BackgroundAnimal models are commonly used in investigating new treatment options for knee joint injuries including injuries to the meniscus. The reliability and applicability of these models to replicate findings in humans depends on determining the most suitable animal proxy. Therefore, this study was designed to compare the wet weight, volume and dimensions of the human meniscus with two commonly used animal models: sheep and pig.MethodsHuman menisci (n = 6 pairs) were obtained from the knee joints of cadaveric male donors. Sheep menisci (n = 6 pairs) and pig menisci (n = 22 pairs) were obtained from the stifle joints of adult sheep and pigs. Meniscal wet weight, volume and dimensions of the body were measured and compared among the species. Anatomical dimensions included circumference, width, peripheral height, articular height and superior articular length.ResultsThe circumference of human menisci (lateral: 84.0 mm, medial: 88.7 mm) was significantly longer than that of sheep (lateral: 50.0 mm, medial: 55.5 mm) and pig (lateral: 66.8 mm, medial: 64.9 mm). The majority of the remaining dimensions of the medial and all of the remaining dimensions of the lateral menisci in sheep showed no statistical difference in comparison to the human menisci. The meniscal weight in pig was significantly larger (lateral: 6.4 g, medial: 5.0 g) than the human (lateral: 4.9 g, medial: 4.4 g) and sheep (lateral: 2.5 g, medial: 2.2 g). Porcine meniscal volume (lateral: 6.5 ml, medial: 5.1 ml) was also larger than the human (lateral: 5.0 ml, medial: 4.5 ml) and sheep (lateral: 2.3 ml, medial: 2.2 ml) menisci. The dimensions measured in the pig meniscus were generally larger than human menisci with statistically significant differences in most categories.ConclusionSheep meniscal dimensions more closely matched human meniscal dimensions than the pig meniscal dimensions. This information may help guide the choice of an animal proxy in meniscal research.

BackgroundMeniscal cartilage displays a poor repair capacity, especially when injury is located in the avascular region of the tissue. Cell-based tissue engineering strategies to generate functional meniscus substitutes is a promising approach to treat meniscus injuries. Meniscus fibrochondrocytes (MFC) can be used in this approach. However, MFC are unable to retain their phenotype when expanded in culture. In this study, we explored the effect of oxygen tension on MFC expansion and on their matrix-forming phenotype.Methodology/Principal FindingsMFC were isolated from human menisci followed by basic fibroblast growth factor (FGF-2) mediated cell expansion in monolayer culture under normoxia (21%O2) or hypoxia (3%O2). Normoxia and hypoxia expanded MFC were seeded on to a collagen scaffold. The MFC seeded scaffolds (constructs) were cultured in a serum free chondrogenic medium for 3 weeks under normoxia and hypoxia. Constructs containing normoxia-expanded MFC were subsequently cultured under normoxia while those formed from hypoxia-expanded MFC were subsequently cultured under hypoxia. After 3 weeks of in vitro culture, the constructs were assessed biochemically, histologically and for gene expression via real-time reverse transcription-PCR assays. The results showed that constructs under normoxia produced a matrix with enhanced mRNA ratio (3.5-fold higher; p<0.001) of collagen type II to I. This was confirmed by enhanced deposition of collagen II using immuno-histochemistry. Furthermore, the constructs under hypoxia produced a matrix with higher mRNA ratio of aggrecan to versican (3.5-fold, p<0.05). However, both constructs had the same capacity to produce a glycosaminoglycan (GAG) –specific extracellular matrix.ConclusionsOur data provide evidence that oxygen tension is a key player in determining the matrix phenotype of cultured MFC. These findings suggest that the use of normal and low oxygen tension during MFC expansion and subsequent neo-tissue formation cultures may be important in engineering different regions of the meniscus.

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Preserving viable articular cartilage is a promising approach to address the shortage of graft tissue and enable the clinical repair of articular cartilage defects in articulating joints, such as the knee, ankle, and hip. In this study, we developed two 2-step, dual-temperature, multicryoprotectant loading protocols to cryopreserve particulated articular cartilage (cubes ~1 mm3 in size) using a mathematical approach, and we experimentally measured chondrocyte viability, metabolic activity, cell migration, and matrix productivity after implementing the designed loading protocols, vitrification, and warming. We demonstrated that porcine and human articular cartilage cubes can be successfully vitrified and rewarmed, maintaining high cell viability and excellent cellular function. The vitrified particulated articular cartilage was stored for a period of 6 months with no significant deterioration in chondrocyte viability and functionality. Our approach enables high-quality long-term storage of viable articular cartilage that can alleviate the shortage of grafts for use in clinically repairing articular cartilage defects.

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