In vitro and in vivo neo-cartilage formation by heterotopic chondrocytes seeded on PGA scaffolds
In vitro and in vivo neo-cartilage formation by heterotopic chondrocytes seeded on PGA scaffolds
复制标题
DOI：
10.1007/s00418-011-0822-2
复制
发表时间：
2011-07-01
影响因子：
2.3
通讯作者：
John, T.
中科院分区：
文献类型：
--
作者：
Lohan, A.;Marzahn, U.;John, T.
组织工程异位软骨植入关节软骨缺损可能是改善关节软骨修复的另一种方法。因此，本研究的目的是在体外和体内利用裸鼠异种移植模型，对异位（耳廓、鼻中隔和关节）软骨细胞植入聚乙二醇酸（PGA）支架制备的组织工程软骨的质量进行表征和比较。采用动态培养方法，将猪关节、耳廓和鼻中隔软骨细胞植入PGA支架。构建体体外预培养3周，然后在裸鼠皮下植入1、6或12周，非种子支架植入作为对照。利用活力测定、宏观和组织学评分系统评估异位新生软骨质量。所有PGA相关异位软骨细胞体外培养均可观察到新软骨形成，体内细胞外软骨基质（ECM）沉积增加。6周的体内培养时间点导致所有软骨种类的结果更加一致，因为除了耳廓软骨细胞PGA培养外，12周的体内构建体大小减少比6周更高。在植入所有软骨细胞亚种的所有构建体中都可以观察到一些退行性组织学变化，例如无细胞ECM区域。特别是，但不限于鼻中隔软骨细胞PGA培养，出现骨化的ECM区域。新生软骨内未发现弹性纤维。关节和非关节软骨细胞构建的新软骨质量没有显著差异。组织工程化异位新软骨植入关节软骨缺损后是否能充分转化还有待进一步研究。
Implantation of tissue-engineered heterotopic cartilage into joint cartilage defects might be an alternative approach to improve articular cartilage repair. Hence, the aim of this study was to characterize and compare the quality of tissue-engineered cartilage produced with heterotopic (auricular, nasoseptal and articular) chondrocytes seeded on polyglycolic acid (PGA) scaffolds in vitro and in vivo using the nude mice xenograft model. PGA scaffolds were seeded with porcine articular, auricular and nasoseptal chondrocytes using a dynamic culturing procedure. Constructs were pre-cultured 3 weeks in vitro before being implanted subcutaneously in nude mice for 1, 6 or 12 weeks, non-seeded scaffolds were implanted as controls. Heterotopic neo-cartilage quality was assessed using vitality assays, macroscopical and histological scoring systems. Neo-cartilage formation could be observed in vitro in all PGA associated heterotopic chondrocytes cultures and extracellular cartilage matrix (ECM) deposition increased in vivo. The 6 weeks in vivo incubation time point leads to more consistent results for all cartilage species, since at 12 weeks in vivo construct size reductions were higher compared with 6 weeks except for auricular chondrocytes PGA cultures. Some regressive histological changes could be observed in all constructs seeded with all chondrocytes subspecies such as cell-free ECM areas. Particularly, but not exclusively in nasoseptal chondrocytes PGA cultures, ossificated ECM areas appeared. Elastic fibers could not be detected within any neo-cartilage. The neo-cartilage quality did not significantly differ between articular and non-articular chondrocytes constructs. Whether tissue-engineered heterotopic neo-cartilage undergoes sufficient transformation, when implanted into joint cartilage defects requires further investigation.