OPTIMIZATION, CHARACTERIZATION, AND IN VIVO HEPATOPROTECTIVE EVALUATION OF NAC-LOADED NANOPARTICLES USING QBD AND IMAGEJ® SOFTWARE
DOI:
https://doi.org/10.22159/ijap.2025v17i2.52384Keywords:
N-acetyl cysteine, Solid lipid nanoparticles, Hepatoprotective analysis, Quality by design (QbD), Image J® softwareAbstract
Objective: This study formulated four novel N-Acetylcysteine (NAC)-lipid nanoparticles (Solid Lipid Nanoparticles [SLN], Nanostructured Lipid Carriers, Nano Drug Conjugate Carriers, Polymer-Lipid Hybrid Nanoparticles) and characterised their physicochemical features and in-vitro drug release pattern to enhance NAC's pharmacological capabilities. The study also designed to test optimised formulation's hepatoprotective efficacy in vivo using Image J®.
Methods: The homogenisation approach was the chosen for nanoparticle preparation, which was then followed by characterisation. The optimisation process was conducted utilising the Box–Behnken design of Quality by Design technique. Hepatoprotective efficacy study using carbon tetrachloride (CCl₄)-induced liver damage stress mouse animal model was utilised to conduct in vivo studies to study the cellular toxicity.
Results: In Fourier Transform Infrared Spectroscopy, X-ray Diffraction, and Differential Scanning Calorimetry studies, the medication and formulative components did not interact, and the lipid structure and NAC structure were unchanged. Scanning Electron Microscopy pictures show the nanoparticles' nearly spherical form and rough surface. The prepared design has an Encapsulation Efficiency (EE) of 62.56 to 86.32%. Polydispersity index ranged from 0.156 to 0.232. The SLN had a mean particle size of 96.23 to 159.10 nm. SLN had the highest release percentage (92.3) among the four varieties. Higuchi model of medication release was observed in optimised NAC-SLNs with spherical and intact chemical structure (88.95% EE, 92.35% drug release, -25.08 mv zeta potential, <200nm of particle size).
Conclusion: Lastly, the 4 types of NAC-nanoparticles showed no statistically significant changes from SLN, making them viable antioxidant drug carriers. This work designed and formulated SLN of the hepatoprotective medication to improve bioavailability and overcome limited solubility, strong protein binding, metabolising enzymes, and efflux mechanisms. The study found that NAC can treat liver ailments if placed into a proper delivery method.
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References
Vida M, Parvaneh A, Maryam S, Seyed MK, Ashraf M. A Review on Various Uses of N-Acetyl Cysteine. Cell J. 2016 Jan;19(1):11–17.
Pardeike J, Hommoss A, Müller RH. Lipid nanoparticles (SLN, NLC) in cosmetic and pharmaceutical dermal products. Eur J Pharm Biopharm. 2009 Jan;71(1):3-9.
Priyanka P, Sri rekha M, Seetha devi A. Review on formulation and evaluation of solid lipid nanoparticles for vaginal application. Int J Pharm Pharm Sci. 2022Jan;14(1):1-8.
Pankaj kumar J, Shikha K, Tapash C. Lipid-polymer hybrid nanocarriers as a novel drug delivery platform. Int J Pharm Pharm Sci. 2022Apr; 14(4): 1-12.
Shah R, Patel M, Bhadra D. Nanostructured lipid carriers for oral bioavailability enhancement of raloxifene: design and in vivo study. J Microencapsul. 2014Aug;31(8):711-718.
Sun TM, Davis ME. Nanodrug delivery systems: nanodrug conjugates. Wiley Interdisciplinary Reviews: Nanomed Nanobiotech. 2011Jan;3(1):6-20.
Gomes MJ, Peixoto D, Teixeira MA. Polymer-lipid hybrid nanoparticles: A synergistic approach for oral bioavailability improvement of poorly soluble drugs. Coll Surf B: Biointerfaces. 2017;159:169-177.
Mohsen S, Gholamhossein Y, Soliman MS, Mozhgan A, Ali MT. Hydrolytic stabilization of irinotecan active metabolite (SN38) against physiologic pH through self-assembly of conjugated poly (2-oxazoline) - poly (L-amino acid) block copolymer: A-synthesis and physicochemical characterization. J Drug Deliv Sci Tech. 2020;60:101933.
Mohsen S, Mahvand SV, Samira SA, Ali MT. Sterically Stabilized Polyionic Complex Nanogels of Chitosan Lysate and PEG-b-Polyglutamic Acid Copolymer for the Delivery of Irinotecan Active Metabolite (SN-38). Curr Drug Deliv. 2021 Jun;18(6):741-752.
Yu LX. Pharmaceutical quality by design: product and process development, understanding, and control. Pharm Res. 2008Apr;25(4):781-791.
Ferreira SL, Bruns RE, Ferreira HS, Matos GD, David JM, Brandao GC, da Silva EG, Portugal LA, dos Reis PS, Souza AS, dos Santos WN. Box-Behnken design: An alternative for the optimization of analytical methods. Analytica Chimica Acta. 2007Feb;597(2):179-186.
Keshav Jindal, Review on solubility: a mandatory tool for pharmaceuticals. Int Res J Pharm. 2017Nov;8(11):11-15.
Kukoc-Modun L, Radić N. Spectrophotometric determination of N-acetyl-L-cysteine and N-(2-mercaptopropionyl)-glycine in pharmaceutical preparations. Int J Anal Chem. 2011;2011:140756.
Georgia IS, Ioanna Z, Hannah B, Fotis S. Formulation design, production and characterisation of solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) for the encapsulation of a model hydrophobic active. Food Hydrocoll Health. 2021;1:100024.
Ayesha J, Ratnamala KV. Formulation and evaluation of sintered gastroretentive tablets of vildagliptin using design of experiment. Asian J Pharm Clin Res. 2023Jun;16(6):46-52.
Mishra V, Bansal KK, Verma A, Yadav N, Thakur S, Sudhakar K, Rosenholm JM. Solid Lipid Nanoparticles: Emerging Colloidal Nano Drug Delivery Systems. Pharmaceutics. 2018Apr;10(4):191.
Madupoju B, Rapaka SR, Malothu N, Desu PK, Areti A. Hepatoprotective activity of QBD-based optimized N-acetyl cysteine solid lipid nanoparticles against CCL4-induced liver injury in mice. Pharmacia. 2023Apr;70(4):1397-1410.
Amit KS. Fabrication of solid lipid nanoparticles by hot high shear homogenization and optimization by Box–Behnken design: An accelerated stability assessment, J Applied Pharm Sci. 2021Sep;11(9):35-47.
Naseri N, Valizadeh H, Zakeri-Milani P. Solid Lipid Nanoparticles and Nanostructured Lipid Carriers: Structure, Preparation and Application. Adv Pharm Bull. 2015Mar;5(3):305-313.
Sayantan P. Formulation and Intracellular Trafficking of Lipid–Drug Conjugate Nanoparticles Containing a Hydrophilic Antitubercular Drug for Improved Intracellular Delivery to Human Macrophages. ACS Omega. 2020Sep;5(9):4433–4448.
Shafique M, Ur Rehman M, Kamal Z, Alzhrani RM, Alshehri S, Alamri AH, Bakkari MA, Sabei FY, Safhi AY, Mohammed AM, Hamd MAE, Almawash S. Formulation development of lipid polymer hybrid nanoparticles of doxorubicin and its in-vitro, in-vivo and computational evaluation. Front Pharmacol. 2023;14:1025013.
Ekambaram P, Abdul HS. Formulation and evaluation of solid lipid nanoparticles of ramipril. J Young Pharm. 2011Mar;3(3):216-220.
Thilak M. Characterization of Nanomaterials: Tools and Challenges, Nanomaterials for Food Applications. Micro and Nano Tech. 2019;313-353.
Stella B, Peira E, Dianzani C, Gallarate M, Battaglia L, Gigliotti CL, Boggio E, Dianzani U, Dosio F. Development and Characterization of Solid Lipid Nanoparticles Loaded with a Highly Active Doxorubicin Derivative. Nanomaterials (Basel). 2018Feb;8(2):110.
Palak G, Hitesh KD. Aceclofenac loaded solid lipid nanoparticles: optimization, in vitro and ex-vivo. Int J App Pharm. 2023Apr;15(4):184-190.
Gill P, Moghadam TT, Ranjbar B. Differential scanning calorimetry techniques: applications in biology and nanoscience. J Biomol Tech. 2010Apr;21(4):167-193
Choubey A, Gilhotra R, Singh SK, Garg G. Formulation and Characterization of Nanomedicine (Solid Lipid Nanoparticle) Associate with the Extract of Pterospermumacerifolium for the Screening of Neurochemicals and Neuroendocrine Effects. Asian J Neurosurg. 2017Apr;12(4):613-619.
Swapnil DP, Vishal B, Reenu Y, Satish P, Maloji R. A systematic review on nano drug delivery system: solid lipid nanoparticles (SLN). Int J Curr Pharm Res. 2024Jan;16(1):10-20.
Devaraj VC, Krishna BG, Viswanatha GL, Kamath JV, Kumar S. Hepatoprotective activity of Hepax-a polyherbal formulation. Asian Pac J Trop Biomed. 2011Feb;1(2):142-146.
Del Rio D, Stewart AJ, Pellegrini N. A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutr Metab Cardiovasc Dis. 2005Apr;15(4):316-328.
Amr A, Mahmoud-Ghoneim D. Texture analysis of liver fibrosis microscopic images: a study on the effect of biomarkers, Acta Biochimica et BiophysicaSinica. 2011Mar;43(3):193-203.
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