alcian-blue and Foreign-Body-Reaction

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.

Topics: Alcian Blue; Alginates; Animals; Artificial Organs; Biodegradation, Environmental; Bone Matrix; Cartilage, Articular; Cattle; Cell Count; Cell Culture Techniques; Cell Differentiation; Cells, Cultured; Chondrocytes; Collagen; Coloring Agents; Foreign-Body Reaction; Gels; Glucuronic Acid; Hexuronic Acids; Lactic Acid; Male; Materials Testing; Mice; Mice, Nude; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Polymers; Prostheses and Implants; Protein Isoforms; Viscosity