Thin magnesium layer confirmed as an antibacterial and biocompatible implant coating in a co‑culture model

Zaatreh,Sarah; Haffner,David; Strauss,Madlen; Dauben,Thomas; Zamponi,Christiane; Mittelmeier,Wolfram; Quandt,Eckhard; Kreikemeyer,Bernd; Bader,Rainer

doi:10.3892/mmr.2017.6218

Thin magnesium layer confirmed as an antibacterial and biocompatible implant coating in a co‑culture model

Authors:
- Sarah Zaatreh
- David Haffner
- Madlen Strauss
- Thomas Dauben
- Christiane Zamponi
- Wolfram Mittelmeier
- Eckhard Quandt
- Bernd Kreikemeyer
- Rainer Bader
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Affiliations: Biomechanics and Implant Technology Research Laboratory, Department of Orthopaedics, University Medicine Rostock, D‑18057 Rostock, Germany, Institute for Materials Science, Faculty of Engineering, University of Kiel, D‑24143 Kiel, Germany, Institute of Medical Microbiology, Virology and Hygiene, University Medicine Rostock, D‑18057 Rostock, Germany
Published online on: February 17, 2017 https://doi.org/10.3892/mmr.2017.6218
Pages: 1624-1630
Copyright: © Zaatreh et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Implant-associated infections commonly result from biofilm‑forming bacteria and present severe complications in total joint arthroplasty. Therefore, there is a requirement for the development of biocompatible implant surfaces that prevent bacterial biofilm formation. The present study coated titanium samples with a thin, rapidly corroding layer of magnesium, which were subsequently investigated with respect to their antibacterial and cytotoxic surface properties using a Staphylococcus epidermidis (S. epidermidis) and human osteoblast (hOB) co‑culture model. Primary hOBs and S. epidermidis were co‑cultured on cylindrical titanium samples (Ti6Al4V) coated with pure magnesium via magnetron sputtering (5 µm thickness) for 7 days. Uncoated titanium test samples served as controls. Vital hOBs were identified by trypan blue staining at days 2 and 7. Planktonic S. epidermidis were quantified by counting the number of colony forming units (CFU). The quantification of biofilm‑bound S. epidermidis on the surfaces of test samples was performed by ultrasonic treatment and CFU counting at days 2 and 7. The number of planktonic and biofilm‑bound S. epidermidis on the magnesium‑coated samples decreased by four orders of magnitude when compared with the titanium control following 7 days of co‑culture. The number of vital hOBs on the magnesium‑coated samples was observed to increase (40,000 cells/ml) when compared with the controls (20,000 cells/ml). The results of the present study indicate that rapidly corroding magnesium‑coated titanium may be a viable coating material that possesses antibacterial and biocompatible properties. A co‑culture test is more rigorous than a monoculture study, as it accounts for confounding effects and assesses additional interactions that are more representative of in vivo situations. These results provide a foundation for the future testing of this type of surface in animals.

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1	Fallpauschalenbezogene Krankenhausstatistik (DRG-Statistik): Operationen und Prozeduren der vollstationären Patientinnen und Patienten der Krankenhäuser (4-Steller). Statistisches Bundesamt (Destatis), Wiesbaden. 2015.
2	Darouiche RO: Treatment of infections associated with surgical implants. N Engl J Med. 350:1422–1429. 2004. View Article : Google Scholar : PubMed/NCBI
3	Walter RP and Blake SM: Re: Two-stage exchange for infected resurfacing arthroplasty use of a novel cement spacer technique. J Arthroplasty. 26:9792011. View Article : Google Scholar : PubMed/NCBI
4	Harris LG and Richards RG: Staphylococci and implant surfaces: A review. Injury. 37 Suppl 2:S3–S14. 2006. View Article : Google Scholar : PubMed/NCBI
5	Casey AL, Lambert PA and Elliott TSJ: Staphylococci. Int J Antimicrob Agents. 29 Suppl 3:S23–S32. 2007. View Article : Google Scholar : PubMed/NCBI
6	Arciola CR, Campoccia D, Speziale P, Montanaro L and Costerton JW: Biofilm formation in Staphylococcus implant infections. A review of molecular mechanisms and implications for biofilm-resistant materials. Biomaterials. 33:5967–5982. 2012. View Article : Google Scholar : PubMed/NCBI
7	Wilde AH and Ruth JT: Two-stage reimplantation in infected total knee arthroplasty. Clin Orthop. 23–35. 1988.PubMed/NCBI
8	Vorndran E, Spohn N, Nies B, Rössler S, Storch S and Gbureck U: Mechanical properties and drug release behavior of bioactivated PMMA cements. J Biomater Appl. 26:581–594. 2012. View Article : Google Scholar : PubMed/NCBI
9	Uchiyama K, Takahira N, Fukushima K, Moriya M, Yamamoto T, Minegishi Y, Sakai R, Itoman M and Takaso M: Two-stage revision total hip arthroplasty for periprosthetic infections using antibiotic-impregnated cement spacers of various types and materials. ScientificWorldJournal. 2013:1472482013. View Article : Google Scholar : PubMed/NCBI
10	Connaughton A, Childs A, Dylewski S and Sabesan VJ: Biofilm disrupting technology for orthopedic implants: What's on the Horizon? Front Med (Lausanne). 1:222014.PubMed/NCBI
11	Staiger MP, Pietak AM, Huadmai J and Dias G: Magnesium and its alloys as orthopedic biomaterials: A review. Biomaterials. 27:1728–1734. 2006. View Article : Google Scholar : PubMed/NCBI
12	Luthringer BJ, Feyerabend F and Willumeit-Römer R: Magnesium-based implants: A mini-review. Magnes Res. 27:142–154. 2014.PubMed/NCBI
13	Robinson DA, Griffith RW, Shechtman D, Evans RB and Conzemius MG: In vitro antibacterial properties of magnesium metal against Escherichia coliPseudomonas aeruginosa and Staphylococcus aureus. Acta Biomater. 6:1869–1877. 2010. View Article : Google Scholar : PubMed/NCBI
14	Zaatreh S, Haffner D, Pasold J, Kreikemeyer B, Mittelmeier W, Podbielski A, Quandt E and Bader R: Track A. Biomaterials and biocompatibility 1. Biomed Tech (Berl). 60 Suppl 1:S1–S30. 2015. View Article : Google Scholar : PubMed/NCBI
15	Cytotoxicity test according to ISO 10993-5. CYTOX. 1999.
16	Zaatreh S, Wegner K, Strauß M, Pasold J, Mittelmeier W, Podbielski A, Kreikemeyer B and Bader R: Co-culture of S. epidermidis and human osteoblasts on implant surfaces: An advanced in vitro model for implant-associated infections. PloS One. 11:e01515342016. View Article : Google Scholar : PubMed/NCBI
17	Schlüter K, Zamponi C, Piorra A and Quandt E: Comparison of the corrosion behaviour of bulk and thin film magnesium alloys. Corros Sci. 52:3973–3977. 2010. View Article : Google Scholar
18	Jonitz A, Lochner K, Lindner T, Hansmann D, Marrot A and Bader R: Oxygen consumption, acidification and migration capacity of human primary osteoblasts within a three-dimensional tantalum scaffold. J Mater Sci Mater Med. 22:2089–2095. 2011. View Article : Google Scholar : PubMed/NCBI
19	Anleitungen-Marienfeld-Superior. [Internet]. [cited 3 Nov 2016]. http://www.marienfeld-superior.com/index.php/anleitungen.html
20	Von Eiff C, Peters G and Heilmann C: Pathogenesis of infections due to coagulasenegative staphylococci. Lancet Infect Dis. 2:677–685. 2002. View Article : Google Scholar : PubMed/NCBI
21	Wang HX, Guan SK, Wang X, Ren CX and Wang LG: In vitro degradation and mechanical integrity of Mg-Zn-Ca alloy coated with Ca-deficient hydroxyapatite by the pulse electrodeposition process. Acta Biomater. 6:1743–1748. 2010. View Article : Google Scholar : PubMed/NCBI
22	Kirkland NT, Lespagnol J, Birbilis N and Staiger MP: A survey of bio-corrosion rates of magnesium alloys. Corros Sci. 52:287–291. 2010. View Article : Google Scholar
23	Fischer J, Prosenc MH, Wolff M, Hort N, Willumeit R and Feyerabend F: Interference of magnesium corrosion with tetrazolium-based cytotoxicity assays. Acta Biomater. 6:1813–1823. 2010. View Article : Google Scholar : PubMed/NCBI
24	Fischer J, Pröfrock D, Hort N, Willumeit R and Feyerabend F: Reprint of: Improved cytotoxicity testing of magnesium materials. Mater Sci Eng B. 176:1773–1777. 2011. View Article : Google Scholar
25	Schlüter K, Zamponi C, Hapke J, Hort N, Kainer KU and Quandt E: Mechanical properties and corrosion behaviour of freestanding, precipitate-free magnesium WE43 thin films. Int J Mater Res. 104:286–292. 2013. View Article : Google Scholar
26	Schlüter K, Shi Z, Zamponi C, Cao F, Quandt E and Atrens A: Corrosion performance and mechanical properties of sputter-deposited MgY and MgGd alloys. Corros Sci. 78:43–54. 2014. View Article : Google Scholar
27	Sanchez AH Martinez, Luthringer BJC, Feyerabend F and Willumeit R: Mg and Mg alloys: How comparable are in vitro and in vivo corrosion rates? A review. Acta Biomater. 13:16–31. 2015. View Article : Google Scholar : PubMed/NCBI
28	Willumeit R, Feyerabend F and Huber N: Magnesium degradation as determined by artificial neural networks. Acta Biomater. 9:8722–8729. 2013. View Article : Google Scholar : PubMed/NCBI
29	Charyeva O, Neilands J, Svensäter G and Wennerberg A: Bacterial biofilm formation on resorbing magnesium implants. Open J Med Microbiol. 5:1–11. 2015. View Article : Google Scholar
30	Gasol JM and Del Giorgio PA: Using flow cytometry for counting natural planktonic bacteria and understanding the structure of planktonic bacterial communities. Sci Mar. 64:197–224. 2000. View Article : Google Scholar
31	Lee JH, Wang H, Kaplan JB and Lee WY: Microfluidic approach to create three-dimensional tissue models for biofilm-related infection of orthopaedic implants. Tissue Eng Part C Methods. 17:39–48. 2011. View Article : Google Scholar : PubMed/NCBI
32	Kaji H, Camci-Unal G, Langer R and Khademhosseini A: Engineering systems for the generation of patterned co-cultures for controlling cell-cell interactions. Biochim Biophys Acta. 1810:239–250. 2011. View Article : Google Scholar : PubMed/NCBI