Biocompatibility and cellular uptake mechanisms of poly(N-isopropylacrylamide) in different cells

Thermosensitive poly(N-isopropylacrylamide) is widely used in various biomedical applications including drug delivery systems, gene delivery systems, switching devices, sensors, and diagnostic assays. To promote these clinical applications, it is essential to have a comprehensive understanding of the biosafety of poly(N-isopropylacrylamide) and the interaction of poly(N-isopropylacrylamide) with different cell lines, which has little research until now. In this work, we evaluated the biocompatibility of poly(N-isopropylacrylamide) including cell viability, nitric oxide production, and apoptosis of macrophages RAW264.7, human bronchial epithelial cells, A549, and human umbilical vein endothelial cells in the presence of poly(N-isopropylacrylamide). We have also examined the cellular uptake mechanisms of poly(N-isopropylacrylamide) using endocytic inhibitors and insighted into the intracellular co-localization of poly(N-isopropylacrylamide) using confocal laser scanning microscope. The results showed that poly(N-isopropylacrylamide) had good biocompatibility and could be internalized by these cells. It is macropinocytosis that poly(N-isopropylacrylamide) could be internalized in RAW264.7 cells and caveolae-mediated endocytosis in human bronchial epithelial cells, A549, and human umbilical vein endothelial cells. In addition, we also evidenced that intracellular poly(N-isopropylacrylamide) was co-localized with lysosome. The study provided important information for the development and clinical applications of poly(N-isopropylacrylamide) in the biomedical field.

[1]  Yi-fang Cheng,et al.  AMPK activation inhibits expression of proinflammatory mediators through downregulation of PI3K/p38 MAPK and NF-κB signaling in murine macrophages. , 2015, DNA and cell biology.

[2]  J. Kizhakkedathu,et al.  Abnormal blood clot formation induced by temperature responsive polymers by altered fibrin polymerization and platelet binding. , 2014, Biomaterials.

[3]  Marco Laurenti,et al.  Structure and polymer dynamics within PNIPAM-based microgel particles. , 2014, Advances in colloid and interface science.

[4]  Kevin Braeckmans,et al.  On the cellular processing of non-viral nanomedicines for nucleic acid delivery: mechanisms and methods. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[5]  Bridgette M Budhlall,et al.  Multicore-shell PNIPAm-co-PEGMa microcapsules for cell encapsulation. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[6]  H. Byrne,et al.  Intracellular localisation, geno- and cytotoxic response of polyN-isopropylacrylamide (PNIPAM) nanoparticles to human keratinocyte (HaCaT) and colon cells (SW 480). , 2010, Toxicology letters.

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

[8]  R. Lundmark,et al.  GRAF1-dependent endocytosis. , 2009, Biochemical Society transactions.

[9]  Joel A. Swanson,et al.  Shaping cups into phagosomes and macropinosomes , 2008, Nature Reviews Molecular Cell Biology.

[10]  Satyajit Mayor,et al.  Pathways of clathrin-independent endocytosis , 2007, Nature Reviews Molecular Cell Biology.

[11]  Arwyn Tomos Jones,et al.  Macropinocytosis: searching for an endocytic identity and role in the uptake of cell penetrating peptides , 2007, Journal of cellular and molecular medicine.

[12]  S. Futaki,et al.  Interaction of arginine-rich peptides with membrane-associated proteoglycans is crucial for induction of actin organization and macropinocytosis. , 2007, Biochemistry.

[13]  Ernst Wagner,et al.  The internalization route resulting in successful gene expression depends on both cell line and polyethylenimine polyplex type. , 2006, Molecular therapy : the journal of the American Society of Gene Therapy.

[14]  En-Tang Kang,et al.  pH- and temperature-responsive hydrogels from crosslinked triblock copolymers prepared via consecutive atom transfer radical polymerizations. , 2006, Biomaterials.

[15]  Shiroh Futaki,et al.  High Density of Octaarginine Stimulates Macropinocytosis Leading to Efficient Intracellular Trafficking for Gene Expression* , 2006, Journal of Biological Chemistry.

[16]  Steven F Dowdy,et al.  Cationic TAT peptide transduction domain enters cells by macropinocytosis. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[17]  J. Gorvel,et al.  Macropinocytosis of polyplexes and recycling of plasmid via the clathrin-dependent pathway impair the transfection efficiency of human hepatocarcinoma cells. , 2004, Molecular therapy : the journal of the American Society of Gene Therapy.

[18]  V. H. Lee,et al.  Clathrin and caveolin-1 expression in primary pigmented rabbit conjunctival epithelial cells: role in PLGA nanoparticle endocytosis. , 2003, Molecular vision.

[19]  Sandra L. Schmid,et al.  Regulated portals of entry into the cell , 2003, Nature.

[20]  Wong,et al.  Biological barriers to cellular delivery of lipid-based DNA carriers. , 1999, Advanced drug delivery reviews.

[21]  H. Radhakrishna,et al.  ADP-Ribosylation Factor 6 Regulates a Novel Plasma Membrane Recycling Pathway , 1997, The Journal of cell biology.

[22]  B. Nichols,et al.  Flotillin-1 defines a clathrin-independent endocytic pathway in mammalian cells , 2006, Nature Cell Biology.

[23]  I. Zuhorn,et al.  Size-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosis. , 2004, The Biochemical journal.

[24]  F. Maxfield,et al.  Endocytic recycling , 2004, Nature Reviews Molecular Cell Biology.