Department of Polymer Engineering, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Taipas, Guimarães, Portugal.
Acta Biomater. 2011 Apr;7(4):1644-52. doi: 10.1016/j.actbio.2010.11.044. Epub 2010 Dec 3.
Cartilage tissue engineering (TE) typically involves the combination of a 3-D biodegradable polymeric support material, with primary chondrocytes or other cell types able to differentiate into chondrocytes. The culture environment in which cell-material constructs are created and stored is an important factor. Various bioreactors have been introduced in TE approaches to provide specific culturing environments that might promote and accelerate cells' potential for chondrogenic differentiation and enhance the production of cartilage extracellular matrix. The aim of the present study was to investigate the chondrogenic differentiation of goat bone marrow cells (GBMCs) under flow perfusion culture conditions. For that purpose, GBMCs were seeded into starch-polycaprolactone fiber mesh scaffolds and cultured in a flow perfusion bioreactor for up to 28 days using culture medium supplemented with transforming growth factor-β1. The tissue-engineered constructs were characterized after several end points (7, 14, 21 and 28 days) by histological staining and immunocytochemistry analysis, as well as by glycosaminoglycan and alkaline phosphatase quantification assays. In addition, the expression of typical chondrogenic markers was assessed by real-time reverse-transcription polymerase chain reaction analysis. In general, the results obtained suggest that a flow perfusion microenvironment favors the chondrogenic potential of GBMCs.
软骨组织工程（TE）通常涉及 3-D 可生物降解的聚合物支撑材料与能够分化为软骨细胞的原代软骨细胞或其他细胞类型的结合。细胞-材料构建体被创建和存储的培养环境是一个重要因素。已经在 TE 方法中引入了各种生物反应器，以提供可能促进和加速细胞软骨分化潜力并增强软骨细胞外基质产生的特定培养环境。本研究的目的是研究在流动灌注培养条件下山羊骨髓细胞（GBMC）的软骨分化。为此，将 GBMC 接种到淀粉-聚己内酯纤维网支架中，并在补充有转化生长因子-β1 的培养基中使用流动灌注生物反应器培养长达 28 天。通过组织学染色和免疫细胞化学分析以及糖胺聚糖和碱性磷酸酶定量测定，在几个终点（7、14、21 和 28 天）后对组织工程构建体进行了表征。此外，通过实时逆转录聚合酶链反应分析评估了典型的软骨形成标志物的表达。总的来说，所得结果表明，流动灌注微环境有利于 GBMC 的软骨形成潜力。
