title: Optimized in vitro procedure for assessing the cytocompatibility of magnesium-based biomaterials.
abstract: Magnesium (Mg) is a promising biomaterial for degradable implant applications that has been extensively studied in vitro and in vivo in recent years. In this study, we developed a procedure that allows an optimized and uniform in vitro assessment of the cytocompatibility of Mg-based materials while respecting the standard protocol DIN EN ISO 10993-5:2009. The mouse fibroblast line L-929 was chosen as the preferred assay cell line and MEM supplemented with 10% FCS, penicillin/streptomycin and 4mM l-glutamine as the favored assay medium. The procedure consists of (1) an indirect assessment of effects of soluble Mg corrosion products in material extracts and (2) a direct assessment of the surface compatibility in terms of cell attachment and cytotoxicity originating from active corrosion processes. The indirect assessment allows the quantification of cell-proliferation (BrdU-assay), viability (XTT-assay) as well as cytotoxicity (LDH-assay) of the mouse fibroblasts incubated with material extracts. Direct assessment visualizes cells attached to the test materials by means of live-dead staining. The colorimetric assays and the visual evaluation complement each other and the combination of both provides an optimized and simple procedure for assessing the cytocompatibility of Mg-based biomaterials in vitro.
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Germany. Electronic address: ol.jung@uke.de.
authors.forename: Ole
authors.initials: O
authors.lastname: Jung
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Smeets
authors.affiliation: Meotec GmbH & Co. KG, Aachen, Germany; Aachen University of Applied Sciences, Biomaterials Laboratory, Aachen, Germany.
authors.forename: Dario
authors.initials: D
authors.lastname: Porchetta
authors.affiliation: Meotec GmbH & Co. KG, Aachen, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Kopp
authors.affiliation: Meotec GmbH & Co. KG, Aachen, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Ptock
authors.affiliation: BMP, Laboratory for Medical Materials Testing, Aachen, Germany.
authors.forename: Ute
authors.initials: U
authors.lastname: Müller
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Germany.
authors.forename: Max
authors.initials: M
authors.lastname: Heiland
authors.affiliation: Laboratory for Machine Tools and Production Engineering (WZL) of RWTH Aachen University, Germany.
authors.forename: Max
authors.initials: M
authors.lastname: Schwade
authors.affiliation: Department of Plastic and Hand Surgery, Burn Center, BG-University Hospital Bergmannsheil, Ruhr University Bochum, Germany.
authors.forename: Björn
authors.initials: B
authors.lastname: Behr
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Germany.
authors.forename: Nadja
authors.initials: N
authors.lastname: Kröger
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Germany.
authors.forename: Lan
authors.initials: L
authors.lastname: Kluwe
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Germany.
authors.forename: Henning
authors.initials: H
authors.lastname: Hanken
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Germany.
authors.forename: Philip
authors.initials: P
authors.lastname: Hartjen
mesh_terms: D000465:Algorithms; D000818:Animals; D001672:Biocompatible Materials / Q000633:toxicity*; D001681:Biological Assay / Q000379:methods / Q000592:standards; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects* / Q000473:pathology; D017408:Guidelines as Topic; D066298:In Vitro Techniques; D038622:Internationality; D008274:Magnesium / Q000633:toxicity*; D008422:Materials Testing / Q000379:methods / Q000592:standards*; D051379:Mice; D018675:Toxicity Tests / Q000379:methods / Q000592:standards*
chemical_list: D001672:Biocompatible Materials; D008274:Magnesium
keywords: Cytocompatibility; In vitro biocompatibility; In vitro cytotoxicity; Magnesium; Plasma electrolytic oxidation (PEO)