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International Journal of Bioprinting Pregabalin impact on 3D neuronal models

peptide hydrogel as bioink, we created constructs with pregabalin-treated and untreated primary mouse embryonic
cortical neurons. This setup allowed us to study the drug’s effects on cell viability, expression of neuronal markers, and
neuron development. Our comparative analysis between 2D and 3D peptide-based cell culture models revealed that
at a therapeutic concentration of 10 µM, pregabalin does not affect neuronal viability or the morphogenesis of cortical
neurons. However, it significantly alters adenosine triphosphate (ATP) release, suggesting potential disruptions in
mitochondrial function. Moreover, gene expression analysis of key genes involved in the development of the forebrain
and the differentiation and maturation of neurons revealed significant alterations, including the downregulation of
Dlx2, Nhlh2, Otp, and Gad67. These findings, together with observed alterations in neuronal activity and oscillations,
emphasize the complex impact of pregabalin on neuronal development and function. They highlight the necessity for
comprehensive clinical evaluations of its use during pregnancy. Furthermore, our research demonstrates the feasibility
and value of integrating 3D cultures with high-throughput 3D bioprinting in neuropharmacology, opening new avenues
for investigating drug effects on neuronal development and function, and contributing to safer clinical practices.

Keywords: Pregabalin; Cortical neurons; Fetal neurodevelopment; Neuropathic pain;
Embryonic neurons; Ultrashort self-assembling peptides; 3D neuronal models

1. Introduction risk was found. Another study reported a higher rate of
12
major birth defects and reduced live births in pregabalin-
Pregabalin is a compound similar to gamma-aminobutyric exposed pregnancies. However, the limitations of the
13
acid (GABA), which acts as an inhibitory neurotransmitter. study precluded forming definitive conclusions. Recently,
This substance functions as a neuromodulator with a an interesting review of five studies suggested there is a
central action that prevents the release of excitatory small increased risk of malformations, but strong evidence
neurotransmitters, possibly reducing excessive excitation 14
in neurons. Pregabalin has been approved by the United for a teratogenic effect is lacking. Another comprehensive
1
States (US) Food and Drug Administration (FDA) for the review indicated pregabalin’s low teratogenic potential,
treatment of fibromyalgia and neuropathic pain caused emphasizing cautious dosing and continual monitoring
2,3
by spinal cord injury, post-herpetic neuralgia (PHN), during pregnancy. Further research on analgesic safety in
3,4
5
15
and diabetic neuropathy. It also has anticonvulsant and pregnancy is warranted. Women who used pregabalin
6
anxiolytic properties, making it a recommended treatment had lower live birth rates and higher rates of CNS
9
for patients with generalized anxiety disorder (GAD) abnormalities than those in a control group. Pregabalin
and partial-onset seizures. Recently, pregabalin use has has been assigned pregnancy category C by the US FDA,
7
expanded to include conditions such as bipolar disorder which indicates that there are insufficient well-controlled
(BPD), social anxiety disorder (SAD), chronic pain, and human trials, but there may be benefits that outweigh any
16
sleep disturbances. A lipophilic analog of pregabalin was concerns. It has also been improperly used, which may
8,9
developed to improve diffusion across the blood–brain have negative consequences. Pregabalin is regarded as the
barrier (BBB). Pregabalin has been shown to bind to the final therapeutic choice for neuropathic pain. Therefore,
α2δ subunit of presynaptic voltage-gated Ca channels in studies on its safety during pregnancy are limited. It is
2+
central nervous system (CNS) tissues in animal models. currently unknown whether pregabalin, at therapeutic
9
As a result, the α2δ subunit reduces the influx of Ca into levels, affects fetal development and causes developmental
2+
17
neurons caused by depolarization, subsequently reducing abnormalities. Pregabalin may affect dopaminergic
the release of excitatory neurotransmitters that are neurons; we have previously demonstrated that it causes
likely linked to the development of anti-convulsant and behavioral sensitization through its effects on the dopamine
analgesic effects. 10,11 reward system. Similarly, one of our previous studies
18
illustrated that pregabalin exposure during the early stages
In animal studies, pregabalin use has been linked of brain development may affect the neurogenesis and
to reproductive damage, including skeletal deformities, morphogenesis of ventral midbrain dopaminergic neurons
neural tube anomalies, and other developmental (VMDNs). In addition, several genes that allow pregabalin
abnormalities. 10,11 However, the use of pregabalin during to affect VMDNs have been identified. 19
human pregnancy has only been partially documented. In
a study assessing first-trimester pregabalin exposure and The two hemispheres of the cerebral cortex are the
major congenital malformations, no significant increase in largest parts of the brain and are loaded with cortical
Volume 10 Issue 4 (2024) 405 doi: 10.36922/ijb.3010