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Mujahid Sher
Ishtiaq Hussain
Farhat Ali Khan
Hamid Hussain Afridi
Wiaam Mujahid Sher
Naila Gulfam
Muhammad Saqib Khalil
Akhtar Aman
Muhammad Sulaiman
Zeeshan Ahmad


silymarin nanoparticles, hepatoprotection, hepatic histopathology, albumin and protein values


Due to the many side effects associated with conventional medicine, people turned their attention towards natural drugs, which have fewer or no side effects. However, their uses are restricted due to lower solubility and bioavailability. Plant-based nanoparticles have been demonstrated as more effective alternatives, and nanoparticles are reported to increase the bioavailability and solubility of natural medicines to make their better use. The nanotechnology system is innovative to enhance the therapeutic effects and bioavailability of naturally occurring drugs. Therefore, silymarin nanoparticles were prepared using two different methods. Characterization analysis confirmed the reduction in particle size and conversion to amorphous states. The achieved nanoparticles have the required size, optimum zeta potential, and PDI. Results from solubility and bioavailability studies are very encouraging. The prepared nanoparticles were tested for hepatoprotection; they proved better than the unprocessed drug in terms of better effects on hepatic marker enzymes, serum total albumin and protein values, and hepatic histopathology. The prepared nano drug is recommended for clinical trials.

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1. Bayda S, Adeel M, Tuccinardi T, Cordani M, Rizzolio F. The history of nanoscience and nanotechnology: from chemical–physical applications to nanomedicine. Molecules, 25 (1), 112 (2019).
2. Bayón-Cordero L, Alkorta I, Arana L. Application of solid lipid nanoparticles to improve the efficiency of anticancer drugs. Nanomaterials, 9(3):474, (2019).
3. Afarid M, Mahmoodi S, Baghban R. Recent achievements in nano-based technologies for ocular disease diagnosis and treatment, review and update. Journal of Nanobiotechnology, 20(1):1- 36, (2022).
4. Ali J, Hussain A, Rehman S, Khan FA, Sher M. Antifungal potential of Mentha piperita leaves and stem extracts against phytopathogenic fungi. Specialty Journal of Biological Sciences, 3, 38-43 (2017).
5. Newman DJ, Cragg GM. Natural products as sources of new drugs over the nearly four decades from 01/1981 to 09/2019. Journal of natural products. 2020 Mar 12;83(3):770-803.
6. Zaman MA, Abbas RZ, Qamar W, Qamar MF, Mehreen U, Shahid Z, Kamran M. Role of secondary metabolites of medicinal plants against Ascaridia galli. World's Poultry Science Journal. 2020 Jul 2; 76 (3):639-55.
7. Cosme P, Rodríguez AB, Espino J, Garrido M. Plant phenolics: Bioavailability as a key determinant of their potential health-promoting applications. Antioxidants, 9(12), 1263 (2020).
8. Gheorghita, G.R.; Paun, V.I.; Neagu, S.; Maria, G.-M.; Enache, M.; Purcarea, C.; Parvulescu, V.I.; Tudorache, M. Cold-Active Lipase-Based Biocatalysts for Silymarin Valorization through Biocatalytic Acylation of Silybin. Catalysts, 11(11), 1390 (2021).
9. Bhattacharya, S. “Nuts and Seeds in Health and Disease Prevention” 2nd ed.; West Bengal Medical Services Corporation Ltd.: Kolkata, India, pp. 429–438 (2020).
10. Shapovalova V. Alcoholic Hepatitis: An experimental meta-analysis. SSP Modern Pharmacy and Medicine. 2023 Feb 16;3(1):1-1.
11. Camini FC, Costa DC. Silymarin: Not just another antioxidant. Journal of basic and clinical physiology and pharmacology, 31(4), 20190206 (2020).
12. Shakya AK. Drug-induced hepatotoxicity and hepatoprotective medicinal plants: a review. Indian Journal of Pharmaceutical Education and Research, 54(2), 234-50 (2020).
13. Marmouzi I, Bouyahya A, Ezzat SM, El Jemli M, Kharbach M. The food plant Silybum marianum (L.) Gaertn.: Phytochemistry, Ethnopharmacology, and clinical evidence. Journal of Ethnopharmacology, 265, 113303-113303 (2021).
14. Adetuyi BO, Omolabi FK, Olajide PA, Oloke JK. Pharmacological, biochemical and therapeutic potential of milk thistle (silymarin): a review. World News of Natural Sciences, 37(2021), 75-91 (2021).
15. Giustarini D, Milzani A, Dalle-Donne I, Rossi R. How to Increase Cellular Glutathione. Antioxidants, 12(5), 1-20 (2023).
16. Samee A, Amir RM, Ahmad A, Watto FM, Ali M, Azam MT, Sheeraz M, Fatima H, Zahoor Z, Zahid M, Ashraf H. Effectiveness of Milk Thistle on Human Body against Diseases: A Comprehensive Review. Scholars Bulletin, 9(2), 8-18 (2023).
17. Assis-Júnior EM, Melo AT, Pereira VB, Wong DV, Sousa NR, Oliveira CM, Malveira LR, Moreira LS, Souza MH, Almeida PR, Lima-Júnior RC. Dual effect of silymarin on experimental non- alcoholic steatohepatitis induced by irinotecan. Toxicology and Applied Pharmacology, 327(2017), 71-9 (2017).
18. Mangwani N, Singh PK, Kumar V. Medicinal plants: adjunct treatment to tuberculosis chemotherapy to prevent hepatic damage. Journal of Ayurveda and Integrative Medicine, 11(4), 522- 28 (2020).
19. Kheong CW, Mustapha NR, Mahadeva S. A randomized trial of silymarin for the treatment of nonalcoholic steatohepatitis. Clinical Gastroenterology and Hepatology. 2017 Dec 1;15(12):1940-9.
20. Colletta C, Colletta A, Placentino G. Lifestyle and silymarin: a fight against liver damage in NAFLD associated-prediabetic disease. Journal of Diabetes & Metabolic Disorders, 19(2020), 883- 94 (2020).
21. Curcio A, Romano A, Cuozzo S, Di Nicola A, Grassi O, Schiaroli D, Nocera GF, Pironti M. Silymarin in combination with vitamin C, vitamin E, coenzyme Q10 and selenomethionine to improve liver enzymes and blood lipid profile in NAFLD patients. Medicina, 56(10), 544 (2020).
22. Beydilli H, Yilmaz N, Cetin ES, Topal Y, Celik OI, Sahin C, Topal H, Cigerci IH, Sozen H. Evaluation of the protective effect of silibinin against diazinon induced hepatotoxicity and free- radical damage in rat liver. Iranian Red Crescent Medical Journal, 17(4), e25310 (2015).
23. Shriram RG, Moin A, Alotaibi HF, Khafagy ES, Al Saqr A, Abu Lila AS, Charyulu RN. Phytosomes as a plausible nano-delivery system for enhanced oral bioavailability and improved hepatoprotective activity of silymarin. Pharmaceuticals, 15(7), 790-809 (2022).
24. El-Far M, Salah N, Essam A, Abd El-Azim AO, El-Sherbiny IM. Silymarin nanoformulation as a potential anticancer agent in experimental Ehrlich ascites carcinoma-bearing animals. Nanomedicine, 13(15), 1865-58 (2018).
25. Anushiravani A, Haddadi N, Pourfarmanbar M, Mohammadkarimi V. Treatment options for nonalcoholic fatty liver disease: a double-blinded randomized placebo-controlled trial. European journal of gastroenterology & hepatology, 31(5), 613-7 (2019).
26. Wang X, Zhang Z, Wu SC. Health benefits of Silybum marianum: Phytochemistry, pharmacology, and applications. Journal of agricultural and food chemistry, 68(42), 11644-64 (2020).
27. Zhu HJ, Brinda BJ, Chavin KD, Bernstein HJ, Patrick KS, Markowitz JS. An assessment of pharmacokinetics and antioxidant activity of free silymarin flavonolignans in healthy volunteers: a dose escalation study. Drug Metabolism and Disposition. 2013 Sep 1;41(9):1679-85.
28. Hosseini S, Rezaei S, Moghaddam MR, Elyasi S, Karimi G. Evaluation of oral nano-silymarin formulation efficacy on prevention of radiotherapy-induced mucositis: A randomized, double- blinded, placebo-controlled clinical trial. PharmaNutrition, 15(2021), 100253 (2021).
29. Sher M, Zahoor M, Shah SW, Khan FA. Is particle size reduction linked to drug efficacy: an overview into nano initiatives in Pharmaceuticals. Zeitschrift für Physikalische Chemie. 2023 Jun 2(0).
30. Khan FA, Zahoor M, Islam NU, Hameed R. Synthesis of Cefixime and Azithromycin Nanoparticles. Journal of Nanomaterials, 2016, 29-37 (2016).
31. Mohamed MS, Abdelhafez WA, Zayed G, Samy AM. Optimization, in-vitro release, and in- vivo evaluation of gliquidone nanoparticles. AAPS PharmSciTech, 21(2), 1-2 (2020).
32. Kakran M, Sahoo NG, Tan IL, Li L. Preparation of nanoparticles of poorly water-soluble antioxidant curcumin by antisolvent precipitation methods. Journal of Nanoparticle Research. 14(2012), 1-11 (2012).
33. Kakran M, Sahoo NG, Li L, Judeh Z. Fabrication of quercetin nanoparticles by anti-solvent precipitation method for enhanced dissolution. Powder Technology, 223(2012), 59-64 (2012).
34. Campodónico A, Collado E, Ricci R, Pappa H, Segall A, Pizzorno MT. Dissolution test for silymarin tablets and capsules. Drug development and industrial pharmacy, 27(3), 261-5 (2001).
35. Sharma S, Jaiswal S, Duffy B, Jaiswal AK. Nanostructured materials for food applications: spectroscopy, microscopy, and physical properties. Bioengineering, 6(1), 26-42 (2019).
36. Racault C, Langlais F, Naslain R. Solid-state synthesis and characterization of the ternary phase Ti 3 SiC 2. Journal of Materials Science, 29(13), 3384-92 (2004).
37. Siekmann B, Westesen K. Thermoanalysis of the recrystallization process of melt- homogenized glyceride nanoparticles. Colloids and surfaces B: Biointerfaces, 3(3):159-75 (2004).
38. Khan BA, Rashid F, Khan MK, Alqahtani SS, Sultan MH, Almoshari Y. Fabrication of capsaicin loaded nanocrystals: Physical characterizations and in vivo evaluation. Pharmaceutics, 13(6), 841-854 (2021).
39. Sahibzada MU, Sadiq A, Khan S, Faidah HS, Naseemullah, Khurram M, Amin MU, Haseeb A. Fabrication, characterization and in vitro evaluation of silibinin nanoparticles: an attempt to enhance its oral bioavailability. Drug Design, Development, and Therapy, 11(2017), 1453-64 (2017).
40. U.S. P. XXII "US Pharmacopeial convention" Rockville, Md 1788-1789 (1990).
41. Wu JW, Lin LC, Hung SC, Chi CW, Tsai TH. Analysis of silibinin in rat plasma and bile for hepatobiliary excretion and oral bioavailability application. Journal of pharmaceutical and biomedical analysis, 45(4), 635-41 (2007).
42. Odiegwu CN, Chianella I, Azubike NC, Odiegwu UO, Ogbuowelu OS. Liver Function Test Values in Albino Wistar Rats Administered with Isolated Nigeria Achatina achatina Snail Lectin. GSC Biological and Pharmaceutical Sciences, 15(2), 92-102 (2021).
43. Roche Diagnostics. Blood Bilirubin and Transaminases Estimations on the Cobas C111 System. Roche Diagnostics Gmbh, Sandhofer Strasse116, D-68305 Mannheim, Germany.2011.
44. Drury RA. A color Atlas of histological staining techniques. Journal of Clinical Pathology, 31(3), 298-298 (1978).
45. Di Costanzo A, Angelico R. Formulation strategies for enhancing the bioavailability of silymarin: the state of the art. Molecules, 24(11), 2155-83 (2019).
46. Németh Z, Csóka I, Semnani Jazani R, Sipos B, Haspel H, Kozma G, Kónya Z, Dobó DG. Quality by Design-Driven Zeta Potential Optimisation Study of Liposomes with Charge Imparting Membrane Additives. Pharmaceutics, 14(9), 1798-1822 (2022).
47. Mahapatra AP, Patil V, Patil R. Solubility enhancement of poorly soluble drugs by using novel techniques: A comprehensive review. International Journal of PharmTech Research, 13(2), 80-93 (2020).
48. Holder CF, Schaak RE. Tutorial on powder X-ray diffraction for characterizing nanoscale materials. Acs Nano, 13(7), 7359-65 (2019).
49. Tavano L, Alfano P, Muzzalupo R, de Cindio B. Niosomes vs microemulsions: new carriers for topical delivery of Capsaicin. Colloids and surfaces B: Biointerfaces, 87(2), 333-9 (2011).
50. Štukelj J, Svanbäck S, Agopov M, Löbmann K, Strachan CJ, Rades T, Yliruusi J. Direct measurement of amorphous solubility. Analytical chemistry, 91(11), 7411-7 (2019).
51. Bremmell KE, Prestidge CA. Enhancing oral bioavailability of poorly soluble drugs with mesoporous silica-based systems: Opportunities and challenges. Drug development and industrial pharmacy, 45(3), 349-58 (2019).
52. Kakran M, Sahoo NG, Li L, Judeh Z, Wang Y, Chong K, Loh L. Fabrication of drug nanoparticles by evaporative precipitation of nanosuspension. International journal of pharmaceutics, 383(1-2), 285-92 (2010).
53. Albariqi AH, Chang RY, Tai W, Ke WR, Chow MY, Tang P, Kwok PC, Chan HK. Inhalable hydroxychloroquine powders for potential treatment of COVID-19. Journal of Aerosol Medicine and Pulmonary Drug Delivery, 34(1), 20-31 (2021).
54. Yingchoncharoen P, Kalinowski DS, Richardson DR. Lipid-based drug delivery systems in cancer therapy: what is available and what is yet to come. Pharmacological Reviews, 68(3), 701-87 (2016).
55. Yousaf AM, Malik UR, Shahzad Y, Mahmood T, Hussain T. Silymarin-laden PVP-PEG polymeric composite for enhanced aqueous solubility and dissolution rate: preparation and in vitro characterization. Journal of Pharmaceutical Analysis, 9(1), 34-9 (2019).
56. Croissant JG, Butler KS, Zink JI, Brinker CJ. Synthetic amorphous silica nanoparticles: toxicity, biomedical and environmental implications. Nature Reviews Materials, 5(12), 886-909 (2020).
57. Ali I, Ullah S, Imran M, Saifullah S, Hussain K, Kanwal T, Nisar J, Shah MR. Synthesis of biocompatible triazole-based non-ionic surfactant and its vesicular drug delivery investigation. Chemistry and Physics of Lipids, 228(2020), 104894-104894 (2020).
58. Shah SM, Ullah F, Khan S, Shah SM, de Matas M, Hussain Z, Minhas MU, AbdEl-Salam NM, Assi KH, Isreb M. Smart nanocrystals of artemether: fabrication, characterization, and comparative in vitro and in vivo antimalarial evaluation. Drug design, development, and therapy, 10(2016), 3837- 50 (2016).
59. Kayaert P, Van den Mooter G. Is the amorphous fraction of a dried nanosuspension caused by milling or by drying? A case study with Naproxen and Cinnarizine. European journal of pharmaceutics and biopharmaceutics, 81(3), 650-6 (2012).
60. Da Silva FL, Marques MB, Kato KC, Carneiro G. Nanonization techniques to overcome poor water-solubility with drugs. Expert opinion on drug discovery, 15(7), 853-64 (2020).
61. Chi Lip Kwok P, Chan HK. Nanotechnology versus other techniques in improving drug dissolution. Current pharmaceutical design, 20(3), 474-82 (2014).
62. Kumar M, Shanthi N, Mahato AK, Soni S, Rajnikanth PS. Preparation of luliconazole nanocrystals loaded hydrogel for improvement of dissolution and antifungal activity. Heliyon, 5(5), e01688-e01688 (2019).
63. Pestovsky YS, Martínez-Antonio A. “Gold Nanoparticles with Immobilized β-cyclodextrin- capsaicin Inclusion Complex for Prolonged Capsaicin Release.” IOP Conference Series: Materials Science and Engineering, (Vol. 389, No. 1, p. 012030) (2018).
64. Upadhyay P, Bhattacharjee M, Bhattacharya S, Ahir M, Adhikary A, Patra P. Silymarin- loaded, lactobionic acid-conjugated porous PLGA nanoparticles induce apoptosis in liver cancer cells. ACS Applied Bio Materials, 3(10), 7178-92 (2020).
65. Piwowarczyk L, Kucinska M, Tomczak S, Mlynarczyk DT, Piskorz J, Goslinski T, Murias M, Jelinska A. Liposomal Nanoformulation as a Carrier for Curcumin and pEGCG—Study on Stability and Anticancer Potential. Nanomaterials, 12(8), 1274 (2022).
66. Pande VV, Abhale VN, Möschwitzer JP. Nanocrystal technology: a particle engineering formulation strategy for the poorly water-soluble drugs. International Journal of Pharmaceutics, 453(1), 126-41 (2016).
67. Voorhees PW. The theory of Ostwald ripening. Journal of Statistical Physics, 38(1/2), 231-52 (1985).
68. Kumar D, Lashari N, Ganat T, Ayoub MA, Soomro AA, Chandio TA. A review on application of nanoparticles in cEOR: Performance, mechanisms, and influencing parameters. Journal of Molecular Liquids, 353(2022), 118821 (2022).
69. Kaushik M, Niranjan R, Thangam R, Madhan B, Pandiyarasan V, Ramachandran C, Oh DH, Venkatasubbu GD. Investigations on the antimicrobial activity and wound healing potential of ZnO nanoparticles. Applied Surface Science, 479(2019), 1169-77 (2019).
70. Kesisoglou F, Panmai S, Wu Y. Nanosizing—oral formulation development and biopharmaceutical evaluation. Advanced drug delivery reviews. 2007 Jul 30;59(7):631-44.
71. Bhujbal SV, Mitra B, Jain U, Gong Y, Agrawal A, Karki S, Taylor LS, Kumar S, Zhou QT. Pharmaceutical amorphous solid dispersion: A review of manufacturing strategies. Acta Pharmaceutica Sinica B, 11(8), 2505-36 (2021).
72. Tvrdý V, Pourová J, Jirkovský E, Křen V, Valentová K, Mladěnka P. Systematic review of pharmacokinetics and potential pharmacokinetic interactions of flavonolignans from silymarin. Medicinal Research Reviews, 41(4), 2195-246 (2021).
73. Abdullah AS, Sayed IE, El-Torgoman AM, Kalam A, Wageh S, Kamel MA. Green Synthesis of silymarin–chitosan nanoparticles as a new nanoformulation with enhanced anti-fibrotic effects against liver fibrosis. International Journal of Molecular Sciences, 23(10), 5420-5420 (2022).
74. Pathak K, Raghuvanshi S. Oral bioavailability: issues and solutions via nanoformulations. Clinical pharmacokinetics, 54(4), 325-57 (2015).
75. Javed S, Kohli K, Ali M. Reassessing bioavailability of silymarin. Alternative medicine review, 16(3), 239-49 (2011).
76. Pradhan R, Lee DW, Choi HG, Yong CS, Kim JO. Fabrication of a uniformly sized fenofibrate microemulsion by membrane emulsification. Journal of Microencapsulation, 30(1), 42-8 (2013).
77. Sahibzada MU, Sadiq A, Zahoor M, Naz S, Shahid M, Qureshi NA. Enhancement of bioavailability and hepatoprotection by silibinin through conversion to nanoparticles prepared by the liquid antisolvent method. Arabian Journal of Chemistry, 13(2), 3682-9 (2020).
78. Hedaya M, Bandarkar F, Nada A. In vitro and in vivo evaluation of ibuprofen nanosuspensions for enhanced oral bioavailability. Medical Principles and Practice, 30(4), 361-8 (2021).
79. Sahibzada MU, Zahoor M, Sadiq A, ur Rehman F, Al-Mohaimeed AM, Shahid M, Naz S, Ullah R. Bioavailability and hepatoprotection enhancement of berberine and its nanoparticles prepared by the liquid antisolvent method. Saudi Journal of Biological Sciences, 28(1), 327-32 (2021).
80. Gupta R, Badhe Y, Mitragotri S, Rai B. Permeation of nanoparticles across the intestinal lipid membrane: dependence on shape and surface chemistry studied through molecular simulations. Nanoscale, 12(11), 6318-33 (2020).
81. Bhattacharyya J, Ahmed AB, Das S. Hepatoprotective function as well as solubility and oral bioavailability of nano-based silymarin: A potential review. International Journal of Pharmaceutical Sciences and Research, 12(10), 1000-11 (2021).
82. Yang KY, Hwang DH, Yousaf AM, Kim DW, Shin YJ, Bae ON, Kim YI, Kim JO, Yong CS, Choi HG. Silymarin-loaded solid nanoparticles provide excellent hepatic protection: physicochemical characterization and in vivo evaluation. International journal of nanomedicine, 8(1), 3333-43 (2013).
83. Liang J, Liu Y, Liu J, Li Z, Fan Q, Jiang Z, Yan F, Wang Z, Huang P, Feng N. Chitosan- functionalized lipid-polymer hybrid nanoparticles for oral delivery of silymarin and enhanced lipid- lowering effect in NAFLD. Journal of Nanobiotechnology, 16(1), 1-2 (2018).
84. El-Nahas AE, Allam AN, Abdelmonsif DA, El-Kamel AH. Silymarin-loaded eudragit nanoparticles: formulation, characterization, and hepatoprotective and toxicity evaluation. AAPS Pharmscitech, 18(8), 3076-86 (2017).
85. Nasr SS, Nasra MM, Hazzah HA, Abdallah OY. Mesoporous silica nanoparticles, a safe option for silymarin delivery: Preparation, characterization, and in vivo evaluation. Drug Delivery and Translational Research, 9(5), 968-79 (2019).
86. Kumar N, Rai A, Reddy ND, Shenoy RR, Mudgal J, Bansal P, Mudgal PP, Arumugam K, Udupa N, Sharma N, Rao CM. Improved in vitro and in vivo hepatoprotective effects of liposomal silymarin in alcohol-induced hepatotoxicity in Wistar rats. Pharmacological Reports, 71(4), 703-12 (2019).
87. Poovi G, Damodharan N. Lipid nanoparticles: A challenging approach for oral delivery of BCS Class-II drugs. Future Journal of Pharmaceutical Sciences, 4(2), 191-205 (2018).
88. Mu H, Holm R. Solid lipid nanocarriers in drug delivery: characterization and design. Expert opinion on drug delivery, 15(8), 771-85 (2018).
89. EL Sayed HE, Morsy LE, Abo Emara TM, Galhom RA. Effect of carbon tetrachloride (CCl4) on the liver in adult albino rats: histological study. The Egyptian Journal of Hospital Medicine, 76(6), 4254-61 (2019).
90. Abdullah AS, El Sayed IE, El-Torgoman AM, Alghamdi NA, Ullah S, Wageh S, Kamel MA. Preparation and characterization of silymarin-conjugated gold nanoparticles with enhanced anti- fibrotic therapeutic effects against hepatic fibrosis in rats: role of MicroRNAs as molecular targets. Biomedicines, 9(12), 1767 (2021).
91. El Rabey HA, Rezk SM, Sakran MI, Mohammed GM, Bahattab O, Balgoon MJ, Elbakry MA, Bakry N. Green coffee methanolic extract and silymarin protect against CCl4-induced hepatotoxicity in albino male rats. BMC complementary medicine and therapies, 21(1), 1-11 (2021).
92. Clichici S, David L, Moldovan B, Baldea I, Olteanu D, Filip M, Nagy A, Luca V, Crivii C, Mircea P, Katona G. Hepatoprotective effects of silymarin coated gold nanoparticles in experimental cholestasis. Materials Science and Engineering: C, 115(2020), 111117 (2020).
93. Abdel-Wahhab MA, El-Nekeety AA, Salman AS, Abdel-Aziem SH, Mehaya FM, Hassan NS. Protective capabilities of silymarin and inulin nanoparticles against hepatic oxidative stress, genotoxicity and cytotoxicity of Deoxynivalenol in rats. Toxicon: Official Journal of the International Society on Toxinology, 142, 1-13 (2018).

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