Poly (N-isopropylacrylamide)-PLA and PLA blend nanoparticles for temperature-controllable drug release and intracellular uptake.

We designed a temperature-responsive and biodegradable novel drug-delivery carrier. A block copolymer, poly (N-isopropylacrylamide-dl-lactide) (PNIPAAm-PLA), was synthesized by the ring-opening polymerization of dl-lactide, and used as a carrier for a drug-delivery system. In this study, temperature-responsive nanoparticles (NPs) encapsulating betamethasone disodium 21-phosphate (BP) were prepared from a blend of PLA homopolymer and block copolymers by an oil-in-water solvent-diffusion method in the presence of zinc ion (PLA/PNIPAAm-PLA (NPs)). The resulting NP size was around 140 nm. The drug release from temperature-responsive NP could be controllable by changing the temperature. Moreover, a murine macrophage-like cell line, RAW 264.7 cells, was used to measure and image the cell uptake of fluorescent PLA/PNIPAAm-PLA NPs at 30 °C and 37 °C on the boundary of LCST (34 °C). Below the LCST, cellular uptake was not observed, but contrary to cellular uptake it was clearly observed above the LCST. Moreover, we found this effect to be useful for controlling the stealthiness by changing the temperature. Present temperature-responsive NPs have successfully exhibited thermo-responsive drug release and intracellular uptake while possessing a biodegradable character.

[1]  Jan Feijen,et al.  Thermosensitive Interpenetrating Polymer Networks: Synthesis, Characterization, and Macromolecular Release , 1994 .

[2]  T. Okano,et al.  Molecular design of outermost surface functionalized thermoresponsive polymeric micelles with biodegradable cores , 2008 .

[3]  T. Okano,et al.  Temperature-responsive liquid chromatography. 2. Effects of hydrophobic groups in N-isopropylacrylamide copolymer-modified silica. , 1997, Analytical chemistry.

[4]  Kazuo Maruyama,et al.  Effective anti-tumor activity of oxaliplatin encapsulated in transferrin-PEG-liposome. , 2008, International journal of pharmaceutics.

[5]  Teruo Okano,et al.  Thermo-responsive polymer nanoparticles with a core-shell micelle structure as site-specific drug carriers , 1997 .

[6]  T. Okano,et al.  Process design for efficient and controlled drug incorporation into polymeric micelle carrier systems. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[7]  T. Ishihara,et al.  Treatment of experimental arthritis with poly(d, l-lactic/glycolic acid) nanoparticles encapsulating betamethasone sodium phosphate , 2005, Annals of the rheumatic diseases.

[8]  R. Müller,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. , 2000, Colloids and surfaces. B, Biointerfaces.

[9]  T. Okano,et al.  Study of temperature-responsibility on the surfaces of a thermo-responsive polymer modified stationary phase. , 2006, Journal of chromatography. A.

[10]  T. Okano,et al.  Preparation and characterization of thermally responsive block copolymer micelles comprising poly(N-isopropylacrylamide-b-DL-lactide). , 1998, Journal of controlled release : official journal of the Controlled Release Society.

[11]  S. Fukushima,et al.  Cyclic RGD Peptide-Conjugated Polyplex Micelles as a Targetable Gene Delivery System Directed to Cells Possessing αvβ3 and αvβ5 Integrins , 2007 .

[12]  M. Jones,et al.  Polymeric micelles - a new generation of colloidal drug carriers. , 1999, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[13]  D. Bazile,et al.  Stealth Me.PEG-PLA nanoparticles avoid uptake by the mononuclear phagocytes system. , 1995, Journal of pharmaceutical sciences.

[14]  E. Ayano,et al.  Polymeric nanoparticles encapsulating betamethasone phosphate with different release profiles and stealthiness. , 2009, International journal of pharmaceutics.

[15]  T. Okano,et al.  Effect of molecular architecture of poly(N-isopropylacrylamide)-trypsin conjugates on their solution and enzymatic properties. , 1996, Bioconjugate chemistry.

[16]  H. Harashima,et al.  Tumor targeting of doxorubicin by anti-MT1-MMP antibody-modified PEG liposomes. , 2007, International journal of pharmaceutics.

[17]  K. Avgoustakis,et al.  Biodistribution properties of nanoparticles based on mixtures of PLGA with PLGA-PEG diblock copolymers. , 2005, International journal of pharmaceutics.

[18]  T. Mizushima,et al.  Prolonging the In Vivo Residence Time of Prostaglandin E1 with Biodegradable Nanoparticles , 2008, Pharmaceutical Research.

[19]  Teruo Okano,et al.  Polymeric micelles as new drug carriers , 1996 .

[20]  T. Okano,et al.  Temperature-Responsive Chromatography Using Poly-(N-isopropylacrylamide) Hydrogel-Modified Silica , 2002, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[21]  K. Avgoustakis,et al.  Effect of dose on the biodistribution and pharmacokinetics of PLGA and PLGA-mPEG nanoparticles. , 2001, International journal of pharmaceutics.

[22]  Michel Vert,et al.  Biodistribution of Long-Circulating PEG-Grafted Nanocapsules in Mice: Effects of PEG Chain Length and Density , 2001, Pharmaceutical Research.

[23]  T. Okano,et al.  Inner core segment design for drug delivery control of thermo-responsive polymeric micelles. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[24]  Hideyoshi Harashima,et al.  A multifunctional envelope type nano device (MEND) for gene delivery to tumours based on the EPR effect: a strategy for overcoming the PEG dilemma. , 2011, Advanced drug delivery reviews.

[25]  Martin C. Garnett,et al.  Physicochemical Evaluation of Nanoparticles Assembled from Poly(lactic acid)−Poly(ethylene glycol) (PLA−PEG) Block Copolymers as Drug Delivery Vehicles , 2001 .

[26]  T. Okano,et al.  Molecular design of biodegradable polymeric micelles for temperature-responsive drug release. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[27]  Chi‐Hwa Wang,et al.  Effect of PEG conformation and particle size on the cellular uptake efficiency of nanoparticles with the HepG2 cells. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[28]  T. Ishida,et al.  Accelerated blood clearance (ABC) phenomenon upon repeated injection of PEGylated liposomes. , 2008, International journal of pharmaceutics.

[29]  T. Okano,et al.  Two-dimensional manipulation of cardiac myocyte sheets utilizing temperature-responsive culture dishes augments the pulsatile amplitude. , 2001, Tissue engineering.

[30]  T. Ishihara,et al.  Efficient encapsulation of a water-soluble corticosteroid in biodegradable nanoparticles. , 2009, International journal of pharmaceutics.

[31]  T. Ishihara,et al.  Role of zinc in formulation of PLGA/PLA nanoparticles encapsulating betamethasone phosphate and its release profile. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[32]  T. Ishida,et al.  Effect of the physicochemical properties of initially injected liposomes on the clearance of subsequently injected PEGylated liposomes in mice. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[33]  K. Avgoustakis,et al.  Effect of copolymer composition on the physicochemical characteristics, in vitro stability, and biodistribution of PLGA-mPEG nanoparticles. , 2003, International journal of pharmaceutics.

[34]  M. Heskins,et al.  Solution Properties of Poly(N-isopropylacrylamide) , 1968 .

[35]  E. Ayano,et al.  Aqueous chromatography system using temperature-responsive polymer-modified stationary phases. , 2006, Journal of separation science.

[36]  G. Kibria,et al.  Dual-ligand modification of PEGylated liposomes shows better cell selectivity and efficient gene delivery. , 2011, Journal of controlled release : official journal of the Controlled Release Society.

[37]  T. Okano,et al.  Temperature-induced intracellular uptake of thermoresponsive polymeric micelles. , 2009, Biomacromolecules.

[38]  T. Okano,et al.  Thermally controlled intracellular uptake system of polymeric micelles possessing poly(N-isopropylacrylamide)-based outer coronas. , 2010, Molecular pharmaceutics.

[39]  T. Okano,et al.  Temperature-Responsive Chromatography Using Poly(N-isopropylacrylamide)-Modified Silica. , 1996, Analytical chemistry.

[40]  Teruo Okano,et al.  Temperature dependence of swelling of crosslinked poly(N,N′-alkyl substituted acrylamides) in water , 1990 .

[41]  T. Okano,et al.  Control of adriamycin cytotoxic activity using thermally responsive polymeric micelles composed of poly(N-isopropylacrylamide-co-N,N-dimethylacrylamide)-b-poly(d,l-lactide) , 1999 .

[42]  P. Gellert,et al.  Polylactide-poly(ethylene Glycol) Micellar-like Particles as Potential Drug Carriers: Production, Colloidal Properties and Biological Performance , 2001, Journal of drug targeting.

[43]  T. Okano,et al.  Reversibly thermo-responsive alkyl-terminated poly(N-isopropylacrylamide) core-shell micellar structures , 1997 .

[44]  Xin Yu Wang,et al.  Accelerated blood clearance of PEGylated liposomes following preceding liposome injection: effects of lipid dose and PEG surface-density and chain length of the first-dose liposomes. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[45]  Wenlei Jiang,et al.  Stabilization and Controlled Release of Bovine Serum Albumin Encapsulated in Poly(D, L-lactide) and Poly(ethylene glycol) Microsphere Blends , 2001, Pharmaceutical Research.

[46]  O. Bourdon,et al.  Relationship between complement activation, cellular uptake and surface physicochemical aspects of novel PEG-modified nanocapsules. , 2001, Biomaterials.

[47]  V. Torchilin,et al.  Biodegradable long-circulating polymeric nanospheres. , 1994, Science.

[48]  Allan G. A. Coombes,et al.  Surface Modification of Poly(lactide-co-glycolide) Nanospheres by Biodegradable Poly(lactide)-Poly(ethylene glycol) Copolymers , 1994, Pharmaceutical Research.

[49]  T. Mizushima,et al.  Preparation and characterization of a nanoparticulate formulation composed of PEG-PLA and PLA as anti-inflammatory agents. , 2010, International journal of pharmaceutics.

[50]  K. Nagayama,et al.  Envelope-type lipid nanoparticles incorporating a short PEG-lipid conjugate for improved control of intracellular trafficking and transgene transcription. , 2009, Biomaterials.

[51]  T. Honda,et al.  Design of Folate-Linked Liposomal Doxorubicin to its Antitumor Effect in Mice , 2008, Clinical Cancer Research.

[52]  H. Ringsdorf,et al.  Interactions of liposomes and hydrophobically-modified poly-(N-isopropylacrylamides): an attempt to model the cytoskeleton. , 1993, Biochimica et biophysica acta.

[53]  T. Okano,et al.  Unique Thermoresponsive Polymeric Micelle Behavior via Cooperative Polymer Corona Phase Transitions , 2008 .

[54]  R. Gref,et al.  MPEO-PLA nanoparticles: effect of MPEO content on some of their surface properties. , 1999, Journal of biomedical materials research.

[55]  Leaf Huang,et al.  Nanoparticles targeted with NGR motif deliver c-myc siRNA and doxorubicin for anticancer therapy. , 2010, Molecular therapy : the journal of the American Society of Gene Therapy.