家兎膝骨軟骨欠損モデルに対する末梢血由来と骨髄由来多血小板フィブリンの効果

外周血和骨髓来源富血小板纤维蛋白对兔膝骨软骨缺损模型的影响

• 批准号: 21K20960
• 负责人: 丸山 真博
• 金额: $ 2万
• 依托单位: Yamagata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Research Activity Start-up
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-08-30 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21K20960/
• 关键词: 多血小板フィブリン; 骨軟骨再生; 骨髄; 再生医療; 家兎

立案した研究実施計画に準じ，家兎膝骨軟骨欠損モデルに対して末梢血由来多血小板フィブリン（P-PRF）と骨髄由来多血小板フィブリン（BM-PRF）の骨軟骨効果について実験を遂行している．まず，予備実験としてニュージーランド白色家兎を3羽用いて，動物実験施設の方々と協力して，手術器具や必要物品の購入をして，右膝蓋骨溝に幅4mm，深さ3mmの骨軟骨欠損を作製できることを確認した．また，P-PRFやBM-PRFの作製可能であることも確認した．次に本実験として，36羽を対照群，P-PRF群，BM-PRF群の3群（各群12羽）に分け手術を施行した．対照群は，右膝蓋骨溝に骨軟骨欠損（幅4mm，深さ3mm）を作製した．P-PRF群では，耳介動脈から採血した動脈血5mlをガラス管に入れ，1640gで10分間遠心分離しP-PRFを作製し，骨軟骨欠損部に充填した．BM-PRF群では，腸骨から採取した骨髄液5mlをガラス管にに入れ，1640gで10分間遠心分離しBM-PRFを作製し，骨軟骨欠損部に充填した．各群とも術後4週（n=6）と術後12週（n=6）時に安楽死させ，骨軟骨修復状況についてマイクロCT解析，病理組織学評価を行った．マイクロCT解析の結果，術後12週時，BM-PRF群では軟骨下骨が他の2群よりも良好に再生されていた．また，組織学的評価（HE染色やサフラニンO染色，コラーゲン1型，2型免疫染色）の結果，BM-PRF群では骨軟骨欠損部は硝子様軟骨組織で修復されていたが，他の２群では線維様軟骨組織で修復されていた．現在，4週時の病理組織学的評価を用いて，TGF-βやVEGFなどの免疫染色を行い，BM-PRF群の修復促進の機序について継続して研究を行っている．

In the case study, the plan was carried out and the peripheral blood was caused by multiple platelets (P-PRF), which was caused by multiple platelets (BM-PRF). As a result, the equipment was used to make sure that the equipment was used for the equipment of the equipment. The essential items of the hand appliance are imported into the machine, the right knee bone is cut into the 4mm, the bone defect of the 3mm bone is deep, and the P-PRF BM-PRF is used to make sure that it is possible to make sure that it is confirmed. This time, this equipment, 36 feather photo group, P-PRF group, BM-PRF group, 3 groups (12 birds in each group), each group is divided into three groups. The right knee patellar bone and bone defect (4mm, deep 3mm) were tested. P-PRF group, the ear was introduced into the exercise group, the heart was separated from the bone in 1640g to 10 minutes, the heart was separated in P-PRF, and the bone was filled in in 1640g to 10 minutes. BM-PRF cluster, bone bone fluid 5ml was collected and BM-PRF was isolated in 1640g to 10 minutes. After 4 years (6 years) of postnatal surgery, euthanasia was performed at 12 (6) years later, CT analysis of bone repair was performed, and histopathology was performed. The results of CT analysis were analyzed. At the last 12 hours, the results of BM-PRF subskeletal allograft were good and regenerated. The results of histology (HE staining, immuno-staining, immunostaining for type 1, type 2) were performed. The results showed that the bone tissue of the bone of the BM-PRF group was repaired with nitrate in the bone tissue of the lower part of the bone, and the bone tissue of the bone tissue of the other two groups was repaired. Now, at 4: 00, the histopathology was used for histology, and TGF- β-VEGF was used for immunostaining. BM-PRF group revision facilitates the study of the order of the machine, the research of the system, and the research of the system.