Synthesis and characterization of Aspartame and Neotame-encapsulated PLGA-TPGS nanoparticles and their modulatory effects on acetylcholinesterase gene expression in male rats
Abstract
This study investigates the synthesis, characterization, and biological impact of novel nanoparticle formulations of two artificial sweeteners Aspartame and Neotame encapsulated in PLGA-TPGS (poly(lactic-co-glycolic acid)-D-α-tocopheryl polyethylene gly col 1000 succinate) matrices. Nanoparticles were developed using a combination of emulsification, sonication, and solvent evapor a tion techniques, followed by detailed physicochemical characterization via UV–Vis spectroscopy, SEM, TEM, DLS, FTIR, and EDX. The in vitro release profiles demonstrated sustained release patterns for both Aspartame (ASPNPs) and Neotame (NEONPs) formulations compared to their free forms. In vivo experiments using male Wistar rats assessed the effects of these sweeteners and their nanoformulations on acetylcholinesterase (AChE) gene expression. Free forms of Aspartame and Neotame significantly el e vated AChE gene expression, suggesting potential neurotoxic effects, while ASPNPs and NEONPs mitigated this elevation, indica t ing a protective role of nanoencapsulation. These findings highlight the potential of PLGA-TPGS nanoparticle systems in reducing the neurotoxic effects associated with chronic intake of synthetic sweeteners through controlled release and improved bioavailability.References
Ahmad, M., Arif, A., Qureshi, S., & Raza, H. (2020). Neurotoxicity of Aspartame and the protective role of antioxidants: A biochemical and molecular study. Toxicology Reports, 7, 1–8.
Ali, H., Hussain, A., & Qamar, W. (2023). Nanoparticles in drug delivery: Recent advances and applications. Nanomedicine Research Journal, 8(2), 99–112.
Aljabbari, A., Alamoudi, K., & Alshahrani, S. (2023). Characterization techniques for PLGA nanoparticles in drug delivery: An updated overview. Pharmaceuticals, 16(4), 499.
Andrabi, S. S., Parvez, S., & Tabassum, H. (2023). Emerging role of nitric oxide in neuroinflammation and oxidative stress: Implications in neurodegenerative diseases. Frontiers in Molecular Neuroscience, 16, 1128547.
Ashok, I., & Sheeladevi, R. (2020). Neurobehavioral changes and increased oxidative stress in brain regions of Aspartame treated Wistar albino rats. Toxicology and Industrial Health, 36(1), 18–29.
Biswas, P., Patel, D., & Singh, M. (2022). PLGA-based nanoformulations for ocular drug delivery: Recent advances and future perspectives. Journal of Controlled Release, 345, 29–45.
Bukhari, S. N. A., Jantan, I., & Haque, S. (2023). Ciprofloxacin-loaded PLGA nanoparticles: Synthesis, optimization, and in vitro gastric release profile. Drug Design, Development and Therapy, 17, 231–242.
Butchko, H. H., Stargel, W. W., & Comer, C. P. (2002). Neotame: A review of the toxicity data. Food and Chemical Toxicology, 40(9), 1325–1339.
Çadırci, E., Altinoz, E., & Büyükokuroğlu, M. E. (2020). Aspartame and oxidative stress: Mechanistic insights. Oxidative Medicine and Cellular Longevity, 2020, 1095383.
Colović, M. B., Krstić, D. Z., Lazarević-Pašti, T. D., Bondžić, A. M., & Vasić, V. M. (2013). Acetylcholinesterase inhibitors: Pharmacology and toxicology. Current Neuropharmacology, 11(3), 315–335.
Danhier, F., Ansorena, E., Silva, J. M., Coco, R., Le Breton, A., & Préat, V. (2012). PLGA-based nanoparticles: An overview of biomedical applications. Journal of Controlled Release, 161(2), 505–522.
Dar, M. A. (2024). Artificial sweeteners and brain health: Revisiting metabolic and neurobehavioral outcomes. Neuroscience Bulletin, 40(2), 165–178.
Fonseca, C., Simões, S., & Gaspar, R. (2002). Paclitaxel-loaded PLGA nanoparticles: Preparation, physicochemical characterization and in vitro anti-tumoral activity. Journal of Controlled Release, 83(2), 273–286.
Iizuka, M. (2020). Excitotoxicity and cholinergic dysfunction: Implications for neurodegenerative diseases. International Journal of Molecular Sciences, 21(5), 1651.
Jiang, Y., Brynskikh, A. M., S-Manickam, D. S., & Kabanov, A. V. (2011). SOD1 nanozyme with reduced toxicity and MPS accumulation for CNS delivery. Journal of Controlled Release, 149(3), 245–251.
Kumar, R., Sharma, A., & Singh, H. (2024). Peptide-modified polymeric nanoparticles in targeted therapy: A review. Advanced Drug Delivery Reviews, 199, 114977.
Li, H., Li, Y., & Zhang, J. (2022). The role of particle size in nanoparticle-based drug delivery systems. Nanomedicine: Nanotechnology, Biology and Medicine, 38, 102449.
Okoro, F. O., & Markus, V. (2025). Artificial sweeteners and type 2 diabetes mellitus: A review of current developments and future research directions. Journal of Diabetes and its Complications, 39, 108954.
Padhi, B. K., Pelletier, G., Singh, M., & Kulkarni, S. (2020). Characterization of the rat acetylcholinesterase readthrough (AChE-R) splice variant: Implications for toxicological studies. Biochemical and Biophysical Research Communications, 532(4), 528–534.
Rathod, S., Kale, P., & Singh, D. (2021). Aspartame-induced oxidative stress in the brain: A dose–response study in rats. Environmental Toxicology and Pharmacology, 82, 103557.
Riordan, D., Alqaisi, B., & Maher, M. (2025). The neuroimmune impact of artificial sweeteners: From gut microbiota to brain function. Brain, Behavior, and Immunity – Health, 32, 100872.
Silman, I., & Sussman, J. L. (2005). Acetylcholinesterase: 'Classical' and 'non-classical' functions and pharmacology. Current Opinion in Pharmacology, 5(3), 293–302.
Sylvetsky, A. C., Jin, Y., Clark, E. J., Welsh, J. A., Rother, K. I., & Talegawkar, S. A. (2017). Consumption of low-calorie sweeteners among children and adults in the United States. Journal of the Academy of Nutrition and Dietetics, 117(3), 441–448.
Wang, Z., Li, Y., Wang, Y., & Zhang, Q. (2023). TPGS-modified nanoparticles for improved drug delivery: Mechanisms and biomedical applications. Drug Delivery, 30(1), 2113–2128.
Zhang, L., Gu, F. X., Chan, J. M., Wang, A. Z., Langer, R. S., & Farokhzad, O. C. (2008). Nanoparticles in medicine: Therapeutic applications and developments. Clinical Pharmacology and Therapeutics, 83(5), 761–769.
Zhang, W., Liu, J., & Wang, Y. (2021). Monodispersity and surface functionalization of PLGA nanoparticles: Key determinants for enhanced cellular uptake. Materials Science and Engineering: C, 118, 111393.
Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons «Attribution» 4.0 License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
