Nanostructured 3D constructs based on chitosan and chondroitin sulphate multilayers for cartilage tissue engineering

• Published in: PloS one Vol. 8; no. 2; p. e55451
• Main Authors: Silva, Joana M, Georgi, Nicole, Costa, Rui, Sher, Praveen, Reis, Rui L, Van Blitterswijk, Clemens A, Karperien, Marcel, Mano, João F
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 20.02.2013; Public Library of Science (PLoS)
• Subjects: Analysis; Animals; Arthritis; Bioengineering; Biology; Biomedical materials; Bone marrow; Cartilage; Cartilage - drug effects; Cartilage - physiology; Cattle; Cell Adhesion - drug effects; Cell culture; Cell Differentiation - drug effects; Cell Proliferation - drug effects; Cell Shape - drug effects; Cell Survival - drug effects; Chickens; Chitosan; Chitosan - pharmacology; Chondrocytes; Chondrocytes - cytology; Chondrocytes - drug effects; Chondrocytes - ultrastructure; Chondroitin; Chondroitin sulfate; Construction engineering; Deoxyribonucleic acid; Differentiation; DNA; DNA - metabolism; Elastic Modulus - drug effects; Electrolytes; Engineering; Extracellular matrix; Gene expression; Genotype & phenotype; Humans; Hydration; Hydrogels; Laboratories; Leaching; Materials Science; Mechanical analysis; Medicine; Microscopy; Multilayers; Nanostructure; Nanostructures - chemistry; Nanostructures - ultrastructure; Paraffin; Polyelectrolytes; Porosity; Quartz Crystal Microbalance Techniques; Scaffolds; Secretion; Spectroscopy, Fourier Transform Infrared; Stromal cells; Sulfates; Technology; Tissue engineering; Tissue Engineering - methods; Tissue Scaffolds - chemistry; Viability; Viscoelasticity; Water uptake; Portugal; Netherlands
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0055451

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 hMSCs.