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International Journal of Bioprinting Pregabalin impact on 3D neuronal models

progenitor cells was gained by specifically addressing the for neuropharmacological research mark a significant
impact of pregabalin on Zic1. advancement in the understanding and application of
The alteration of Zic1 gene expression observed in this this technology. We have demonstrated the versatility and
study suggests the potential disruption of regulatory pathways efficacy of 3D bioprinting in creating complex neuronal
governing neural development. Zic1, as a critical player in architectures. The ability to maintain high viability in both
the early stages of brain development, can influence the pregabalin-treated and untreated cortical neurons post-
expression of downstream target genes, thereby modulating bioprinting is a promising outcome. This suggests that
the bioprinting process does not seem to adversely affect
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the fate and differentiation of neural progenitor cells. On the fundamental viability or morphology of the neurons.
the other hand, pregabalin exposure induced non-significant The successful expression of neuronal markers TUJ1 and
alteration in the expression of Lhx6, Emx2, Gsx2, and Nkx2- TBR1 post-printing further supports the potential of
1 in ECNs. In earlier research, it was found that the functional this technology to study drug interactions with neurons,
traits of the neuronal progeny of progenitors in the adult offering a more nuanced understanding than traditional
CNS are heavily influenced by the genetic specifications of 2D cultures. The distribution of cells across different
the progenitors’ embryos by Nkx2-1. 70
layers within the 3D structures, as observed through
Pregabalin’s impact on neural activity and oscillations Z-stack imaging, highlights the technique’s capability to
can be understood through its interaction with various replicate the spatial heterogeneity of neural tissue. This
neural pathways and receptors. Our MEA data indicated aspect is particularly relevant for neuropharmacological
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that upon initial treatment of primary cortical neurons applications, where the 3D context can significantly
with pregabalin, there is a transient increase in neural influence drug efficacy and neuron–drug interactions.
activity and oscillations, followed by a gradual decrease Therefore, our study extends the utility of high-throughput
over time. This phenomenon can be attributed to several 3D bioprinting beyond simple model creation, suggesting
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factors related to the drug’s mechanism of action. its potential for exploring complex neuropharmacological
questions, such as drug penetration, diffusion, and the
The initial reduction in calcium influx caused
by pregabalin binding to the α2δ subunit 72,73 may specific targeting of neuronal subtypes.
paradoxically trigger a compensatory response in the Our study not only demonstrates the successful
neural network, resulting in increased activity and application of high-throughput 3D bioprinting in
oscillations. Neurons might attempt to counteract the neuropharmacology but also highlights the potential of
decrease in neurotransmitter release by increasing their this technology to revolutionize our approach to drug
firing rate, which could contribute to the observed initial discovery and evaluation. By continuing to refine and
increase in neural activity. However, as the drug’s effects expand upon this method, we can anticipate significant
become more established and calcium channel inhibition strides in our understanding and treatment of neurological
continues, the neural network adapts to the altered calcium conditions, moving closer to the realization of more
dynamics. This adaptation involves the normalization of effective and personalized therapeutic strategies.
neurotransmitter release and synaptic activity, ultimately Our research resulted in significant insights into
leading to a decline in neuronal activity and oscillations the consequences of pregabalin exposure during early
observed shortly after treating the neurons with the drug. brain development, addressing a crucial gap in the
Additionally, pregabalin’s modulation of potassium literature. It also demonstrates that a 3D in vitro model
channels and other neurotransmitter systems, such as based on ultrashort self-assembling peptides and high
the inhibition of excitatory amino acid transporters, 72,73 throughput robotic 3D bioprinting technology can
could also play a role in the initial increase and subsequent be efficiently used in the field of neuropharmacology.
decrease of neural activity. These secondary mechanisms Maintenance of regional identity in the forebrain requires
might contribute to the initial response and the subsequent transcription factors encoded by Gad67, Otp, Zic1, Nkx2-
adaptation observed over time. 1, NhIh2, Emx2, Gsx2, Dlx2, Olig2, and Lhx6. Pregabalin
administration to ECNs significantly and non-significantly
Notably, the response of neurons to pregabalin can altered the expression of some of these genes via either
vary based on the specific neural circuitry and dosage of their upregulation or downregulation. These early results
pregabalin, warranting further research to gain a more warrant more research to validate the safety of pregabalin
comprehensive understanding of the temporal dynamics during pregnancy. By elucidating pregabalin’s impact
of pregabalin’s effects on neuronal activity.
on gene regulation and neuronal activity, we provide
The results of our study employing high-throughput essential knowledge for clinicians. Awareness of these
3D robotic bioprinting to fabricate cortical neuron models risks is pivotal for informed decision-making when

Volume 10 Issue 4 (2024) 420 doi: 10.36922/ijb.3010