Three-dimensional cartilage tissue engineering using adult stem cells from osteoarthritis patients

Southmead Hospital, and University of Bristol, Bristol, UK.
Arthritis & Rheumatology (Impact Factor: 7.87). 01/2007; 56(1):177-87. DOI: 10.1002/art.22285
Source: PubMed

ABSTRACT To determine whether it is possible to engineer 3-dimensional hyaline cartilage using mesenchymal stem cells derived from the bone marrow (BMSCs) of patients with osteoarthritis (OA).
Expanded BMSCs derived from patients with hip OA were seeded onto polyglycolic acid scaffolds and differentiated using transforming growth factor beta3 in the presence or absence of parathyroid hormone-related protein (PTHrP) to regulate hypertrophy. Micromass pellet cultures were established using the same cells for comparison. At the end of culture, the constructs or pellets were processed for messenger RNA (mRNA) analysis by quantitative real-time reverse transcription-polymerase chain reaction. Matrix proteins were analyzed using specific assays.
Cartilage constructs engineered from BMSCs were at least 5 times the weight of equivalent pellet cultures. Histologic, mRNA, and biochemical analyses of the constructs showed extensive synthesis of proteoglycan and type II collagen but only low levels of type I collagen. The protein content was almost identical to that of cartilage engineered from bovine nasal chondrocytes. Analysis of type X collagen mRNA revealed a high level of mRNA in chondrogenic constructs compared with that in undifferentiated BMSCs, indicating an increased risk of hypertrophy in the tissue-engineered cells. However, the inclusion of PTHrP at a dose of 1 microM or 10 microM during the culture period resulted in significant suppression of type X collagen mRNA expression and a significant decrease in alkaline phosphatase activity, without any loss of the cartilage-specific matrix proteins.
Three-dimensional hyaline cartilage can be engineered using BMSCs from patients with OA. This method could thus be used for the repair of cartilage lesions.


Available from: Trevor J Sims, Oct 14, 2014
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Lumican is a glycoprotein that is found in the extracellular matrix of many connective tissues, including cartilage. It is a member of the small leucine-rich repeat proteoglycans family and along with two others, decorin and fibromodulin, has the capacity to bind to fibrillar collagens and limit their growth. Cartilage tissue engineering provides a potential method for the production of three-dimensional tissue for implantation into eroded joints. Many studies have demonstrated the growth of cartilage in vitro. However in all cases, biochemical analysis of the tissue revealed a significant deficit in the collagen content. We have now tested the hypothesis that the reduced collagen accumulation in engineered cartilage is a result of over-expression of decorin, fibromodulin or lumican. We have found that the lumican gene and protein are both over-expressed in engineered compared to natural cartilage whereas this is not the case for decorin or fibromodulin. Using a small hairpin lumican antisense sequence we were able to knockdown the lumican gene and protein expression in chondrocytes being used for tissue engineering. This resulted in increased accumulation of type II collagen (the major collagen of cartilage) whilst there was no significant alteration in the proteoglycan content. Furthermore, the antisense knockdown of lumican resulted in an increase in the average collagen fibril diameter measured by transmission electron microscopy. These results suggest that lumican plays a pivotal role in the development of tissue engineered cartilage and that regulation of this protein may be important for the production of high-quality implants.
    Matrix Biology 05/2008; 27(6):526-34. DOI:10.1016/j.matbio.2008.04.002 · 3.65 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Due to the attractive properties of poly(L-lactic acid) (PLLA) and chitosan (CHT) for tissue engineering applications, this study is aimed at analyzing the chondrogenic potential of human bone marrow derived-mesenchymal stem cells (BM-MSCs) derived from osteoarthritic patients (OA), in pure or CHT-coated PLLA, using different coating methodologies. Whilst PLLA scaffolds coated in one-step (PLLA-CHT1) yielded CHT smooth pellicles filling the PLLA macropores, a two-step strategy resulted in a CHT fiber-like thin coating covering PLLA pore walls (PLLA-CHT2). Both strategies led to the incorporation of similar content of CHT and a 2-fold increase in the scaffolds water uptake capacity, providing elastic moduli values comparable to the ones found for human articular cartilage. After confirming OA derived-BM-MSCs, metabolic activity in the scaffolds, the chondrogenic potential was tested at 30 and 60 days, in a chondrogenic differentiation medium. PLLA scaffolds improved the chondrogenic differentiation of BM-MSCs, regarding cell pellet conventional culture and presented a typical cartilage zonal distribution, although was not able to revert a terminal differentiation. In PLLA-CHT1, on a short term, a rather heterogeneous tissue was formed, with confined areas of either slower cell infiltration or a faster maturation, with enhanced chondrogenic phenotype. In PLLA-CHT2, a similar tissue to PLLA was obtained, albeit on the long term, these scaffolds helped to maintain a hyaline-like phenotype and prevented the advance of the hypertrophic process. These results demonstrate the importance of the scaffolds microenvironment on the cellular events of chondrogenesis.
    Tissue Engineering Part A 10/2014; 21(3-4). DOI:10.1089/ten.TEA.2014.0133 · 4.70 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Restoration of articular cartilage function and structure following pathological or traumatic damage is still considered a challenging problem in the orthopaedic field. Currently, tissue engineering-based reconstruction of articular cartilage is a feasible and continuously developing strategy to restore structure and function. Successful articular cartilage tissue engineering strategy relies largely on several essential components including cellular component, supporting 3D carrier scaffolding matrix, bioactive agents, proper physical stimulants, and safe gene delivery. Designing the right formulations from these components remain the main concern of the orthopaedic community. Utilization of mesenchymal stem cells (MSCs) for articular cartilage tissue engineering is continuously increasing compared to use of chondrocytes. Various sources of MSCs have been investigated including adipose tissue, amniotic fluid, blood, bone marrow, dermis, embryonic stem cells, infrapatellar fat pad, muscle, periosteum, placenta, synovium, trabecular bone, and umbilical cord. MSCs derived from bone marrow and umbilical cord are currently in different phases of clinical trials. A wide range of matrices have been investigated to develop tissue engineering - based strategies including carbohydrate-based scaffolds (agarose, alginate, chitosan/chitin, and hyaluronate), protein-based scaffolds (collagen, fibrin, and gelatin), and artificial polymers (polyglycolic acid, polylactic acid, poly(lactic-co-glycolic acid), polyethylene glycol, and polycaprolactone). Collagen - based scaffolds and photopolymerizable PEG - based scaffolds are currently in different phases of clinical trials. TGF-β1, TGF-β3, BMP-2, and hypoxic environment are the recommended bioactive agents to induce optimum chondrogenesis of MSCs, while TGF-β1, TGF-β3, SOX-9, BMP-2, and BMP-7 genes are the best candidate for gene delivery to MSCs. Electromagnetic field and the combination of shear forces/dynamic compression are the best maturation-promoting physical stimulants.
    Histology and histopathology 01/2014; · 2.24 Impact Factor