Serum-derived nanoparticles: de novo generation and growth in vitro, and internalization by mammalian cells in culture.

AIM While nanoparticles (NPs) have been shown to form spontaneously in body fluids such as serum, the possible implications of these NPs for cell cultures that use supporting media containing serum remain unclear. To understand the de novo formation of NPs, we delineated their growth characteristics, chemical composition and interaction with cells in culture. MATERIALS & METHODS Serum-derived particles were analyzed using a combination of dynamic light scattering, turbidity measurements, spectroscopic techniques and optical/electron microscopies. RESULTS NPs were found in serum and in serum-containing medium and they increased in size and number during incubation. The mineral particles, consisting mainly of calcium carbonate phosphate bound to organics such as proteins, underwent an amorphous-to-crystalline transformation with time. Serum-derived particles were internalized by the cells tested, eventually reaching lysosomal compartments. CONCLUSION The spontaneous formation of serum-derived NPs and their internalization by cells may have overlooked effects on cultured cells in vitro as well as potential pathophysiological consequences in vivo.

[1]  J. Martel,et al.  Critical Evaluation of Gamma-Irradiated Serum Used as Feeder in the Culture and Demonstration of Putative Nanobacteria and Calcifying Nanoparticles , 2010, PloS one.

[2]  M. Orellana,et al.  Spontaneous assembly of marine dissolved organic matter into polymer gels , 1998, Nature.

[3]  Gaurav Sahay,et al.  Endocytosis of nanomedicines. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[4]  Tadashi Kokubo,et al.  Apatite formation on surfaces of ceramics, metals and polymers in body environment , 1998 .

[5]  R. Ryall The future of stone research: rummagings in the attic, Randall’s plaque, nanobacteria, and lessons from phylogeny , 2008, Urological Research.

[6]  Bengt Fadeel,et al.  Close encounters of the small kind: adverse effects of man-made materials interfacing with the nano-cosmos of biological systems. , 2010, Annual review of pharmacology and toxicology.

[7]  Jeremy J. W. Chen,et al.  Titanium dioxide nanoparticles induce emphysema‐like lung injury in mice , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[8]  D. Mckay,et al.  Nanobacteria: Fact or Fiction? Characteristics, Detection, and Medical Importance of Novel Self-Replicating, Calcifying Nanoparticles , 2006, Journal of investigative medicine : the official publication of the American Federation for Clinical Research.

[9]  P. Dieppe,et al.  The effects of serum and human albumin on calcium hydroxyapatite crystal growth. , 1990, The Biochemical journal.

[10]  Jan Martel,et al.  Putative Nanobacteria Represent Physiological Remnants and Culture By-Products of Normal Calcium Homeostasis , 2009, PloS one.

[11]  David J. Young,et al.  Fetuin-A/Albumin-Mineral Complexes Resembling Serum Calcium Granules and Putative Nanobacteria: Demonstration of a Dual Inhibition-Seeding Concept , 2009, PloS one.

[12]  David J. Young,et al.  Characterization of Granulations of Calcium and Apatite in Serum as Pleomorphic Mineralo-Protein Complexes and as Precursors of Putative Nanobacteria , 2009, PloS one.

[13]  Parag Aggarwal,et al.  Interaction of colloidal gold nanoparticles with human blood: effects on particle size and analysis of plasma protein binding profiles. , 2009, Nanomedicine : nanotechnology, biology, and medicine.

[14]  J. Lingeman,et al.  Mechanism of Formation of Human Calcium Oxalate Renal Stones on Randall's Plaque , 2007, Anatomical record.

[15]  P. Couvreur,et al.  Analysis of plasma protein adsorption onto PEGylated nanoparticles by complementary methods: 2‐DE, CE and Protein Lab‐on‐chip® system , 2007, Electrophoresis.

[16]  J. Thompson,et al.  An alternative interpretation of nanobacteria-induced biomineralization. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[17]  Jean-Christophe Leroux,et al.  Effect of poly(N-vinyl-pyrrolidone)-block-poly(D,L-lactide) as coating agent on the opsonization, phagocytosis, and pharmacokinetics of biodegradable nanoparticles. , 2009, Biomacromolecules.

[18]  M. Stoller,et al.  Association between Randall’s plaque and calcifying nanoparticles , 2008, International journal of nanomedicine.

[19]  Feng Zhang,et al.  Quantitative analysis of the protein corona on FePt nanoparticles formed by transferrin binding , 2010, Journal of The Royal Society Interface.

[20]  H. Cheung,et al.  Calcium phosphate particle induction of metalloproteinase and mitogenesis: effect of particle sizes. , 1997, Osteoarthritis and cartilage.

[21]  Neva Ciftcioglu,et al.  Nanobacteria: An alternative mechanism for pathogenic intra- and extracellular calcification and stone formation , 1998 .

[22]  D. Hukins,et al.  Seeded growth of hydroxyapatite in the presence of dissolved albumin. , 1994, Journal of inorganic biochemistry.

[23]  Martin Raff,et al.  Intracellular Vesicular Traffic , 2002 .

[24]  J. Ashwell,et al.  Activation of the Lck tyrosine kinase targets cell surface T cell antigen receptors for lysosomal degradation. , 1997, Immunity.

[25]  P. Price,et al.  Biochemical Characterization of the Serum Fetuin-Mineral Complex* , 2003, Journal of Biological Chemistry.

[26]  R. Mendelsohn,et al.  Thermal denaturation of globular proteins. Fourier transform-infrared studies of the amide III spectral region. , 1987, Biophysical journal.

[27]  D. Sparks,et al.  Nanominerals, Mineral Nanoparticles, and Earth Systems , 2008, Science.

[28]  Joachim Mayer,et al.  Ultrastructural analysis of vascular calcifications in uremia. , 2010, Journal of the American Society of Nephrology : JASN.

[29]  M. McKee,et al.  Calcium oxalate crystals in fetal bovine serum: Implications for cell culture, phagocytosis and biomineralization studies in vitro , 2008, Journal of cellular biochemistry.

[30]  Jan Martel,et al.  Purported nanobacteria in human blood as calcium carbonate nanoparticles , 2008, Proceedings of the National Academy of Sciences.

[31]  J. Lieske Can biologic nanoparticles initiate nephrolithiasis? , 2008, Nature Clinical Practice Nephrology.

[32]  J. Skepper,et al.  Multifunctional roles for serum protein fetuin-a in inhibition of human vascular smooth muscle cell calcification. , 2005, Journal of the American Society of Nephrology : JASN.

[33]  Costanza Miliani,et al.  The application of in situ mid-FTIR fibre-optic reflectance spectroscopy and GC-MS analysis to monitor and evaluate painting cleaning. , 2009, Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy.

[34]  Xinggui Zhou,et al.  Electrochemical Impedance Spectroscopy Study of the Nucleation and Growth of Apatite on Chemically Treated Titanium , 2002 .

[35]  P. Dieppe,et al.  Hydroxyapatite and urate crystal induced cytokine release by macrophages. , 1989, Annals of the rheumatic diseases.

[36]  W. Jahnen-Dechent,et al.  The serum protein alpha2-HS glycoprotein/fetuin inhibits apatite formation in vitro and in mineralizing calvaria cells. A possible role in mineralization and calcium homeostasis. , 1996, The Journal of biological chemistry.

[37]  H. Anderson,et al.  The role of matrix vesicles in growth plate development and biomineralization. , 2005, Frontiers in bioscience : a journal and virtual library.

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

[39]  J. Lingeman,et al.  Apatite plaque particles in inner medulla of kidneys of calcium oxalate stone formers: osteopontin localization. , 2005, Kidney international.

[40]  M. Björklund,et al.  Nanobacteria: an infectious cause for kidney stone formation. , 1999, Kidney international.

[41]  Craig A. Poland,et al.  Asbestos, carbon nanotubes and the pleural mesothelium: a review of the hypothesis regarding the role of long fibre retention in the parietal pleura, inflammation and mesothelioma , 2010, Particle and Fibre Toxicology.

[42]  D. Raoult,et al.  Nanobacteria Are Mineralo Fetuin Complexes , 2008, PLoS pathogens.

[43]  J. Martel,et al.  The rise and fall of nanobacteria. , 2010, Scientific American.

[44]  T. Xia,et al.  Toxic Potential of Materials at the Nanolevel , 2006, Science.

[45]  Y. Totsuka,et al.  Material nanosizing effect on living organisms: non-specific, biointeractive, physical size effects , 2009, Journal of The Royal Society Interface.

[46]  J. Lieske,et al.  Characterization of biofilm formed by human-derived nanoparticles. , 2009, Nanomedicine.

[47]  W. Semmler,et al.  Determination of Plasma Protein Adsorption on Magnetic Iron Oxides: Sample Preparation , 1997, Pharmaceutical Research.

[48]  M. McKee,et al.  Mineral chaperones: a role for fetuin-A and osteopontin in the inhibition and regression of pathologic calcification , 2008, Journal of Molecular Medicine.

[49]  E. Kajander Nanobacteria – propagating calcifying nanoparticles , 2006, Letters in applied microbiology.

[50]  F. Brodsky,et al.  T Cell Receptor Engagement Leads to Phosphorylation of Clathrin Heavy Chain during Receptor Internalization , 2004, The Journal of experimental medicine.

[51]  Tadashi Kokubo,et al.  Spontaneous Formation of Bonelike Apatite Layer on Chemically Treated Titanium Metals , 1996 .

[52]  J. Lieske,et al.  Biologic nanoparticles and platelet reactivity. , 2009, Nanomedicine.

[53]  N. Fujii,et al.  Fully phosphorylated fetuin-A forms a mineral complex in the serum of rats with adenine-induced renal failure. , 2009, Kidney international.

[54]  S. Grinstein,et al.  Differential Role of Actin, Clathrin, and Dynamin in Fcγ Receptor-mediated Endocytosis and Phagocytosis* , 2003, The Journal of Biological Chemistry.

[55]  R. Müller,et al.  Protein adsorption patterns on poloxamer- and poloxamine-stabilized solid lipid nanoparticles (SLN). , 2005, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[56]  P. Price,et al.  Bone Origin of the Serum Complex of Calcium, Phosphate, Fetuin, and Matrix Gla Protein: Biochemical Evidence for the Cancellous Bone‐Remodeling Compartment , 2002, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[57]  F. N. Ghadially As You Like It, Part 3: A Critique and Historical Review of Calcification as Seen with the Electron Microscope , 2001, Ultrastructural pathology.