Machado C B, Ventura J M G, Lemos A F, Ferreira J M F, Leite M F, Goes A M
Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais, Brazil.
Biomed Mater. 2007 Jun;2(2):124-31. doi: 10.1088/1748-6041/2/2/010. Epub 2007 Apr 17.
A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation.
开发了一种多孔3D支架，以支持和增强间充质干细胞（MSC）在体外分化为成骨细胞的过程。该3D支架由壳聚糖、明胶和软骨素制成，并通过EDAC交联。对支架的物理化学性质进行了评估。扫描电子显微镜（SEM）显示支架具有高孔隙率和相互连通的孔隙；流变学测量表明，该支架表现出强凝胶的特征行为。交联支架压缩试验中的弹性模量比未交联支架高约50倍。21天后，经历水解降解的3D基质重量损失超过40%。从大鼠骨髓中收集MSC，并接种到壳聚糖-明胶-软骨素3D支架以及2D培养板中。将MSC分化为成骨细胞21天。在成骨过程中每周监测细胞增殖和碱性磷酸酶活性。MTT法和碱性磷酸酶活性表明，3D培养中MSC的成骨分化得到改善。在第21天，通过PCR分析检测骨标志物骨桥蛋白和骨钙素。这项研究表明，壳聚糖-明胶-软骨素3D结构为成骨过程提供了良好的环境，并增强了细胞增殖。
