Meniscus cells seeded in type I and type II collagen-GAG matrices in vitro.
The objective of this study was to determine the proliferative and biosynthetic activity of calf meniscus cells seeded in type I and type II collagen-glycosaminoglycan (GAG) copolymers with the overall goal to develop a cell-seeded implant for future investigations to improve the regeneration of the knee meniscus. The cell-seeded matrices were digested in protease and analyzed for GAG by a modification of the dimethyl-methylene blue method and assayed for DNA content. Other specimens were evaluated histologically after 1, 7, 14 and 21 days. Contraction of the same types of matrices, seeded with adult canine meniscus cells, was measured at the same time points. After three weeks, cells were observed throughout the type II matrix, whereas the type I matrix was densely populated at the margins. The cell morphology and the cell density after three weeks in both matrices was consistent with the normal meniscus. DNA assay for the type I matrix showed a 40% decrease over the first week and a final amount of DNA that was not significantly different from the initial value, whereas the type II matrix doubled its DNA content over the same time period. The cells continued their biosynthesis of GAG and type I collagen. GAG content of the type II matrix increased by 50% more than the type I matrix after three weeks. Over the same time period, the type I matrix displayed a significant shrinkage to approximately 50% of its initial value whereas in contrast, the type II matrix and the unseeded controls showed no significant shrinkage. The number of cells and the higher GAG synthesis in the type II matrix, and its resistance to cell-mediated contracture, commend it for future investigation of the regeneration of meniscus in vivo.
Mueller
S M
SM
Department of Orthopedic Surgery, Brigham & Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Shortkroff
S
S
Schneider
T O
TO
Breinan
H A
HA
Yannas
I V
IV
Spector
M
M
D000818:Animals; D001672:Biocompatible Materials; D001696:Biomechanical Phenomena; D002417:Cattle; D002455:Cell Division; D003094:Collagen / Q000096:biosynthesis; D003470:Culture Media; D004247:DNA / Q000378:metabolism; D004285:Dogs; D006025:Glycosaminoglycans / Q000096:biosynthesis; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008422:Materials Testing; D008592:Menisci, Tibial / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D012038:Regeneration
D001672:Biocompatible Materials; D003470:Culture Media; D006025:Glycosaminoglycans; D003094:Collagen; D004247:DNA