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Development of a three dimensional in vitro model system for nondestructive investigation of osteoblast cell growth in scaffolds
|Claudia Bergemann 1, Friederike Kunz 1, Ernst-Dieter Klinkenberg 2, Volker Weißmann 3, Stefan Ofe 3, Sebastian Spath 4, Hermann Seitz 4, Regina Lange 5, Patrick Elter 5, Ulrich Beck 5, Rainer Bader 6, Barbara J. Nebe 1|
1. University of Rostock, Medical Faculty, Dept. of Cell Biology, Schillingallee 69, Rostock 18057, Germany
Three dimensional cell cultures for tissue engineering are possibly limited concerning nutrition and hypoxia inside the scaffolds which may lead to changes in cell behavior. The aim of the present study was to create an in vitro three dimensional (3D) model system to investigate the interaction of human osteoblasts with porous scaffolds in the depth of the corpus. A clamping ring was designed, in which two porous discs can be horizontally fixed. Thereby a 3D module with 4 levels with a maximal distance of 10 mm is generated. Porous discs could be made from different materials, e.g. porous tantalum (Ta) (Zimmer, Inc.), ceramic, polymer and composite scaffolds manufactured by rapid prototyping. Human MG63 osteoblastic cells (ATCC) were seeded apical onto the 3D model and placed in DMEM with 10 % FCS. After 7 and 14 days the cell ingrowth from level 1 to level 2-4 by field emission scanning electron microscopy (FESEM) and mRNA expression for proliferative and osteogenic marker genes was analyzed. Preliminary results demonstrated a cell migration through the scaffolds after 7 and 14 days on Ta and ceramic scaffolds. Cell morphology was similar in the different levels of the model system, but mRNA expression indicated an influence of the corpus depth on cell differentiation. Ta scaffolds for in vitro cell culture studies were normally used with a maximal height of 5 mm for the ingrowth of cells. In this study we present a 3D model with 4 levels for nondestructive cell analysis, where the maximal distance for ingrowths is 10 mm. This model is a well suited system for the investigation of structurally new designed composite implant materials. Examination of cultured cells from distinct levels will improve our knowledge about the cell growth inside a 3D corpus.
Presentation: Poster at E-MRS Fall Meeting 2009, Symposium I, by Claudia Bergemann
See On-line Journal of E-MRS Fall Meeting 2009
Submitted: 2009-05-13 12:32 Revised: 2009-08-14 15:41