Skip to main content
SpringerLink
Log in

Methotrexate-loaded PEGylated gold nanoparticles as hemocompatible and pH-responsive anticancer drug nanoconjugate

  • Research paper
  • Published:
Journal of Nanoparticle Research Aims and scope Submit manuscript

Abstract

Methotrexate (MTX) is a well-known chemotherapeutic agent for solid tumor. However, its clinical usage is limited due to low permeability, cellular drug efflux, and non-specific drug delivery. These constraints require dose dumping and result in dose-dependent toxic effects. In the current study, MTX-loaded PEGylated gold nanoparticles (PEG-MTX-AuNPs) exposed to acidic pH (tumor cell) trigger a significant drug release profile. MTX was conjugated to citrate functionalized AuNP and stabilized by thiolated methyl polyethylene glycol (mPEG-SH). PEG-MTX-AuNP was cationic with 39.5 ± 1.2 mV zeta potential and a particle size of 39 nm, as revealed by DLS and TEM. AuNP size ranging from 40 to 50 nm was favorable for maximum endosomal uptake and fast drug release. MTX-AuNP showed 39.4% (22.28 µg/ml) drug loading efficiency. Spectroscopic examinations confirmed MTX chemisorption onto AuNPs via carboxyl (–COOH) and gold dative bond. In vitro release study indicated a 6.5-fold increased MTX release in the acidic environment (pH 4.5) compared to physiological pH. The terminal mPEG provided stability in a biological medium, refrained from protein deactivation, and exhibited significant hemocompatibility (less than 5% hemolysis up to 10 μM concentration). In vitro cytotoxicity against human rhabdomyosarcoma (RD) cells indicated significant (p < 0.0001) anticancer activity of PEG-MTX-AuNPs with low median growth inhibition concentration (GI50 = 3.61 ± 0.94 μM) compared to MTX (13.12 ± 0.98 μM) and AuNPs (5.24 ± 0.98 μM) alone. In conclusion, PEG-MTX-AuNPs offer higher stability in ionic and biological media, hemocompatibility, and acidic pH-dependent MTX release, and mediate enhanced in vitro anticancer activity that makes them a potential candidate for skeletal muscle sarcomas.

Graphic abstract

Methotrexate (MTX) is a weak dicarboxylic acid which become ionized at pH 7.4 (PBS) and unionized at endosomal pH 4.5. In ionized state MTX exchanges the negatively charged citrate ions and chemisorbed on AuNPs surface to stabilize and cap AuNPs. While in acidic media MTX become unionized and released in media. The enhanced intracellular release improves therapeutic outcome of MTX. The chemisorption of MTX was further confirmed with FTIR, supported by XRD and UV results.

Methotrexate binding mechanism to AuNP surface

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Ajmal M, Yunus U, Matin A, Haq NU (2015) Synthesis, characterization and in vitro evaluation of methotrexate conjugated fluorescent carbon nanoparticles as drug delivery system for human lung cancer targeting. J Photochem Photobiol B 153:111–120

    CAS  Google Scholar 

  • Ao LM, Gao F, Pan BF, Cui DX, Gu HC (2006) Interaction between gold nanoparticles and bovine serum albumin or sheep antirabbit immunoglobulin G. Chin J Chem 24(2):253–256

    CAS  Google Scholar 

  • Arakelova E, Grigoryan S, Arsenyan F, Babayan N, Grigoryan R, Sarkisyan N (2014) In vitro and in vivo anticancer activity of nanosize zinc oxide composites of doxorubicin. Int J Med Heal Pharm Biomed Eng 8:33–38

    Google Scholar 

  • Aseichev A, Azizova O, Beckman E, Skotnikova O, Dudnik L, Shcheglovitova O, Sergienko V (2014) Effects of gold nanoparticles on erythrocyte hemolysis. Bull Exp Biol Med 156(4):495–498

    CAS  Google Scholar 

  • Barabadi H, Vahidi H, Mahjoub MA, Kosar Z, Kamali KD, Ponmurugan K, Hosseini O, Rashedi M, Saravanan M (2019) Emerging antineoplastic gold nanomaterials for cervical cancer therapeutics: a systematic review. J Clust Sci 15:1–12

    Google Scholar 

  • Barabadi H, Vahidi H, Kamali KD, Hosseini O, Mahjoub MA, Rashedi M, Shoushtari FJ, Saravanan M (2020) Emerging theranostic gold nanomaterials to combat lung cancer: a systematic review. J Clust Sci 31(2):323–330

    CAS  Google Scholar 

  • Beidokhti HRN, Ghaffarzadegan R, Mirzakhanlouei S, Ghazizadeh L, Dorkoosh FA (2017) Preparation, characterization, and optimization of folic acid-chitosan-methotrexate core-shell nanoparticles by box-behnken design for tumor-targeted drug delivery. AAPS PharmSciTech 18(1):115–129

    Google Scholar 

  • Bertino JR (2009) Cancer research: from folate antagonism to molecular targets. Best Pract Res Clin Haematol 22(4):577–582

    Google Scholar 

  • Boomi P, Poorani GP, Palanisamy S, Selvam S, Ramanathan G, Ravikumar S, Barabadi H, Prabu HG, Jeyakanthan J, Saravanan M (2019) Evaluation of antibacterial and anticancer potential of polyaniline-bimetal nanocomposites synthesized from chemical reduction method. J Clust Sci 30(3):715–726

    CAS  Google Scholar 

  • Caputo F, Clogston J, Calzolai L, Rösslein M, Prina-Mello A (2019) Measuring particle size distribution of nanoparticle enabled medicinal products, the joint view of EUNCL and NCI-NCL. A step by step approach combining orthogonal measurements with increasing complexity. J Controlled Release 299:31–43

    CAS  Google Scholar 

  • Chen Y-H, Tsai C-Y, Huang P-Y, Chang M-Y, Cheng P-C, Chou C-H, Chen D-H, Wang C-R, Shiau A-L, Wu C-L (2007) Methotrexate conjugated to gold nanoparticles inhibits tumor growth in a syngeneic lung tumor model. Mol Pharm 4(5):713–722

    CAS  Google Scholar 

  • Deol S, Weerasuriya N, Shon Y-S (2015) Stability, cytotoxicity and cell uptake of water-soluble dendron–conjugated gold nanoparticles with 3, 12 and 17 nm cores. J Mater Chem B 3(29):6071–6080

    CAS  Google Scholar 

  • Dong YC, Hajfathalian M, Maidment PS, Hsu JC, Naha PC, Si-Mohamed S, Breuilly M, Kim J, Chhour P, Douek P (2019) Effect of gold nanoparticle size on their properties as contrast agents for computed tomography. Sci Rep 9(1):1–13

    Google Scholar 

  • Du S, Kendall K, Toloueinia P, Mehrabadi Y, Gupta G, Newton J (2012) Aggregation and adhesion of gold nanoparticles in phosphate buffered saline. J Nanopart Res 14(3):758

    Google Scholar 

  • Durgadas C, Sharma C, Paul W, Rekha M, Sreenivasan K (2012) Aggregation of gold nanoparticles followed by methotrexate release enables Raman imaging of drug delivery into cancer cells. J Nanopart Res 14(9):1127

    Google Scholar 

  • Elahi N, Kamali M, Baghersad MH (2018) Recent biomedical applications of gold nanoparticles: a review. Talanta 184:537–556

    CAS  Google Scholar 

  • Fan J, Hung W, Li W, Yeh J (2009) Biocompatibility study of gold nanoparticles to human cells. 13th international conference on biomedical engineering. Springer, pp 870–873

    Google Scholar 

  • Fuliaş A, Popoiu C, Vlase G, Vlase T, Oneţiu D, Săvoiu G, Simu G, Pătruţescu C, Ilia G, Ledeţi I (2014) Thermoanalytical and spectroscopic study on methotrexate–active substance and tablet. Dig J Nanomater Biostruct 9(1):93–98

    Google Scholar 

  • Gao W, Chan JM, Farokhzad OC (2010) pH-responsive nanoparticles for drug delivery. Mol Pharm 7(6):1913–1920

    CAS  Google Scholar 

  • Gao J, Huang X, Liu H, Zan F, Ren J (2012) Colloidal stability of gold nanoparticles modified with thiol compounds: bioconjugation and application in cancer cell imaging. Langmuir 28(9):4464–4471

    CAS  Google Scholar 

  • Ghorbani M, Zarei M, Mahmoodzadeh F, Ghorbani M (2020) Targeted delivery of methotrexate using a new PEGylated magnetic/gold nanoplatform covered with pH-responsive shell. Int J Polym Mater Polym Biomater 70:1–10

    Google Scholar 

  • Guerrini L, Alvarez-Puebla R, Pazos-Perez N (2018) Surface modifications of nanoparticles for stability in biological fluids. Materials 11(7):1154

    Google Scholar 

  • Held J, Mosheimer-Feistritzer B, Gruber J, Mur E, Weiss G (2018) Methotrexate therapy impacts on red cell distribution width and its predictive value for cardiovascular events in patients with rheumatoid arthritis. BMC Rheumatol 2(1):6

    Google Scholar 

  • Holmboe L, Andersen AM, Mørkrid L, Slørdal L, Hall KS (2012) High dose methotrexate chemotherapy: pharmacokinetics, folate and toxicity in osteosarcoma patients. Br J Clin Pharmacol 73(1):106–114

    CAS  Google Scholar 

  • Jia H, Gao X, Chen Z, Liu G, Zhang X, Yan H, Zhou H, Zheng L (2012) The high yield synthesis and characterization of gold nanoparticles with superior stability and their catalytic activity. CrystEngComm 14(22):7600–7606

    CAS  Google Scholar 

  • Jokerst JV, Lobovkina T, Zare RN, Gambhir SS (2011) Nanoparticle PEGylation for imaging and therapy. Nanomedicine 6(4):715–728

    CAS  Google Scholar 

  • Khatua A, Priyadarshini E, Rajamani P, Patel A, Kumar J, Naik A, Saravanan M, Barabadi H, Prasad A, Paul B (2020) Phytosynthesis, characterization and fungicidal potential of emerging gold nanoparticles using Pongamia pinnata leave extract: a novel approach in nanoparticle synthesis. J Clust Sci 31(1):125–131

    CAS  Google Scholar 

  • Kimling J, Maier M, Okenve B, Kotaidis V, Ballot H, Plech A (2006) Turkevich method for gold nanoparticle synthesis revisited. J Phys Chem B 110(32):15700–15707

    CAS  Google Scholar 

  • Krishnamurthy S, Esterle A, Sharma NC, Sahi SV (2014) Yucca-derived synthesis of gold nanomaterial and their catalytic potential. Nanoscale Res Lett 9(1):627–635

    Google Scholar 

  • Kumari SDC, Tharani C, Narayanan N, Kumar CS (2013) Formulation and characterization of methotrexate loaded sodium alginate chitosan Nanoparticles. Indian J Res Pharm Biotechnol 1(6):915

    Google Scholar 

  • Lin L, Xu W, Liang H, He L, Liu S, Li Y, Li B, Chen Y (2015) Construction of pH-sensitive lysozyme/pectin nanogel for tumor methotrexate delivery. Colloids Surf B 126:459–466

    CAS  Google Scholar 

  • Liu X, Atwater M, Wang J, Huo Q (2007) Extinction coefficient of gold nanoparticles with different sizes and different capping ligands. Colloids Surf B 58(1):3–7

    CAS  Google Scholar 

  • Madhusudhan A, Reddy GB, Venkatesham M, Veerabhadram G, Kumar DA, Natarajan S, Yang M-Y, Hu A, Singh SS (2014) Efficient pH dependent drug delivery to target cancer cells by gold nanoparticles capped with carboxymethyl chitosan. Int J Mol Sci 15(5):8216–8234

    CAS  Google Scholar 

  • Manson J, Kumar D, Meenan BJ, Dixon D (2011) Polyethylene glycol functionalized gold nanoparticles: the influence of capping density on stability in various media. Gold Bull 44(2):99–105

    CAS  Google Scholar 

  • Moore TL, Rodriguez-Lorenzo L, Hirsch V, Balog S, Urban D, Jud C, Rothen-Rutishauser B, Lattuada M, Petri-Fink A (2015) Nanoparticle colloidal stability in cell culture media and impact on cellular interactions. Chem Soc Rev 44(17):6287–6305

    CAS  Google Scholar 

  • Muhammad Z, Raza A, Ghafoor S, Naeem A, Naz SS, Riaz S, Ahmed W, Rana NF (2016) PEG capped methotrexate silver nanoparticles for efficient anticancer activity and biocompatibility. Eur J Pharm Sci 91:251–255

    CAS  Google Scholar 

  • Muthukumar T, Prabhavathi S, Chamundeeswari M, Sastry TP (2014) Bio-modified carbon nanoparticles loaded with methotrexate possible carrier for anticancer drug delivery. Mater Sci Eng C 36:14–19

    CAS  Google Scholar 

  • Nadhman A, Nazir S, Khan MI, Ayub A, Muhammad B, Khan M, Shams DF, Yasinzai M (2015) Visible-light-responsive ZnCuO nanoparticles: benign photodynamic killers of infectious protozoans. Int J Nanomed 10:6891

    CAS  Google Scholar 

  • Rahman S (2016) Size and concentration analysis of gold nanoparticles with ultraviolet-visible spectroscopy. Undergrad J Math Model 7(1):2

    Google Scholar 

  • Rahme K, Nolan MT, Doody T, McGlacken GP, Morris MA, O’Driscoll C, Holmes JD (2013) Highly stable PEGylated gold nanoparticles in water: applications in biology and catalysis. RSC Adv 3(43):21016–21024. https://doi.org/10.1039/c3ra41873a

    Article  CAS  Google Scholar 

  • Saravanan M, Barabadi H, Ramachandran B, Venkatraman G, Ponmurugan K (2019) Emerging plant-based anti-cancer green nanomaterials in present scenario. In: Barcelo D, Petrovic B (eds) Comprehensive analytical chemistry. Elsevier, Amsterdam, pp 291–318

    Google Scholar 

  • Saravanan M, Vahidi H, Cruz DM, Vernet-Crua A, Mostafavi E, Stelmach R, Webster TJ, Mahjoub MA, Rashedi M, Barabadi H (2020) Emerging antineoplastic biogenic gold nanomaterials for breast cancer therapeutics: a systematic review. Int J Nanomed 15:3577

    CAS  Google Scholar 

  • Sengani M, Grumezescu AM, Rajeswari VD (2017) Recent trends and methodologies in gold nanoparticle synthesis–a prospective review on drug delivery aspect. OpenNano 2:37–46

    Google Scholar 

  • Stapf M, Teichgräber U, Hilger I (2017) Methotrexate-coupled nanoparticles and magnetic nanochemothermia for the relapse-free treatment of T24 bladder tumors. Int J Nanomed 12:2793

    CAS  Google Scholar 

  • Strober W (2015) Trypan blue exclusion test of cell viability. Curr Protoc Immunol. https://doi.org/10.1002/0471142735.ima03bs111

    Article  Google Scholar 

  • Tran NTT, Wang T-H, Lin C-Y, Tai Y (2013) Synthesis of methotrexate-conjugated gold nanoparticles with enhanced cancer therapeutic effect. Biochem Eng J 78:175–180

    CAS  Google Scholar 

  • Woolley PV III, Sacher RA, Priego VM, Schanfield MS, Bonnem EM (1983) Methotrexate-induced immune haemolytic anaemia. Br J Haematol 54(4):543–552

    Google Scholar 

  • Xie J, Fan Z, Li Y, Zhang Y, Yu F, Su G, Xie L, Hou Z (2018) Design of pH-sensitive methotrexate prodrug-targeted curcumin nanoparticles for efficient dual-drug delivery and combination cancer therapy. Int J Nanomed 13:1381

    CAS  Google Scholar 

Download references

Acknowledgements

We are indebted to the NILOP Nanotheragnostic Research Laboratories of Pakistan Atomic Energy Commission for technical support. We are also thankful to Prof. Dr. N.M. Butt, Chairman, Preston Institute of Nano Science and Technology, for the provision of analytical support in UV-Spectrum analysis.

Funding

This work is funded by PAK-NORWAY Institutional Cooperation Program, PK3004.

Author information

Authors and Affiliations

  1. NILOP Nanomedicine Research Laboratories, National Institute of Lasers and Optronics College, PIEAS, Lehtrar Road Nilore, Islamabad, 44000, Pakistan

    Mehreen Rahman, Ummarah Kanwal, Uzma Azeem Awan & Abida Raza

  2. Department of Pharmacy, University of Peshawar, Peshawar, 25000, Pakistan

    Mehreen Rahman & Jamshaid Ali Khan

Authors
  1. Mehreen Rahman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jamshaid Ali Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ummarah Kanwal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Uzma Azeem Awan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Abida Raza
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Jamshaid Ali Khan or Abida Raza.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Rahman, M., Khan, J.A., Kanwal, U. et al. Methotrexate-loaded PEGylated gold nanoparticles as hemocompatible and pH-responsive anticancer drug nanoconjugate. J Nanopart Res 23, 195 (2021). https://doi.org/10.1007/s11051-021-05296-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11051-021-05296-0

Keywords

Search

Navigation