可視光硬化型ゼラチンを用いた革新的軟骨再生療法

使用可见光固化明胶的创新软骨再生疗法

• 批准号: 25462334
• 负责人: 山根 健太郎
• 金额: $ 3.16万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2013
• 资助国家: 日本
• 起止时间: 2013-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-25462334/
• 关键词: 生体材料; 軟骨再生療法; 可視光硬化型ゼラチン; 結合性改変成長因子

軟骨再生に必要な細胞や成長因子を患部へ効果的に集積させる方法として、可視光硬化型ゼラチンと細胞、結合性改変成長因子を用いた併用療法を検討した。成長因子としてコラーゲンへの強い結合能を有する結合改変成長因子を開発し、可視光硬化型ゼラチンからの徐放能を検討した結果、結合改変成長因子はゼラチンへの優れた結合能を有し、より長期間ゼラチン内に留まることがわかった。また、骨髄間質細胞（BMSC）を用いたゼラチン内での細胞培養で、使用組成における同ゼラチンの毒性はなく、長期間にわたり細胞が生存できることを確認した。以上のin vitro実験の結果を踏まえ、可視光硬化型ゼラチンと細胞、結合性改変成長因子併用療法の有効性についてウサギ大腿骨内側顆骨軟骨欠損モデルを用いて検討した。ウサギ大腿骨内側顆に径3mmの骨軟骨欠損を作成し、同部位に結合性改変成長因子（本研究ではBMP4を使用）を混和したゼラチンを充填し、可視光によって硬化させた。4週後に同部位でのmRNA発現量（type-2 collagen、aggrecan、SOX9）をRT-PCRにて定量した結果、併用群で有意な上昇を認めた。12週後の同部位の組織学的検討では、併用群で欠損部に硝子軟骨様細胞の増生を認め、Wayne scoreでも有意に優れた結果が得られた。今後は、ゼラチンに細胞（BMSC）を含有させた併用療法を検討し、最終的にはゼラチンに細胞と成長因子を同時に含有させた併用療法を検討する予定である。これらにおいて良好な結果が得られることは、再生医療において可視光硬化型ゼラチンが優れたscaffoldとなりうることを証明するものである。

The cell growth factors necessary for bone regeneration were used to determine the results of the affected parts of the bone, the sclerosing cells and the combinative growth factors were used. The combination of growth factor and growth factor can be combined with the improvement of growth factor, the results of light hardening, the combination of growth factor and growth factor, and the combination of growth factor and growth factor. Bone marrow cell culture, bone marrow cell culture (BMSC), cell culture, long-term cell culture and long-term cell culture. The results of the above in vitro tests showed that there were significant differences in the growth factors of osteosclerotic bone, smooth sclerosing type and combined growth factors. The intrathighbone diameter of the thighs and the bone defects of the 3mm thighs were formed, and the growth factors of bone defects in the same site (which was used in this study) were mixed with the growth factors of the thighs, which could be cured by light exposure. After 4 weeks, the results of mRNA quantitative analysis (type-2 collagen, aggrecan, SOX9) and RT-PCR quantitative analysis were performed in the same site. After 12-year-old, the histology of histology in the same part of the body, the bone cells of the saltpetre group and the Wayne score group were detected and the results were satisfactory. In the future, there is a method for determining the cell growth factor (BMSC), and the most effective growth factor is the same as that for the cell growth factor. The results of the good results showed that there was no significant difference between the two groups, and the regenerative medicine could be used to detect light sclerosing scaffold disease.