Evaluation of Neuroprotective Efficacy of a Specialized Enteral Product Based on a Model of Traumatic Brain Injury in Drosophila melanogaster

Introduction. Traumatic brain injuries are a serious medical and social problem that requires the development of effective neuro-rehabilitation strategies, including specialised nutritional support. The Drosophila melanogaster model with experimental TBI is a highly effective tool for screening potential neuroprotective agents due to the conservativeness of fundamental biological mechanisms in drosophila and humans.

The purpose of the study. To study the ability of a specialised enteral product to improve survival and functional status in Drosophila melanogaster flies after modelling a traumatic brain injury.

Materials and Methods. The study was conducted on 6 groups of flies: injured and intact flies that received a standard nutrient medium with the addition of 1% of the experimental or control product, as well as groups on a standard medium without additives. TBI was modelled using the original high-impact injury setup. The mortality index after 24 hours, survival according to the Kaplan—Meyer curve, motor activity in the negative geotaxis test for 4 days after injury, and brain histopathology were evaluated.

Results. The introduction of the experimental product led to a decrease in acute mortality in the group of injured flies with the experimental product (TM-O) by 22.9% and 24.0% relative to the groups with injury on the standard medium and with the control product, respectively. Flies of the TM-O group demonstrated higher motor activity in the long-term period and the maximum life expectancy among the injured groups, comparable to intact flies. Histological analysis revealed less pronounced pathological changes in the brains of TM-O flies compared to the rest of the injured groups.

Conclusions. It has been established that the EN product from the meat of reconvalescent pigs has a pronounced neuroprotective and functional-restorative effect in the model of TBI in D. melanogaster. The scientific novelty lies in the primary experimental confirmation of the effectiveness of a fundamentally new approach to the creation of specialised products — the use of life-modified meat from animals adapted to the pathology of the central nervous system. The results obtained substantiate the prospect of further preclinical and clinical studies of this product as a component of metabolic support for patients with brain damage.

1. Abdulbaki, H. H., & Al-Deeb, M. A. (2022). Research Article Chlorpyrifos-induced dopaminergic damage in <i>Drosophila</i> <i>melanogaster</i> assessed by gene expression, AChE assay, and negative geotaxis using a new feeding device. Genetics and Molecular Research, 21(3). https://doi.org/10.4238/gmr19056

2. Ankrah, N. Y. D., Barker, B. E., Song, J., Wu, C., McMullen, J. G., & Douglas, A. E. (2021). Predicted Metabolic Function of the Gut Microbiota of Drosophila melanogaster. MSystems, 6(3). https://doi.org/10.1128/mSystems.01369-20

3. Barekat, A., Gonzalez, A., Mauntz, R. E., Kotzebue, R. W., Molina, B., El-Mecharrafie, N., Conner, C. J., Garza, S., Melkani, G. C., Joiner, W. J., Lipinski, M. M., Finley, K. D., & Ratliff, E. P. (2016). Using Drosophila as an integrated model to study mild repetitive traumatic brain injury. Scientific Reports, 6(1), 25252. https://doi.org/10.1038/srep25252

4. Blommer, J., Fischer, M. C., Olszewski, A. R., Katzenberger, R. J., Ganetzky, B., & Wassarman, D. A. (2021). Ketogenic diet reduces early mortality following traumatic brain injury in Drosophila via the PPARγ ortholog Eip75B. PLOS ONE, 16(10), e0258873. https://doi.org/10.1371/journal.pone.0258873

5. Bolus, H., Crocker, K., Boekhoff-Falk, G., & Chtarbanova, S. (2020). Modeling Neurodegenerative Disorders in Drosophila melanogaster. International Journal of Molecular Sciences, 21(9), 3055. https://doi.org/10.3390/ijms21093055

6. Buhlman, L. M., Krishna, G., Jones, T. B., & Thomas, T. C. (2021). Drosophila as a model to explore secondary injury cascades after traumatic brain injury. Biomedicine & Pharmacotherapy, 142, 112079. https://doi.org/10.1016/j.biopha.2021.112079

7. Cassotta, M., Cianciosi, D., Elexpuru‐Zabaleta, M., Pascual, I. E., Cano, S. S., Giampieri, F., & Battino, M. (2024). Human‐based new approach methodologies to accelerate advances in nutrition research. Food Frontiers, 5(3), 1031–1062. https://doi.org/10.1002/fft2.369

8. Cesur, M. F., Basile, A., Patil, K. R., & Çakır, T. (2023). A new metabolic model of Drosophila melanogaster and the integrative analysis of Parkinson’s disease. Life Science Alliance, 6(8), e202201695. https://doi.org/10.26508/lsa.202201695

9. Ciancarelli, I., Morone, G., Iosa, M., Cerasa, A., Calabrò, R. S., & Tozzi Ciancarelli, M. G. (2024). Neuronutrition and Its Impact on Post-Stroke Neurorehabilitation: Modulating Plasticity Through Diet. Nutrients, 16(21), 3705. https://doi.org/10.3390/nu16213705

10. Eickelberg, V., Lüersen, K., Staats, S., & Rimbach, G. (2022). Phenotyping of Drosophila Melanogaster—A Nutritional Perspective. Biomolecules, 12(2), 221. https://doi.org/10.3390/biom12020221

11. Fedulova, L., Vasilevskaya, E., Tikhonova, O., Kazieva, L., Tolmacheva, G., & Makarenko, A. (2022). Proteomic Markers in the Muscles and Brain of Pigs Recovered from Hemorrhagic Stroke. Genes, 13(12), 2204. https://doi.org/10.3390/genes13122204

12. Flatt, T. (2020). Life-History Evolution and the Genetics of Fitness Components in Drosophila melanogaster. Genetics, 214(1), 3–48. https://doi.org/10.1534/genetics.119.300160

13. Giansanti, M. G., Frappaolo, A., & Piergentili, R. (2025). Drosophila melanogaster: How and Why It Became a Model Organism. International Journal of Molecular Sciences, 26(15), 7485. https://doi.org/10.3390/ijms26157485

14. Katanaev, V. L. (2023). Humanization for neurological disease modeling: A roadmap to increase the potential of Drosophila model systems. Animal Models and Experimental Medicine, 6(3), 230–236. https://doi.org/10.1002/ame2.12322

15. Katchur, N. J., & Notterman, D. A. (2024). Recent insights from non-mammalian models of brain injuries: an emerging literature. Frontiers in Neurology, 15. https://doi.org/10.3389/fneur.2024.1378620

16. Katzenberger, R. J., Ganetzky, B., & Wassarman, D. A. (2023). Lissencephaly-1 mutations enhance traumatic brain injury outcomes in Drosophila. GENETICS, 223(3). https://doi.org/10.1093/genetics/iyad008

17. Katzenberger, R. J., Loewen, C. A., Wassarman, D. R., Petersen, A. J., Ganetzky, B., & Wassarman, D. A. (2013). A Drosophila model of closed head traumatic brain injury. Proceedings of the National Academy of Sciences, 110(44). https://doi.org/10.1073/pnas.1316895110

18. Li, T., Shi, W., Ho, M. S., & Zhang, Y. Q. (2024). A Pvr–AP-1–Mmp1 signaling pathway is activated in astrocytes upon traumatic brain injury. ELife, 12. https://doi.org/10.7554/eLife.87258.3

19. Nitta, Y., & Sugie, A. (2022). Studies of neurodegenerative diseases using Drosophila and the development of novel approaches for their analysis. Fly, 16(1), 275–298. https://doi.org/10.1080/19336934.2022.2087484

20. Obafemi, O. T., Ayeleso, A. O., Adewale, O. B., Unuofin, J., Ekundayo, B. E., Ntwasa, M., & Lebelo, S. L. (2025). Animal models in biomedical research: Relevance of Drosophila melanogaster. Heliyon, 11(1), e41605. https://doi.org/10.1016/j.heliyon.2024.e41605

21. Poblete, R. A., Yaceczko, S., Aliakbar, R., Saini, P., Hazany, S., Breit, H., Louie, S. G., Lyden, P. D., & Partikian, A. (2023). Optimization of Nutrition after Brain Injury: Mechanistic and Therapeutic Considerations. Biomedicines, 11(9), 2551. https://doi.org/10.3390/biomedicines11092551

22. Saikumar, J., Byrns, C. N., Hemphill, M., Meaney, D. F., & Bonini, N. M. (2020). Dynamic neural and glial responses of a head-specific model for traumatic brain injury in Drosophila. Proceedings of the National Academy of Sciences, 117(29), 17269–17277. https://doi.org/10.1073/pnas.2003909117

23. Saikumar, J., Kim, J., Byrns, C. N., Hemphill, M., Meaney, D. F., & Bonini, N. M. (2021). Inducing different severities of traumatic brain injury in Drosophila using a piezoelectric actuator. Nature Protocols, 16(1), 263–282. https://doi.org/10.1038/s41596-020-00415-y

24. Shah, E. J., Gurdziel, K., & Ruden, D. M. (2019). Mammalian Models of Traumatic Brain Injury and a Place for Drosophila in TBI Research. Frontiers in Neuroscience, 13. https://doi.org/10.3389/fnins.2019.00409

25. Shah, E. J., Gurdziel, K., & Ruden, D. M. (2020). Drosophila Exhibit Divergent Sex-Based Responses in Transcription and Motor Function After Traumatic Brain Injury. Frontiers in Neurology, 11. https://doi.org/10.3389/fneur.2020.00511

26. Shestopalov, A. E., Yakovleva, A. V., Yadgarov, M. Ya., Sergeev, I. V., & Kuzovlev, A. N. (2024). Prevalence and Impact of Malnutrition Risk on Outcomes in Critically Ill Patients with Traumatic Brain Injury and Stroke: A Retrospective Cohort Study Using Electronic Health Records. Nutrients, 16(15), 2396. https://doi.org/10.3390/nu16152396

27. Swanson, L. C., Rimkus, S. A., Ganetzky, B., & Wassarman, D. A. (2020). Loss of the Antimicrobial Peptide Metchnikowin Protects Against Traumatic Brain Injury Outcomes in Drosophila melanogaster. G3 Genes|Genomes|Genetics, 10(9), 3109–3119. https://doi.org/10.1534/g3.120.401377

28. van Alphen, B., Stewart, S., Iwanaszko, M., Xu, F., Li, K., Rozenfeld, S., Ramakrishnan, A., Itoh, T. Q., Sisobhan, S., Qin, Z., Lear, B. C., & Allada, R. (2022). Glial immune-related pathways mediate effects of closed head traumatic brain injury on behavior and lethality in Drosophila. PLOS Biology, 20(1), e3001456. https://doi.org/10.1371/journal.pbio.3001456

29. Victor Atoki, A., Aja, P. M., Shinkafi, T. S., Ondari, E. N., Adeniyi, A. I., Fasogbon, I. V., Dangana, R. S., Shehu, U. U., & Akin-Adewumi, A. (2025). Exploring the versatility of Drosophila melanogaster as a model organism in biomedical research: a comprehensive review. Fly, 19(1). https://doi.org/10.1080/19336934.2024.2420453

30. Ye, C., Behnke, J. A., Hardin, K. R., & Zheng, J. Q. (2023). Drosophila melanogaster as a model to study age and sex differences in brain injury and neurodegeneration after mild head trauma. Frontiers in Neuroscience, 17. https://doi.org/10.3389/fnins.2023.1150694