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Tissue engineering using hydroxyapatite fiber sheet as a scaffold for bone regeneration culturing mesenchymal stem cells, and application for posterolateral spine fusion as a bone graft

使用羟基磷灰石纤维片作为骨再生培养间充质干细胞支架的组织工程及其作为骨移植物在后外侧脊柱融合中的应用

基本信息

批准号：
14571403
负责人：
MATSUMOTO Morio
金额：
$ 1.6万
依托单位：
KEIO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571403/
关键词：
hydroxyapatite spine fusion stem cell bone regeneration rhBMP-2 tissue engineering rat athymic mouse 骨芽細胞 BMP

项目摘要

According to the results of in vitro studies using MC3-T3 cells and bone marrow cells in 2002, hydroxyapatite fibers (HAF) might be useful either as a carrier of rhBMP-2 and a scaffold for bone regeneration to support 3D cell proliferation. In 2002, we have made posterolateral fusion model of rats. Using these models, in 2003, we made two studies, 1) to evaluate usefulness of HAF as a carrier of rhBMP-2 in vivo with future implication of clinical use for posterolateral spinal fusion surgery, 2) to evaluate the biological property and usufulness of hydroxyapatite fiber as a scaffold of osteoblast using rat osteoblastic cells in vivo. Experiment 1) Bilateral posterolateral fusion between L5 and 6 was performed to 14week old Wistar rats using HAF containing rhBMP-2 (5, 10, 16μg). Specimens were retrieved and spinal bone fusion was assessed after implantation. Experiment 2) Osteoblastic cells were isolated from skulls of 1day old wistar rats. Diameter of pores and porosity of HAFs used in … More this study were controlled to be 5μm and 95% (S0), 250μm and 99%(52000) respectively. 5×10^6 cells were seeded in each scaffolds. The subcultures were performed for 2weeks. Scaffolds with osteoblastic cells were implanted in the back of 4-week-old athymic mouse at the subcutaneous site. Specimens were retrieved after implantation. Bone formation was evaluated by pathologic findings. Result 1) Bone fusion was observed at 4 weeks in all specimens in the group of 15μg rhBMP-2. Result 2) Enchondral bone formation was found at all the cells cultured in the scaffolds implanted in vivo. S2000 scaffolds were porose and able to support 3D cell proloferation comparing with S0. They could introduce new bone formation inside the scaffolds in vivo. But they decreased in size due to their mechanical fragility. Thus we conclude that HAF is not only an acceptable material for delivering rhBMP-2 to a fusion site, thereby enhancing solid posterolateral spinal fusion, but also a scaffold for bone regeneration to support 3D cell proliferation. Less

根据2002年使用MC 3-T3细胞和骨髓细胞的体外研究结果，羟基磷灰石纤维（HAF）可能用作rhBMP-2的载体和骨再生支架，以支持三维细胞增殖。2002年我们制作了大鼠后外侧融合模型。2003年，我们利用这些模型进行了两项研究，1）评价HAF作为rhBMP-2载体的体内有效性，以及将来在脊柱后外侧融合手术中的临床应用意义; 2）评价羟基磷灰石纤维作为成骨细胞支架的生物学特性和体内使用大鼠成骨细胞的可用性。实验1）用含rhBMP-2（5、10、16μg）的HAF对14周龄Wistar大鼠进行L5、6节段的后外侧融合。取出样本，植入后评估脊柱骨融合情况。实验2）取1日龄Wistar大鼠颅骨分离成骨细胞。所用HAF的孔径和孔隙率 ...更多信息本研究分别控制在5μm和95%（S 0），250μm和99%（52000）。在每个支架中接种5×10^6个细胞。传代培养2周。将含有成骨细胞的支架植入4周龄无胸腺小鼠背部皮下。植入后取出样本。骨形成的病理结果进行了评价。结果1）15μg rhBMP-2组4周时所有标本均出现骨性融合。结果2）在体内植入的支架中培养的细胞均能形成软骨样骨。与S 0相比，S2000支架多孔，能够支持细胞的三维增殖。它们可以在体内在支架内引入新的骨形成。但由于其机械脆弱性，它们的尺寸减小了。因此，我们得出结论，HAF不仅是一种可接受的材料，用于将rhBMP-2递送到融合部位，从而增强坚固的后外侧脊柱融合，而且还是一种用于骨再生的支架，以支持3D细胞增殖。少