Silva J.M. - Nanostructured 3D Constructs Based on Chitosan and Chondroitin Sulphate Multilayers for Cartilage Tissue Engineering

Joana M. Silva, Nicole Georgi, Rui Costa, Praveen Sher, Rui L. Reis, Clemens A. Van Blitterswijk, Marcel Karperien, Joao F. Mano

PLOS 2013


Nanostructured three-dimensional constructs combining layer-by-layer technology (LbL) and template leaching were
processed and evaluated as possible support structures for cartilage tissue engineering. Multilayered constructs were
formed by depositing the polyelectrolytes chitosan (CHT) and chondroitin sulphate (CS) on either bidimensional glass
surfaces or 3D packet of paraffin spheres. 2D CHT/CS multi-layered constructs proved to support the attachment and
proliferation of bovine chondrocytes (BCH). The technology was transposed to 3D level and CHT/CS multi-layered
hierarchical scaffolds were retrieved after paraffin leaching. The obtained nanostructured 3D constructs had a high porosity
and water uptake capacity of about 300%. Dynamical mechanical analysis (DMA) showed the viscoelastic nature of the
scaffolds. Cellular tests were performed with the culture of BCH and multipotent bone marrow derived stromal cells (hMSCs)
up to 21 days in chondrogenic differentiation media. Together with scanning electronic microscopy analysis, viability tests
and DNA quantification, our results clearly showed that cells attached, proliferated and were metabolically active over the
entire scaffold. Cartilaginous extracellular matrix (ECM) formation was further assessed and results showed that GAG
secretion occurred indicating the maintenance of the chondrogenic phenotype and the chondrogenic differentiation of