Department of Periodontology, Faculty of Dentistry, Hacettepe University, Beytepe, Ankara, Turkey.
Artif Organs. 2010 Jan;34(1):65-74. doi: 10.1111/j.1525-1594.2009.00798.x. Epub 2009 Oct 11.
The purpose of this study is to investigate the convenience of bone morphogenetic protein-6 (BMP-6)-loaded chitosan scaffolds with preosteoblastic cells for bone tissue engineering. MC3T3-E1 cells were seeded into three different groups: chitosan scaffolds, BMP-6-loaded chitosan scaffolds, and chitosan scaffolds with free BMP-6 in culture medium. Tissue-engineered constructs were characterized by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide assay, scanning electron microscopy (SEM), mineralization assay (von Kossa), alkaline phosphatase (ALP) activity, and osteocalcin (OCN) assays. BMP-6-loaded chitosan scaffolds supported proliferation of the MC3T3-E1 mouse osteogenic cells in a similar pattern as the unloaded chitosan scaffolds group and as the chitosan scaffolds with free BMP-6 group. SEM images of the cell-seeded scaffolds revealed significant acceleration of extracellular matrix synthesis in BMP-6-loaded chitosan scaffolds. Both levels of ALP and OCN were higher in BMP-6-loaded chitosan scaffold group compared with the other two groups. In addition, BMP-6-loaded scaffolds showed strong staining in mineralization assays. These findings suggest that BMP-6-loaded chitosan scaffold supports cellular functions of the osteoblastic cells; therefore, this scaffold is considered as a new promising vehicle for bone tissue engineering applications.
本研究旨在探讨载有骨形态发生蛋白-6（BMP-6）的壳聚糖支架与前成骨细胞在骨组织工程中的便利性。将 MC3T3-E1 细胞接种到三组不同的细胞中：壳聚糖支架、载有 BMP-6 的壳聚糖支架和含有游离 BMP-6 的壳聚糖支架在培养基中。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐测定法、扫描电子显微镜（SEM）、矿化测定法（von Kossa）、碱性磷酸酶（ALP）活性和骨钙素（OCN）测定法来对组织工程构建体进行特征描述。载有 BMP-6 的壳聚糖支架以与未负载的壳聚糖支架组和具有游离 BMP-6 的壳聚糖支架组相似的方式支持 MC3T3-E1 小鼠成骨细胞的增殖。细胞接种支架的 SEM 图像显示，载有 BMP-6 的壳聚糖支架中外细胞外基质合成得到显著加速。与其他两组相比，载有 BMP-6 的壳聚糖支架组的 ALP 和 OCN 水平均更高。此外，载有 BMP-6 的支架在矿化测定中显示出强烈的染色。这些发现表明，载有 BMP-6 的壳聚糖支架支持成骨细胞的细胞功能；因此，该支架被认为是骨组织工程应用的一种有前途的新型载体。
