Establishing an in Vitro Model of Astrocyte Activation in Ischemic Traumatic Brain Injury: Therapeutic Synergy of Melatonin and Oxygen Nanobubbles
4 affiliations
- 1University of Illinois at Urbana−champaign , , , ,
- 2Carle Foundation Hospital , , , ,
- 3University of Illinois at Urbana−Champaign , , , ,
- 4Carle-Illinois College of Medicine , , ,
Abstract
Secondary injury following ischemic traumatic brain injury (TBI) is heavily driven by neurovascular dysfunction involving endothelial damage, astrocyte reactivity, inflammation, and hypoxic stress, among other contributing pathophysiological processes. Here, we establish a mechanistically informative in vitro ischemic TBI model that isolates endothelial-astrocyte crosstalk using human brain microvascular endothelial cells (HBMECs) and astrocytes. Using this platform, we evaluated melatonin and oxygen nanobubbles (ONBs) as a combinatorial therapeutic strategy. Fabricated ONBs exhibited a mean hydrodynamic diameter of 78.35 ± 0.42 nm, a negative surface charge of −37.86 ± 1.32 mV, and sustained oxygen retention (56.38 ± 0.86 mg/L initially). Mechanical needle-scratch injury combined with hypoxia induced primary endothelial injury characterized by reduced viability and increased expression of inflammatory and hypoxia-associated markers, quantified by quantitative real-time PCR (qRT-PCR). Injured HBMECs subsequently triggered astrocyte reactivity marked by elevated GFAP (1.93-fold; p < 0.0001), NLRP3 (3.21-fold; p < 0.0001), NF-κβ signaling (1.62-fold; p < 0.0001), and HIF-1α (2.28-fold; p < 0.0001), while reactive astrocytes propagated a secondary wave of endothelial degeneration, establishing a bidirectional injury loop. Melatonin alone only partially mitigated injury-associated signaling, whereas the melatonin-ONBs combination restored astrocytic and endothelial gene expression to near-baseline levels. These findings highlight a multimodal therapeutic approach that effectively suppresses inflammatory, apoptotic, and hypoxic signaling in ischemic TBI.
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BibTeX
@article{igbalaye2026establishing,
title = {{Establishing an in Vitro Model of Astrocyte Activation in Ischemic Traumatic Brain Injury: Therapeutic Synergy of Melatonin and Oxygen Nanobubbles}},
author = {Jimoh Olamilekan Igbalaye and Shreyaan Jain and Elizabeth Wenning and Anika Bushra and Rhea Nagabhyru and Zsaleigh Dagdagan and Joseph Irudayaraj},
journal = {ACS Omega},
year = {2026},
month = oct,
doi = {10.1021/acsomega.6c06656},
url = {https://pubs.acs.org/acsodf/article/11/41/63265/5444942/Establishing-an-in-Vitro-Model-of-Astrocyte},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - JOUR
TI - Establishing an in Vitro Model of Astrocyte Activation in Ischemic Traumatic Brain Injury: Therapeutic Synergy of Melatonin and Oxygen Nanobubbles
AU - Jimoh Olamilekan Igbalaye
AU - Shreyaan Jain
AU - Elizabeth Wenning
AU - Anika Bushra
AU - Rhea Nagabhyru
AU - Zsaleigh Dagdagan
AU - Joseph Irudayaraj
PY - 2026
DA - 2026/10/06
JO - ACS Omega
DO - 10.1021/acsomega.6c06656
UR - https://pubs.acs.org/acsodf/article/11/41/63265/5444942/Establishing-an-in-Vitro-Model-of-Astrocyte
AB - Secondary injury following ischemic traumatic brain injury (TBI) is heavily driven by neurovascular dysfunction involving endothelial damage, astrocyte reactivity, inflammation, and hypoxic stress, among other contributing pathophysiological processes. Here, we establish a mechanistically informative in vitro ischemic TBI model that isolates endothelial-astrocyte crosstalk using human brain microvascular endothelial cells (HBMECs) and astrocytes. Using this platform, we evaluated melatonin and oxygen nanobubbles (ONBs) as a combinatorial therapeutic strategy. Fabricated ONBs exhibited a mean hydrodynamic diameter of 78.35 ± 0.42 nm, a negative surface charge of −37.86 ± 1.32 mV, and sustained oxygen retention (56.38 ± 0.86 mg/L initially). Mechanical needle-scratch injury combined with hypoxia induced primary endothelial injury characterized by reduced viability and increased expression of inflammatory and hypoxia-associated markers, quantified by quantitative real-time PCR (qRT-PCR). Injured HBMECs subsequently triggered astrocyte reactivity marked by elevated GFAP (1.93-fold; p < 0.0001), NLRP3 (3.21-fold; p < 0.0001), NF-κβ signaling (1.62-fold; p < 0.0001), and HIF-1α (2.28-fold; p < 0.0001), while reactive astrocytes propagated a secondary wave of endothelial degeneration, establishing a bidirectional injury loop. Melatonin alone only partially mitigated injury-associated signaling, whereas the melatonin-ONBs combination restored astrocytic and endothelial gene expression to near-baseline levels. These findings highlight a multimodal therapeutic approach that effectively suppresses inflammatory, apoptotic, and hypoxic signaling in ischemic TBI.
SN - 2470-1343
ER -