Intracisternal administration of tanshinone IIA-loaded nanoparticles leads to reduced tissue injury and functional deficits in a porcine model of ischemic stroke

Elizabeth S Waters; Erin E Kaiser; Xueyuan Yang; Madison M Fagan; Kelly M Scheulin; Julie H Jeon; Soo K Shin; Holly A Kinder; Anil Kumar; Simon R Platt; Kylee J Duberstein; Hea Jin Park; Jin Xie; Franklin D West

doi:10.1016/j.ibneur.2020.11.003

Intracisternal administration of tanshinone IIA-loaded nanoparticles leads to reduced tissue injury and functional deficits in a porcine model of ischemic stroke

IBRO Neurosci Rep. 2021 Jan 5:10:18-30. doi: 10.1016/j.ibneur.2020.11.003. eCollection 2021 Jun.

Authors

Elizabeth S Waters^{1

2

3}, Erin E Kaiser^{1

2}, Xueyuan Yang⁴, Madison M Fagan^{1

3}, Kelly M Scheulin^{1

2

3}, Julie H Jeon⁵, Soo K Shin^{1

3

6}, Holly A Kinder^{1

2

3}, Anil Kumar⁴, Simon R Platt^{1

7}, Kylee J Duberstein^{1

3}, Hea Jin Park⁵, Jin Xie^{1

4}, Franklin D West^{1

2

3

6}

Affiliations

¹ Regenerative Bioscience Center, University of Georgia, Athens, GA 30602, United States.
² Biomedical and Health Sciences Institute, University of Georgia, Athens, GA 30602, United States.
³ Department of Animal and Dairy Science, College of Agricultural and Environmental Sciences, University of Georgia, Athens, GA 30602, United States.
⁴ Department of Chemistry, Franklin College of Arts and Sciences, University of Georgia, Athens, GA 30602, United States.
⁵ Department of Foods and Nutrition, College of Family and Consumer Sciences, University of Georgia, Athens, GA 30602, United States.
⁶ Interdisciplinary Toxicology Program, College of Pharmacy, University of Georgia, Athens, GA 30602, United States.
⁷ Department of Small Animal Medicine and Surgery, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, United States.

Abstract

Background: The absolute number of new stroke patients is annually increasing and there still remains only a few Food and Drug Administration (FDA) approved treatments with significant limitations available to patients. Tanshinone IIA (Tan IIA) is a promising potential therapeutic for ischemic stroke that has shown success in pre-clinical rodent studies but lead to inconsistent efficacy results in human patients. The physical properties of Tan-IIA, including short half-life and low solubility, suggests that Poly (lactic-co-glycolic acid) (PLGA) nanoparticle-assisted delivery may lead to improve bioavailability and therapeutic efficacy. The objective of this study was to develop Tan IIA-loaded nanoparticles (Tan IIA-NPs) and to evaluate their therapeutic effects on cerebral pathological changes and consequent motor function deficits in a pig ischemic stroke model.

Results: Tan IIA-NP treated neural stem cells showed a reduction in SOD activity in in vitro assays demonstrating antioxidative effects. Ischemic stroke pigs treated with Tan IIA-NPs showed reduced hemispheric swelling when compared to vehicle only treated pigs (7.85 ± 1.41 vs. 16.83 ± 0.62%), consequent midline shift (MLS) (1.72 ± 0.07 vs. 2.91 ± 0.36 mm), and ischemic lesion volumes (9.54 ± 5.06 vs. 12.01 ± 0.17 cm³) when compared to vehicle-only treated pigs. Treatment also lead to lower reductions in diffusivity (-37.30 ± 3.67 vs. -46.33 ± 0.73%) and white matter integrity (-19.66 ± 5.58 vs. -30.11 ± 1.19%) as well as reduced hemorrhage (0.85 ± 0.15 vs 2.91 ± 0.84 cm³) 24 h post-ischemic stroke. In addition, Tan IIA-NPs led to a reduced percentage of circulating band neutrophils at 12 (7.75 ± 1.93 vs. 14.00 ± 1.73%) and 24 (4.25 ± 0.48 vs 5.75 ± 0.85%) hours post-stroke suggesting a mitigated inflammatory response. Moreover, spatiotemporal gait deficits including cadence, cycle time, step time, swing percent of cycle, stride length, and changes in relative mean pressure were less severe post-stroke in Tan IIA-NP treated pigs relative to control pigs.

Conclusion: The findings of this proof of concept study strongly suggest that administration of Tan IIA-NPs in the acute phase post-stroke mitigates neural injury likely through limiting free radical formation, thus leading to less severe gait deficits in a translational pig ischemic stroke model. With stroke as one of the leading causes of functional disability in the United States, and gait deficits being a major component, these promising results suggest that acute Tan IIA-NP administration may improve functional outcomes and the quality of life of many future stroke patients.

Keywords: ADC, Apparent Diffusion Coefficient; ANOVA, analysis of variance; AU, arbitrary units; BBB, blood brain barrier; Baic, Baicalin; CNS, central nervous system; CSF, cerebral spinal fluid; DAMPS, damaged-associated molecular patterns; DLS, dynamic light scattering; DTI, Diffusion Tensor Imaging; DWI, Diffusion-Weighted Imaging; Edar, Edaravone; FA, fractional anisotropy; FDA, Food and Drug Administration; GABA, γ-aminobutyric acid; GM, gray matter; IC, inhibitory concentration; ICH, intracerebral hemorrhage; IL-6, interleukin 6; IM, intramuscular; Ischemic stroke; LPS, lipopolysaccharide; MCA, middle cerebral artery; MCAO, middle cerebral artery occlusion; MLS, midline shift; NP, nanoparticle; NSCs, neural stem cells; Nanomedicine; PBS, phosphate buffered saline; PEG–PLGA, polyethyleneglycol–polylactic-co-glycolic acid; PLGA nanoparticle; PLGA, Poly (lactic-co-glycolic acid); PLGA-b-PEG-OH, poly (lactide-co-glycolide)-b-poly (ethylene glycol)-maleimide; Pig stroke model; Piog, Pioglitazone; Puer, Puerarin; ROS, reactive oxygen species; Resv, Resveratrol; SOD, superoxide dismutase; STAIR, Stroke Therapy Academic and Industry Roundtable; T2*, T2Star; T2FLAIR, T2 Fluid Attenuated Inversion Recovery; T2W, T2Weighted; TD, transdermal; TEM, transmission electron microscopy; TNF-α, tumor necrosis factor α; Tan IIA, Tanshinone IIA; Tan IIA-NPs, Tan IIA PLGA NPs; Tan IIA-NPs, Tan IIA-loaded nanoparticles; Tanshinone IIA; UGA, University of Georgia; WM, white matter; ddH2O, double-distilled water; tPA, Tissue plasminogen activator.

Abstract

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