Effects of Cell Seeding Methods on Chondrogenic Differentiation of Rat Mesenchymal Stem Cells in Polyhydroxybutyrate/Chitosan Scaffolds

Abstract

The aim of our study was to examine the effects of passive and active cell seeding techniques on in vitro chondrogenic differentiation of mesenchymal stem cells (MSC) isolated from rat bone marrow and seeded on porous biopolymer scaffolds based on polyhydroxybutyrate/chitosan (PCH) blends. This paper is focused on the distribution of the cells on and in the scaffolds, since it influences the uniformity of the created extracellular matrix (ECM), as well as the homogenity of the distribution of chondrogenic markers in vitro which ultimately affects the quality of the newly created tissue after in vivo implantation. The three types of cell-scaffold constructs were examined by: fluorescence microscopy, SEM, histology and quantitative analysis of the glycosaminoglycans after chondrogenic cultivation. The results demonstrated that the active cells seeded via the centrifugation of the cell suspension onto the scaffold guaranteed an even distribution of cells on the bulk of the scaffold and the uniform secretion of the ECM products by the differentiated cells.

Keywords

• bone marrow
• glycosaminoglycans
• ecm
• mesenchymal stem cells
• polyhydroxybutyrate/chitosan scaffold