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.
Marijnissen
W J
WJ
Department of Orthopaedics, Head and Neck Surgery, Erasmus University Medical Center, Rotterdam, The Netherlands.
van Osch
G J
GJ
Aigner
J
J
Verwoerd-Verhoef
H L
HL
Verhaar
J A
JA
D000423:Alcian Blue; D000464:Alginates; D000818:Animals; D001187:Artificial Organs / Q000009:adverse effects; D001673:Biodegradation, Environmental; D001857:Bone Matrix; D002358:Cartilage, Articular / Q000166:cytology*; D002417:Cattle; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002478:Cells, Cultured / Q000637:transplantation; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D003094:Collagen / Q000032:analysis; D004396:Coloring Agents; D005549:Foreign-Body Reaction / Q000209:etiology; D005782:Gels; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D019344:Lactic Acid; D008297:Male; D008422:Materials Testing; D051379:Mice; D008819:Mice, Nude; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers; D019736:Prostheses and Implants; D020033:Protein Isoforms / Q000032:analysis; D014783:Viscosity
D000464:Alginates; D004396:Coloring Agents; D005782:Gels; D006603:Hexuronic Acids; D011108:Polymers; D020033:Protein Isoforms; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D020723:Glucuronic Acid; D003094:Collagen; D000423:Alcian Blue