Infrared spectroscopic analysis of human and bovine articular cartilage proteoglycans using carbohydrate peak or its second derivative.

Fourier transform infrared (FTIR) microspectroscopy has been used to estimate the spatial proteoglycan (PG) and collagen contents in articular cartilage (AC). However, it is not clear whether the results of FTIR analyses are consistent between different species. Our aim was to clarify how three different FTIR PG parameters in use, i.e., the integrated absorbance in the carbohydrate region, the carbohydrate/amide I ratio, and the second derivative peak at 1062 cm-1, can indicate the densitometrically assessed (reference method) spatial PG content in a sample set consisting of osteoarthritic human and bovine AC samples. The results show that all the parameters can accurately reflect the PG content, when the species are analyzed separately. When all samples are pooled, the correlation with the reference method is high (r=0.760, n=104) for the second derivative peak at 1062 cm-1 and is significantly lower (p<0.05) for the carbohydrate region (r=0.587, n=104) and for the carbohydrate/amide I ratio (r=0.579, n=104). Therefore, the analysis of the carbohydrate region may provide inconsistent results, if the cartilage samples from different species are in use. Based on the present study, second derivative analysis yields more consistent results for human and bovine cartilages.

Minor influence of lifelong voluntary exercise on composition, structure, and incidence of osteoarthritis in tibial articular cartilage of mice compared with major effects caused by growth, maturation, and aging.

We investigated the effects of lifelong voluntary exercise on articular cartilage of mice. At the age of 4 weeks C57BL mice (n = 152) were divided into two groups, with one group serving as a sedentary control whereas the other was allowed free access to a running wheel from the age of 1 month onward. Mice were euthanized at four different time points (1, 2, 6, and 18 months of age). Articular cartilage samples were gathered from the load-bearing area of the tibial medial plateaus, and osteoarthritis was graded. Additionally, the proteoglycan content distribution was assessed using digital densitometry, collagen fibril orientation, and parallelism with polarized light microscopy, and collagen content using Fourier transform infrared imaging spectroscopy. The incidence of osteoarthritis increased with aging, but exercise had no effect on this trend. Furthermore, the structure and composition revealed significant growth, maturation, and age-dependent properties. Exercise exerted a minor effect on collagen fibril orientation in the superficial zone. Fibril orientation at 2 months of age was more perpendicular to surface (p < 0.05) in controls compared with runners, whereas the situation was reversed at the age of 18 months (p < 0.05). The collagen content of the superficial zone was higher (p < 0.01) at the age of 18 months in controls compared with runners but the proteoglycan content did not display any exercise-dependent changes. In conclusion, growth, maturation, and aging exerted a clear effect on integrity, structure, and composition of medial tibial plateau articular cartilage in mice, whereas lifelong voluntary exercise had only a minor effect on collagen architecture and content.

Effects of growth and exercise on composition, structural maturation and appearance of osteoarthritis in articular cartilage of hamsters.

Articular cartilage composition and structure are maintained and remodeled by chondrocytes under the influence of loading. Exercise-induced changes in the composition, structure, mechanical properties and tissue integrity of growing and aging hamster articular cartilage were investigated. Articular cartilage samples (n = 191) were harvested from the proximal tibiae of hamsters aged 1, 3, 6, 12 and 15 months. The hamsters were divided into runners and controls. The runners had free access to a running wheel between 1 and 3 months (runner groups 3-, 12- and 15-month-old hamsters) or 1 and 6 months (runner group 6-month-old hamsters) of age. Control animals were subjected to a sedentary lifestyle. Mechanical indentation tests and depth-wise compositional and structural analyses were performed for the cartilage samples. Furthermore, the integrity of articular cartilage was assessed using histological osteoarthritis grading. Exercise affected the collagen network organization after a 5-month exercise period, especially in the middle and deep zones. However, no effect on the mechanical properties was detected after exercise. Before the age of 12 months, the runners showed less osteoarthritis than the controls, whereas at 15 months of age the situation was reversed. It is concluded that, in hamsters, physical exercise at a young age enhances cartilage maturation and alters the depth-wise cartilage structure and composition. This may be considered beneficial. However, exercise at a young age demonstrated adverse effects on cartilage at a later age with a significant increase in the incidence of osteoarthritis.