Cellular uptake, biodistribution and protection against oxidative damage by Withania somnifera leaf extract-loaded PCL and MPEG-PCL nanoparticles
DOI:
https://doi.org/10.21814/jus.5628Keywords:
Withania somnifera, zebrafish embryos, cellular uptake, neuroprotection, PEGylated nanoparticlesAbstract
Withania somnifera (WS) or Ashwagandha is a well-known medicinal plant, cultivated in dry areas of India and Pakistan, where it represents an important resource as a widely used medicinal crop. Because of its anti-inflammatory and immunomodulatory effects, its extract is used, alone or in combination with other herbal extracts, in the treatment of age-related and neurodegenerative disorders. For tapping on the important therapeutic potential of this resource for biomedicine, strategies for controlled delivery and biodistribution improvement are necessary to guarantee treatment efficacy. Pharmacological properties of WS leaf extract (WSE) are mainly attributed to the presence of withanolides. In the present study, WSE was encapsulated in nanoparticles composed of biodegradable polymers as poly-ε-caprolactone (PCL) and methoxy poly-ethylene glycol poly-ε-caprolactone (MPEG-PCL) di-block copolymer. Laser doppler anemometry (LDA), X-Ray Diffraction (XRD) and Transmission Electron Microscopy (TEM) were used to analyze their size and shape. The particle size distribution of WSE-loaded PCL and MPEG-PCL nanoparticles was measured as 214-268 and 30-62 nm, respectively, presenting in both cases a spherical shape. U251 glioma cells, representative of the most common and lethal type of intracranial tumor which is glioblastoma, were exposed to these nanoparticles, demonstrating their efficient uptake. It was showed that MPEG-PCL nanoparticles containing WSE offered better protection to U251 cells against tert-butyl hydroperoxide (tBHP)-induced oxidative damage (95.1%), compared to PCL nanoparticles with WSE (56.4%) and free WSE (39.0%). In vivo distribution of these nanoparticles was further analyzed using zebrafish embryos to validate their biocompatibility in a relevant vertebrate neurodevelopment model.
References
T. A. Oyedepo and S. Palai, "Herbal remedies, toxicity, and regulations. reparation of phytopharmaceuticals for the management of disorders," in Academic Press, 2021, pp. 89-127. DOI: 10.1016/B978-0-12-820284-5.00014-9.
A. G. Atanasov et al., "Natural products in drug discovery: advances and opportunities," Nat Rev Drug Discov, vol. 20, pp. 200–216, 2021. DOI: 10.1038/s41573-020-00114-z.
S. Paul et al., "Withania somnifera (L.) Dunal (Ashwagandha): A comprehensive review on ethnopharmacology, pharmacotherapeutics, biomedicinal and toxicological aspects," Biomedicine & Pharmacotherapy, vol. 143, p. 112175, 2021. DOI: 10.1016/j.biopha.2021.112175.
T. K. Smith et al., "Herbal supplement sales in US increase by record-breaking 17.3% in 2020" HerbalGram, vol. 131, pp. 52-65, 2021.
A. L. Lopresti and S. J. Smith, "Ashwagandha (Withania somnifera) for the treatment and enhancement of mental and physical conditions: A systematic review of human trials," Journal of Herbal Medicine, vol. 28, p. 100434, 2021. DOI: 10.1016/j.hermed.2021.100434.
N. Alam et al., "High catechin concentrations detected in Withania somnifera (Ashwagandha) by high performance liquid chromatography analysis," BMC Complement Altern. Med., vol. 11, 2011, Art. no. 65. DOI: 10.1186/1472-6882-11-65.
V. Pandey et al., "Withania somnifera: Advances and implementation of molecular and tissue culture techniques to enhance its application," Front. Plant Sci., vol. 8, 2017, Art. no. 1390. DOI: 10.3389/fpls.2017.01390.
S. Paroha et al., "Conventional and nanomaterial-based techniques to increase the bioavailability of therapeutic natural products: a review," Environ Chem Lett, vol. 18, pp. 1767–1778, 2020. DOI: 10.1007/s10311-020-01038-1.
C. Hu et al., "Micelle or polymersome formation by PCL-PEG-PCL copolymers as drug delivery systems," Chinese Chemical Letters, vol. 28, no. 9, pp. 1905-1909, 2017. DOI: 10.1016/j.cclet.2017.07.020.
R. Saka and N. Chella, "Nanotechnology for delivery of natural therapeutic substances: a review," Environ Chem Lett, vol. 19, pp. 1097–1106, 2021. DOI: 10.1007/s10311-020-01103-9.
N. P. Aditya et al., "Fabrication of amorphous curcumin nanosuspensions using beta-lactoglobulin to enhance solubility, stability, and bioavailability," Colloids Surf. B Biointerfaces, vol. 127, pp. 114-121, 2015. DOI: 10.1016/j.colsurfb.2015.01.027.
B. Ray et al., "Neuroprotective and neurorescue effects of a novel polymeric nanoparticle formulation of curcumin (NanoCurc) in the neuronal cell culture and animal model: implications for Alzheimer's disease," J. Alzheimers Dis., vol. 23, pp. 61-77, 2011. DOI: 10.3233/JAD-2010-101374.
S. Dhawan et al., "Formulation development and systematic optimization of solid lipid nanoparticles of quercetin for improved brain delivery," J. Pharm. Pharmacol., vol. 63, pp. 342-351, 2011. DOI: 10.1111/j.2042-7158.2010.01225.x.
G. Marslin et al., "Curcumin encapsulated into methoxy poly(ethylene glycol) poly(epsilon-caprolactone) nanoparticles increases cellular uptake and neuroprotective effect in glioma cells," Planta Med., vol. 83, pp. 434-444, 2017. DOI: 10.1055/s-0042-112030.
D. M. Teleanu et al., "Blood-Brain Delivery Methods Using Nanotechnology," Pharmaceutics, vol. 10, no. 4, p. 269, Dec. 2018. DOI: 10.3390/pharmaceutics10040269.
J. Ribeiro et al., "A New Perspective for the Treatment of Alzheimer’s Disease: Exosome-like Liposomes to Deliver Natural Compounds and RNA Therapies," Molecules, vol. 28, no. 16, p. 6015, 2023. DOI: 10.3390/molecules28166015.
M. N. Ragnaill et al., "Internal benchmarking of a human blood-brain barrier cell model for screening of nanoparticle uptake and transcytosis," Eur. J. Pharm. Biopharm., vol. 77, pp. 360-367, 2011. DOI: 10.1016/j.ejpb.2010.12.024.
H. Danafar and U. Schumacher, "MPEG–PCL copolymeric nanoparticles in drug delivery systems," Cogent Medicine, vol. 3, 2016, Art. no. 1142411. DOI: 10.1080/2331205X.2016.1142411.
OECD, "Test No. 203: Fish, Acute Toxicity Test," in OECD Guidelines for the Testing of Chemicals, Section 2, OECD Publishing, Paris, 2019. DOI: 10.1787/9789264070144-en.
A. C. N. Oliveira et al., "Counter ions and constituents combination affect DODAX : MO nanocarriers toxicity in vitro and in vivo," Toxicol. Res. (Camb.), vol. 5, pp. 1244-1255, 2016. DOI: 10.1039/C6TX00074F.
H. Danafar et al., "Biodegradable m-PEG/PCL core-shell micelles: preparation and characterization as a sustained release formulation for curcumin," Adv. Pharm. Bull., vol. 4, pp. 501-510, 2014. DOI: 10.5681/apb.2014.074.
H. Gao et al., "Ligand modified nanoparticles increases cell uptake, alters endocytosis and elevates glioma distribution and internalization," Sci. Rep., vol. 3, 2013, Art. no. 2534. DOI: 10.1038/srep02534.
P. Dumont et al., "Overexpression of apolipoprotein J in human fibroblasts protects against cytotoxicity and premature senescence induced by ethanol and tert-butylhydroperoxide," Cell Stress Chaperones, vol. 7, pp. 23-35, 2002. DOI: 10.1379/1466-1268(2002)007<0023:ooajih>2.0.co;2.
Y. J. Shin et al., "Protective effect of clusterin on oxidative stress-induced cell death of human corneal endothelial cells," Mol. Vis., vol. 15, pp. 2789-2795, 2009. PMID: 20019877; PMCID: PMC2793897.
K. T. Savjani et al., "Drug solubility: importance and enhancement techniques," ISRN Pharm., 2012, Art. no. 195727. DOI: 10.10.5402/2012/195727.
A. Kumari et al., "Biodegradable polymeric nanoparticles based drug delivery systems," Colloids Surf. B Biointerfaces, vol. 75, pp. 1-18, 2010. DOI: 10.1016/j.colsurfb.2009.09.001.
V. Karanam et al., "Poly (varepsilon-caprolactone) nanoparticles of carboplatin: Preparation, characterization and in vitro cytotoxicity evaluation in U-87 MG cell lines," Colloids Surf. B Biointerfaces, vol. 130, pp. 48-52, 2015. DOI: 10.1016/j.colsurfb.2015.04.005.
P. W. Sylvester, "Optimization of the tetrazolium dye (MTT) colorimetric assay for cellular growth and viability," in Methods Mol. Biol, 2011, vol. 716, pp. 157-168. DOI: 10.1007/978-1-61779-012-6_9.
R. T. Carroll et al., "Brain-targeted delivery of Tempol-loaded nanoparticles for neurological disorders," J. Drug Target, vol. 18, pp. 665-674, 2010. DOI: 10.3109/10611861003639796.
D. Ding et al., "Cisplatin-loaded gelatin-poly(acrylic acid) nanoparticles: synthesis, antitumor efficiency in vivo and penetration in tumors," Eur. J. Pharm. Biopharm., vol. 79, pp. 142-149, 2011. DOI: 10.1016/j.ejpb.2011.01.008.
A. C. Carvalho et al., "Methanolic extract of Hypericum perforatum cells elicited with Agrobacterium tumefaciens provides protection against oxidative stress induced in human HepG2 cells," Ind. Crops Prod., vol. 59, pp. 177-183, 2014. DOI: 10.1016/j.indcrop.2014.05.018.
A. Oliveira et al., "Development, characterization, antioxidant and hepatoprotective properties of poly(Ɛ-caprolactone) nanoparticles loaded with a neuroprotective fraction of Hypericum perforatum," International Journal of Biological Macromolecules, vol. 110, pp. 185-196, 2018. DOI: 10.1016/j.ijbiomac.2017.10.103.
S. Doggui et al., "Neuroprotective efficacy of curcumin-loaded nanoparticles in Parkinson's disease model of zebrafish," Mol. Neurodegener., vol. 6, 2011, Art. no. 28. DOI: 10.1186/1750-1326-6-28.
M. N. Tiwari et al., "Nicotine-encapsulated poly(lactic-co-glycolic) acid nanoparticles improve neuroprotective efficacy against MPTP-induced parkinsonism," Free Radic. Biol. Med., vol. 65, pp. 704-718, 2013. DOI: 10.1016/j.freeradbiomed.2013.07.042.
A. K. Sachdeva et al., "Neuroprotective potential of sesamol and its loaded solid lipid nanoparticles in ICV-STZ-induced cognitive deficits: behavioral and biochemical evidence," Eur. J. Pharmacol., vol. 747, pp. 132-140, 2015. DOI: 10.1016/j.ejphar.2014.11.014.
C. B. Kimmel et al., "Stages of embryonic development of the zebrafish," Dev. Dyn., vol. 203, pp. 253-310, 1995. DOI: 10.1002/aja.1002030302.
G. Kari et al., "Zebrafish: an emerging model system for human disease and drug discovery," Clin. Pharmacol. Ther., vol. 82, pp. 70-80, 2007. DOI: 10.1038/sj.clpt.6100223.
N. S. Sipes et al., "Zebrafish: as an integrative model for twenty-first century toxicity testing," Birth Defects Res. C Embryo Today, vol. 93, pp. 256-267, 2011. DOI: 10.1002/bdrc.20214.
W. Peng et al., "Oral delivery of capsaicin using MPEG-PCL nanoparticles," Acta Pharmacol. Sin., vol. 36, pp. 139-148, 2015. DOI: 10.1038/aps.2014.113.
K. Wang et al., "Preparation and in vitro release of buccal tablets of naringenin-loaded MPEG-PCL nanoparticles," RSC Advances, vol. 4, pp. 33672-33679, 2014. DOI: 10.1039/C4ra04920a.
T. W. Shen et al., "Distribution and cellular uptake of pegylated polymeric particles in the lung towards cell-specific targeted delivery," Pharm. Res., vol. 32, pp. 3248–3260, 2015. DOI: 10.1007/s11095-015-1701-7.
J. V. Jokerst et al., "Nanoparticle PEGylation for imaging and therapy," Nanomedicine, vol. 6, pp. 715-728, 2011. DOI: 10.2217/Nnm.11.19.
N. Vij et al., "Development of PEGylated PLGA nanoparticle for controlled and sustained drug delivery in cystic fibrosis," J. Nanobiotechnology, vol. 8, 2010, Art. no. 22. DOI: 10.1186/1477-3155-8-22.
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