Tissue-engineered cartilage using serially passaged articular chondrocytes. Chondrocytes in alginate, combined in vivo with a synthetic (E210) or biologic biodegradable carrier (DBM)
In vitro multiplication of isolated autologous chondrocytes is required to obtain an adequate number of cells to generate neo-cartilage, but is known to induce cell-dedifferentiation. The aim of this study was to investigate whether multiplied chondrocytes can be used to generate neo-cartilage in vivo. Adult bovine articular chondrocytes, of various differentiation stages, were suspended in alginate at densities of 10 or 50 million/ml, either directly after isolation (P0) or after multiplication in monolayer for one (P1) or three passages (P3). Alginate with cells was seeded in demineralized bovine bone matrix (DBM) or a fleece of polylactic/polyglycolic acid (E210) and implanted in nude mice for 8 weeks. The newly formed tissue was evaluated by Alcian Blue and immunohistochemical staining for collagen type-II and type-I. Structural homogeneity of the tissue, composed of freshly isolated as well as serially passaged cells, was found to be enhanced by high-density seeding (50 million/ml) and the use of E210 as a carrier. The percentage of collagen type-II positive staining P3-cells was generally higher when E210 was used as a carrier. Furthermore, seeding P3-chondrocytes at the highest density (50 million/ml) enhanced collagen type-II expression. This study shows promising possibilities to generate structurally regular neo-cartilage using multiplied chondrocytes in alginate in combination with a fleece of polylactic/polyglycolic acid.
|Keywords||*Artificial Organs/adverse effects, Alcian Blue, Alginates, Animals, Biodegradation, Environmental, Bone Matrix, Cartilage, Articular/*cytology, Cattle, Cell Count, Cell Culture Techniques/methods, Cell Differentiation, Cells, Cultured/transplantation, Chondrocytes/*cytology/metabolism/transplantation, Collagen/analysis, Coloring Agents, Foreign-Body Reaction/etiology, Gels, Glucuronic Acid, Hexuronic Acids, Lactic Acid, Male, Materials Testing, Mice, Mice, Nude, Polyglycolic Acid, Polymers, Prostheses and Implants, Protein Isoforms/analysis, Viscosity|
|Persistent URL||dx.doi.org/10.1016/S0142-9612(99)00218-5, hdl.handle.net/1765/15602|
Marijnissen, W.J.C.M., van Osch, G.J.V.M., Aigner, J., Verwoerd-Verhoef, H.L., & Verhaar, J.A.N.. (2003). Tissue-engineered cartilage using serially passaged articular chondrocytes. Chondrocytes in alginate, combined in vivo with a synthetic (E210) or biologic biodegradable carrier (DBM). Biomaterials, 21(6), 571–580. doi:10.1016/S0142-9612(99)00218-5