Member of the Research Training Group, 1.09.2012 - 31.12.2013
PhD Defence 30.09.2014, Summa Cum Laude

Interaktionen von humanen Osteoblasten mit geometrisch strukturierten Implantatoberflächen

Interaktionen von humanen Osteoblasten mit geometrisch strukturierten Implantatoberflächen

Micro- and nanotopography as well as the surface chemistry of biomaterials affect cell adhesion, proliferation and cell differentiation. Furthermore, the organization and localization of intracellular adhesion components such as the actin cytoskeleton are also altered dependent on the material surface topography. However, the detailed influence of the material micro-structure on cellular mechanisms on the molecular level is still unknown. This study is intended to elucidate such effects using regular pillar structures to characterize the modulation of cell responses like the regulation of voltage-sensitive calcium channels as well as signaling molecules in human osteoblasts. To analyze cell behavior on defined biomaterial surfaces, human osteoblastic MG-63 cells were cultured on geometrically micro textured titanium coated silicon wafers, as opposed to planar titanium references. Samples were fabricated by a photolithographic process using the negative tone resist SU-8 and sputter-coated with 100 nm titanium. Immunofluorescence staining and flow cytometry are used to detect the expression levels and the function of T type calcium channels. Knowledge about the biocomplexity of cell behavior dependent on topographical characteristics is of clinical relevance for the development of implant designs in tissue engineering.

• Analysis of spatial localization of intracellular adhesion structures in human osteoblasts by LSM and FACS
• Expression of Ca2+ channels in human osteoblasts by LSM and FACS
• Electrophysiological characterization of Ca²⁺ channels by patch clamp techniques
• Time dependent phosphorylation of signaling proteins by Bio-Plex xMAP™-Technology

Peer-reviewed journal papers:

• NEBE, J.B., MOERKE, C., STAEHLKE, S., FINKE, B., SCHNABELRAUCH, M., ANSELME, K., HELM, C.A., FRANK, M. and REBL, H., 2017. Complex cell physiology on topographically and chemically designed material surfaces. Material Science Forum, 879, pp. 78-83. DOI: 10.4028/www.scientific.net/MSF.879.78
• FINKE, B., TESTRICH, H., REBL, H., WALSCHUS, U., SCHLOSSER, M., ZIETZ, C., STAEHLKE, S., NEBE, J.B., WELTMANN, K.D., MEICHSNER, J. and POLAK, M. 2016. Plasma-deposited fluorocarbon polymer films on titanium for preventing cell adhesion: a surface finishing for temporarily used orthopaedic implants.  Journal of Physics: Applied Physics, 49 (23), DOI: 10.1088/0022-3727/49/23/234002.
• STAEHLKE, S., KOERTGE, A. and  NEBE, J.B., 2015. Intracellular calcium dynamics dependent on defined microtopographical features of titanium. Biomaterials 46. pp. 48-57.
• NEBE, J.B., FINKE, B., KOERTGE, A., REBL, H. and STAEHLKE, S., 2014. Geometrical Micropillars Combined with Chemical Surface Modifications – Independency of Actin Filament Spatial Distribution in Primary Osteoblasts. Material Science Forum, 783-786, pp.1320-1325.
• BITTIG, A.T., MATSCHEGEWSKI, C., NEB,E J.B., STAEHLKE, S. and UHRMACHER, A.M., 2014. Membrane related dynamics and the formation of actin in cells growing on micro-topographies: a spatial computational model. BMC Systems Biology, 8, (106), pp.1-35.
• STAEHLKE, S., KUNZ, F., LOEFFLER, R., MATSCHEGEWSKI, C., FLEISCHER, M., KERN, D. P. and NEBE J. B., 2013. Regulation of T-type calcium channels in osteoblasts on micro-structured surface topography. Advanced Materials Research, 647, pp.  476-481, DOI: 10.4028/www.scientific.net/AMR.647.476
• MATSCHEGEWSKI, C., STAEHLKE, S., BIRKHOLZ, H., LANGE, R., BECK, U., ENGEL, K. and NEBE, JB., 2012. Automatic Actin Filament Quantification of Osteoblasts and their Morphometric Analysis on Microtextured Silicon-titanium Arrays. Materials, 5, pp. 1176-1195. DOI:10.3390/ma50711762012.
• LOEFFLER, R., FLEISCHER, M., MATSCHEGEWSKI, C., STAEHLKE, S., LANGE, R., NEBE, JB., KERN, DP., 2012. Pyramid Array Substrates for Biomedical Studies. Journal of Vacuum Science and Technology.  B 30. doi.org/10.1116/1.4757109.
• MATSCHEGEWSKI, C., STÄHLKE, S., BIRKHOLZ, H., LÖFFLER, R., KERN, D., ENGEL, K., NEBE, B., 2012. Quantitative analysis of the cellular actin cytoskeleton on geometrically designed surface topography. Materials Science Forum, 706-709, pp. 543-548.
• LANGE, R., ELTER, P., BIALA, K., MATSCHEGEWSKI, C., STÄHLKE, S., LÖFFLER, R., FLEISCHER, M., NEBE, J.B., KERN, D. and BECK, U., 2010. Titanium surfaces structured with regular geometry-material investigations and cell morphology. Surface and Interface Analysis, 42(6), pp 497-501.
• MATSCHEGEWSKI, C., STAEHLKE, S., LÖFFLER, R., LANGE, R., CHAI, F., KERN, D., BECK, U. and NEBE, J.B., 2010. Cell architecture-cell function dependencies on titanium arrays with regular geometry.  Biomaterials, 31(22), pp. 5729-5740. doi:10.1016/j.biomaterials.2010.03.073.
• NEBE, J. B., JESSWEIN, H., WEIDEMANN, A., FINKE, B., LANGE, R., BECK, U., STÄHLKE, S. and SCHRÖDER, K., 2010. Osteoblast's sensitivity to topographical and chemical features of titanium. Material Science Forum, 638-642, pp. 652-657.

Peer-reviewed paper important congresses:

• MÖRKE, C., STÄHLKE, S. and NEBE, B., 2014. Influence of geometric micro-pillared structures on the cell physiology of MG-63 osteoblast-like cells. International Meeting of the German Society for Cell Biology, Regensburg, pp. 81.
• STAEHLKE, S., KUNZ, F., NEBE, J.B., 2013. Intracellular calcium mobilization in osteoblasts in dependance on defined surface micro structures. BioInterface, Minneapolis, USA --> Ausgewähltes Poster
• STÄHLKE, S., KUNZ, F., MATSCHEGEWSKI, C., NEBE, J.B., 2013. Analyse der intrazellulären Kalzium Mobilisierung in Osteoblasten auf definierter Mikrostruktur. Deutsche Gesellschaft für Biomaterialien (DGBM) ISSN 2193-0651 BIONANO-MATERIALS, 14, p.81, Erlangen --> DGBM Posterpreis
• STAEHLKE, S., KUNZ, F., LOEFFLER, R., MATSCHEGEWSKI, C., FLEISCHER, M., KERN, D.P., NEBE, J.B., 2013. Intracellular signaling in osteoblasts on regularly structured titanium surfaces. 6th Annual meeting Scandinavian Society for Biomaterials (ScSB) Materials for Tissue Engineerings, Hafjell, Norway.
• STEFFEN, J., STAEHLKE, S., MATSCHEGEWSKI, C., KUNZ, F., LOEFFLER, R. and NEBE, JB., 2012.  Correlation of calcium channel expression and actin organization – bone cells on microstructures. BioNanoMaterials, 13 (1-4), pp. 219.
• LÖFFLER, R., FLEISCHER, M., KERN, D.P., MATSCHEGEWSKI, C., STAEHLKE, S., NEBE, J.B., 2012. Pyramid Array Substrates for Biomedical Studies. 56th International Conference on Electron, Ion, and Photon Beam Technology and Nanofabrication (EIPBN), Hawaii, USA.
• KUNZ, F., QUADE, A., MATSCHEGEWSKI, C., STAEHLKE, S., BIRKHOLZ, H., REBL, H., STEFFEN, J., NEBE, JB., 2012. Actin cytoskeleton mediated cell response of osteoblasts to plasma amino functionalised titanium surfaces. Poster, 4th International Symposium Interface Biology of Implants, p.49.
• LANGE, R., LÖFFLER, R., STÄHLKE, S., ELTER, P., KÖRTGE, A., NEBE; JB., KERN, D., BECK, U., 2012. Electrochemical and scanning electron microscopic characterization of biomaterial surfaces with regular geometry dry etched in Titanium. ISBN: 978-3-00-038220-8, 4th International Symposium Interface Biology of Implants, p.51.
• STAEHLKE, S., KUNZ, F., LOEFFLER, R., MATSCHEGEWSKI, C., FLEISCHER, M., KERN, D. P. and NEBE J. B., 2012. Regulation of T-type Calcium Channels in Osteoblasts on Micro-Structured Surface Topography. 2012 International Conference on Biomaterial and Bioengineering (ICBB), Hong Kong, China. pp. 21.
• STAEHLKE, S., MATSCHEGEWSKI, C., LOEFFLER, R.,  LANGE, R., BECK, U., KERN, D. and NEBE, J.B., 2011.  Analyse der Interaktion von Osteoblasten auf mikrostrukturierten Oberflächen in Bezug auf die Organisation des Aktinzytoskeletts. BIOmaterialien 12(1-4), pp. 115.
• MATSCHEGEWSKI, C., BIRKHOLZ,  H., STAEHLKE, S., LANGE, R., BECK, U., ENGEL, K. and NEBE, J.B., 2011. Determination of cellular phenotype and actin quantification of osteoblasts on geometrically defined microtextured titanium arrays. BIOmaterialien 12(1-4), pp. 113.
• MATSCHEGEWSKI, C., BIRKHOLZ,  H., STAEHLKE, S., LOEFFLER, R., KERN, D., ENGEL, K. and NEBE, J.B., 2011. Quantification of intracellular structures as a tool for the correlation of material vs. cellular parameters. BMT 2011, Freiburg
• MATSCHEGEWSKI, C., BIRKHOLZ,  H., STAEHLKE, S., LANGE, R., BECK, U.,  ENGEL, K. and NEBE, J.B., 2011. Cell morphometric analysis of osteoblasts on geometrically microstructured titanium surfaces – basic research for orthopedic implants. World Congress on regenerative medicine (WCRM) 2011 Leipzig.
• REBL, H., FINKE, B., MATSCHEGEWSKI, C., STAEHLKE, S., LANGE, R., KERN, D.P., BECK, U., SCHROEDER, K., NEBE, J.B., 2010. Cell architecture on plasmafunctionalized, structured metal surfaces. ESB, CD abstract, no. H 3492, Tampere, Finland
• MATSCHEGEWSKI, C., STAEHLKE, S., LOEFFLER, R., LANGE, R., CHAI, F., KERN, D.P., BECK, U. and NEBE, J.B., 2010. Cell Architecture-Cell Function Dependencies on Micro-Arrays with Cubic Pillar Structures. Deutsche Gesellschaft für Materialkunde, DGM, Material Science and Engineering, Topic F, Biomat- F3, Interfaces, Darmstadt.
• BIALA, K., LANGE, R., STÄHLKE, S., NEBE, JB., und BECK, U., 2010. Physical and electrochemical characterisation of regular microstructred titanium and their influence on cell morphology. Biomedizinische Technik/ Biomedical Engineering, 55 (1), pp. 169. DOI:10.1515/bmt.2010.712.
• NEBE, B., REBL, H., FINKE, B., MATSCHEGEWSKI, C., STÄHLKE, S., BECK, U., KERN, D., WELTMANN, K.D. and SCHRÖDER, K., 2010. Plasmachemisch modifizierte Metalloberflächen und ihr Einfluss auf die Zellphysiologie. Workshop on Metallic Biomaterials, abstract Booklet p.7, Geesthacht.

• Prof. Dr. Barbara Nebe
• Dr. Friederike Kunz