derived from blood, we performed a rabbit femur transplant experiment. We changed granule size and the mixed ratio in the range that could gelate and evaluated bone formation after transplantation and the absorption of the artificial bone. We learned the most effective combination was that of 0.5 mg/ml of β-TCP granules until 6 month. It was recognized that the combination rate of Tetrabone of 0.4-0.7 mg/ml was most suitable for bone cartilage regeneration like β-TCP.
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