Pharmaceutical Research

, 35:200 | Cite as

Radiolabeled PLGA Nanoparticles for Effective Targeting of Bendamustine in Tumor Bearing Mice

  • Iliyas Khan
  • Avinash Gothwal
  • Ankur Kaul
  • Rashi Mathur
  • Anil Kumar Mishra
  • Umesh GuptaEmail author
Research Paper



Bendamustine is an important drug for the treatment of chronic lymphatic leukaemia (CLL), non-Hodgkin lymphoma (NHL). However, its delivery is challenging due to its instability. Current approach reports the development and characterization of bendamustine encapsulated PLGA nanoparticles for the effective targeting to leukemic cells.


The prepared, bendamustine loaded PLGA nanoparticles (BLPNP) were developed and characterized for particle size, zeta potential and polydispersity index. The formed nanoparticles were further characterized with the help of electron microscopy for surface morphology. The formed nanoparticles were evaluated for cytotoxicity, cell uptake, ROS and cell apoptosis against THP-1 leukemic cells as a part of in vitro evaluation. In vivo organ bio-distribution and tumor regression studies were performed to track in vivo behaviour of BLPNP.


The average particle size was 138.52 ± 3.25 nm, with 0.192 ± 0.036 PDI and − 25.4 ± 1.38 mV zeta potential. TEM images revealed the homogeneous particle size distribution with uniform shape. In vitro release exhibited a sustained drug-release behaviour up to 24 h. Cytotoxicity against THP-1 cells through MTT assay observed IC50 value of 27.8 ± 2.1 μM for BLPNP compared to pure drug, which was 50.42 ± 3.4 μM. Moreover, in vitro studies like cell-uptake and cell apoptosis studies further confirmed the higher accumulation of BLPNP in comparison to the pure drug. Organ distribution and tumor regression studies were performed to track in vivo behaviour of bendamustine loaded nanoparticles.


The overall study described a promising approach in terms of safety, least erythrocytic toxicity, better IC50 value with enhance tumor targeting and regression.


bendamustine eat nanoparticles PLGA THP-1 



Bendamustine loaded PLGA nanoparticles


Controlled drug delivery systems


Chronic lymphatic leukaemia




Ehrlich ascites tumor


Enhanced permeability and retention


Fluorescence assisted cell sorting


Fluorescein isothiocyanate


Instant thin layer chromatography


Non-Hodgkin lymphoma




Propidium iodide


Poly lactide-co-glycolic acid


Blank PLGA nanoparticles


Polyvinyl alcohol


Transmission electron microscope


  1. 1.
    Zhang Y, Chan HF, Leong KW. Advanced material and processing for drug delivery: the past and future. Adv Drug Deliv Rev. 2013;65:104–20.CrossRefGoogle Scholar
  2. 2.
    Safari J, Zarnegar Z. Advanced drug delivery systems: nanotechnology of health design a review. J Saudi Chem Soc. 2014;18:85–99.CrossRefGoogle Scholar
  3. 3.
    Naahidi S, Jafari M, Edalat F, Raymond K, Khademhosseini A, Chen P. Biocompatibility of engineered nanoparticles for drug delivery. J Control Release. 2013;166:182–94.CrossRefGoogle Scholar
  4. 4.
    Danhier F, Feron O, Preat V. To exploit the tumor microenvironment: passive and active tumor targeting of nanocarriers for anti-cancer drug delivery. J Control Release. 2010;148:135–46.CrossRefGoogle Scholar
  5. 5.
    Danhier F, Ansorena E, Silva JM, Coco R, Breton AL, Preat V. PLGA-based nanoparticles: an overview of biomedical applications. J Control Release. 2012;161:505–22.CrossRefGoogle Scholar
  6. 6.
    Khan I, Gothwal A, Sharma AK, Kesharwani P, Gupta L, Iyer A, et al. PLGA nanoparticles and their versatile role in anticancer drug delivery. Critical Rev Therap Drug Carr System. 2016;33(2):159–93.CrossRefGoogle Scholar
  7. 7.
    Acharya S, Sahoo SK. PLGA nanoparticles containing various anticancer agents and tumour delivery by EPR effect. Adv Drug Deliv Rev. 2011;62:170–83.CrossRefGoogle Scholar
  8. 8.
    Khan I, Gothwal A, Sharma AK, Qayum A, Singh SK, Gupta U. Biodegradable nano-architectural PEGylated approach for the improved stability and anticancer efficacy of bendamustine. Int J Bio Macromol. 2016;92:1242–51.CrossRefGoogle Scholar
  9. 9.
    Gothwal A, Khan I, Kumar P, Raza K, Kaul A, Mishra AK, et al. Bendamustine-PAMAM conjugates for improved apoptosis, efficacy, and in vivo pharmacokinetics: a sustainable delivery tactic. Mol Pharm. 2018;15:2084–97.CrossRefGoogle Scholar
  10. 10.
    Thomas SC, Sharma H, Rawat P, Verma AK, Leekha A, Kumaar V, et al. Synergistic anticancer efficacy of Bendamustine hydrochloride loaded bioactive hydroxyapatite nanoparticles: in-vitro, ex-vivo and in-vivo evaluation. Colloids Surfaces B: Biointerfaces. 2016;146:852–60.Google Scholar
  11. 11.
    Bhandari J, Mishra H, Mishra PK, Wimmer R, Ahmad FJ, Talegaonkar S. Cellulose nanofiber aerogel as a promising biomaterial for customized oral drug delivery. Int J Nanomedicine. 2017;12:2021–31.CrossRefGoogle Scholar
  12. 12.
    Kalimuthu K, Lubin BC, Bazylevich A, Gellerman G, Shpilberg O, Luboshits G, et al. Gold nanoparticles stabilize peptide-drug-conjugates for sustained targeted drug delivery to cancer cells. J Nanobiotech. 2018:16–34.Google Scholar
  13. 13.
    Thomas SC, Madaan T, Iqbal Z, Talegaonkar S. Box-Behnken Design of Experiment Assisted Development and Optimization of Bendamustine HCl loaded hydroxyapatite nanoparticles. Current Drug Deliv. 2018;15:000–0.CrossRefGoogle Scholar
  14. 14.
    Singh Y, Chandrashekhar A, Meher JG, Durga KK, Viswanadham R, Pawar VK, et al. Nanosized complexation assemblies housed inside reverse micelles churn out monocytic delivery cores for bendamustine hydrochloride. Euro J Pharm Biopharm. 2017;113:198–10.CrossRefGoogle Scholar
  15. 15.
    Pencheva E, Bogomilova A, Koseva N, Obreshkova D, Troev K. HPLC study on the stability of bendamustine hydrochloride immobilized onto polyphosphoesters. J Pharm Biomed Anal. 2008;48:1143–50.CrossRefGoogle Scholar
  16. 16.
    Singhai AK, Jain S, Jain NK. Evaluation of an aqueous injection of Ketoprofen. Pharmazie. 1997;52:149–51.PubMedGoogle Scholar
  17. 17.
    Agarwal P, Gupta U, Jain NK. Glycoconjugated peptide dendrimers-based nanoparticulate system for the delivery of chloroquine phosphate. Biomaterials. 2007;28:3349–59.CrossRefGoogle Scholar
  18. 18.
    ashe HB, Babbar AK, Jain S, Sharma RK, Mishra AK, Asthana A, et al. Investigations on biodistribution of technetium-99m-labeled carbohydrate-coated poly(propylene imine) dendrimers. Nanomedicine: Nanotech Biol Medicine. 2007;3:120–7.CrossRefGoogle Scholar
  19. 19.
    Naik S, Patel D, Chuttani K, Mishra AK, Misra AN. In vitro mechanistic study of cell death and in vivo performance evaluation of RGD grafted PEGylated docetaxel liposomes in breast cancer. Nanomedicine: Nanotech Bio Medicine. 2012;8:951–62.CrossRefGoogle Scholar
  20. 20.
    Pathak A, Kumar P, Chuttani K, Jain S, Mishra AK, Vyas SP, et al. Gene expression, biodistribution, and Pharmacoscintigraphic evaluation of chondroitin sulfate PEI Nanoconstructs mediated tumor gene therapy. ACS Nano. 2009;3:1493–505.CrossRefGoogle Scholar
  21. 21.
    Wang H, Zhao Y, Wu Y, Hu YL, Nan K, Nie G, et al. Enhanced anti-tumor efficacy by co-delivery of doxorubicin and paclitaxel with amphiphilic methoxy PEG-PLGA copolymer nanoparticles. Biomaterials. 2011;32:8281–90.CrossRefGoogle Scholar
  22. 22.
    Gupta U, Sharma S, Khan I, Gothwal A, Sharma AK, Singh Y, et al. Enhanced apoptotic and anticancer potential of paclitaxel loaded biodegradable nanoparticles based on chitosan. Int J Biol Macromol. 2017;98:810–9.CrossRefGoogle Scholar
  23. 23.
    Upadhyay S, Khan I, Gothwal A, Pachouri P, Bhaskar N, Gupta UD, et al. Conjugated and entrapped HPMA-PLA nano-polymeric micelles based dual delivery of first line anti TB drugs: improved and safe drug delivery against sensitive and resistant Mycobacterium Tuberculosis. Pharm Res. 2017;34:1944–55.CrossRefGoogle Scholar
  24. 24.
    Singh P, Gupta U, Asthana A, Jain NK. Folate and folate-PEG-PAMAM dendrimers: synthesis, characterization and targeted anticancer drug delivery potential in tumor induced mice. Bioconjug Chem. 2008;19(19):2239–52.CrossRefGoogle Scholar
  25. 25.
    Gupta M, Chashoo G, Sharma PR, Saxena AK, Gupta PN, Agarwal GP, et al. Dual targeted polymeric nanoparticles based on tumor endothelium and tumor cells for enhanced antitumor drug deliver. Mol Pharm. 2014;11:697–715.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Pharmacy, School of Chemical Sciences and PharmacyCentral University of RajasthanAjmerIndia
  2. 2.Division of Cyclotron and Radiopharmaceutical SciencesInstitute of Nuclear Medicine and Allied SciencesNew DelhiIndia

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