Development of an electrode cell impedance method to measure osteoblast cell activity in magnesium-conditioned media
Magnesium (Mg) as a biodegradable metal has potential advantages as an implant material. This paper studies the effect of magnesium ions on osteoblast (U2-OS) behavior since magnesium implants mainly dissolve as divalent magnesium ions (Mg2+). A real-time monitoring technique based on electric cell-substrate impedance sensing (ECIS) was used for measuring cell proliferation, migration, adhesion, and cytotoxicity in magnesium-conditioned media. The impedance results show that U2-OS proliferation and adhesion were inhibited in not only a magnesium-free medium but also in a medium with a high concentration of magnesium. The impedance method produced more sensitive results than the output of an MTT assay. Other standard bioanalytical tests were conducted for comparison with the ECIS method. Immunochemistry was carried out to study cell adhesion in magnesium-conditioned media by staining using F-actin and α-tubulin and correlated cell density on the electrode with impedance. Bone tissue formation was studied using von Kossa staining and indicated the mineralization level of cells in magnesium-conditioned media decreased with the increase of magnesium ion concentration. Real-time PCR provided gene expression indicators of cell growth, apoptosis, inflammation, and migration. Compared to the bioanalytical methods of immunochemistry and MTT assays, which need preparation time and post-washing step, ECIS was able to measure cell activity in real time without any cell culture modification. In summary, ECIS might be an effective way to study biodegradable magnesium implants.
KeywordsBiodegradable magnesium implant U2-OS cells Electric cell-substrate impedance sensing (ECIS) Cytotoxicity MTT assay Real-time PCR
This work was sponsored by the NSF ERC for Revolutionizing Metallic Biomaterials, http://erc.ncat.edu/.
- 4.Zeng R, Dietzel W, Witte F, Hort N (2008) Blawert C 10:B3–B14Google Scholar
- 6.Song G (2007) Song S 9:298–302Google Scholar
- 7.Lorenz C, Brunnera J, Kollmannsberger P, Jaafar L, Fabry B, Virtanena S (2009) Acta Biomaterialia 5:2783–2789Google Scholar
- 12.Günther T (2008) Magnes Res 21:185–187Google Scholar
- 13.Hornby J (1973) Embryol Exp Morph 30:511–518Google Scholar
- 20.Abed E, Moreau R (2010) Am J Physiol Cell Physiol 297:C360–368Google Scholar
- 22.Maier JA, Bernardini D, Rayssiguier Y, Mazur A (2004) Biochim Biophys Acta 24:6–12Google Scholar
- 23.Moomaw AS, Maguire ME (2008) Physiology (Bethesda) 23:275–285Google Scholar
- 25.Günther T (2006) Magnes Res 19:225–36Google Scholar