Characterization of oriented protein-ceramic and protein-polymer-composites for cartilage tissue engineering using synchrotron μ-CT

Abstract In this paper we report on the synthesis of three different gelatine based scaffold materials for the reconstruction of articular cartilage defects. The first scaffold design is based on an unmodified, oriented gelatine network, while the second design further comprises an attached inorganic hydroxyapatite layer and the third design includes poly(l-lactide) microspheres as a model material for future drug-release applications. All three scaffold designs were characterized and imaged using synchrotron μ-CT, obtaining a complete volumetric reconstruction of a previously defined sample region. Furthermore, two unmodified scaffolds were cultivated for one week with porcine chondrocytes. Afterwards the attached cells were labelled using a combination of Au-lysine and silver enhancer. In synchrotron μ-CT analysis we were thus able to map the cell distribution due to the difference in X-ray absorption of the labelled cells and the non labelled scaffolds in a volume of several millimetres.

[1]  R. Zehbe,et al.  Oriented Collagen-Based/Hydroxyapatite Matrices for Articular Cartilage Replacement , 2003 .

[2]  W. Knauf,et al.  Culture of haematopoietic cells in a 3-D bioreactor made of Al2O3 or apatite foam , 2004, Journal of materials science. Materials in medicine.

[3]  J C Fiala,et al.  Reconstruct: a free editor for serial section microscopy , 2005, Journal of microscopy.

[4]  Wei Sun,et al.  3D microtomographic characterization of precision extruded poly-epsilon-caprolactone scaffolds. , 2004, Journal of biomedical materials research. Part B, Applied biomaterials.

[5]  W M Lai,et al.  Some surface characteristics of articular cartilage. I. A scanning electron microscopy study and a theoretical model for the dynamic interaction of synovial fluid and articular cartilage. , 1974, Journal of biomechanics.

[6]  R. Zehbe,et al.  Short-term human chondrocyte culturing on oriented collagen coated gelatine scaffolds for cartilage replacement. , 2005, Bio-medical materials and engineering.

[7]  Angela Lin,et al.  Quantitative microcomputed tomography analysis of mineralization within three-dimensional scaffolds in vitro. , 2004, Journal of biomedical materials research. Part A.

[8]  N. Dagalakis,et al.  Design of an artificial skin. Part III. Control of pore structure. , 1980, Journal of biomedical materials research.

[9]  Felix Beckmann,et al.  Osteoconductive modifications of Ti-implants in a goat defect model: characterization of bone growth with SR muCT and histology. , 2005, Biomaterials.

[10]  Yoram Bresler,et al.  O(N2log2N) filtered backprojection reconstruction algorithm for tomography , 2000, IEEE Trans. Image Process..

[11]  Dietmar W Hutmacher,et al.  A comparison of micro CT with other techniques used in the characterization of scaffolds. , 2006, Biomaterials.

[12]  J. Roth,et al.  Cationic colloidal gold--a new probe for the detection of anionic cell surface sites by electron microscopy. , 1986, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[13]  Savio L-Y Woo,et al.  Cell orientation determines the alignment of cell-produced collagenous matrix. , 2003, Journal of biomechanics.

[14]  C. Kirkpatrick,et al.  Cyclooxygenases and prostaglandin E2 receptors in growth plate chondrocytes in vitro and in situ – prostaglandin E2 dependent proliferation of growth plate chondrocytes , 2006, Arthritis research & therapy.

[15]  R. Robb,et al.  Optimal segmentation of microcomputed tomographic images of porous tissue-engineering scaffolds. , 2005, Journal of biomedical materials research. Part A.