Abstract
In this study, the processing conditions for fabricating bovine serum albumin (BSA)-loaded poly(d,l-lactide-co-glycolide) (PLGA) nanoparticles via a water/oil/water double emulsion technique were adjusted and release profiles were studied. Particle size and surface morphology of the BSA-loaded PLGA nanoparticles were comprehensively controlled as a function of processing determinants. The nanoparticles were intended as a carrier for controlled delivery of therapeutic proteins; however, BSA was chosen as a hydrophilic model protein encapsulated within PLGA nanoparticles to investigate the effective formulation parameters. Several key processing parameters were changed including surfactant(s) concentration in the internal and external aqueous phases, BSA concentration, poly(vinyl alcohol) (PVA) characteristics, and power of ultrasonicator probe to investigate their effects on the morphological characteristics and size distribution of the nanoparticles (NPs). The prepared NPs showed spherical shape with smooth and pore-free surfaces along with a relatively narrow particle size distribution. The mean particle size of the optimized formulation was 251.3 ± 8.5 nm, which is ideal for drug delivery applications. Our results demonstrate that using PVA with Mw 13–23 kDa and degree of hydrolysis approximately 87–89 % yields better results than PVA of higher molecular weight and higher degree of hydrolysis. Surfactants concentrations in internal (Span 60) and external phase (Tween 80) of the emulsions, which play a key role in determining NP characteristics and cumulative percentage BSA released, were optimized at 14 % (w/w) and 4 % (w/v), respectively. Optimal level of ultrasonication power (50 W) was also determined. According to the results, the optimized protein-loaded NPs with proper shape, size, and surface properties were prepared and these may act as a good candidate for protein delivery.
Similar content being viewed by others
References
Lu Y, Chen SC (2004) Micro and nano-fabrication of biodegradable polymers for drug delivery. Adv Drug Deliv Rev 56:1621–1633
Halliday A, Wallace GG, Cook M (2012) Novel methods of antiepileptic drug delivery–polymer-based implants. Adv Drug Deliv Rev 64:953–964
Winzenburg G, Schmidt C, Fuchs S, Kissel T (2004) Biodegradable polymers and their potential use in parenteral veterinary drug delivery systems. Adv Drug Deliv Rev 56:1453–1466
Budhian A, Siegel SJ, Winey KI (2007) Haloperidol-loaded PLGA nanoparticles: systematic study of particle size and drug content. Int J Pharm 336:367–375
Thompson CJ, Hansford D, Higgins S, Rostron C, Hutcheon GA, Mundaya DL (2007) Evaluation of ibuprofen-loaded microspheres prepared from novel copolyesters. Int J Pharm 329:53–61
Bysell H, Månsson R, Hansson P, Malmsten M (2011) Microgels and microcapsules in peptide and protein drug delivery. Adv Drug Deliv Rev 63:1172–1185
Tan LM, Choong PFM, Dass CR (2010) Review, recent developments in liposomes, microparticles and nanoparticles for protein and peptide drug delivery. Peptides 31:184–193
Floy BJ, Visor GC, Sanders LM (1993) Design of biodegradable polymer systems for controlled release of bioactive agents. In: El-Nokaly MA, Piatt DM, Charpentier BA (eds) Polymeric delivery systems. ACS, Washington, pp 154–167
Niwa T, Takeuchi H, Hino T, Kunou N, Kawashima Y (1993) Preparations of biodegradable nanospheres of water-soluble and insoluble drugs with D,L-lactide/glycolide copolymer by a novel spontaneous emulsification solvent diffusion method, and the drug release behavior. J Control Release 25:89–98
Tabata Y, Gutta S, Langer R (1993) Controlled delivery systems for proteins using polyanhydride microspheres. Pharm Res 10:487–496
Johnson OL, Cleland JL, Lee HJ, Jones AJS, Putney SD (1996) A month-long effect from a single injection of microencapsulated human growth hormone. Nat Med 7:795–799
Feirong K, Jagdish S (2001) Effects of additives on release of a model protein from PLGA microspheres. AAPS Pharmsci Tech 2:1–7
Ravivarapu HB, Burton K, Deluca PP (2000) Polymer and microspher blending to alter the release of peptide from PLGA microspheres. Eur J Pharm Biopharm 50:263–270
Torchilin VP (2007) Micellar nanocarriers: pharmaceutical perspectives. Pharm Res 24:1–16
Cohen-Sela E, Chorny M, Koroukhov N, Danenberg HD, Golomb G (2009) A new double emulsion solvent diffusion technique for encapsulating hydrophilic molecules in PLGA nanoparticles. J Control Release 133(2):90–95
Wu L, Ding J (2004) In vitro degradation of three-dimensional porous poly(d,l-lactide-co-glycolide) scaffolds for tissue engineering. Biomaterials 25:5821–5830
DellaPorta G, Castaldo F, Scognamiglio M, Paciello L, Parascandola P, Reverchona E (2012) Bacteria microencapsulation in PLGA microdevices by supercritical emulsion extraction. J Supercrit Fluids 63:1–7
Lu JM, Wang XW, Marin-Muller C, Wang H, Lin PH, Yao QZ, Chen CY (2009) Current advances in research and clinical applications of PLGA-based nanotechnology. Expert Rev Mol Diagn 9:325–341
Schwendeman SP, Cardamone M, Klibanov A, Langer R (1996) In: Cohen S, Bernstein H (eds) Microparticulate systems for the delivery of proteins and vaccines. Marcel Dekker, New York, pp 1–49
Berkland C, Kim KK, Pack DW (2001) Fabrication of PLG microspheres with precisely controlled and monodisperse size distributions. J Control Release 73:59–74
Amidi M, Mastrobattista E, Jiskoot W, Hennink WE (2010) Chitosan-based delivery systems for protein therapeutics and antigens. Adv Drug Deliv Rev 62(1):59–82
Shim MS, Kwon YJ (2012) Stimuli-responsive polymers and nanomaterials for gene delivery and imaging applications. Adv Drug Deliv Rev 64(11):1046–1059
Prabha S, Zhou WZ, Panyam J, Labhasetwar V (2002) Size-dependency of nanoparticle-mediated gene transfection studies with fractionated nanoparticles. Int J Pharm 244:105–115
Hans ML, Lowman AM (2002) Biodegradable nanoparticles for drug delivery and imaging. Curr Opin Solid State Mater Sci 6:319–327
Kobsa S, Saltzman WM (2008) Bioengineering approaches to controlled protein delivery. Pediatr Res 63(5):513–519
Rosenoer VM (1977) Albumin structure, function and uses. Pergamon, Oxford
Liu Y, Deng X (2002) Influences of preparation conditions on particle size and DNA loading efficiency for poly (DL-lactic acid-polyethylene glycol) microspheres entrapping free DNA. J Control Release 83:147–155
Márquez AL, Palazolo GG, Wagner JR (2007) Water in oil (w/o) and double (w/o/w) emulsions prepared with spans: microstructure, stability, and rheology. Colloid Polym Sci 285:1119–1128
Khoee S, Yaghoobian M (2009) An investigation into the role of surfactants in controlling particle size of polymeric nanocapsules containing penicillin-G in double emulsion. Eur J Med Chem 44:2392–2399
Mitchell DJ, Ninham BW (1981) Micelles, vesicles and microemulsions. Chem Soc Faraday Trans II 77:601–629
Mondal N, Samanta A, Pal TK, Ghosal SK (2008) Effect of different formulation variables on some particle characteristics of poly(DL-lactide-co-glycolide) nanoparticles. Yakugaku Zasshi 128(4):595–601
Kibbe AH (2000) Handbook of pharmaceutical excipients, 3rd edn. Pharmaceutical Press, Washington
Zhua Y, Zhang G, Yang H, Hong X (2005) Surfactant Deterg 8(4):353–358
Arshady R (1991) Preparation of biodegradable microspheres and microcapsules: 2. Polylactides and related polyesters. J Control Release 17:1–22
Feczkó T, Tóth J, Gyenis J (2008) Comparison of the preparation of PLGA–BSA nano- and microparticles by PVA, poloxamer and PVP. Colloids Surf A Physicochem Eng Asp 39(1–3):188–195
Feng SS (2004) Nanoparticle of biodegradable polymer for new concept chemotherapy. Expert Rev Med Devices 1(1):115–125
Jeong Y, Cho C, Kim S, Ko K, Kim S, Shim Y, Nah J (2001) Preparation of poly(DL-lactide-co-glycolide) nanoparticles without surfactant. Appl Polym Sci 80:2228–2236
Zhang X, Jackson JK, Burt HM (1996) Development of amphiphilic diblock copolymers as micellar carriers of taxol. Int J Pharm 132(1–2):195–206
Boury F, Ivanova T, Panaiotov I, Proust JE, Bois A, Richou J (1995) Dynamic properties of poly(D,L-lactide) and PVA monolayers at the air/water and dichloromethane air/water interfaces. J Colloid Interf Sci 169:380–392
Murakami H, Kawashima Y, Niwa T, Hino T, Takeuchi H, Kobayashi M (1997) Influence of the degrees of hydrolyzation and polymerization of PVA on the preparation and properties of PLGA nanoparticles. Int J Pharm 149:43–49
Zambaux MF, Bonneaux F, Gref R, Maincent P, Dellacherie E, Alonso MJ, Labrude P, Vigneron C (2000) Influence of experimental parameters on the characteristics of poly(lactid acid) nanoparticles prepared by double emulsion method. J Control Release 50:31–40
Konan YN, Cerny R, Favet J, Berton M, Gurny R, Allemann E (2003) Preparation and characterization of sterile sub-200 nm meso-tetra(4-hydroxylphenyl)porphyrin-loaded nanoparticles for photodynamic therapy. Eur J Pharm Biopharm 55:115–124
Feczkó T, Tóth J, Dósa G, Gyenis J (2011) Influence of process conditions on the mean size of PLGA nanoparticles. Chem Eng Process Process Intensif 50(8):846–853
Yan C, Resau JH, Hewetson J, West M, Rill WL, Kende M (1994) Characterization and morphological analysis of protein-loaded PLGA microparticles prepared by water/oil/water emulsion technique. J Control Release 32:231–241
Koppolu B, Rahimi M, Nattama S, Wadajkar A, Nguyen KT (2010) Development of multiple-layer polymeric particles for targeted and controlled drug delivery. Nanomedicine 6(2):355–361
Kang F, Singh J (2003) Conformational stability of a model protein (bovine serum albumin) during primary emulsification process of PLGA microspheres synthesis. Int J Pharm 260:149–156
Jalil R, Nixon JR (1990) Microencapsulation using poly(L-lactic acid). 2. Preparative variables affecting microcapsule properties. J Microencapsul 7:25–39
Arakawa T, Kita Y (2000) Stabilizing effects of caprylate and acetyltryptophanate on heat-induced aggregation of bovine serum albumin. Biochim Biophys Acta 1479:32–36
Clark AH, Saunderson DHP, Suggett A (1981) Infrared and laser-Raman spectroscopic studies of thermally-induced globular protein gels. Int J Pept Protein Res 17:353–364
Yang A, Yang L, Liu W, Li Z, Xu H, Yang X (2007) Tumor necrosis factor alpha blocking peptide loaded PEG-PLGA nanoparticles: preparation and in vitro evaluation. Int J Pharm 331:123–132
Peng ZG, Hidajat K, Uddin MS (2004) Adsorption of bovine serum albumin on nanosized magnetic particles. Colloid Interf Sci 271:277–283
Roach P, Farrar D, Perry CC (2006) Surface tailoring for controlled protein adsorption: effect of topography at the nanometer scale and chemistry. J Am Chem Soc 128(12):3939–3945
Bellusci M, Barbera AL, Padella F, Secci D (2011) Multifunctional ferrite-albumin nano particles in nanomedicine. Energ Ambiente Inneovazione 4–5:80–87
Nakamoto K (2009) Infrared and Raman spectra of inorganic and coordination compounds, 5th edn. Wiley-Interscience, New York
Acknowledgments
The authors are grateful to the Iran National Science Foundation (INSF) for the financial assistance (grant no. 89003650) and Iran Polymer and Petrochemical Institute for technical support. We would especially like to thank Dr Manizheh Motavalian for kindly providing freeze-drying facilities during the particle fabrication, and Mrs Khosravi and Mr Pirhajatie for performing SEM and TEM analyses.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Azizi, M., Farahmandghavi, F., Joghataei, M. et al. Fabrication of protein-loaded PLGA nanoparticles: effect of selected formulation variables on particle size and release profile. J Polym Res 20, 110 (2013). https://doi.org/10.1007/s10965-013-0110-z
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10965-013-0110-z