Liese Juliane, Marzahn Ulrike, El Sayed Karym, Pruss Axel, Haisch Andreas, Stoelzel Katharina
Department of General and Visceral Surgery, Johann Wolfgang Goethe University, Theodor-Stern-Kai 7, 60590, Frankfurt, Germany.
Cell Tissue Bank. 2013 Jun;14(2):255-66. doi: 10.1007/s10561-012-9322-4. Epub 2012 Jun 20.
Tissue Engineering is an important method for generating cartilage tissue with isolated autologous cells and the support of biomaterials. In contrast to various gel-like biomaterials, human demineralized bone matrix (DBM) guarantees some biomechanical stability for an application in biomechanically loaded regions. The present study combined for the first time the method of seeding chondrocyte-macroaggregates in DBM for the purpose of cartilage tissue engineering. After isolating human nasal chondrocytes and creating a three-dimensional macroaggregate arrangement, the DBM was cultivated in vitro with the macroaggregates. The interaction of the cells within the DBM was analyzed with respect to cell differentiation and the inhibitory effects of chondrocyte proliferation. In contrast to chondrocyte-macroaggregates in the cell-DBM constructs, morphologically modified cells expressing type I collagen dominated. The redifferentiation of chondrocytes, characterized by the expression of type II collagen, was only found in low amounts in the cell-DBM constructs. Furthermore, caspase 3, a marker for apoptosis, was detected in the chondrocyte-DBM constructs. In another experimental setting, the vitality of chondrocytes as related to culture time and the amount of DBM was analyzed with the BrdU assay. Higher amounts of DBM tended to result in significantly higher proliferation rates of the cells within the first 48 h. After 96 h, the vitality decreased in a dose-dependent fashion. In conclusion, this study provides the proof of concept of chondrocyte-macroaggregates with DBM as an interesting method for the tissue engineering of cartilage. The as-yet insufficient redifferentiation of the chondrocytes and the sporadic initiation of apoptosis will require further investigations.
组织工程是利用分离的自体细胞和生物材料支架来生成软骨组织的重要方法。与各种凝胶状生物材料不同，人脱矿骨基质（DBM）可为生物力学负荷区域的应用提供一定的生物力学稳定性。本研究首次将软骨细胞大聚集体接种于DBM的方法用于软骨组织工程。分离出人鼻软骨细胞并形成三维大聚集体排列后，将DBM与大聚集体进行体外培养。分析了DBM内细胞在细胞分化和软骨细胞增殖抑制作用方面的相互作用。与细胞-DBM构建体中的软骨细胞大聚集体相比，表达I型胶原的形态学修饰细胞占主导。以II型胶原表达为特征的软骨细胞再分化在细胞-DBM构建体中仅少量发现。此外，在软骨细胞-DBM构建体中检测到凋亡标志物半胱天冬酶3。在另一个实验设置中，用BrdU检测法分析了软骨细胞活力与培养时间和DBM量的关系。较高量的DBM在前48小时往往导致细胞增殖率显著更高。96小时后，活力呈剂量依赖性下降。总之，本研究提供了以DBM为载体的软骨细胞大聚集体作为软骨组织工程有趣方法的概念验证。软骨细胞再分化尚不充分以及凋亡的零星启动需要进一步研究。
