Cell culture of human nasoseptal chondrocytes using bioflow reaction

利用生物流反应培养人鼻中隔软骨细胞

基本信息

批准号：
15591911
负责人：
ISOGAI Noritaka
金额：
$ 2.3万
依托单位：
Kinki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

This study evaluated the effectiveness of b-FGF impregnated in gelatin microspheres to achieve slow growth factor release for augmenting the in vivo chondrogenic response. Whereas ^<125>I-labeled b-FGF injected in solution showed rapid in vivo clearance from the injection site (only 3% residual after 24 hours), when incorporated into gelatin microspheres, 44% and 18% of the b-FGF remained at 3 and 14 days, respectively. Canine chondrocytes were isolated and grown in vitro onto ear-shaped poly-lactide/capro-lactone copolymers for one week, then implanted into the dorsal subcutaneous tissue of nude mice ; implants contained b-FGF either in free solution or in gelatin microspheres. A third group underwent pre-injection of b-FGF in gelatin microspheres four days before chondrocyte-copolymer implantation. The implants with b-FGF-incorporated microspheres showed the greatest chondrogenic characteristics at 5 and 10 weeks postoperatively : good shape and biomechanical trait retention, strong (histologic) metachromasia, rich vascularization of surrounding tissues, and increased gene expression for type II collagen (cartilage marker) and factor VIII-related antigen (vascular marker). In the case of implant site pre-administration with b-FGF-impregnated microspheres, the implant architecture was not maintained as well, and reduced vascularization and metachromasia was also apparent. In conclusion, these findings indicate that a sustained release of b-FOE augments neovascularization and chondrogenesis in a tissue-engineered cartilage construct.

本研究评估了浸渍在明胶微球中的 b-FGF 实现生长因子缓慢释放以增强体内软骨形成反应的有效性。然而注射到溶液中的125 I标记的b-FGF显示出从注射位点的体内快速清除(24小时后仅残留3％)，而当掺入明胶微球中时，保留了44％和18％的b-FGF分别在第 3 天和第 14 天。分离犬软骨细胞，体外在耳形聚丙交酯/己内酯共聚物上生长1周，然后植入裸鼠背部皮下组织；植入物含有游离溶液或明胶微球中的b-FGF。第三组在软骨细胞-共聚物植入前四天预先注射明胶微球中的 b-FGF。含有 b-FGF 微球的植入物在术后 5 周和 10 周时表现出最大的软骨形成特征：良好的形状和生物力学特征保留、强烈的（组织学）异色性、周围组织丰富的血管化以及 II 型胶原（软骨）基因表达增加标记）和因子 VIII 相关抗原（血管标记）。在植入部位预施用 b-FGF 浸渍微球的情况下，植入物结构也没有得到维持，并且血管化和异色性的减少也很明显。总之，这些发现表明，b-FOE 的持续释放可增强组织工程软骨结构中的新血管形成和软骨形成。