Geometrical Micropillars Combined with Chemical Surface Modifications – Independency of Actin Filament Spatial Distribution in Primary Osteoblasts

Nebe, J. Barbara; Finke, Birgit; Koertge, Andreas; Rebl, Henrike; Staehlke, Susanne

doi:10.4028/www.scientific.net/msf.783-786.1320

Cited by 6 publications

(5 citation statements)

References 10 publications

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“…As it was previously shown that biological cues are highly affected by topographical and biochemical changes of biomaterials, we experimentally determined the optimal L-HT-EphA2 concentration to be 5 μg mL –1 ( Figure S3A,B ). 93 It is also known that this family of tyrosine kinase proteins influences the actin cytoskeleton, affecting cell shape and adhesion. 94 Thus, we evaluated morphology, cell adhesion and spreading as a response to microenvironmental cues as an experimental readout.…”

Section: Resultsmentioning

confidence: 99%

Geometric Control of Cell Behavior by Biomolecule Nanodistribution

Pospíšil

Hrabovský

Bohačiaková

et al. 2022

ACS Biomater. Sci. Eng.

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Many dynamic interactions within the cell microenvironment modulate cell behavior and cell fate. However, the pathways and mechanisms behind cell−cell or cell−extracellular matrix interactions remain understudied, as they occur at a nanoscale level. Recent progress in nanotechnology allows for mimicking of the microenvironment at nanoscale in vitro; electron-beam lithography (EBL) is currently the most promising technique. Although this nanopatterning technique can generate nanostructures of good quality and resolution, it has resulted, thus far, in the production of only simple shapes (e.g., rectangles) over a relatively small area (100 × 100 μm), leaving its potential in biological applications unfulfilled. Here, we used EBL for cell-interaction studies by coating cell-culture-relevant material with electron-conductive indium tin oxide, which formed nanopatterns of complex nanohexagonal structures over a large area (500 × 500 μm). We confirmed the potential of EBL for use in cell-interaction studies by analyzing specific cell responses toward differentially distributed nanohexagons spaced at 1000, 500, and 250 nm. We found that our optimized technique of EBL with HaloTags enabled the investigation of broad changes to a cell-culturerelevant surface and can provide an understanding of cellular signaling mechanisms at a single-molecule level.

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Section: Resultsmentioning

confidence: 99%

Geometric Control of Cell Behavior by Biomolecule Nanodistribution

Pospíšil

Hrabovský

Bohačiaková

et al. 2022

ACS Biomater. Sci. Eng.

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“…In a general comparison with adherent human primary osteoblasts (HOB), the cell size of the MG-63 cells is up to six times smaller (Pautke et al, 2004) (Figure 6). Comparative studies of the MG-63 cells and HOB cells show no morphological differences regarding actin cytoskeleton organization on different topographically modified biomaterials (L€ uthen et al, 2005;Nebe et al, 2014).…”

Section: Morphologymentioning

confidence: 92%

“…The morphology of MG-63 cells growing in T 75 -culture flasks was quite similar over the passage range P5-P30 ( Figure 6A). Comparative studies of the MG-63 cells and HOB cells show no morphological differences regarding actin cytoskeleton organization on different topographically modified biomaterials (L€ uthen et al, 2005;Nebe et al, 2014). In a general comparison with adherent human primary osteoblasts (HOB), the cell size of the MG-63 cells is up to six times smaller (Pautke et al, 2004) (Figure 6).…”

Section: Morphologymentioning

confidence: 99%

Phenotypic stability of the human MG‐63 osteoblastic cell line at different passages

Staehlke

Rebl

Nebe

2018

Cell Biology International

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One of the most popular cell lines in osteogenesis studies is the human osteoblastic line MG-63. For cell biological investigation, it is important that the cells remain stable in their phenotype over several passages in cell culture. MG-63 cells can be used to provide fundamental insights into cell--material interaction. The aim of this study is to present a systematic characterization of the physiological behavior of MG-63 cells in the range of passages 5-30. Significant cell physiology processes during the first 24 h, including cell morphology, availability of adhesion receptors, cell cycle phases, as well as the expression of the signaling proteins Akt, GSK3a/b, IkB-a, ERK1/2, p38-MAPK, and intracellular calcium ion mobilization, remained stable over the entire range of passages P5-P30. Due to these stable characteristics in a wide range of cell culture passages, MG-63 cells can be considered as a suitable in vitro model to analyze the biocompatibility and biofunctionality of implant materials.

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“…The cell behavior on a biomaterial's surface is highly complex, and all the adhesion and signaling processes occurring at the interface are not fully understood until now. We revealed on a micro-pillared surface that osteoblasts developed an actin pattern adequate to the underlying surface -in mimicry fashion [1,2].…”

Section: Introductionmentioning

confidence: 94%

Complex Cell Physiology on Topographically and Chemically Designed Material Surfaces

Nebe

Moerke

Staehlke

et al. 2016

MSF

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A crucial factor for ingrowth of permanent implants in the bone is the rapid cellular acceptance. The topographical features often follow mechanical aspects for implant stability. But several of these implants fail due to insufficient cell adhesion. Cells are able to perceive the physico-chemical properties of their surrounding and to pass these signals into the cell to modulate their adhesion structures, growth or production of extracellular matrix. However, the complex cell physiology at the material interface is not yet fully understood, particular on stochastically structured topographies resulting from industrial production. We could find out that corundum blasted titanium hampered the organization of actin filaments inside the cells, clustered adhesion components, e. g. beta-1 integrins and tensin, and the cells bridged the valleys which reduces cell-substrate contacts. These morphological changes strongly diminished the mineralization of osteoblasts. To shed light on cause and effect we reduced the physical complexity of the material surface by introduction of regular micro-structures (pillars, grooves) using deep reactive ion etching. Now it was more obvious what cells are doing on sharp edged topographies ‒ the actin filaments of our cells were clustered around the pillars. As a result the intracellular calcium signaling and the protein synthesis were impaired. Our recent findings indicated an attempted phagocytosis of the micro-pillars by osteoblasts. Therefore we conclude that implants used in orthopedic surgery should avoid any sharp-edged topographical features that could induce phagocytosis by the surrounding cells, which is an unnecessarily energy consuming process.

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Geometrical Micropillars Combined with Chemical Surface Modifications – Independency of Actin Filament Spatial Distribution in Primary Osteoblasts

Cited by 6 publications

References 10 publications

Geometric Control of Cell Behavior by Biomolecule Nanodistribution

Geometric Control of Cell Behavior by Biomolecule Nanodistribution

Phenotypic stability of the human MG‐63 osteoblastic cell line at different passages

Complex Cell Physiology on Topographically and Chemically Designed Material Surfaces

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