Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Dec;29(4):528-536.
doi: 10.1111/jns.12659. Epub 2024 Sep 27.

Oxygen matters: Unraveling the role of oxygen in the neuronal response to cisplatin

Jose Crugeiras  1   2 , Aina Calls  1   2 , Estefanía Contreras  1   2 , Montse Alemany  3 , Xavier Navarro  1   2 , Victor J Yuste  4 , Oriol Casanovas  5 , Esther Udina  1   2 , Jordi Bruna  2   3
Affiliations

Oxygen matters: Unraveling the role of oxygen in the neuronal response to cisplatin

Jose Crugeiras et al. J Peripher Nerv Syst. 2024 Dec.

Abstract

Background and aims: Cell culture is a fundamental experimental tool for understanding cell physiology. However, translating these findings to in vivo settings has proven challenging. Replicating donor tissue conditions, including oxygen levels, is crucial for achieving meaningful results. Nevertheless, oxygen culture conditions are often overlooked, particularly in the context of chemotherapy-induced neurotoxicity.

Methods: In this study, we investigated the role of oxygen levels in primary neuronal cultures by comparing neuronal performance under cisplatin exposure (1 μg/mL) in supraphysiological normoxia (representing atmospheric conditions in a standard incubator; 18.5% O2) and physioxia (representing physiologic oxygen conditions in nervous tissue; 5% O2). Experiments were also conducted to assess survival, neurite development, senescence marker expression, and proinflammatory cytokine secretion.

Results: Under control conditions, both oxygen concentration conditions exhibited similar behaviors. However, after cisplatin administration, sensory neurons cultured under supraphysiological normoxic conditions show higher mortality, exhibit an evolutionarily proinflammatory cytokine profile over time, and activate apoptotic-regulated neuron death markers. In contrast, under physiological conditions, neurons treated with cisplatin exhibited senescence marker expression and an attenuated inflammatory secretome.

Interpretation: These results underscore the critical role of oxygen in neuronal culture, particularly in studying compounds where neuronal damage is mechanistically linked to oxidative stress. Even at identical doses of evaluated neurotoxic drugs, distinct cellular phenotypic fates can emerge, impacting translatability to the in vivo setting.

Keywords: cell culture; chemotherapy‐induced neuropathy; cisplatin; neurotoxicity; oxygen.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

FIGURE 1
FIGURE 1
Neuronal survival after CDDP administration in DRG cells cultured under different atmospheric conditions (supraphysiological normoxia (18.5% O2) and physioxia (5% O2)). The percentage of surviving cells after CDDP treatment was calculated with respect to the basal concentration for each condition. A significant loss of neurons over time was observed in CDDP‐treated cultures, and this loss was greater under normoxic (18.5% O2) conditions than under physioxic (5% O2) conditions. n = 4 cultures. Statistical analyses were performed using repeated measures two‐way ANOVA followed by the post hoc Bonferroni correction. ****p < .001 CDDP Normoxia (18.5% O2) versus Control Normoxia (18.5% O2). % p < .001 CDDP Physioxia (5% O2) versus Control Physioxia (5% O2). &p < .001 CDDP Physioxia (5% O2) 4th day versus CDDP Physioxia (5% O2) 1st day. €p < .001 CDDP Normoxia (18.5% O2) 4th day versus CDDP Normoxia (18.5% O2) 1st day. *p < .05 CDDP Physioxia (5% O2) versus CDDP Normoxia (18.5% O2). The data are presented as the means ± SDs.
FIGURE 2
FIGURE 2
Neurite extension after CDDP administration in DRG cell cultures under different atmospheric conditions (supraphysiological normoxia (18.5% O2) and physioxia (5% O2)). Quantification of the longest neurites in each condition (n = 3–4 experiments per condition). Two‐way ANOVA followed by post hoc Bonferroni correction was used for multiple comparisons. *p < .05 CDDP Normoxia (18.5% O2) versus Control Normoxia (18.5% O2). **p < .005 CDDP Physioxia (5% O2) versus Control Physioxia (5% O2). The data are expressed as the mean ± SD.
FIGURE 3
FIGURE 3
Senescence hallmarks after CDDP administration in DRG cells cultured under different atmospheric conditions. The expression of different markers was analyzed via RT‐qPCR. (A) Expression level of Lamin B1. (n = 5–6 experiments per condition. *p < .01 CDDP Normoxia (18.5% O2) versus CDDP Physioxia (5% O2). **p < .05 CDDP Physioxia (5% O2) versus Control Physioxia (5% O2)). (B) Expression levels of Cdkn1a, a senescence inducer (n = 5–7 experiments per condition). *p < .05 CDDP Physioxia (5% O2) versus Control Physioxia (5% O2). ****p < .0001 CDDP Normoxia (18.5% O2) versus Control Normoxia (18.5% O2). (C) Expression levels of Cdkn2a, a senescence inducer (n = 5–7 experiments per condition). **p < .05 CDDP Physioxia (5% O2) versus Control Physioxia (5% O2). (D) Expression level of caspase‐3 (n = 3 experiments per condition). **p < .05 CDDP Normoxia (18.5% O2) versus Control Normoxia (18.5% O2). ***p < .0001 CDDP Physioxia (5% O2) versus CDDP Normoxia (18.5% O2)). (E) Expression levels of IL‐6 (n = 3 experiments per condition). (A–E) Two‐way ANOVA followed by post hoc Bonferroni correction for multiple comparisons. The data are expressed as the fold change versus the control condition and are presented as the mean ± SD. RT‐qPCR, real‐time quantitative polymerase chain reaction.
FIGURE 4
FIGURE 4
Secretome cytokine composition in the supernatant of DRG cultures under different atmospheric conditions after CDDP administration. Measurement of the fold change in the expression of each cytokine in physioxia and normoxia CDDP cultures at two time points. n = 3–8 independent experiments for each condition and time point. (A) IL‐6. *p < .05 CDDP Physioxia (5% O2) 24 h versus CDDP Normoxia (18.5% O2) 24 h). (B) CXCL10. (C) CXCL1 (GROα). (D) CCL5 (RANTES) *p < .05 CDDP Physioxia (5% O2) 24 h versus CDDP Normoxia (18.5% O2) 24 h). (E) CCL3 (MIP‐1a) ***p < .005 CDDP Physioxia (5% O2) 24 h versus CDDP Normoxia (18.5% O2) 24 h). (F) CCL11 (EOTAXIN) **p < .001 CDDP Normoxia (18.5% O2) 72 h versus CDDP Normoxia (18.5% O2) 24 h. *** < 0.005 CDDP Normoxia (18.5% O2) 72 h versus Control Normoxia (18.5% O2) 72 h. ****p < .0001 CDDP Physioxia (5% O2) 72 h versus CDDP Normoxia (18.5% O2) 72 h). (A–F) Two‐way ANOVA followed by post hoc Bonferroni correction for multiple comparisons. The data are expressed as the fold change versus the control condition and are presented as the mean ± SD.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Smith PA. Neuropathic pain; what we know and what we should do about it. Front Pain Res (Lausanne). 2023;4. doi:10.3389/FPAIN.2023.1220034 - DOI - PMC - PubMed
    1. Khodosevich K, Dragicevic K, Howes O. Drug targeting in psychiatric disorders—how to overcome the loss in translation? Nat Rev Drug Discov. 2024;23(3):218‐231. doi:10.1038/S41573-023-00847-7 - DOI - PubMed
    1. Seyhan AA. Lost in translation: the valley of death across preclinical and clinical divide—identification of problems and overcoming obstacles. Transl Med Commun. 2019;4(1):1‐19. doi:10.1186/S41231-019-0050-7 - DOI
    1. Calls A, Carozzi V, Navarro X, Monza L, Bruna J. Pathogenesis of platinum‐induced peripheral neurotoxicity: insights from preclinical studies. Exp Neurol. 2020;325. doi:10.1016/J.EXPNEUROL.2019.113141 - DOI - PubMed
    1. Staff NP, Fehrenbacher JC, Caillaud M, Damaj MI, Segal RA, Rieger S. Pathogenesis of paclitaxel‐induced peripheral neuropathy: a current review of in vitro and in vivo findings using rodent and human model systems. Exp Neurol. 2020;324:113121. doi:10.1016/J.EXPNEUROL.2019.113121 - DOI - PMC - PubMed

LinkOut - more resources