Despite years of research, treatment of traumatic brain injury (TBI) remains challenging. Considerable data exists that some volatile anesthetics might be neuroprotective. However, several studies have also revealed a rather neurotoxic profile of anesthetics. In this study, we investigated the effects of argon 50%, desflurane 6% and their combination in an in vitro TBI model with incubation times similar to narcotic time slots in a daily clinical routine. Organotypic hippocampal brain slices of 5- to 7-day-old mice were cultivated for 14 days before TBI was performed. Slices were eventually incubated for 2 hours in an atmosphere containing no anesthetic gas, argon 50% or desflurane 6% or both. Trauma intensity was evaluated via fluorescent imagery. Our results show that neither argon 50% nor desflurane 6% nor their combination could significantly reduce the trauma intensity in comparison to the standard atmosphere. However, in comparison to desflurane 6%, argon 50% displayed a rather neuroprotective profile within the first 2 hours after a focal mechanical trauma (P = 0.015). A 2-hour incubation in an atmosphere containing both gases, argon 50% and desflurane 6%, did not result in significant effects in comparison to the argon 50% group or the desflurane 6% group. Our findings demonstrate that within a 2-hour incubation time neither argon nor desflurane could affect propidium iodide-detectable cell death in an in vitro TBI model in comparison to the standard atmosphere, although cell death was less with argon 50% than with desflurane 6%. The results show that within this short time period processes concerning the development of secondary injury are already taking place and may be manipulated by argon.
Introduction: Organotypic Hippocampal Brain Slices (OHBS) provide an advantageous alternative to in vivo models to scrutinize Traumatic Brain Injury (TBI). We followed a wellestablished TBI protocol, but noticed that several factors may influence the results in such a setup. Here, we describe a structured approach to generate more comparable results and discuss why specific eligibility criteria should be applied. Methods:We defined necessary checkpoints and developed inclusion and exclusion criteria that take the observed variation in such a model into consideration. Objective measures include the identification and exclusion of pre-damaged slices and outliers. Six steps were outlined in this study. Results:A six-step approach to enhance comparability is proposed and summarized in a flowchart. We applied the suggested measures to data derived from our TBI-experiments examining the impact of three different interventions in 1459 OHBS. Our exemplary results show that through equal requirements set for all slices more precise findings are ensured. Conclusion:Results in a TBI experiment on OHBS should be analyzed critically as inhomogeneities may occur. In other words, in order to ensure more precise findings, a structured approach of comparing the results should be followed. Further research is recommended to confirm and further develop this framework. Highlights• Several factors may influence the results in an organotypic hippocampal brain slice model scrutinizing traumatic brain injury.• In order to enhance comparability, a structured approach comprising inclusion and exclusion criteria is outlined.• The proposed approach is applied to data derived from a traumatic brain injury experiment and exemplary results show that through equal requirements set for all slices more precise findings are ensured. Plain Language SummarySeveral researchers utilized organotypic hippocampal slice (OHBS) cultures to scrutinize processes and possible treatment approaches for traumatic brain injury (TBI). Following a well-established protocol for a TBI-experiment we noticed that several factors may influence the results in such a set-up. We describe a structured approach to generate more precise results and discuss why specific eligibility criteria should be applied. We propose 6 steps comprising checkpoints, inclusion-and exclusion criteria in order to enhance comparability. The proposed approach is applied to data derived from our TBI experiment. The exemplary results demonstrate how the outcome can differ when a measure is applied vs. when it is not applied. BGrüßer L., et al. (2019). A 6-step Approach to Gain Higher Quality Results From Organotypic Hippocampal Brain Slices. BCN, 10(5), 485-496.
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