Fibrin gel improved the spatial uniformity and phenotype of human chondrocytes seeded on collagen scaffolds.
A scaffold made of equine collagen type I based material has been assessed for its use in the preparation of tissue-engineered cartilage implants with human articular chondrocytes. Improvements of cell-seeding efficiency and specific gene expression were studied by combining solid scaffold with fibrin glue or human blood plasma. Following 3 weeks of static culture, mRNA expression levels of collagen type I, collagen type II, aggrecan and versican were analyzed by real-time quantitative PCR and compared to those in native cartilage and monolayer cell cultures. Constructs prepared with fibrin glue or plasma showed higher cell seeding efficiencies than those prepared without gel. Chondrocytes seeded directly onto a collagen scaffold appeared fibroblastic in shape while those encapsulated in fibrin gel were spherical. The presence of fibrin glue positively influences on mRNA levels of collagen type II and aggrecan, while blood plasma enhanced only the level of collagen type II expression. Levels of collagen type I and versican decreased in presence of fibrin glue. In orthopaedics, the combination of solid collagen fleece with fibrin gel for implant preparation is seen to be preferred over solid material or even cells in a suspension, since fibrin gel improves seeding capacity of the scaffold, supports equal distribution of cells and stimulates higher chondrogenic phenotype expression.
Malicev
Elvira
E
Educell d.o.o., Letaliska c. 33, 1000 Ljubljana, Slovenia. elvira.malicev@educell.si
Radosavljevic
Damjan
D
Velikonja
Nevenka Kregar
NK
D000328:Adult; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000737:chemistry* / Q000187:drug effects; D003094:Collagen / Q000737:chemistry* / Q000235:genetics; D015718:Fibrin Tissue Adhesive / Q000494:pharmacology; D006801:Humans; D008297:Male; D010641:Phenotype; D012313:RNA / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods
D015718:Fibrin Tissue Adhesive; D012313:RNA; D003094:Collagen