BMP activation and Wnt-signalling affect biochemistry and functional biomechanical properties of cartilage tissue engineering constructs

Summary Objectives Bone morphogenetic protein (BMP-) and Wnt-signalling play crucial roles in cartilage homeostasis. Our objective was to investigate whether activation of the BMP-pathway or stimulation of Wnt-signalling cascades effectively enhances cartilage-specific extracellular matrix (ECM) accumulation and functional biomechanical parameters of chondrocyte-seeded tissue engineering (TE)-constructs. Design Articular chondrocytes were cultured in collagen-type-I/III-matrices over 6 weeks to create a biomechanical standard curve. Effects of stimulation with 100 ng/mL BMP-4/-7 heterodimer or 10 mM lithium chloride (LiCl) on ECM-deposition was quantified and characterized histologically. Biomechanical parameters were determined by the Very Low Rubber Hardness (VLRH) method and under confined compression stress relaxation. Results BMP-4/-7 treatment resulted in stronger collagen type-II staining and significantly enhanced glycosaminoglycan (GAG) deposition (3.2-fold; * P P  = 0.001) reaching 83% of native cartilage values after 28 days, a value not reached before 9 weeks without stimulation. LiCl treatment enhanced VLRH slightly, but significantly (∼1.3-fold; * P  = 0.016) with a trend to more ECM-deposition. BMP-4/-7 treatment significantly enhanced the E Modulus (105.7 ± 34.1 kPa; * P  = 0.000001) compared to controls (8.0 ± 4.2 kPa). Poisson's ratio was significantly improved by BMP-4/-7 treatment (0.0703 ± 0.0409; * P  = 0.013) vs controls (0.0432 ± 0.0284) and a significantly lower permeability (5.8 ± 2.1056 × 10 −14  mˆ4/N.s; * P  = 0.00001) was detected compared to untreated scaffolds (4.4 ± 3.1289 × 10 −13  mˆ4/N.s). Conclusions While Wnt-activation is less effective, BMP-4/-7 heterodimer stimulation approximated native cartilage features in less than 50% of standard culture time representing a promising strategy for functional cartilage TE to improve biomechanical parameters of engineered cartilage.