Differentiation of mesenchymal stem cells in chitosan scaffolds with double micro and macroporosity

Abstract

Bone Marrow mesenchymal stem cells can be induced to differentiate into osteoblasts to regenerate damaged bone tissue using tissue engineering techniques. In this study, we examine the use of chitosan scaffolds with double pore structure prepared by an innovative method that combines freeze gelation (that produces micropores) and particle leaching out technique (that produces interconnected spherical macropores) seeking to enhance the osteogenic differentiation of goat bone marrow stromal cells (GBMSCs). The double pore architecture of the scaffold was characterized by scanning electron microscopy (SEM), microcomputed tomography and confocal laser scanning microscopy. The obtained hierarchical pore structure allowed very efficient seeding of GBMSCs that are able to occupy the whole volume of the scaffold, showing good adhesion and proliferation. GBMSCs were differentiated into osteoblasts as indicated by alkaline phosphatase activity and osteocalcin expression. The results of this study demonstrate that chitosan scaffold may be promising biomaterial for bone regeneration.

Poly(3-hydroxybutyrate)/poly(ethylene glycol) scaffolds with different microstructure: the effect on growth of mesenchymal stem cells

“…Scaffolds S2 showed also the lowest growth of 3T3 fibroblasts on 6th day of cell cultivation. The values of cell viability estimated by XTT were similar to the values obtained in other experiments for study of cell growth in polymer scaffolds by other colorimetric assays for assessing cell metabolic activity (Zhang et al 2010;Cruz et al 2010;Ribeiro-Samy et al 2013).…”

“…The observed data on the growth of cells on polymer scaffolds can be connected both with the properties of PHB/PEG composite and the morphology of these scaffolds. The physicochemical (e.g., mechanical) properties of polymer materials have a great influence on growth and differentiation of stem cells grown on them (Cruz et al 2010;Raucci et al 2012;Rumian et al 2013;Humpolicek et al 2015;Fraioli et al 2016;Zhang et al 2016;Kuznetsova et al 2016). Moreover, apparently PHB is an example of such biomaterial possessing osteoinductive properties: scaffolds based on PHB can induce spontaneous differentiation of stem cells and stimulate bone tissue regeneration in vivo (Wang et al 2004;Bonartsev et al 2008;Shishatskaya et al 2014;Lim et al 2017).…”

“…Scaffolds with mean pore sizes ranging from 20 to 1500 µm were used in bone tissue engineering applications (Richardson et al 2006;Oh et al 2007;Zhang et al 2010;Moisenovich et al 2015;Rumian et al 2013;Kuznetsova et al 2016). The minimum pore size for a significant bone growth and MSC cultivation is 40 µm with an optimal range of 100-200 µm (Klawitter et al 1976;Richardson et al 2006;Cruz et al 2010;Misra et al 2010;Zhang et al 2010;Murphy et al 2010;Ribeiro-Samy et al 2013;Shishatskaya et al 2014;Hadjicharalambous et al 2015). However, as shown in the study of growth of different kinds of cells (chondrocytes, osteoblasts, and fibroblasts) in cylindrical polycaprolactone scaffolds with gradually increasing pore size (from 88 to 405 µm) and porosity (from 80 to 94%), the difference in scaffold microstructure is relevant only for long-term cultivation of cells, while the growth of cells with a duration of their cultivation of less than 14 days was the same and did not depended on pore size and porosity (Oh et al 2007).…”

Chitosan-based scaffolds for bone tissue engineering

“…Chitosan solution concentration affects pore diameter, pore wall thickness and thus scaffold mechanical properties whereby higher concentrations correlate with smaller pores and thicker pore walls. 84 Whereas lyophilization can be time and energy intensive, freeze gelation can be a more efficient process that also minimizes the presence of residual acetic acid in scaffolds. 85 Care must be taken, however, to optimize the freeze gelation system to minimize local melting during gelation.…”

Stirred flow bioreactor modulates chondrocyte growth and extracellular matrix biosynthesis in chitosan scaffolds

“…Chitosan scaffolds with hierarchical pore structure were prepared by a combination of freeze gelation and leaching out technique of PEMA microspheres, used as porogen as described in previous work 23. Briefly, CHT solutions with concentration of 4 wt % were poured into a cylindrical mold (7 mm diameter and 25 mm thickness), mixed with 70 wt % of PEMA microspheres and frozen in liquid nitrogen (−196°C).…”