Non-immortalized human tenocyte cultures as a vehicle for understanding cellular aspects to tendinopathy.

The biochemical mechanisms underlying tendinopathy are obscure. We briefly describe preliminary observations of human tenocyte behaviour in culture as a vehicle for determining the role of reactive oxygen in tendon pathology.

Phenotypic drift in human tenocyte culture.

Tendon ruptures are increasingly common, repair can be difficult, and healing is poorly understood. Tissue engineering approaches often require expansion of cell numbers to populate a construct, and maintenance of cell phenotype is essential for tissue regeneration. Here, we characterize the phenotype of human Achilles tenocytes and assess how this is affected by passaging. Tenocytes, isolated from tendon samples from 6 patients receiving surgery for rupture of the Achilles tendon, were passaged 8 times. Proliferation rates and cell morphology were recorded at passages 1, 4, and 8. Total collagen, the ratio of collagen types I and III, and decorin were used as indicators of matrix formation, and expression of the integrin beta1 subunit as a marker of cell-matrix interactions. With increasing passage number, cells became more rounded, were more widely spaced at confluence, and confluent cell density declined from 18,700/cm2 to 16,100/cm2 ( p = 0.009). No change to total cell layer collagen was observed but the ratio of type III to type I collagen increased from 0.60 at passage 1 to 0.89 at passage 8 ( p < 0.001). Decorin expression significantly decreased with passage number, from 22.9 +/- 3.1 ng/ng of DNA at passage 1, to 9.1 +/- 1.8 ng/ng of DNA at passage 8 ( p < 0.001). Integrin expression did not change. We conclude that the phenotype of tenocytes in culture rapidly drifts with progressive passage.

The nutritive value of stackburned yellow maize for livestock: tests in vitro and in broiler chicks.

Difference in the nutritive value of four grades of stackburned yellow maize, obtained from a single storage unit in Mozambique, were examined. Samples were analysed for chemical composition, and subjected to the following in vitro assays for estimating digestibility: total dietary fibre and pancreatin for non-ruminants, and gas production using sheep rumen fluid for ruminant livestock. Samples were also fed to broiler chicks at 600 g/kg diet in a growth trial. There were no significant differences in crude protein contents of the maize samples, but there was evidence for the development of Maillard reaction products. Detectable amino acids were lower in discoloured maize, with decreases of 52% in lysine, 35% in arginine, and 15% in glycine concentration in the most severely discoloured sample compared with control. Total starch, reducing sugar, acid-detergent fibre and amylase-neutral-detergent fibre values increased, while total non-reducing sugar content decreased with increased discolouration. Total dietary fibre and pancreatin assays indicated a lowering in digestibility of maize with increasing discolouration. Weight gain of chicks (P = 0.0228), efficiency of feed utilization (P = 0.0009) and the metabolizable energy value of diets decreased (P < 0.0001) with increasing stackburn discolouration. There were no significant effects on N retention of diets. In vitro fermentation using sheep rumen fluid showed a linear decrease in gas production with increasing maize discolouration, indicating a reduction in rumen degradability with stackburn.