title: Adenoviral transduction supports matrix expression of alginate cultured articular chondrocytes.
abstract: The present study examines the effects of adenoviral (Ad) transduction of human primary chondrocyte on transgene expression and matrix production. Primary chondrocytes were isolated from healthy articular cartilage and from cartilage with mild osteoarthritis (OA), transduced with an Ad vector and either immediately cultured in alginate or expanded in monolayer before alginate culture. Proteoglycan production was measured using dimethylmethylene blue (DMMB) assay and matrix gene expression was quantified by real-time PCR. Viral infection of primary chondrocytes results in a stable long time transgene expression for up to 13 weeks. Ad transduction does not significantly alter gene expression and matrix production if chondrocytes are immediately embedded in alginate. However, if expanded prior to three dimension (3D) culture in alginate, chondrocytes produce not only more proteoglycans compared to non-transduced controls, but also display an increased anabolic and decreased catabolic activity compared to non-transduced controls. We therefore suggest that successful autologous chondrocyte transplantation (ACT) should combine adenoviral transduction of primary chondrocytes with expansion in monolayer followed by 3D culture. Future studies will be needed to investigate whether the subsequent matrix production can be further improved by using Ad vectors bearing genes encoding matrix proteins.
authors.affiliation: University of Rostock, Medical Faculty, Institute of Immunology, Rostock, Germany. dpohle@gmx.net
authors.forename: D
authors.initials: D
authors.lastname: Pohle
authors.forename: R
authors.initials: R
authors.lastname: Kasch
authors.forename: P
authors.initials: P
authors.lastname: Herlyn
authors.forename: R
authors.initials: R
authors.lastname: Bader
authors.forename: T
authors.initials: T
authors.lastname: Mittlmeier
authors.forename: B M
authors.initials: BM
authors.lastname: Pützer
authors.forename: B
authors.initials: B
authors.lastname: Müller-Hilke
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000368:Aged; D000369:Aged, 80 and over; D000464:Alginates / Q000737:chemistry*; D002358:Cartilage, Articular / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D005260:Female; D005270:Femur Head / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D005819:Genetic Markers; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D018709:Statistics, Nonparametric; D014161:Transduction, Genetic; D014162:Transfection / Q000379:methods
chemical_list: D000464:Alginates; D016326:Extracellular Matrix Proteins; D005819:Genetic Markers; D006603:Hexuronic Acids; D020723:Glucuronic Acid