Hollow-layered nanoparticles for therapeutic delivery of peptide prepared using electrospraying

  • Manoochehr RasekhEmail author
  • Christopher Young
  • Marta Roldo
  • Frédéric Lancien
  • Jean-Claude Le Mével
  • Sassan Hafizi
  • Zeeshan Ahmad
  • Eugen Barbu
  • Darek Gorecki
Delivery Systems Original Research
Part of the following topical collections:
  1. Delivery Systems


The viability of single and coaxial electrospray techniques to encapsulate model peptide—angiotensin II into near mono-dispersed spherical, nanocarriers comprising N-octyl-O-sulphate chitosan and tristearin, respectively, was explored. The stability of peptide under controlled electric fields (during particle generation) was evaluated. Resulting nanocarriers were analysed using dynamic light scattering and electron microscopy. Cell toxicity assays were used to determine optimal peptide loading concentration (~1 mg/ml). A trout model was used to assess particle behaviour in vivo. A processing limit of 20 kV was determined. A range of electrosprayed nanoparticles were formed (between 100 and 300 nm) and these demonstrated encapsulation efficiencies of ~92 ± 1.8 %. For the single needle process, particles were in matrix form and for the coaxial format particles demonstrated a clear core–shell encapsulation of peptide. The outcomes of in vitro experiments demonstrated triphasic activity. This included an initial slow activity period, followed by a rapid and finally a conventional diffusive phase. This was in contrast to results from in vivo cardiovascular activity in the trout model. The results are indicative of the substantial potential for single/coaxial electrospray techniques. The results also clearly indicate the need to investigate both in vitro and in vivo models for emerging drug delivery systems.


Encapsulation Dynamic Light Scattering Tristearin Outer Needle Base Drug Delivery System 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



The authors would like to acknowledge PeReNE (EU-INTERREG) for supporting this study. The authors also thank Professor Simon Cragg for assistance with the SEM and TEM and Dr Simone Elgass for developing the HPLC method.


  1. 1.
    Ferrari M. Cancer nanotechnology: opportunities and challenges. Nat Rev Cancer. 2005;5(3):161–71.CrossRefGoogle Scholar
  2. 2.
    Bertling J, Blomer J, Kummel R. Hollow microspheres. Chem Eng Technol. 2004;27:829–37.CrossRefGoogle Scholar
  3. 3.
    Mathiowitz E, Jacob JS, Jong YS, Carino GP, Chickering DE, Chaturvedi P, Santos CA, Vijayaraghavan K, Montgomery S, Bassett M, Morrell C. Biologically erodable microsphere as potential oral-drug delivery system. Nature. 1997;386:410–4.CrossRefGoogle Scholar
  4. 4.
    Panyam J, Labhasetwar J. Biodegradable nanoparticles for drug and gene delivery to cells and tissue. Adv Drug Deliv Rev. 2003;55(3):329–47.CrossRefGoogle Scholar
  5. 5.
    Luan X, Skupin M, Siepmann J, Bodmeier R. Key parameters affecting the initial release (burst) and encapsulation efficiency of peptide-containing poly (lactide-co-glycolide) microparticles. Int J Pharm. 2006;324:168–75.CrossRefGoogle Scholar
  6. 6.
    Yang YY, Chung TS, Ng NP. Morphology drug distribution and in vitro release profiles of biodegradable polymeric microspheres containing protein fabricated by double-emulsion solvent extraction/evaporation method. Biomaterials. 2001;22(3):231–41.CrossRefGoogle Scholar
  7. 7.
    Lemoine D, Preat V. Polymeric nanoparticles as delivery system for influenza virus glycoproteins. J Control Release. 1998;54:15–27.CrossRefGoogle Scholar
  8. 8.
    Song CX, Labhasetwar V, Murphy H, Qu X, Humphrey WR, Shebuski RJ, et al. Formulation and characterization of biodegradable nanoparticles for intravascular local drug delivery. J Control Release. 1997;43:197–212.CrossRefGoogle Scholar
  9. 9.
    Fessi H, Puisieux F, Devissaguet JP, Ammoury N, Benita S. Nanocapsule formation by interfacial deposition following solvent displacement. Int J Pharm. 1989;55:R1–4.CrossRefGoogle Scholar
  10. 10.
    Barichello JM, Morishita M, Takayama K, Nagai T. Encapsulation of hydrophilic and lipophilic drugs in PLGA nanoparticles by the nanoprecipitation method. Drug Dev Ind Pharm. 1999;25:471–6.CrossRefGoogle Scholar
  11. 11.
    Perez C, Sanchez A, Putnam D, Ting D, Langer R, Alonso MJ. Poly(lactic acid)-poly(ethylene glycol) nanoparticles as new carriers for the delivery of plasmid DNA. J Control Release. 2001;75:211–24.CrossRefGoogle Scholar
  12. 12.
    Chronopoulou L, Fratoddi I, Palocci C, Venditti I, Russo MV. Osmosis based method drives the self-assembly of polymeric chains into micro and nanostructures. Langmuir. 2009;25:119406.CrossRefGoogle Scholar
  13. 13.
    York P. Strategies for particle design using supercritical fluid technologies. Pharm Sci Technol Today. 1999;2:430–40.CrossRefGoogle Scholar
  14. 14.
    Gu F, Zhang L, Teply BA, Mann N, Wang A, Radovic-Moreno AF, Langer R, Farokhzad OC. Precise engineering of targeted nanoparticles by using self-assembled biointegrated block copolymers. Proc Natl Acad Sci USA. 2008;105:2586–91.CrossRefGoogle Scholar
  15. 15.
    Ye A. Surface protein composition and concentration of whey protein isolate-stabilized oil-in-water emulsions: effect of heat treatment. Colloids Surf B. 2010;78(1):24–9.CrossRefGoogle Scholar
  16. 16.
    Yeo Y, Park K. Control of encapsulation efficiency and initial burst in polymeric microparticle systems. Arch Pharm Res. 2004;27(1):1–12.CrossRefGoogle Scholar
  17. 17.
    Gulfam M, Kim JE, Lee JM, Ku B, Chung BH, Chung BG. Anticancer drug-loaded gliadin nanoparticles induce apoptosis in breast cancer cells. Langmuir. 2012;28(21):8216–23.CrossRefGoogle Scholar
  18. 18.
    Valo H, Peltonen L, Vehviläinen S, Karjalainen M, Kostiainen R, Laaksonen T, Hirvonen J. Electrospray encapsulation of hydrophilic and hydrophobic drugs in poly(l-lactic acid) nanoparticles. Small. 2009;5(15):1791–8.CrossRefGoogle Scholar
  19. 19.
    Zamani M, Prabhakaran MP, Ramakrishna S. Advances in drug delivery via electrospun and electrosprayed nanomaterials. Int J Nanomed. 2013;8:2997–3017.Google Scholar
  20. 20.
    Hartman RPA, Brunner DJ, Camelot DMA, Marijnissen JCM, Scarlett B. Jet break-up in electrohydrodynamic atomization in the cone-jet mode. J Aerolsol Sci. 2000;31:65–95.CrossRefGoogle Scholar
  21. 21.
    Wu Y, MacKay JA, McDaniel JR, Chilkoti A, Clark RL. Fabrication of elastin-like polypeptide nanoparticles for drug delivery by electrospraying. Biomacromolecules. 2009;10:19–24.CrossRefGoogle Scholar
  22. 22.
    Guarino V, Cirillo V, Altobelli R, Ambrosio L. Polymer-based platforms by electric field-assisted techniques for tissue engineering and cancer therapy. Expert Rev Med Devices. 2015;12(1):113–29.CrossRefGoogle Scholar
  23. 23.
    Ruiz-Ortega M, Lorenzo O, Rupérez M, Esteban V, Suzuki Y, Mezzano S, Plaza JJ, Egido J. Role of the renin-angiotensin system in vascular diseases: expanding the field. Hypertension. 2001;38(6):1382–7.CrossRefGoogle Scholar
  24. 24.
    Bunjes H, Drechsler M, Koch MHJ, Westesen K. Incorporation of the model drug ubidecarenone into solid lipid nanoparticles. Pharm Res. 2001;18:287–93.CrossRefGoogle Scholar
  25. 25.
    Westesen K, Siekmann B. Biodegradable colloidal drug carrier systems based on solid lipids. In: Benita S, editor. Microencapsulation. Marcel Dekker: New York; 1996. p. 213–58.Google Scholar
  26. 26.
    Muller RH, Mader K, Gohla S. Solid lipid nanoparticles (SLN) for controlled delivery-a review of the state of the art. Eur J Pharm Biopharm. 2000;50:161–77.CrossRefGoogle Scholar
  27. 27.
    Jenning V, Lippacher A, Gohla S. Medium scale production of solid lipid nanoparticles (SLN) by high pressure homogenisation. J Microencapsul. 2002;19:1–10.CrossRefGoogle Scholar
  28. 28.
    Lippacher A, Muller RH, Mader K. Preparation of semisolid drug carriers for topical application based on solid lipid nanoparticles. Int J Pharm. 2001;214:9–12.CrossRefGoogle Scholar
  29. 29.
    Zhang C, Qu G, Sun Y, Yang T, Yao Z, Shen W, Shen Z, Ding Q, Zhou H, Ping Q. Biological evaluation of N-octyl-O-sulfate chitosan as a new nano-carrier of intravenous drugs. Eur J Pharm Sci. 2008;33:415–23.CrossRefGoogle Scholar
  30. 30.
    Benigni A, Cassis P, Remuzzi G. Angiotensin II revisited: new roles in inflammation, immunology and aging. EMBO Mol Med. 2010;2:247–57.CrossRefGoogle Scholar
  31. 31.
    Green S, Roldo M, Douroumis D, Bouropoulos N, Lamprou D, Fatouros DG. Chitosan derivatives alter release profiles of model compounds from calcium phosphate implants. Carbohydr Res. 2009;344(7):901–7.CrossRefGoogle Scholar
  32. 32.
    López-Herrera JM, Barrero A, Lopez A, Loscertales IG, Marquez M. Scaling laws. Aerosol Sci. 2003;34(5):535–52.CrossRefGoogle Scholar
  33. 33.
    Taylor G. Disintegration of water drops in an electric field. Proc R Soc Lond Ser A. 1964;280(1382):383–97.CrossRefGoogle Scholar
  34. 34.
    Montesano R, et al. Increased proteolytic activity is responsible for the aberrant morphogenetic behavior of endothelial cells expressing the middle T oncogene. Cell. 1990;62:435–45.CrossRefGoogle Scholar
  35. 35.
    Le Mével JC, Olson KR, Conklin D, Waugh D, Smith DD, Vaudry H, Conlon JM. Cardiovascular actions of trout urotensin II in the conscious trout, oncorhynchus mykiss. Am J Physiol Regul Integr Comp Physiol. 1996;271:1335–43.Google Scholar
  36. 36.
    Lancien F, Wong M, Al Arab A, Mimassi N, Takei Y, Le Mével JC. Central ventilatory and cardiovascular actions of angiotensin peptides in trout. Am J Physiol Regul Integr Comp Physiol. 2012;303:311–20.CrossRefGoogle Scholar
  37. 37.
    Barrero A, Ganan-Calvo AM, Davila J, Palacio A, Gomez-Gonzalez E. Low and high Reynolds number flows inside Taylor cones. Phys Rev E. 1998;58(6):7309.CrossRefGoogle Scholar
  38. 38.
    Ku BK, Kim SS. Electrospray characteristics of highly viscous liquids. Aerosol Sci. 2002;33:1361–78.CrossRefGoogle Scholar
  39. 39.
    Lastow O, Balachandran W. Novel low voltage EHD spray nozzle for atomization of water in the cone jet mode. J Electrostat. 2007;65:490–9.CrossRefGoogle Scholar
  40. 40.
    Loscertales IG, Barrero A, Guerrero I, Cortijo R, Marquez M, Ganan-Calvo AM. Micro/nano encapsulation via electriped coaxial liquid jets. Science. 2002;295:1695–8.CrossRefGoogle Scholar
  41. 41.
    Xie J, Ng WJ, Lee LY, Xie Jingwei, Wang CH. Encapsulation of protein drugs in biodegradable microparticles by coaxial electrospray. J Colloid Interface Sci. 2008;317:469–76.CrossRefGoogle Scholar
  42. 42.
    Jiang H, Hu Y, Li Y, Zhao P, Zhu K, Chen W. A facile technique to prepare biodegradable coaxial electrospun nanofibers for controlled release of bioactive agents. J Control Release. 2005;108:237–43.CrossRefGoogle Scholar
  43. 43.
    Kim SY, Lee H, Cho S, Park JW, Park J, Hwang J. Size control of chitosan capsules containing insulin for oral drug delivery via a combined process of ionic gelation with electrohydrodynamic atomization. Ind Eng Chem Res. 2011;50:13762–70.CrossRefGoogle Scholar
  44. 44.
    Park I, Kim W, Kim SS. Multi-jet mode electrospray for non-conducting fluids using two fluids and a coaxial grooved nozzle. Aerosol Sci Technol. 2011;45:629–34.CrossRefGoogle Scholar
  45. 45.
    Raiche AT, Puleo DA. Triphasic release model for multilayered gelatin coatings that can recreate growth factor profiles during wound healing. J Drug Target. 2001;9(6):449–60.CrossRefGoogle Scholar
  46. 46.
    Christophersen PC, Zhang L, Yang M, Nielsen HM, Müllertz A, Mu H. Solid lipid particles for oral delivery of peptide and protein drugs I–elucidating the release mechanism of lysozyme during lipolysis. Eur J Pharm Biopharm. 2013;85(3A):473–80.CrossRefGoogle Scholar
  47. 47.
    Scalia S, Mezzena M. Incorporation of quercetin in lipid microparticles: effect on photo- and chemical-stability. J Pharm Biomed Anal. 2009;49(1):90–4.CrossRefGoogle Scholar
  48. 48.
    Christophersen PC, Zhang L, Müllertz A, Nielsen HM, Yang M, Mu H. Solid lipid particles for oral delivery of peptide and protein drugs II-the digestion of trilaurin protects desmopressin from proteolytic degradation. Pharm Res. 2014;31(9):2420–8.CrossRefGoogle Scholar
  49. 49.
    Balls AK, Matlack MB, Tucker IW. The hydrolysis of glycerides by crude pancrease lipase. J Biol Chem. 1937;122:125–38.Google Scholar
  50. 50.
    Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65:55–63.CrossRefGoogle Scholar
  51. 51.
    Altman FP. Tetrazolium salts and formazans. Prog Histochem Cytochem. 1976;9:1–56.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Manoochehr Rasekh
    • 1
    Email author
  • Christopher Young
    • 1
  • Marta Roldo
    • 1
  • Frédéric Lancien
    • 2
  • Jean-Claude Le Mével
    • 2
  • Sassan Hafizi
    • 1
  • Zeeshan Ahmad
    • 3
  • Eugen Barbu
    • 1
  • Darek Gorecki
    • 1
  1. 1.School of Pharmacy and Biomedical SciencesUniversity of PortsmouthPortsmouthUK
  2. 2.Neurophysiology Laboratory, LaTIM UMR 1101University of BrestBrest Cedex 3France
  3. 3.Leicester School of PharmacyDe Montfort UniversityLeicesterUK

Personalised recommendations