ABSTRACT
As subchondral bone is often affected during cartilage injuries, the aim of research is to generate osteochondral implants in vitro using tissue engineering techniques. These constructs consist of a cartilage layer grown on top of a bone phase. In clinical applications, phosphate ceramics have gained acceptance as bone substitute materials because of their great affinity to natural bone. Furthermore, the interaction between cartilage and the underlying bone equivalent is essential for the development and success of osteochondral implants. Here, the influence of a carrier containing hydroxyapatite on the quality of cartilage constructs generated in vitro is investigated. Attempts are made to explain the effects described, by considering chemical and physical properties of the biomaterial.
Subject(s)
Bone Substitutes/chemistry , Cartilage, Articular/growth & development , Ceramics/chemistry , Chondrocytes/cytology , Chondrocytes/physiology , Durapatite/chemistry , Prostheses and Implants , Animals , Cartilage, Articular/cytology , Cell Adhesion/physiology , Cell Proliferation , Cells, Cultured , Surface Properties , SwineABSTRACT
OBJECTIVE: The aim of the current study was to identify molecular markers for articular cartilage (AC) that can be used as tools for the quality control of tissue engineered (TE) cartilage. DESIGN: A genome-wide expression analysis was performed using RNA isolated from articular and growth plate (GP) cartilage, both extracted from the knee joints of 6 weeks old minipigs. After confirming the specific expression for selected genes by RT-PCR, these were used as molecular markers for the quality control of TE cartilage. RESULTS: Albeit several known chondrocyte markers were expressed to a similar extent in articular and GP cartilage, our genome-wide expression analysis led us to identify genes being selectively expressed in either GP or articular chondrocytes. These findings led us to perform a RT-PCR expression analysis for the corresponding genes to demonstrate the absence of GP-specific markers in TE cartilage, while common or AC markers were expressed. CONCLUSIONS: Taken together, these results provide important novel insights into chondrocyte biology in general and AC in particular. In addition, it is reasonable to speculate, that some of the identified genes play distinct roles in the regulation of articular chondrocyte differentiation and/or function, thereby raising the possibility that they may serve as targets for non-operative therapies of osteoarthritis (OA).
Subject(s)
Cartilage, Articular/cytology , Chondrocytes/metabolism , Tissue Engineering/methods , Animals , Biomarkers , Cartilage, Articular/anatomy & histology , Cartilage, Articular/metabolism , Gene Expression , Gene Expression Profiling/methods , Genetic Markers , Genome-Wide Association Study/methods , Growth Plate/metabolism , Reverse Transcriptase Polymerase Chain Reaction/methods , Swine , Swine, MiniatureABSTRACT
Technical aspects play an important role in tissue engineering. Especially an improved design of bioreactors is crucial for cultivation of artificial three-dimensional tissues in vitro. Here formation of cartilage-carrier-constructs is used to demonstrate that the quality of the tissue can be significantly improved by using optimized culture conditions (oxygen concentration, growth factor combination) as well as special bioreactor techniques to induce fluid-dynamic, hydrostatic or mechanical load during generation of cartilage.
Subject(s)
Bioreactors , Cartilage/physiology , Chondrocytes/physiology , Tissue Culture Techniques/instrumentation , Tissue Engineering/instrumentation , Alginates/chemistry , Animals , Cartilage/cytology , Cartilage/drug effects , Cell Differentiation , Chondrocytes/cytology , Chondrocytes/drug effects , Equipment Design , Gels , Humans , Intercellular Signaling Peptides and Proteins/pharmacology , Oxygen/metabolism , Rheology , Tissue Engineering/methods , Tissue ScaffoldsABSTRACT
Owing to the growing demand of cartilage tissue repair and transplants, engineered cartilage cells have emerged as a prospective solution. Several bioreactors were built for artificially grown cartilage cells. In this work, a recently designed flow bed bioreactor is numerically investigated and compared with experimental results. The flow field inside the bioreactor was modelled using the lattice Boltzmann method. The flow consists of two phases which are the liquid component (nutrition supply) and gas component (oxygen supply). The flow field is simulated using the multi-phase lattice Boltzmann method, whilst the cell activity is modelled using Michaelis-Menten kinetics. The oxygen diffusion level at the exit of the nutrition phase is used as an evaluation process between the numerical and experimental results reporting the possibility of using the proposed model to fully simulate such bioreactors, though greatly saving time and money. Shear stress and pressure distributions are as well compared with published human cartilage load measurements to estimate the dynamic similarity between the bioreactor and the human knee. The predicted oxygen levels proved consistent trends with the experimental work with a 7% difference after 1h measuring time. The shear stress levels recorded 10-11 orders of magnitude lower than in humans and also one order of magnitude lower in the pressure distribution.
