Toward scaffold-based meniscus repair: effect of human serum, hyaluronic acid and TGF-ß3 on cell recruitment and re-differentiation

• Published in: Osteoarthritis and cartilage 2013-05, Vol.21 (5), p.773-781
• Main Authors: Freymann, U, Endres, M, Goldmann, U, Sittinger, M, Kaps, C
• Format: Article
• Language: English
• Subjects: 3D micro-mass cultures; Cell Culture Techniques - methods; Cell Differentiation - drug effects; Cell re-differentiation; Chemotaxis - drug effects; Chondrocytes - cytology; Chondrocytes - drug effects; Chondrocytes - physiology; Collagen - biosynthesis; Collagen - genetics; Culture Media, Conditioned; Dose-Response Relationship, Drug; Gene Expression Regulation - drug effects; Human serum; Humans; Hyaluronic acid; Hyaluronic Acid - administration & dosage; Hyaluronic Acid - pharmacology; Menisci, Tibial - cytology; Menisci, Tibial - drug effects; Meniscus repair; Rheumatology; Serum; Tibial Meniscus Injuries; Tissue Engineering - methods; Tissue Scaffolds; Transforming growth factor (TGF-β3); Transforming Growth Factor beta3 - pharmacology
• ISSN: 1063-4584; 1522-9653
• DOI: 10.1016/j.joca.2013.02.655

Summary Objective Repair approaches for the non-vascular meniscus are rarely developed. Recent strategies use scaffold-based techniques and inducing factors. The aim of the study was the investigation of cell recruitment and re-differentiation inducing factors for a scaffold-based meniscus repair approach. Method 3D cultivation of in vitro expanded human meniscus-derived cells was performed in high-density cultures supplemented with 25% hyaluronic acid (HA), 10% human serum (HS) or 10 ng/ml transforming growth factor (TGF-ß3) compared to untreated controls. The in vitro cell recruitment potential of different HS concentrations was tested by chemotaxis assay. Analysis of chondrocytic markers (type I, II, IX collagen and proteoglycans) was performed on protein and gene expression level. Results Cells were attracted by 1–20% HS. 3D cultures supplemented with 10% HS and 25% HA showed meniscus-like gene expression profiles at day 7 with significantly increased cartilage oligomeric matrix protein (COMP) and aggrecan expression levels in the HS group and a slightly increased profile in the HA group compared to control. The TGF-ß3 group showed an additional induction of gene expression levels for type II and type IX collagen . Histological findings confirmed these results by proteoglycan and type I collagen staining in all groups and type II collagen staining only in the TGF-ß3 group. Conclusion This study demonstrates that human meniscus cells are attracted by HS and allow for meniscal matrix formation in 3D culture in the presence of HA and HS, whereas TGF-ß3 additive does not initiate meniscal tissue. Regarding non-vascular meniscus repair, results of this study encourage scaffold-based repair approaches.