Comparison of marker gene expression in chondrocytes from patients receiving autologous chondrocyte transplantation versus osteoarthritis patients.

NMI Natural and Medical Sciences Institute at the University of Tübingen, Markwiesenstrasse, 72770 Reutlingen, Germany.
Arthritis research & therapy (Impact Factor: 4.12). 02/2007; 9(3):R60. DOI: 10.1186/ar2218
Source: PubMed

ABSTRACT Currently, autologous chondrocyte transplantation (ACT) is used to treat traumatic cartilage damage or osteochondrosis dissecans, but not degenerative arthritis. Since substantial refinements in the isolation, expansion and transplantation of chondrocytes have been made in recent years, the treatment of early stage osteoarthritic lesions using ACT might now be feasible. In this study, we determined the gene expression patterns of osteoarthritic (OA) chondrocytes ex vivo after primary culture and subculture and compared these with healthy chondrocytes ex vivo and with articular chondrocytes expanded for treatment of patients by ACT. Gene expression profiles were determined using quantitative RT-PCR for type I, II and X collagen, aggrecan, IL-1beta and activin-like kinase-1. Furthermore, we tested the capability of osteoarthritic chondrocytes to generate hyaline-like cartilage by implanting chondrocyte-seeded collagen scaffolds into immunodeficient (SCID) mice. OA chondrocytes ex vivo showed highly elevated levels of IL-1beta mRNA, but type I and II collagen levels were comparable to those of healthy chondrocytes. After primary culture, IL-1beta levels decreased to baseline levels, while the type II and type I collagen mRNA levels matched those found in chondrocytes used for ACT. OA chondrocytes generated type II collagen and proteoglycan-rich cartilage transplants in SCID mice. We conclude that after expansion under suitable conditions, the cartilage of OA patients contains cells that are not significantly different from those from healthy donors prepared for ACT. OA chondrocytes are also capable of producing a cartilage-like tissue in the in vivo SCID mouse model. Thus, such chondrocytes seem to fulfil the prerequisites for use in ACT treatment.


Available from: Reinout Stoop, May 28, 2015
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    ABSTRACT: Topics: Aufbau und Funktion des hyalinen Knorpels; Prinzip des Regenerationsgewebes; Genese der traumatischen Gelenkdegeneration und Arthrose (OA); Genese der nicht-traumatischen Gelenkdegeneration und Arthrose (OA); Inzidenz von Schäden des hyalinen Knorpels; Bedeutung der OA für den Patienten und das Gesundheitssystem; Welche Gelenke können therapiert werden?; Chirurgische Methoden zur Restauration des hyalinen Knorpels; MACT: Wirkungs-Prinzip und klinische Indikationen; Vergleich MAC mit anderen Verfahren: Evidenzlage; Vergleich MACT und Mikrofrakturierung: Defektgröße; Vergleich MACT und Mikrofrakturierung: Rückkehr zum Sport, klinische Nachhaltigkeit; MACT: Korrelation von Gewebequalität und klinischem Verlauf; MACT: Langfristiger klinischer Erfolg; MACT und OA; MACT: Verbesserungskonzepte; AMIC: Wirkungs-Prinzip; AMIC: klinische Indikationen und Hinweise zur Mikrofrakturierung; AMIC: Klinische Ergebnisse; AMIC: Zusamenfassung; Knorpel-Rekonstruktion: neue Impulse, neue Produkte; Nationale Firmen und Produkte - Amedrix GmbH; Arthro-Kinetics AG; BioTissueTechnologies GmbH; CellGenix GmbH; Co.don AG; TETEC AG; TiGenix N.V.; Internationale Firmen und Produkte - Genzyme Biosurgery/ Sanofi-Aventis; Fidia Advanced Biopolymers; CellMatrix Cartilix; Tissue Bank of France (TBF); Histogenics; DePuy Mitek / Johnson & Johnson Regenerative Therapeutics LLC; ISTO Technologies, Inc. / Zimmer, Inc.; Zukünftige Therapiekonzepte: Identifizierung und Modulation geeigneter Stoffwechselwege; Zukünftige Therapiekonzepte: Transplantation adulter mesenchymaler Stammzellen; Fazit
    „Was gibt es Neues in der Chirurgie? Jahresband 2013: Berichte zur chirurgischen Fort- und Weiterbildung“, 2013 edited by Joachim Jähne, Alfred Königsrainer, Wolfgang Schröder, Norbert P. Südkamp, 01/2013: chapter Was gibt es Neues in der Knorpelchirurgie?: pages 35-47; HJR Verlagsgruppe Hüthig, Jehle, Rehm.., ISBN: ISBN 978-3609769912
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    ABSTRACT: BACKGROUND:Several patient- and defect-specific factors influencing clinical outcomes after matrix-associated chondrocyte transplantation (MACT) have been identified, including the patient's age, location of the defect, or duration of symptoms before surgery. Little is known, however, about the influence of cell-specific characteristics on clinical results after transplantation. PURPOSE:The aim of the present study was to investigate the influence of cell differentiation and interleukin-1 β (IL-1β) expression on clinical outcomes up to 5 years after MACT. STUDY DESIGN:Case series; Level of evidence, 4. METHODS:Twenty-seven patients who underwent MACT of the tibiofemoral joint area of the knee were included in this study. Clinical assessments were performed preoperatively as well as 6, 12, 24, and 60 months after transplantation by using the following scores: the Knee injury and Osteoarthritis Outcome Score (KOOS), the International Knee Documentation Committee (IKDC) Subjective Knee Form, the Noyes sports activity rating scale, the Brittberg clinical score, and a visual analog scale (VAS) for pain. The quality of repair tissue was assessed by magnetic resonance imaging using the magnetic resonance observation of cartilage repair tissue (MOCART) score at 1 and 5 years. Cell differentiation (defined as collagen type II:type I expression ratio), aggrecan, and IL-1β expression were determined by real-time polymerase chain reaction in transplant residuals and were correlated with clinical outcomes. RESULTS:The largest improvements in clinical scores were found during the first year. Two years postoperatively, a stable improvement was reached until 5 years after transplantation, with a mean IKDC score of 34.4 ± 8.6 preoperatively to 77.9 ± 16 after 24 months (P < .001). Cell differentiation showed a significant positive correlation with nearly all clinical scores at different time points, especially after 12 months (P < .05). IL-1β expression negatively influenced clinical outcomes at 24 months (Brittberg score) and 60 months (Brittberg and VAS scores) after surgery (P < .05). No correlation was found between the MOCART score and clinical outcomes or gene expression. CONCLUSION:Our data demonstrate that cell differentiation and IL-1β expression influence clinical outcomes up to 5 years after MACT.
    The American Journal of Sports Medicine 11/2013; 42(1). DOI:10.1177/0363546513507543 · 4.70 Impact Factor
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    ABSTRACT: IntroductionThis study aimed to evaluate whether profiles of several soluble mediators in synovial fluid and cartilage tissue are pathology-dependent and how their production is related to in vitro tissue formation by chondrocytes from diseased and healthy tissue.Methods Samples were obtained from donors without joint pathology (n¿=¿39), with focal defects (n¿=¿65) and osteoarthritis (n¿=¿61). A multiplex bead assay (Luminex) was performed measuring up to 21 cytokines: Interleukin (IL)-1¿, IL-1ß, IL-1RA, IL-4, IL-6, IL-6R¿, IL-7, IL-8, IL-10, IL-13, tumor necrosis factor (TNF)¿, Interferon (IFN)¿, oncostatin M (OSM), leukemia inhibitory factor (LIF), adiponectin, leptin, monocyte chemotactic factor (MCP)1, RANTES, basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), vascular growth factor (VEGF).ResultsIn synovial fluid of patients with cartilage pathology, IL-6, IL-13, IFN¿ and OSM levels were higher than in donors without joint pathology (P ¿0.001). IL-13, IFN¿ and OSM were also different between donors with cartilage defects and OA (P <0.05). In cartilage tissue from debrided defects, VEGF was higher than in non-pathological or osteoarthritic joints (P ¿0.001). IL-1¿, IL-6, TNF¿ and OSM concentrations (in ng/ml) were markedly higher in cartilage tissue than in synovial fluid (P <0.01). Culture of chondrocytes generally led to a massive induction of most cytokines (P <0.001). Although the release of inflammatory cytokines was also here dependent on the pathological condition (P <0.001) the actual profiles were different from tissue or synovial fluid and between non-expanded and expanded chondrocytes. Cartilage formation was lower by healthy unexpanded chondrocytes than by osteoarthritic or defect chondrocytes.Conclusions Several pro-inflammatory, pro-angiogenic and pro-repair cytokines were elevated in joints with symptomatic cartilage defects and/or osteoarthritis, although different cytokines were elevated in synovial fluid compared to tissue or cells. Hence a clear molecular profile was evident dependent on disease status of the joint, which however changed in composition depending on the biological sample analysed. These alterations did not affect in vitro tissue formation with these chondrocytes, as this was at least as effective or even better compared to healthy chondrocytes.