Abstract
Purpose
We evaluated the role of a poly(ethylene glycol) (PEG) surface coating to increase residence times and alter the cellular fate of nano- and microparticles delivered to the lung.
Methods
Three sizes of PRINT hydrogel particles (80 × 320 nm, 1.5 and 6 μm donuts) with and without a surface PEG coating were instilled in the airways of C57/b6 mice. At time points of 1, 7, and 28 days, BALF and whole lungs were evaluated for the inflammatory cytokine Il-6 and chemokine MIP-2, histopathology, cellular populations of macrophages, dendritic cells (DCs), and granulocytes, and particulate uptake within these cells through flow cytometry, ELISAs, and fluorescent imaging.
Results
Particles of all sizes and surface chemistries were readily observed in the lung with minimal inflammatory response at all time points. Surface modification with PEGylation was found to significantly increase lung residence times and homogeneous lung distribution, delaying macrophage clearance of all sizes, with the largest increase in residence time observed for 80 × 320 nm particles. Additionally, it was observed that DCs were recruited to the airway following administration of unPEGylated particles and preferentially associated with these particles.
Conclusions
Pulmonary drug delivery vehicles designed with a PEG surface coating can be used to delay particle uptake and promote cell-specific targeting of therapeutics.
Similar content being viewed by others
Abbreviations
- AEM:
-
2-aminoethyl methacrylate
- AM:
-
Alveolar macrophages
- BALF:
-
Broncheoalveolar lavage fluid
- COPD:
-
Chronic obstructive pulmonary disease
- DC:
-
Dendritic cells
- Dpi:
-
Dry powder inhaler
- DPPC:
-
1,2-dipalmitoylphosphatidylcholine
- ELISA:
-
Enzyme-linked immuno assay
- HP4A:
-
Tetra(ethylene glycol) monoacrylate
- IACUC:
-
Institutional animal care and use committee
- IV:
-
Intravenous
- LPPs:
-
Large porous particles
- LPS:
-
Lipopolysaccharide
- MΦ:
-
Macrophages
- PEG:
-
Poly(ethylene glycol)
- PEG700DA:
-
Poly(ethylene glycol)700
- PRINT:
-
Particle Replication In Non-wetting Templates
- SEM:
-
Scanning electron microscopy
- TPO:
-
Diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide
References
Patton JS, Byron PR. Inhaling medicines: delivering drugs to the body through the lungs. Nat Rev Drug Discov. 2007;6:67–74.
Pulliam B, Sung JC, Edwards DA. Design of nanoparticle-based dry powder pulmonary vaccines. Expert Opin Drug Deliv. 2007;4:651–63.
Chow AH, Tong HH, Chattopadhyay P, Shekunov BY. Particle engineering for pulmonary drug delivery. Pharm Res. 2007;24:411–37.
Azarmi S, Roa WH, Lobenberg R. Targeted delivery of nanoparticles for the treatment of lung diseases. Adv Drug Deliv Rev. 2008;60:863–75.
Garcia A, Mack P, Williams S, Fromen C, Shen T, Tully J, et al. Microfabricated engineered particle systems for respiratory drug delivery and other pharmaceutical applications. J Drug Delivery. 2012;2012:941243.
Mansour HM, Rhee Y, Wu X. Nanomedicine in pulmonary delivery. Int J Nanomedicine. 2009;4:299–319.
Newmanand SP, Busse WW. Evolution of dry powder inhaler design, formulation, and performance. Respir Med. 2002;96:293–304.
Lombry C, Edwards DA, Preat V, Vanbever R. Alveolar macrophages are a primary barrier to pulmonary absorption of macromolecules. Am J Physiol Lung Cell Mol Physiol. 2004;286:L1002–8.
Lehnert BE. Pulmonary and thoracic macrophage subpopulations and clearance of particles from the lung. Environ Health Perspect. 1992;97:17–46.
Sharma R, Saxena D, Dwivedi AK, Misra A. Inhalable microparticles containing drug combinations to target alveolar macrophages for treatment of pulmonary tuberculosis. Pharm Res. 2001;18:1405–10.
Champion JA, Walker A, Mitragotri S. Role of particle size in phagocytosis of polymeric microspheres. Pharm Res. 2008;25:1815–21.
Edwards DA, Hanes J, Caponetti G, Hrkach J, Ben-Jebria A, Eskew ML, et al. Large porous particles for pulmonary drug delivery. Science. 1997;276:1868–971.
El-Sherbiny IM, McGill S, Smyth HD. Swellable microparticles as carriers for sustained pulmonary drug delivery. J Pharm Sci. 2010;99:2343–56.
Moon JJ, Huang B, Irvine DJ. Engineering nano- and microparticles to tune immunity. Adv Mater. 2012;24:3724–46.
Nembrini C, Stano A, Dane KY, Ballester M, van der Vlies AJ, Marsland BJ, et al. Nanoparticle conjugation of antigen enhances cytotoxic T-cell responses in pulmonary vaccination. Proc Natl Acad Sci U S A. 2011;108:E989–97.
Steinman RM. Dendritic cells: versatile controllers of the immune system. Nat Med. 2007;13:1155–9.
Naito T, Suda T, Suzuki K, Nakamura Y, Inui N, Sato J, et al. Lung dendritic cells have a potent capability to induce production of immunoglobulin A. Am J Respir Cell Mol Biol. 2008;38:161–7.
Hubbell JA, Thomas SN, Swartz MA. Materials engineering for immunomodulation. Nature. 2009;462:449–60.
Guilliams M, Lambrecht BN, Hammad H. Division of labor between lung dendritic cells and macrophages in the defense against pulmonary infections. Mucosal Immunol. 2013;6:464–73.
Ahsan F, Rivas IP, Khan MA, Torres Suarez AI. Targeting to macrophages: role of physicochemical properties of particulate carriers–liposomes and microspheres–on the phagocytosis by macrophages. J Control Release : Off J Control Release Soc. 2002;79:29–40.
Perry JL, Reuter KG, Kai MP, Herlihy KP, Jones SW, Luft JC, et al. PEGylated PRINT nanoparticles: the impact of PEG density on protein binding, macrophage association, biodistribution, and pharmacokinetics. Nano Lett. 2012;12:5304–10.
Liand SD, Huang L. Nanoparticles evading the reticuloendothelial system: role of the supported bilayer. Biochim Biophys Acta. 2009;1788:2259–66.
Owens 3rd DE, Peppas NA. Opsonization, biodistribution, and pharmacokinetics of polymeric nanoparticles. Int J Pharm. 2006;307:93–102.
Knop K, Hoogenboom R, Fischer D, Schubert US. Poly(ethylene glycol) in drug delivery: pros and cons as well as potential alternatives. Angew Chem Int Ed Engl. 2010;49:6288–308.
Muralidharan P, Mallory E, Malapit M, Hayes Jr D, Mansour HM. Inhalable PEGylated phospholipid nanocarriers and PEGylated therapeutics for respiratory delivery as aerosolized colloidal dispersions and dry powder inhalers. Pharmaceutics. 2014;6:333–53.
Bayard FJ, Thielemans W, Pritchard DI, Paine SW, Young SS, Backman P, et al. Polyethylene glycol-drug ester conjugates for prolonged retention of small inhaled drugs in the lung. J Control Release : Off J Control Release Soc. 2013;171:234–40.
Koussoroplis SJ, Paulissen G, Tyteca D, Goldansaz H, Todoroff J, Barilly C, et al. PEGylation of antibody fragments greatly increases their local residence time following delivery to the respiratory tract. J Control Release : Off J Control Release Soc. 2014;187:91–100.
Youn YS, Kwon MJ, Na DH, Chae SY, Lee S, Lee KC. Improved intrapulmonary delivery of site-specific PEGylated salmon calcitonin: optimization by PEG size selection. J Control Release : Off J Control Release Soc. 2008;125:68–75.
Merkel OM, Beyerle A, Librizzi D, Pfestroff A, Behr TM, Sproat B, et al. Nonviral siRNA Delivery to the Lungs: Investigation of PEG-PEI Polyplexes and Their In Vivo Performance. Mol Pharm. 2009;6:1246–60.
Meenach SA, Anderson KW, Zach Hilt J, McGarry RC, Mansour HM. Characterization and aerosol dispersion performance of advanced spray-dried chemotherapeutic PEGylated phospholipid particles for dry powder inhalation delivery in lung cancer. Eur J Pharm Sci. 2013;49:699–711.
Patel B, Gupta V, Ahsan F. PEG-PLGA based large porous particles for pulmonary delivery of a highly soluble drug, low molecular weight heparin. J Control Release : Off J Control Release Soc. 2012;162:310–20.
Fromen CA, Robbins GR, Shen TW, Kai MP, Ting JPY, and DeSimone JM. Controlled analysis of nanoparticle charge on mucosal and systemic antibody responses following pulmonary immunization. Proc Natl Acad Sci. 2015;112:488–93.
Ibricevic A, Guntsen SP, Zhang K, Shrestha R, Liu Y, Sun JY, et al. PEGylation of cationic, shell-crosslinked-knedel-like nanoparticles modulates inflammation and enhances cellular uptake in the lung. Nanomed Nanotechnol Biol Med. 2013;9:912–22.
Fu J, Fiegel J, Krauland E, Hanes J. New polymeric carriers for controlled drug delivery following inhalation or injection. Biomaterials. 2002;23:4425–33.
Lai SK, O’Hanlon DE, Harrold S, Man ST, Wang YY, Cone R, et al. Rapid transport of large polymeric nanoparticles in fresh undiluted human mucus. Proc Natl Acad Sci U S A. 2007;104:1482–7.
Suk JS, Kim AJ, Trehan K, Schneider CS, Cebotaru L, Woodward OM, et al. Lung gene therapy with highly compacted DNA nanoparticles that overcome the mucus barrier. J Control Release : Off J Control Release Soc. 2014;178:8–17.
Evora C, Soriano I, Rogers RA, Shakesheff KN, Hanes J, Langer R. Relating the phagocytosis of microparticles by alveolar macrophages to surface chemistry: the effect of 1,2-dipalmitoylphosphatidylcholine. J Control Release : Off J Control Release Soc. 1998;51:143–52.
Rolland JP, Maynor BW, Euliss LE, Exner AE, Denison GM, DeSimone JM. Direct fabrication and harvesting of monodisperse shape-specific nanobiomaterials. J Am Chem Soc. 2005;127:10069–100.
Fromen CA, Shen TW, Larus AE, Mack P, Maynor BW, Luft JC, et al. Synthesis and characterization of monodisperse uniformly shaped respirable aerosols. AICHE J. 2013;59:3184–94.
Roberts RA, Shen T, Allen IC, Hasan W, DeSimone JM, Ting JP. Analysis of the murine immune response to pulmonary delivery of precisely fabricated nano- and microscale particles. PLoS One. 2013;8:e62115.
Guzman J, Iglesias MT, Riande E. Synthesis and polymerization of acrylic monomers with hydrophilic long side groups. Polymer. 1997;38:5227–32.
Tabataand Y, Ikada Y. Effect of the size and surface charge of polymer microspheres on their phagocytosis by macrophage. Biomaterials. 1988;9:356–62.
Salvi S, Blomberg A, Rudell B, Kelly F, Sandstrom T, Holgate ST, et al. Acute inflammatory responses in the airways and peripheral blood after short-term exposure to diesel exhaust in healthy human volunteers. Am J Respir Crit Care Med. 1999;159:702–9.
Oberdörster G, Oberdörster E, Oberdörster J. Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect. 2005;113:823–39.
Alexis F, Pridgen E, Molnar LK, Farokhzad OC. Factors affecting the clearance and biodistribution of polymeric nanoparticles. Mol Pharm. 2008;5:505–15.
de Gennes PG. Polymers at an interface: a simplified view. Adv Colloid Interface Sci. 1987;27:189–209.
Gref R, Lück M, Quellec P, Marchand M, Dellacherie E, Harnisch S, et al. ‘Stealth’ corona-core nanoparticles surface modified by polyethylene glycol (PEG): influences of the corona (PEG chain length and surface density) and of the core composition on phagocytic uptake and plasma protein adsorption. Colloids Surf B Biointerfaces. 2000;18:301–13.
Chu KS, Schorzman AN, Finniss MC, Bowerman CJ, Peng L, Luft JC, et al. Nanoparticle drug loading as a design parameter to improve docetaxel pharmacokinetics and efficacy. Biomaterials. 2013;34:8424–9.
Wright JR. Immunoregulatory functions of surfactant proteins. Nat Rev Immunol. 2005;5:58–68.
Haczku A. Protective role of the lung collectins surfactant protein A and surfactant protein D in airway inflammation. J Allergy Clin Immunol. 2008;122:861–79.
Young HW, Sun CX, Evans CM, Dickey BF, Blackburn MR. A3 adenosine receptor signaling contributes to airway mucin secretion after allergen challenge. Am J Respir Cell Mol Biol. 2006;35:549–58.
ACKNOWLEDGMENTS AND DISCLOSURES
We thank R. Roberts, K. Reuter, J. Perry, S. Tian, J. P.Y. Ting, A. Pandya, B. Udis, N. Fisher, S. Coquery, L. Wai, and, J. Weaver for useful discussions and technical assistance. We acknowledge Liquidia Technologies for providing PRINT molds, and the core facilities at UNC, including the Nucleic Acids Core Facility, CHANL imaging facility, the LCCC Histopathology Core, the Histology Facility of the Department of Cell and Molecular Physiology, the Department of Microbiology and Immunology Flow Cytometry Core Facility, and DLAM facility. This work was funded in part by the NIH Pioneer Award to J.M.D. (1DP1OD006432), DTRA award (HDTRA1-13-1-0045), and the NSF Graduate Research Fellowship, as well as NCI Center Core Support Grant CA016086. J.M.D. is a founder and maintains a financial interest in Liquidia Technologies.
Author information
Authors and Affiliations
Corresponding author
Additional information
Tammy W. Shen and Catherine A. Fromen contributed equally to this work.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Supplemental Figure 1
Representative flow cytometric analysis of BALF and Lung cells. (A) Gating scheme used to define three major lung cell populations: Granulocytes CD45 + CD11c-Ly6G+, Macrophages (Mφs) CD45+CD11c+MHCIIlo, Dendritic cells (DCs) CD45+CD11c+MHCIIhi. (B) Representative gating of particle positive populations of Granulocytes, AMs and DCs. Blue curves representative data from 80 × 320 day 1 BALF. Red curves represent fluorescence minus one (FMO) controls used to set gates. Black curves represent saline controls. (JPEG 92 kb)
Rights and permissions
About this article
Cite this article
Shen, T.W., Fromen, C.A., Kai, M.P. et al. Distribution and Cellular Uptake of PEGylated Polymeric Particles in the Lung Towards Cell-Specific Targeted Delivery. Pharm Res 32, 3248–3260 (2015). https://doi.org/10.1007/s11095-015-1701-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11095-015-1701-7