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- Title
Extracellular matrix of meniscal cartilage is enhanced by co-culture of meniscus and stem cells under low oxygen tension.
- Authors
Matthies, Norah-Faye; Adesida, Adetola; Jomha, Nadr
- Abstract
Background: Meniscus is a fibrocartilaginous tissue in the knee joint where it protects articular cartilage, distributes load, provides shock absorption and joint stability. Meniscus injury can lead to poor biomechanical function and degenerative joint changes such as osteoarthritis. The avascular area of the tissue limits its reparative capacity. Cell-based tissue engineering can aid meniscus repair. Meniscus' biomechanical role is due to its extra-cellular matrix (ECM); mainly composed of proteoglycans, collagens I and II. ECM formation is influenced by many factors; we hypothesized that co-culture of meniscus cells (MEN) and mesenchymal stem cells will enhance ECM formation and that matrix formation will be further enhanced by a low O2 tension due to meniscus' avascular nature and low O2 tension within the knee joint. Methods: Tissue specimens of bone marrow (BMSC) and menisci were obtained, digested, expanded (BMSC only) and mixed at MEN/BMSC ratios of 5/95, 10/90, and 25/75, respectively. Controls were pure BMSC or MEN. Cells were spun to form spherical pellets and cultured for 3 weeks at 37°C in a humidified chamber at either 21% (normoxia) or 3% (hypoxia) O2 tension. At least two independent pellets from each group were processed for DNA, glycosaminoglycans (GAG), histochemical/ immunohistochemistry (IHC) analysis for Alcian blue staining of sulphated GAG, collagens I and II, and qRT-PCR for aggrecan, collagens I and II. Results: Co-cultures of MEN and BMSCs had higher GAG contents than all controls, regardless of O2 tension. Alcian blue staining correlated with GAG data. With a few exceptions, hypoxia potentiated ECM formation that was greater than in normoxia. Hypoxia-generated co-cultures showed both increased total GAG content and GAG per cell (GAG/DNA). IHC and qRT-PCR showed higher gene expression of aggrecan, collagens I and II in co-cultures relative to controls; hypoxia further enhanced this phenotype. Conclusions: Our results demonstrated that co-culture of MEN and BMSCs is an effective way of inducing ECM formation for cell-based meniscus tissue engineering; this process is further potentiated by low O2 tension. Clinical application of these results suggests that co-delivery of MEN and BMSCs on scaffolds may result in the generation of functional meniscus grafts with improved biomechanical properties.
- Subjects
MENISCUS (Anatomy); CARTILAGE; STEM cells; BONE marrow; EXTRACELLULAR matrix; TISSUE engineering
- Publication
UBC Medical Journal, 2011, Vol 2, Issue 2, p45
- ISSN
1920-7425
- Publication type
Abstract