title: Chondrocyte-seeded collagen matrices implanted in a chondral defect in a canine model.
abstract: The objective of our study was to evaluate reparative tissues formed in chondral defects in an adult canine model implanted with cultured autologous articular chondrocytes seeded in type I and II collagen GAG matrices. Two defects were produced in the trochlea grooves of the knees of 21 dogs, with cartilage removed down to the tidemark. This study includes the evaluation of 36 defects distributed among five treatment groups: Group A, type II collagen matrix seeded with autologous chondrocytes under a sutured type II collagen flap; Group B, type I collagen matrices seeded with chondrocytes under a sutured fascia flap; Group C, unseeded type I collagen matrix implanted under a sutured fascia flap; Group D, fascia lata flap alone; and Group E, untreated defects. All animals were killed 15 weeks after implantation. Six other defects were created at the time of death and evaluated immediately after production as 'acute defect controls'. In three additional defects, unseeded matrices were sutured to the defect and the knee closed and reopened after 30 min to determine if early displacement of the graft was occurring; these defects served as 'acute implant controls'. The areal percentages of four tissue types in the chondral zone of the original defect were determined histomorphometrically: fibrous tissue (FT); hyaline cartilage (HC); transitional tissue (TT, including fibrocartilage); and articular cartilage (AC). New tissue formed in the remodeling subchondral bone underlying certain defects was also assessed. Bonding of the repair tissue to the subchondral plate and adjacent cartilage, and degradation of the adjacent tissues were evaluated. There were no significant differences in the tissues filling the original defect area of the sites treated with chondrocyte-seeded type I and type II matrices. Most of the tissue in the area of the original defect in all of the groups was FT and TT. The areal percentage of HC plus AC was highest in group E, with little such tissue in the cell-seeded groups, and none in groups C and D. The greatest total amount of reparative tissue, however, was found in the cell-seeded type II matrix group. Moreover, examination of the reparative tissue formed in the subchondral region of defects treated with the chondrocyte-seeded collagen matrices (Groups A and B) demonstrated that the majority of the tissue was positive for type II collagen and stained with safranin O. These results indicate an influence of the exogenous chondrocytes on the process of chondrogenesis in this site. In all groups with implants (A-D), 30(50% of the FT and TT was bonded to the adjacent cartilage. Little of this tissue (6-22%) was attached to the subchondral plate, which was only about 50% intact. Remarkable suture damage was found in sections from each group in which sutures were used. Harvest sites showed no regeneration of normal articular cartilage, 18 weeks after the biopsy procedure. Future studies need to investigate other matrix characteristics, and the effects of cell density and incubation of the seeded sponges prior to implantation on the regenerative response.
authors.affiliation: Department of Orthopedic Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA. stefan.nehrer@akh-wien.ac.at
authors.forename: S
authors.initials: S
authors.lastname: Nehrer
authors.forename: H A
authors.initials: HA
authors.lastname: Breinan
authors.forename: A
authors.initials: A
authors.lastname: Ramappa
authors.forename: H P
authors.initials: HP
authors.lastname: Hsu
authors.forename: T
authors.initials: T
authors.lastname: Minas
authors.forename: S
authors.initials: S
authors.lastname: Shortkroff
authors.forename: C B
authors.initials: CB
authors.lastname: Sledge
authors.forename: I V
authors.initials: IV
authors.lastname: Yannas
authors.forename: M
authors.initials: M
authors.lastname: Spector
mesh_terms: D000818:Animals; D001705:Bioprosthesis; D002358:Cartilage, Articular / Q000166:cytology / Q000293:injuries / Q000637:transplantation*; D017690:Cell Transplantation / Q000379:methods; D003094:Collagen; D003240:Connective Tissue Diseases / Q000601:surgery*; D004285:Dogs; D006025:Glycosaminoglycans; D007592:Joint Diseases / Q000601:surgery*; D011108:Polymers; D013264:Stifle; D014182:Transplantation, Autologous
chemical_list: D006025:Glycosaminoglycans; D011108:Polymers; C058506:collagen-glycosaminoglycan copolymer; D003094:Collagen