Untersuchung zum 3-D-Einwachsverhalten von Osteoblasten like cells (MG63) in resorbierbare Gelatine-Schwämme

Osseous defects still remain a challenge to modern traumatology and tumor surgery. The use of autologous bone – gained from the iliac crest – is regarded as golden standard for the treatment of osseous defects. The treatment contains the possibility of complications such as the risk of a second surgical intervention and a further infection as a result. Therefore the fundamental research of alternative bone materials is of special interest. The study examines the three-dimensional ingrowth of Osteoblast like cells (MG63) in hematostatic Spongostan® which is used in clinical practice for several years. In the first place this study is supposed to show a descriptive approach and not to demonstrate an indicator of clinical application or treatment of osseous defects. Thereby observation of cell ingrowth in Spongostan® was the main aspect. The MG63 cells used in the study have a transformed phenotype. Compared to primary native cultures transformed cells are characterized by unlimited growth and missing of contact inhibition. The trial contains the comparison of two different cell culture dishes by otherwise identical test conditions. With the objective of offering valuable clues to the impact of an improved oxygen supply by using a cell culture dish with a gas–permeable film bottom compared to a cell culture dish with a regular bottom. During the three week observation period medium was changed every 48 hours. Over the whole period several parameters were regularly taken and analysed. In order to evaluate the calcification osteogenic marker such as osteocalcin and calcium were meseared. As proinflammatory cytokines who facilitate bone destruction through direct or indirect osteoclasts activity were IL–6, IL–8 and TNF–α identified. The assessment of ingrowth development of the paraffin–bedded and cut Spongostan®- sponges was made after the observation period relating to light microscope. Investigating these light microscopic pictures a difference between the in two different cell culture dishes cultivated test serious could be identified. The under surface of the sponges lying on regular film bottom showed no ingrowth of cells whereas the sponges on gas–permeable film developed cells. A calcification was not detected relating to concentration of osteocalcin in any of cell cultures. All cytokines reached a plateau within half time of the study period and could be related to as an indicator of continuing cell growth. Spongostan® is generally applicable as scaffold for cell ingrowth and offers a high level of biocompatibility. In comparison to the control cultures the cells reacted neither with a higher release of cytokines in form of a proinflammatory reaction nor with a increased demise of cells under the use of Spongostan®. In terms of ingrowth and behaviour on biomaterials MG63 cells are not a good indicator of clinical performance.