title: Primary neuronal-astrocytic co-culture platform for neurotoxicity assessment of di-(2-ethylhexyl) phthalate.
abstract: Plastics such as polyvinyl chlorides (PVC) are widely used in many indoor constructed environments; however, their unbound chemicals, such as di-(2-ethylhexyl) phthalates (DEHP), can leach into the surrounding environment. This study focused on DEHP's effect on the central nervous system by determining the precise DEHP content in mice brain tissue after exposure to the chemical, to evaluate the specific exposure range. Primary neuronal-astrocyte co-culture systems were used as in vitro models for chemical hazard identification of DEHP. Oxidative stress was hypothesized as a probable mechanism involved, and therefore the total reactive oxygen species (ROS) concentration was determined as a biomarker of oxidative stress. In addition, NeuriteTracer, a neurite tracing plugin with ImageJ, was used to develop an assay for neurotoxicity to provide quantitative measurements of neurological parameters, such as neuronal number, neuron count and neurite length, all of which could indicate neurotoxic effects. The results showed that with 1 nmol/L DEHP exposure, there was a significant increase in ROS concentrations, indicating that the neuronal-astrocyte cultures were injured due to exposure to DEHP. In response, astrocyte proliferation (gliosis) was initiated, serving as a mechanism to maintain a homeostatic environment for neurons and protect neurons from toxic chemicals. There is a need to assess the cumulative effects of DEHP in animals to evaluate the possible uptake and effects on the human neuronal system from exposure to DEHP in the indoor environment.
authors: Section of Environmental Biomedicine, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, Central China Normal University, Wuhan 430079, China; Department of Molecular and Medical Pharmacology, University of California Los Angeles (UCLA), Los Angeles, CA 90095, USA. Electronic address: wuyang886@gmail.com.
authors: Yang
authors: Y
authors: Wu
authors: Wuhan Institute of Drug and Food Control, Wuhan 430012, China.
authors: Ke
authors: K
authors: Li
authors: Section of Environmental Biomedicine, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, Central China Normal University, Wuhan 430079, China.
authors: Haoxiao
authors: H
authors: Zuo
authors: Section of Environmental Biomedicine, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, Central China Normal University, Wuhan 430079, China.
authors: Ye
authors: Y
authors: Yuan
authors: Department of Molecular and Medical Pharmacology, University of California Los Angeles (UCLA), Los Angeles, CA 90095, USA.
authors: Yi
authors: Y
authors: Sun
authors: Section of Environmental Biomedicine, Hubei Key Laboratory of Genetic Regulation and Integrative Biology, Central China Normal University, Wuhan 430079, China. Electronic address: yangxu@mail.ccnu.edu.cn.
authors: Xu
authors: X
authors: Yang
mesh_terms: D000818:Animals; D018920:Coculture Techniques / Q000379:methods; D004051:Diethylhexyl Phthalate / Q000008:administration & dosage / Q000737:chemistry / Q000633:toxicity*; D006801:Humans; D007928:Lethal Dose 50; D051379:Mice; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000166:cytology* / Q000187:drug effects*; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species; D013047:Specific Pathogen-Free Organisms
chemical_list: D017382:Reactive Oxygen Species; D004051:Diethylhexyl Phthalate
keywords: astrocyte gliosis; di-(2-ethylhexyl)phthalate; immunostaining; neurite length; neuron count; neurotoxicity; oxidative stress