title: Use of cortical neuronal networks for in vitro material biocompatibility testing.
abstract: Neural interfaces aim to restore neurological function lost during disease or injury. Novel implantable neural interfaces increasingly capitalize on novel materials to achieve microscale coupling with the nervous system. Like any biomedical device, neural interfaces should consist of materials that exhibit biocompatibility in accordance with the international standard ISO10993-5, which describes in vitro testing involving fibroblasts where cytotoxicity serves as the main endpoint. In the present study, we examine the utility of living neuronal networks as functional assays for in vitro material biocompatibility, particularly for materials that comprise implantable neural interfaces. Embryonic mouse cortical tissue was cultured to form functional networks where spontaneous action potentials, or spikes, can be monitored non-invasively using a substrate-integrated microelectrode array. Taking advantage of such a platform, we exposed established positive and negative control materials to the neuronal networks in a consistent method with ISO 10993-5 guidance. Exposure to the negative controls, gold and polyethylene, did not significantly change the neuronal activity whereas the positive controls, copper and polyvinyl chloride (PVC), resulted in reduction of network spike rate. We also compared the functional assay with an established cytotoxicity measure using L929 fibroblast cells. Our findings indicate that neuronal networks exhibit enhanced sensitivity to positive control materials. In addition, we assessed functional neurotoxicity of tungsten, a common microelectrode material, and two conducting polymer formulations that have been used to modify microelectrode properties for in vivo recording and stimulation. These data suggest that cultured neuronal networks are a useful platform for evaluating the functional toxicity of materials intended for implantation in the nervous system.
authors.affiliation: Electrical and Computer Engineering Department, George Mason University, 4400 University Dr. MSN 1G5, Fairfax, VA 22030, USA. Electronic address: hcharkhk@masonlive.gmu.edu.
authors.forename: Hamid
authors.initials: H
authors.lastname: Charkhkar
authors.forename: Christopher
authors.initials: C
authors.lastname: Frewin
authors.forename: Maysam
authors.initials: M
authors.lastname: Nezafati
authors.forename: Gretchen L
authors.initials: GL
authors.lastname: Knaack
authors.forename: Nathalia
authors.initials: N
authors.lastname: Peixoto
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Saddow
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Pancrazio
mesh_terms: D000200:Action Potentials; D000818:Animals; D001672:Biocompatible Materials / Q000633:toxicity*; D015374:Biosensing Techniques / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004594:Electrophysiology; D005347:Fibroblasts; D051379:Mice; D009420:Nervous System / Q000187:drug effects; D009474:Neurons / Q000187:drug effects*; D020959:Polyethylene / Q000302:isolation & purification* / Q000633:toxicity; D011108:Polymers / Q000633:toxicity
chemical_list: D001672:Biocompatible Materials; D011108:Polymers; D020959:Polyethylene
keywords: Biomaterials; Cytotoxicity; Extracellular recordings; Functional assay; Microelectrode array; Primary neuronal cultures