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Nanoformulated fingolimod attenuates NLRP3 inflammasome activation and promotes functional recovery in a rat model of spinal cord injury

Spinal Cord (2026)Cite this article

Subjects

Abstract

Study Design

Animal study.

Objectives

The NLRP3 inflammasome is a key mediator in secondary cascade of spinal cord injury (SCI), making it a potential therapeutic target. This study aimed to develop a nanoformulated version of fingolimod and investigate its effects on neuroinflammation, NLRP3 inflammasome activity, glial activation, lesion volume, and functional outcomes in a rat model of moderate contusive SCI.

Setting

Experimental animal research laboratory, AlSafwa University College, Karbala, Iraq.

Methods

Fingolimod-loaded PLGA nanoparticles were synthesized using the emulsion solvent evaporation method and characterized for size, morphology, and drug release. Adult male Wistar rats underwent a standardized contusion SCI at T9 and were randomized into three groups (n = 13): Sham, SCI + Vehicle, and SCI + Nano-Fingolimod (10 ng/ml). Behavioral assessments using the BBB locomotor test were performed on days 1, 3, 5, 7, 10, 12, and 14 post-injuries. Histological analysis quantified lesion volume and inflammatory cell infiltration. Immunofluorescence (IF) and Western blotting were used to evaluate the expression of NLRP3, ASC, cleaved caspase-1, IL-1β, GFAP, and Iba-1.

Results

Nano-fingolimod significantly improved BBB locomotor scores compared to the SCI group (p < 0.0001), indicating enhanced motor recovery. Histological examination revealed reduced lesion volume and inflammatory cell density in the treatment group. IF and Western blot analyses showed marked suppression of NLRP3 inflammasome signaling components (NLRP3, ASC, cleaved caspase-1, and IL-1β) and reduced glial activation (GFAP, Iba-1).

Conclusion

Nanoformulated fingolimod provides neuroprotection and improves functional recovery after SCI by attenuating NLRP3 inflammasome activation and glial reactivity. Targeted nanodelivery enhances its CNS bioavailability and reduces toxicity, positioning it as a promising candidate for SCI therapy.

Sponsorship

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Fig. 1: Characterization and therapeutic effects of Fingolimod-loaded nanoparticles in a rat model of spinal cord injury (SCI).
Fig. 2: Nano-Fingolimod improves locomotor recovery and attenuates lesion severity and inflammatory infiltration following spinal cord injury.
Fig. 3: Nano-Fingolimod attenuates glial activation and NLRP3 inflammasome signaling following spinal cord injury (SCI).

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Data availability

The datasets generated analysed during the current study are available from the corresponding author on reasonable request.

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Funding

The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

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Authors and Affiliations

  1. Branch of Prosthodontics, College of Dentistry, University of Al-Ameed, Karbala, Iraq

    Mohammed Hussein M. Alsharbaty

  2. Department of Biological Sciences, College of Science, University of Jeddah, Jeddah, Saudi Arabia

    Reem S. Alazragi

  3. College of Medical and Health Technologies, Al-Zahraa University for Women, Karbala, Iraq

    Siba Mekaael yaseen

  4. College of Pharmacy, University of Al-Ameed, Karbala, Iraq

    Uday Abdul-Reda Hussein

  5. Centre for Research on Environment and Renewable Energy, University of Karbala, 56001, Karbala, Iraq

    Mustafa Mudhafar

  6. Department of Anesthesia Techniques and Intensive Care, Al-Taff university college, 56001, Kerbala, Iraq

    Mustafa Mudhafar

  7. Faculty of Dentistry, University of Kerbala, Karbala, 56001, Iraq

    Hasan Ali Alsailawi

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  1. Mohammed Hussein M. Alsharbaty
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Contributions

All authors contributed to the overall conceptualization and study design. Mohammed Hussein M. Alsharbaty, Reem S. Alazragi, and Uday Abdul-Reda Hussein performed the experimental work, data acquisition, and initial data analysis. Siba Mekaael Yaseen assisted in histological assessment and interpretation of imaging data. Mustafa Mudhafar drafted the initial version of the manuscript and contributed to data visualization. Hasan Ali Alsailawi supervised the project, secured resources, and performed the critical revision of the manuscript for important intellectual content. All authors reviewed, edited, and approved the final manuscript and agree to be accountable for the integrity of the work.

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Correspondence to Hasan Ali Alsailawi.

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All experimental procedures were approved by the Institutional Animal Ethics Committee of AlSafwa University College, Karbala, Iraq (Approval No. H15REA195).

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M. Alsharbaty, M.H., Alazragi, R.S., Mekaael yaseen, S. et al. Nanoformulated fingolimod attenuates NLRP3 inflammasome activation and promotes functional recovery in a rat model of spinal cord injury. Spinal Cord (2026). https://doi.org/10.1038/s41393-026-01177-2

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