Using 5-fluorouracil-encored plga nanoparticles for the treatment of colorectal cancer: the in-vitro characterization and cytotoxicity studies

Document Type : Research Paper


Institute of Pharmaceutical Sciences, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur (C.G.) 495 009, INDIA


Objective(s): Colorectal cancer (CRC) is a prevalent cancer worldwide. The present study aimed to synthesize and investigate the potential of wheat germ agglutinin (WGA) conjugated with polylactic-co-glycolic acid (PLGA) nanoparticles (NPs) incorporating 5-fluorouracil (5-FU).
Materials and Methods: The NPs were investigated in terms of various characteristics, such as the particle size, surface charge, surface morphology, entrapment efficiency rate, and in-vitro drug release profile in simulated gastric and intestinal fluids. The optimized NPs were conjugated with WGA and characterized for the WGA conjugation efficiency, mucoadhesion, and cytotoxicity studies.
Results: The zeta potential of the WGA-conjugated NPs decreased (-17.9±1.4 mV) possibly due to the conjugation of the NPs with WGA, which reduced the zeta potential. The WGA-conjugated NPs exhibited sustained drug release effects (p<0.05) compared to the marketed formulation containing 5-FU after 24 hours. In addition, the optimized NPs followed the Higuchi kinetics, showing diffusion-controlled drug release mechanisms. Finally, the WGA-conjugated PLGA NPs could significantly inhibit the growth of colon cancer cells (HT-29 and COLO-205) compared to the non-conjugated NPs and pure drug solution (P<0.05).
Conclusion: According to the results, the WGA-conjugated NPs could be potential carrier systems compared to the non-conjugated NPs for the effective management of CRC.


1.Siegel RL, Miller KD, and Jemal A. Cancer Statistics, 2017. CA Cancer J Clin 2017;67:7-30.
2.Longley DB, Harkin DP, and Johnston PG. 5-fluorouracil: mechanisms of action and clinical strategies. Nat Rev Cancer 2003;3:330-338.
3.Diasio RB, and Harris BE. Clinical pharmacology of 5-fluorouracil. Clin Pharmacokinet 1989;16:215-237.
4.El-Hammadi MM, Delgado AV, Melguizo C, Prados JC, and Arias JL. Folic acid-decorated and PEGylated PLGA nanoparticles for improving the antitumour activity of 5-fluorouracil. Int J Pharm 2017;516:61-70.
5.van Kuilenburg AB, and Maring JG. Evaluation of 5-fluorouracil pharmacokinetic models and therapeutic drug monitoring in cancer patients. Pharmacogenomics 2013;14:799-811.
6.Mattos AC, Altmeyer C, Tominaga TT, Khalil NM, and Mainardes RM. Polymeric nanoparticles for oral delivery of 5-fluorouracil: Formulation optimization, cytotoxicity assay and pre-clinical pharmacokinetics study. Eur J Pharm Sci 2016;84:83-91.
7.Garg A, Patel V, Sharma R, Jain A, and Yadav AK. Heparin-appended polycaprolactone core/corona nanoparticles for site specific delivery of 5-fluorouracil. Artif Cells Nanomed Biotechnol 2017;45:1-10.
8.Jain SK, Patel K, Rajpoot K, and Jain A. Development of a Berberine Loaded Multifunctional Design for the Treatment of Helicobacter pylori Induced Gastric Ulcer. Drug Deliv Lett 2019;9:50-57.
9.Patrey NK, Rajpoot K, Jain AK, and Jain SK. Diltiazem loaded floating microspheres of Ethylcellulose and Eudragit for gastric delivery: in vitro evaluation. AJBR 2016;2:71-77.
10.Jain SK, Kumar A, Kumar A, Pandey AN, and Rajpoot K. Development and in vitro characterization of a multiparticulate delivery system for acyclovir-resinate complex. Artif Cells Nanomed Biotechnol 2016;44:1266-1275.
11.Jain SK, Prajapati N, Rajpoot K, and Kumar A. A novel sustained release drug-resin complex-based microbeads of ciprofloxacin HCl. Artif Cells Nanomed Biotechnol 2016;44:1891-1900.
12.Rajpoot K. Solid Lipid Nanoparticles: A Promising Nanomaterial in Drug Delivery. Curr Pharm Des 2019;25:1-16.
13.El-Hammadi MM, and Arias JL. Advanced Engineering Approaches in the Development of PLGA-Based Nanomedicines. In Handbook of Nanoparticles: Springer International Publishing, 2016, 1009-1039.
14.Figueiredo M, and Esenaliev R. PLGA Nanoparticles for Ultrasound-Mediated Gene Delivery to Solid Tumors. J Drug Deliv 2012;2012:767839.
15.Shakeri-Zadeh A, Khoee S, Shiran M-B, Sharifi AM, and Khoei S. Synergistic effects of magnetic drug targeting using a newly developed nanocapsule and tumor irradiation by ultrasound on CT26 tumors in BALB/c mice. J Mater Chem B 2015;3:1879-1887.
16.Tang Q, Wang Y, Huang R, You Q, Wang G, Chen Y, Jiang Z, Liu Z, Yu L, Muhammad S, and Wang X. Preparation of anti-tumor nanoparticle and its inhibition to peritoneal dissemination of colon cancer. PLoS One 2014;9:e98455.
17.Wang Y, Li P, Chen L, Gao W, Zeng F, and Kong LX. Targeted delivery of 5-fluorouracil to HT-29 cells using high efficient folic acid-conjugated nanoparticles. Drug Deliv 2015;22:191-198.
18.Badran MM, Mady MM, Ghannam MM, and Shakeel F. Preparation and characterization of polymeric nanoparticles surface modified with chitosan for target treatment of colorectal cancer. Int J Biol Macromol 2017;95:643-649.
19.Rajpoot K, and Jain SK. Colorectal cancer-targeted delivery of oxaliplatin via folic acid-grafted solid lipid nanoparticles: preparation, optimization, and in vitro evaluation. Artif Cells Nanomed Biotechnol 2018;46:1236-1247.
20.Rajpoot K, and Jain SK. Irinotecan hydrochloride trihydrate loaded folic acid-tailored solid lipid nanoparticles for targeting colorectal cancer: development, characterization, and in vitro cytotoxicity study using HT-29 cells. J Microencapsul 2019;36:659-676.
21.Rajpoot K, and Jain SK. Oral delivery of pH-responsive alginate microbeads incorporating folic acid-grafted solid lipid nanoparticles exhibits enhanced targeting effect against colorectal cancer: A dual-targeted approach. Int J Biol Macromol 2020. [Online ahead of print] DOI: 10.1016/j.ijbiomac.2020.02.132
22.Gabor F, Schwarzbauer A, and Wirth M. Lectin-mediated drug delivery: binding and uptake of BSA-WGA conjugates using the Caco-2 model. Int J Pharm 2002;237:227-239.
23.Yin Y, Chen D, Qiao M, Lu Z, and Hu H. Preparation and evaluation of lectin-conjugated PLGA nanoparticles for oral delivery of thymopentin. J Control Release 2006;116:337-345.
24.Zou W, Liu C, Chen Z, and Zhang N. Studies on bioadhesive PLGA nanoparticles: A promising gene delivery system for efficient gene therapy to lung cancer. Int J Pharm 2009;370:187-195.
25.Olde Damink LHH, Dijkstra PJ, van Luyn MJA, van Wachem PB, Nieuwenhuis P, and Feijen J. Cross-linking of dermal sheep collagen using a water-soluble carbodiimide. Biomaterials 1996;17:765-773.
26.Ponchel G, and Irache J. Specific and non-specific bioadhesive particulate systems for oral delivery to the gastrointestinal tract. Adv Drug Deliv Rev 1998;34:191-219.
27.Anande NM, Jain SK, and Jain NK. Con-A conjugated mucoadhesive microspheres for the colonic delivery of diloxanide furoate. Int J Pharm 2008;359:182-189.
28.Jain SK, Haider T, Kumar A, and Jain A. Lectin-Conjugated Clarithromycin and Acetohydroxamic Acid-Loaded PLGA Nanoparticles: a Novel Approach for Effective Treatment of H. pylori. AAPS PharmSciTech 2016;17:1131-1140.
29.Montisci MJ, Giovannuci G, Duchene D, and Ponchel G. Covalent coupling of asparagus pea and tomato lectins to poly(lactide) microspheres. Int J Pharm 2001;215:153-161.
30.Kim JS, Cho KJ, Tran TH, Nurunnabi M, Moon TH, Hong SM, and Lee YK. In vivo NIR imaging with CdTe/CdSe quantum dots entrapped in PLGA nanospheres. J Colloid Interface Sci 2011;353:363-371.
31.Paharia A, Yadav AK, Rai G, Jain SK, Pancholi SS, and Agrawal GP. Eudragit-coated pectin microspheres of 5-fluorouracil for colon targeting. AAPS PharmSciTech 2007;8:12.
32.Rai G, Jain SK, Agrawal S, Bhadra S, Pancholi SS, and Agrawal GP. Chitosan hydrochloride based microspheres of albendazole for colonic drug delivery. Pharmazie 2005;60:131-134.
33.Subudhi MB, Jain A, Jain A, Hurkat P, Shilpi S, Gulbake A, and Jain SK. Eudragit S100 Coated Citrus Pectin Nanoparticles for Colon Targeting of 5-Fluorouracil. Materials (Basel) 2015;8:832-849.
34.Vichai V, and Kirtikara K. Sulforhodamine B colorimetric assay for cytotoxicity screening. Nat Protoc 2006;1:1112-1116.
35.Yin Y, Chen D, Qiao M, Wei X, and Hu H. Lectin-conjugated PLGA nanoparticles loaded with thymopentin: ex vivo bioadhesion and in vivo biodistribution. J Control Release 2007;123:27-38.
36.Hariharan S, Bhardwaj V, Bala I, Sitterberg J, Bakowsky U, and Ravi Kumar MN. Design of estradiol loaded PLGA nanoparticulate formulations: a potential oral delivery system for hormone therapy. Pharm Res 2006;23:184-195.
37.Shenoy DB, and Amiji MM. Poly(ethylene oxide)-modified poly(epsilon-caprolactone) nanoparticles for targeted delivery of tamoxifen in breast cancer. Int J Pharm 2005;293:261-270.
38.Li X, Xu Y, Chen G, Wei P, and Ping Q. PLGA nanoparticles for the oral delivery of 5-Fluorouracil using high pressure homogenization-emulsification as the preparation method and in vitro/in vivo studies. Drug Dev Ind Pharm 2008;34:107-115.
39.Mo Y, and Lim LY. Preparation and in vitro anticancer activity of wheat germ agglutinin (WGA)-conjugated PLGA nanoparticles loaded with paclitaxel and isopropyl myristate. J Control Release 2005;107:30-42.
40.Mo Y, and Lim LY. Paclitaxel-loaded PLGA nanoparticles: potentiation of anticancer activity by surface conjugation with wheat germ agglutinin. J Control Release 2005;108:244-262.
41.Grabarek Z, and Gergely J. Zero-length crosslinking procedure with the use of active esters. Anal Biochem 1990;185:131-135.
42.Staros JV, Wright RW, and Swingle DM. Enhancement by N-hydroxysulfosuccinimide of water-soluble carbodiimide-mediated coupling reactions. Anal Biochem 1986;156:220-222.
43.Jain SK, Gupta M, Sahoo AK, Pandey AN, and Jain AK. Lectin conjugated gastro-retentive microspheres of amoxicillin for effective treatment of Helicobacter pylori. Curr Sci 2014;106:267-276.
44.Araujo J, Vega E, Lopes C, Egea MA, Garcia ML, and Souto EB. Effect of polymer viscosity on physicochemical properties and ocular tolerance of FB-loaded PLGA nanospheres. Colloids Surf B Biointerfaces 2009;72:48-56.
45.Bogataj M, Vovk T, Kerec M, Dimnik A, Grabnar I, and Mrhar A. The correlation between zeta potential and mucoadhesion strength on pig vesical mucosa. Biol Pharm Bull 2003;26:743-746.
46.Liu Y, Li K, Pan J, Liu B, and Feng SS. Folic acid conjugated nanoparticles of mixed lipid monolayer shell and biodegradable polymer core for targeted delivery of Docetaxel. Biomaterials 2010;31:330-338.
47.Xie J, and Wang CH. Self-assembled biodegradable nanoparticles developed by direct dialysis for the delivery of paclitaxel. Pharm Res 2005;22:2079-2090.
48.Ocal H, Arica-Yegin B, Vural I, Goracinova K, and Calis S. 5-Fluorouracil-loaded PLA/PLGA PEG-PPG-PEG polymeric nanoparticles: formulation, in vitro characterization and cell culture studies. Drug Dev Ind Pharm 2014;40:560-567.
49.Mu L, and Feng SS. Fabrication, characterization and in vitro release of paclitaxel (Taxol®) loaded poly (lactic-co-glycolic acid) microspheres prepared by spray drying technique with lipid/cholesterol emulsifiers. J Control Release 2001;76:239-254.
50.Mohammadi G, Nokhodchi A, Barzegar-Jalali M, Lotfipour F, Adibkia K, Ehyaei N, and Valizadeh H. Physicochemical and anti-bacterial performance characterization of clarithromycin nanoparticles as colloidal drug delivery system. Colloids Surf B Biointerfaces 2011;88:39-44.
51.Siepmann J, and Peppas NA. Higuchi equation: derivation, applications, use and misuse. Int J Pharm 2011;418:6-12.
52.Desai KG, and Park HJ. Encapsulation of vitamin C in tripolyphosphate cross-linked chitosan microspheres by spray drying. J Microencapsul 2005;22:179-192.
53.Venkatraman SS, Jie P, Min F, Freddy BY, and Leong-Huat G. Micelle-like nanoparticles of PLA-PEG-PLA triblock copolymer as chemotherapeutic carrier. Int J Pharm 2005;298:219-232.
54.Kalam MA, Humayun M, Parvez N, Yadav S, Garg A, Amin S, Sultana Y, and Ali A. Release kinetics of modified pharmaceutical dosage forms: a review. Cont J Pharm Sci 2007;1:30-35.
55.Fojo T, and Coley HM. The role of efflux pumps in drug-resistant metastatic breast cancer: new insights and treatment strategies. Clin Breast Cancer 2007;7:749-756.
56.Garg A, Rai G, Lodhi S, Jain AP, and Yadav AK. Hyaluronic acid embedded cellulose acetate phthlate core/shell nanoparticulate carrier of 5-fluorouracil. Int J Biol Macromol 2016;87:449-459.
57.Irache JM, Durrer C, Duchêne D, and Ponchel G. In vitro study of lectin-latex conjugates for specific bioadhesion. J Control Release 1994;31:181-188.
58.Khin MM, Hua JS, Ng HC, Wadstrom T, and Bow H. Agglutination of Helicobacter pylori coccoids by lectins. World J Gastroenterol 2000;6:202-209.
59.Irache JM, Durrer C, Duchene D, and Ponchel G. Bioadhesion of lectin-latex conjugates to rat intestinal mucosa. Pharm Res 1996;13:1716-1719.