ABSTRACT
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 < 0.01) correlating with improved hardness (â¼1.7-fold; *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 (â¼1.3-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) m4/N.s; *P = 0.00001) was detected compared to untreated scaffolds (4.4 ± 3.1289 × 10(-13) m4/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.
Subject(s)
Bone Morphogenetic Proteins/physiology , Cartilage, Articular/physiology , Tissue Engineering/methods , Wnt Signaling Pathway/physiology , Animals , Biomechanical Phenomena , Bone Morphogenetic Protein 4/pharmacology , Bone Morphogenetic Protein 7/pharmacology , Cartilage, Articular/cytology , Cartilage, Articular/metabolism , Cells, Cultured , Chondrocytes/drug effects , Chondrocytes/metabolism , Collagen Type II/metabolism , Extracellular Matrix/physiology , Glycosaminoglycans/metabolism , Hardness , Lithium Chloride/pharmacology , Sus scrofaABSTRACT
Specific biomechanical properties represent important quality markers of cartilage tissue engineering (TE) constructs. The aim of the study was to identify a sensitive biomechanical test to assess mechanical properties of cartilage TE constructs. Biomechanical testing of in vitro cultivated constructs following the very low rubber hardness (VLRH) principle illustrated significant differences between constructs cultured under chondrogenic conditions over various periods of time. An increase in proteoglycan and collagen type II deposition corresponded to increasing VLRH hardness values. Although a decrease in proteoglycan was detected after ectopic implantation of constructs into SCID mice, no reduction in biomechanical hardness values was observed. A functional estimation of TE constructs requires determination of biomechanical and biochemical parameters as quality features.
Subject(s)
Biocompatible Materials/chemistry , Fractures, Cartilage/physiopathology , Fractures, Cartilage/surgery , Guided Tissue Regeneration/instrumentation , Regeneration/physiology , Tissue Scaffolds , Animals , Biomechanical Phenomena , Equipment Design , Equipment Failure Analysis/methods , Fractures, Cartilage/pathology , Humans , Materials Testing/methods , Swine , Treatment OutcomeABSTRACT
Values for the friction coefficient of articular cartilage are given in ranges of percentage and lower and are calculated as a quotient of the friction force and the perpendicular loading force acting on it. Thus, a sophisticated system has to be provided for analysing the friction coefficient under different conditions in particular when cartilage should be coupled as friction partner. It is possible to deep-freeze articular cartilage before measuring the friction coefficient as the procedure has no influence on the results. The presented tribological system was able to distinguish between altered and native cartilage. Furthermore, tissue engineered constructs for cartilage repair were differentiated from native cartilage probes by their friction coefficient. In conclusion a tribological equipment is presented to analyze the friction coefficient of articular cartilage, in vivo generated cartilage regenerates and in vitro tissue engineered constructs regarding their biomechanical properties for quality assessment.
