Adult cartilage has a limited healing capacity. Damages resulting from disease or injury increase over time and cause severe pain. One approach to reinstate the cartilage function is tissue engineering (TE). However, the generation of TE cartilage is time consuming and expensive and its properties are so far suboptimal. As in vivo cartilage is subject to loading, it is assumed that mechanical stimulation may enhance the quality of TE cartilage. In this study the short-term influence of variable compressive strain amplitudes on mechanical and biochemical properties of scaffold-free TE cartilage was investigated. Primary porcine chondrocytes were isolated, proliferated, redifferentiated, and transferred onto hydroxyapatite carriers, resulting in scaffold-free cartilage-carrier constructs. These constructs were placed in a custom-made bioreactor. Compression amplitudes of 5%, 10%, and 20% were applied. In each experiment four constructs were loaded with dynamic compression (3000 cycles/day, 1 Hz) for 14 days and four constructs served as unloaded control. The cartilage was evaluated biochemically, histological, and mechanically. No difference in glycosaminoglycan or collagen content between the loaded and the control groups was found. However, a positive correlation between compression amplitude and normalized Young's modulus was detected (R(2)=0.59, p<0.001). The highest compression amplitude of 20% had the strongest positive effect on the mechanical properties of the TE cartilage (Young's modulus increase of 241±28% compared to unloaded control). The data presented suggest that preconditioning with higher load amplitudes might be an attractive way of generating stiffer tissue and may help accelerating the cultivation of mechanically competent TE cartilage.

Technical aspects play an important role in tissue engineering. Especially an improved design of bioreactors is crucial for cultivation of artificial three-dimensional tissues in vitro. Here formation of cartilage-carrier-constructs is used to demonstrate that the quality of the tissue can be significantly improved by using optimized culture conditions (oxygen concentration, growth factor combination) as well as special bioreactor techniques to induce fluid-dynamic, hydrostatic or mechanical load during generation of cartilage.

The probability of fractures of the cortical shell of vertebral bodies increases as ageing progresses. Ageing involves all the spinal component changes. However, the effect of the spinal component ageing on the fracture risk of the cortical shell remains poorly understood. In this study, the influence of the ageing of the spinal components on cortical shell strain was investigated. A lumbar spinal specimen (L3-L5) was mechanically tested under a quasi-static axial compressive load. Clinical computed tomography images of the same specimen were used to create a corresponding finite element model. The material properties were determined by calibrating the finite element model using the L4 cortical shell strains of the anterior centre measurement site. The remaining experiment data (axial displacement, the intra-discal pressures, L4 cortical shell strain on the lateral measurement site) were used to evaluate the model. The individual ageing process of the six spinal components (cortical shell, cancellous bone, bony endplate, posterior elements, nucleus pulposus and annulus matrix) was simulated by changing their Young's moduli and Poisson's ratios, and the effect on cortical shell strain was investigated. Results show that the cortical shell strain is more sensitive to the ageing of the cortical shell and the cancellous bone than to the ageing of the nucleus pulposus, the annulus matrix, and the bony endplates and of the posterior elements. The results can help the clinicians focus on the aspects that mainly influence the vertebral cortex fracture risk factor.

For long term durability of tissue-engineered cartilage implanted in vivo, the development of the collagen fi bre network orientation is essential as well as the distribution of collagen, since expanded chondrocytes are known to synthesise collagen type I. Typically, these properties differ strongly between native and tissue-engineered cartilage. Nonetheless, the clinical results of a pilot study with implanted tissue-engineered cartilage in pigs were surprisingly good. The purpose of this study was therefore to analyse if the structure and composition of the artifi cial cartilage tissue changes in the fi rst 52 weeks after implantation. Thus, collagen network orientation and collagen type distribution in tissue-engineered cartilage-carrier-constructs implanted in the knee joints of Göttinger minipigs for 2, 26 or 52 weeks have been further investigated by processing digitised microscopy images of histological sections. The comparison to native cartilage demonstrated that fibre orientation over the cartilage depth has a clear tendency towards native cartilage with increasing time of implantation. After 2 weeks, the collagen fi bres of the superfi cial zone were oriented parallel to the articular surface with little anisotropy present in the middle and deep zones. Overall, fi bre orientation and collagen distribution within the implants were less homogenous than in native cartilage tissue. Despite a relatively low number of specimens, the consistent observation of a continuous approximation to native tissue is very promising and suggests that it may not be necessary to engineer the perfect tissue for implantation but rather to provide an intermediate solution to help the body to heal itself. Keywords:Collagen fi bre organisation development in vivo; collagen distribution development in vivo; tissueengineered cartilage; cartilage implant; polarisation microscopy; digital image analysis

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Technical aspects play an important role in tissue engineering. Especially an improved design of bioreactors is crucial for cultivation of artificial threedimensional tissues in vitro. Here formation of cartilage-carrier-constructs is used to demonstrate that the quality of the tissue can be significantly improved by using optimized culture conditions (oxygen concentration, growth factor combination) as well as special bioreactor techniques to induce fluid-dynamic, hydrostatic or mechanical load during generation of cartilage .

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