Chondrocyte differentiation for auricular cartilage reconstruction using a chitosan based hydrogel
J. García-López1,2, D. Garciadiego-Cázares2, Y. Melgarejo-Ramírez1, R. Sánchez-Sánchez1, L. Solís-Arrieta1, Z. García-Carvajal2, J.I. Sánchez-Betancourt4, C. Ibarra2, G. Luna-Bárcena3* and C. Velasquillo1*
1Biotecnology Laboratory, 2Tissue Engineering, Cell Therapy and Regenerative Medicine Unit. National Institute of Rehabilitation, Mexico City, 3Polymer and Biopolymer Research Group, CINVESTAV, Queretaro and 4Faculty of Veterinary Medicine, National Autonomous University of Mexico, Mexico City, Mexico
*These authors contributed equally to this work
Offprint requests to: Cristina Velasquillo PhD., Laboratorio de Biotecnologia, Instituto Nacional de Rehabilitación, Calz, México-Xochimilco 289, Arenal de Guadalupe. Tlalpan CP 14389, México Distrito Federal, México. e-mail: mvelasquillo@inr.gob.mx
Summary. Tissue engineering with the use of biodegradable and biocompatible scaffolds is an interesting option for ear repair. Chitosan-Polyvinyl alcohol-Epichlorohydrine hydrogel (CS-PVA-ECH) is biocompatible and displays appropriate mechanical properties to be used as a scaffold. The present work, studies the potential of CS-PVA-ECH scaffolds seeded with chondrocytes to develop elastic cartilage engineered-neotissues. Chondrocytes isolated from rabbit and swine elastic cartilage were independently cultured onto CS-PVA-ECH scaffolds for 20 days to form the appropriate constructs. Then, in vitro cell viability and morphology were evaluated by calcein AM and EthD-1 assays and Scanning Electron Microscopy (SEM) respectively, and the constructs were implanted in nu/nu mice for four months, in order to evaluate the neotissue formation. Histological analysis of the formed neotissues was performed by Safranin O, Toluidine blue (GAG's), Verhoeff-Van Gieson (elastic fibers), Masson's trichrome (collagen) and Von Kossa (Calcium salts) stains and SEM. Results indicate appropriate cell viability, seeded with rabbit or swine chondrocyte constructs; nevertheless, upon implantation the constructs developed neotissues with different characteristics depending on the animal species from which the seeded chondrocytes came from. Neotissues developed from swine chondrocytes were similar to auricular cartilage, while neotissues from rabbit chondrocytes were similar to hyaline cartilage and eventually they differentiate to bone. This result suggests that neotissue characteristics may be influenced by the animal species source of the chondrocytes isolated. Histol Histopathol 30, 1477-1485 (2015)
Key words: Auricular cartilage, Microtia, Chondrocyte differentiation, Chitosan hydrogel, Endochondral ossification
DOI: 10.14670/HH-11-642