Effect of flow perfusion on the osteogenic differentiation of bone marrow stromal cells cultured on starch‐based three‐dimensional scaffolds

Abstract

This study aims to investigate the effect of culturing conditions (static and flow perfusion) on the proliferation and osteogenic differentiation of rat bone marrow stromal cells seeded on two novel scaffolds exhibiting distinct porous structures. Specifically, scaffolds based on SEVA-C (a blend of starch with ethylene vinyl alcohol) and SPCL (a blend of starch with polycaprolactone) were examined in static and flow perfusion culture. SEVA-C scaffolds were formed using an extrusion process, whereas SPCL scaffolds were obtained by a fiber bonding process. For this purpose, these scaffolds were seeded with marrow stromal cells harvested from femoras and tibias of Wistar rats and cultured in a flow perfusion bioreactor and in 6-well plates for 3, 7, and 15 days. The proliferation and alkaline phosphatase activity patterns were similar for both types of scaffolds and for both culture conditions. However, calcium content analysis revealed a significant enhancement of calcium deposition on both scaffold types cultured under flow perfusion. This observation was confirmed by Von Kossa-stained sections and tetracycline fluorescence. Histological analysis and confocal images of the cultured scaffolds showed a much better distribution of cells within the SPCL scaffolds than the SEVA-C scaffolds, which had limited pore interconnectivity, under flow perfusion conditions. In the scaffolds cultured under static conditions, only a surface layer of cells was observed. These results suggest that flow perfusion culture enhances the osteogenic differentiation of marrow stromal cells and improves their distribution in three-dimensional, starch-based scaffolds. They also indicate that scaffold architecture and especially pore interconnectivity affect the homogeneity of the formed tissue.

Cell colonization in degradable 3D porous matrices

“…135 The presence of flow within a reactor also affects the production of ECM elements, for example rat bone marrow cells produce greater mineralization in scaffolds under direct perfusion. 136 Once the scaffold is placed into a flow system (either implanted in vivo or grown in vitro in a bioreactor) the effect of loading from both external forces and fluid flow can affect cell colonization. While the scaffold itself will be subjected to the bulk forces supplied by the tissue and fluid flow, the cells will experience the micromechanical properties of the individual fibers and local shear stresses within the porous structure (Fig.…”

Quantification of fluid shear stress in bone tissue engineering scaffolds with spherical and cubical pore architectures

“…However, direct measurement of fluid-induced shear stress within scaffolds is not feasible, which prevents researchers from correlating the levels of fluid shear 5 stress to biochemical responses, see in particular (Gomes et al 2003;Jagodzinski et al 2008;Liu et al 2012a) and Table 1. Therefore, researchers are required to employ analytical predictions, based on idealised flow through a cylinder or two plates (Blecha et al 2010;Goldstein et al 2001), or estimate wall shear stress (WSS) magnitudes from existing computational models (Bancroft et al 2002;Grayson et al 2008;Vance et al 2005;Yu et al 2004), see Table 1.…”

Nano-ceramic Composite Scaffolds for Bioreactor-based Bone Engineering

“…MSCs of different species might exhibit peak marker expression at different times; however, the general pattern of production is relatively conserved because it is related to each marker's function. ALP usually reaches a maximum that coincides with early osteoblastic differentiation and decreases afterward when matrix maturation and mineralization begin [28]. Additionally, OCN and OPN characterization by enzymelinked immunosorbent assay further confirmed the osteoblastic differentiation state of HMSCs (data not shown).…”