title: Temporal matrix synthesis and histologic features of a chondrocyte-laden porous collagen cartilage analogue.
abstract: Cartilage resurfacing by chondrocyte transplantation, using porous collagen matrices as a vehicle to secure the cells in cartilage defects, has been used experimentally in animals. This in vitro study evaluated the temporal morphologic features and proteoglycan synthesis of chondrocyte-laden collagen matrices. Forty-two porous collagen disks were implanted with a minimum of 6 x 10(6) viable chondrocytes, covered by a polymerized collagen gel layer, and 6 disks were harvested after 0, 3, 7, 10, 14, 18, or 22 days of incubation in supplemented Ham's F12 medium at 37 C and 5% CO2. Histologic and histochemical evaluation of formalin-fixed segments of the cultured disks indicated that the chondrocytes proliferated in the implant, producing small groups and linear segments of cells by day 14. The collagen framework remained intact over the course of the study with thick areas attributable to depositions of matrix material after day 10. Alcian blue-stained matrix was evident in the pericellular region of chondrocytes in sections of disks harvested on days 14, 18, and 22. Glycosaminoglycan (GAG) assay by dimethylmethylene blue dye binding after papain digestion of the disk segments revealed negligible amounts of GAG at day 0. Significant (P < or = 0.0001) increase in total GAG content was observed by day 3 (0.329 micrograms/mg of disk) and further increases were observed until a plateau in GAG quantity was seen on day 14. Mean peak GAG content was 0.553 +/- 0.062 micrograms/mg. Secondary treatment of the papain-digested implants with keratanase and chondroitinase ABC yielded similar trends in chondroitin sulfate (CS) and keratan sulfate (KS) concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.
authors.forename: A J
authors.initials: AJ
authors.lastname: Nixon
authors.forename: A E
authors.initials: AE
authors.lastname: Sams
authors.forename: G
authors.initials: G
authors.lastname: Lust
authors.forename: D
authors.initials: D
authors.lastname: Grande
authors.forename: H O
authors.initials: HO
authors.lastname: Mohammed
mesh_terms: D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D002455:Cell Division; D002478:Cells, Cultured; D002809:Chondroitin Sulfates / Q000378:metabolism; D003094:Collagen; D005109:Extracellular Matrix / Q000378:metabolism; D006025:Glycosaminoglycans / Q000096:biosynthesis; D006736:Horses; D007632:Keratan Sulfate / Q000378:metabolism
chemical_list: D006025:Glycosaminoglycans; D002809:Chondroitin Sulfates; D003094:Collagen; D007632:Keratan Sulfate