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Development of a three dimensional in vitro model system for nondestructive investigation of osteoblast cell growth in scaffolds   

Claudia Bergemann 1Friederike Kunz 1Ernst-Dieter Klinkenberg 2Volker Weißmann 3Stefan Ofe 3Sebastian Spath 4Hermann Seitz 4Regina Lange 5Patrick Elter 5Ulrich Beck 5Rainer Bader 6Barbara J. Nebe 1

1. University of Rostock, Medical Faculty, Dept. of Cell Biology, Schillingallee 69, Rostock 18057, Germany
2. DOT GmbH, Charles-Darwin-Ring 1a, Rostock 18059, Germany
3. Institute for Polymer Technologies, Alter Holzhafen 19, Wismar 23966, Germany
4. University of Rostock, Dept. of Mechanical Engineering and Marine Technology, Justus-von-Liebig-Weg 6, Rostock 18059, Germany
5. University of Rostock, Dept. of Electrical Engineering and Informatics, Albert-Einstein-Str. 2, Rostock 18059, Germany
6. University of Rostock, Dept. of Orthopaedics, Doberaner Str. 142, Rostock 18055, 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.


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Related papers

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