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RESEARCH ARTICLE

Investigating the effect of pregabalin on neuronal development using ultrashort self-assembling peptides: Assessing 3D neuronal cultures with high throughput robotic 3D bioprinting

Walaa F. Alsanie1,2* ,  Sherin Abdelrahman3,4,5 ,  Majid Alhomrani1,2 ,  Alexander U. Valle-Pérez3,4,5 ,  Ebtisam Abdulah Alosimi2 ,  Hamza Habeeballah6 ,  Heba A. Alkhatabi7,8,9 ,  Raed I. Felimban7,10 ,  Abdulhakeem S. Alamri1,2 ,  Abdulaziz Alsharif1 ,  Bassem M. Raafat11 ,  Yusuf S. Althobaiti12,13 ,  Ahmed Gaber2,14 ,  Charlotte A. E. Hauser3,4,5*
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1 Department of Clinical Laboratories Sciences, The Faculty of Applied Medical Sciences, Taif University, Taif, Saudi Arabia
2 Centre of Biomedical Sciences Research (CBSR), Deanship of Scientific Research, Taif University, Taif, Saudi Arabia
3 Laboratory for Nanomedicine, Division of Biological and Environmental Science and Engineering (BESE), King Abdullah University of Science and Technology (KAUST), Jeddah, Saudi Arabia
4 Computational Bioscience Research Center (CBRC), KAUST, Jeddah, Saudi Arabia
5 Red Sea Research Center (RSRC), KAUST, Jeddah, Saudi Arabia
6 Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences in Rabigh, King Abdulaziz University, Jeddah, Saudi Arabia
7 Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia
8 Center of Excellence in Genomic Medicine Research (CEGMR), King Abdulaziz University, Jeddah, Saudi Arabia
9 King Fahd Medical Research Centre, Hematology Research Unit, King Abdulaziz University, Jeddah, Saudi Arabia
10 Center of Innovation in Personalized Medicine (CIPM), 3D Bioprinting Unit, King Abdulaziz University, Jeddah, Saudi Arabia
11 Department of Radiological Sciences, College of Applied Medical Sciences, Taif University, Taif, Saudi Arabia
12 Department of Pharmacology and Toxicology, College of Pharmacy, Taif University, Taif, Saudi Arabia
13 Addiction and Neuroscience Research Unit, Taif University, Taif, Saudi Arabia
14 Department of Biology, College of Science, Taif University, Taif, Saudi Arabia
IJB 2024, 10(4), 3010 https://doi.org/10.36922/ijb.3010
Submitted: 25 February 2024 | Accepted: 13 May 2024 | Published: 18 July 2024
© 2024 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Pregabalin is a widely prescribed drug for various neurological disorders, yet its effects on embryonic cortical neuron development when given to pregnant women remain inadequately explored. In this study, we employed advanced three-dimensional (3D) culturing and in-house developed high-throughput robotic 3D bioprinting technologies to evaluate their potential in neuropharmacology applications, using pregabalin as a model compound. Using a robotic 3D bioprinter and tetrameric IIZK 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
Funding
This project was financially support by Ministry of education under project number 1-441-120.
Conflict of interest
The authors declare no conflicts of interest.
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