title: In vivo continuous and simultaneous monitoring of brain energy substrates with a multiplex amperometric enzyme-based biosensor device.
abstract: Enzyme-based amperometric biosensors are widely used for monitoring key biomarkers. In experimental neuroscience there is a growing interest in in vivo continuous and simultaneous monitoring of metabolism-related biomarkers, like glucose, lactate and pyruvate. The use of multiplex biosensors will provide better understanding of brain energy metabolism and its role in neuropathologies such as diabetes, ischemia, and epilepsy. We have developed and characterized an implantable multiplex microbiosensor device (MBD) for simultaneous and continuous in vivo monitoring of glucose, lactate, and pyruvate. First, we developed and characterized amperometric microbiosensors for monitoring lactate and pyruvate. In vitro evaluation allowed us to choose the most suitable biosensors for incorporation into the MBD, along with glucose and background biosensors. Fully assembled MBDs were characterized in vitro. The calculated performance parameters (LOD, LR, LRS, IMAX and appKM) showed that the multiplex MBD was highly selective and sensitive (LRS≥100 nA/mM) for each analyte and within an adequate range for in vivo application. Finally, MBDs were implanted in the mPFC of anesthetized adult male Wistar rats for in vivo evaluation. Following an equilibration period, baseline brain levels of glucose (1.3±0.2 mM), lactate (1.5±0.4 mM) and pyruvate (0.3±0.1 mM) were established. Subsequently, the MBDs recorded the responses of the animals when submitted to hyperglycemic (40% glucose i.v.) and hypoglycemic (5 U/kg insulin i.v.) challenges. Afterwards, MBDs were recalibrated to convert electrochemical readings into accurate substrate concentrations and to assess biofouling. The presented MBD can monitor simultaneously multiple biomarkers in vivo.
authors.affiliation: Brains On-Line BV, Groningen, the Netherlands; University of Groningen Institute of Pharmacy, Groningen, the Netherlands. Electronic address: carlos.cordeiro@brainsonline.org.
authors.forename: C A
authors.initials: CA
authors.lastname: Cordeiro
authors.affiliation: Brains On-Line BV, Groningen, the Netherlands.
authors.forename: M G
authors.initials: MG
authors.lastname: de Vries
authors.affiliation: University of Groningen Institute of Pharmacy, Groningen, the Netherlands.
authors.forename: W
authors.initials: W
authors.lastname: Ngabi
authors.affiliation: Brains On-Line BV, Groningen, the Netherlands.
authors.forename: P E
authors.initials: PE
authors.lastname: Oomen
authors.affiliation: Brains On-Line BV, Groningen, the Netherlands; University of Groningen Institute of Pharmacy, Groningen, the Netherlands; Brains On-Line LLC, San Francisco, California, USA.
authors.forename: T I F H
authors.initials: TI
authors.lastname: Cremers
authors.affiliation: Brains On-Line BV, Groningen, the Netherlands; University of Groningen Institute of Pharmacy, Groningen, the Netherlands.
authors.forename: B H C
authors.initials: BH
authors.lastname: Westerink
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D005947:Glucose / Q000302:isolation & purification* / Q000378:metabolism; D019344:Lactic Acid / Q000302:isolation & purification* / Q000378:metabolism; D008297:Male; D008991:Monitoring, Physiologic; D019289:Pyruvic Acid / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats
chemical_list: D019344:Lactic Acid; D019289:Pyruvic Acid; D005947:Glucose
keywords: Amperometric; Continuous biomonitoring; Enzyme-based; In vivo; Multiplex; Real-time