Publikationsserver der Universitätsbibliothek Marburg

Titel:Novel Hybrid Polymeric and Inorganic Structures for Applications in Nanobiotechnology
Autor:Valdeperez Toledo, Daniel
Weitere Beteiligte: Parak, Wolfgang J. (Prof. Dr.)
Veröffentlicht:2017
URI:https://archiv.ub.uni-marburg.de/diss/z2017/0227
URN: urn:nbn:de:hebis:04-z2017-02275
DOI: https://doi.org/10.17192/z2017.0227
DDC: Physik
Titel(trans.):Neuartige hybride Polymerstrukturen und anorganische Strukturen zur Anwendung in der Nanobiotechnologie
Publikationsdatum:2017-12-14
Lizenz:https://creativecommons.org/licenses/by-nc-sa/4.0

Dokument

Schlagwörter:
Biotechnologie, Nanopartikel, Nanoparticles, biotechnology, Proteine, polymere, Polymere, proteine

Summary:
This cumulative doctoral dissertation deals with the use of diverse polymers in different applications within nanoscience. The synthesis and characterization of several nano and microstructures is also explained, focusing on the later surface modification via the use of different polymers. Polymers are chemical compounds formed by the combination of several repeating structural units (monomers) in a process called polymerization. These structures are assembled following a specific pattern and their subsequent properties are given by the monomers added in the polymerization process. Several uses of polymers have been reported, being their use in the process of engineering novel composite materials for applications within fields like aerospace industry, biotechnology or medicine. The work shown in this thesis aimed to implement novel applications for some of the general polymer uses found in literature and the employment of amphiphilic zwitterionic polymers to test their stability for different biological applications. The dissertation is first focused on the study of three different applications of polymers inside nanotechnology. One of the most common applications of the use of amphiphilic polymers is the coating of inorganic NPs initially synthesized in organic solutions, transferring them into aqueous solutions. The resulting polymer coated NPs count on functional groups on their surface allowing further modifications for new functionalities. This procedure is applied to NPs with different size (ranging from 4 to 29 nm core size) and material (gold and iron oxide). A second application of the polymers is the protection of highly unstable, water and oxygen-sensitive clusters from degradation in aqueous environments. For that purpose gold NPs (Au NPs) of 4 nm were used as template and the clusters were collected between the surface of the NP and the amphiphilic polymer shell. The kinetic activity of the clusters was studied in aqueous environment, obtaining signal in at least the first 24 hours after the coating. As a complementary study inside this dissertation, different amphiphilic zwitterionic polymers were synthesized and optimized for a correct stabilization of NPs in water. The influence of parameters like pH, protein concentration and ionic strength was studied to obtain a complete description of the stability of the different zwitterionic polymer-coated NPs, comparing them to the single charge polymer coated NPs (e.g. fully positive or fully negative). A third application involves the self-assembly of alternating-charge polyelectrolyte layers deposited via adsorption on sacrificial calcium carbonate cores, yielding polymeric hollow microstructures able to be provided with physical and biological properties. Both properties are obtained via the accumulation of iron oxide nanoparticles between the polymer layers and the attachment of specific antibodies vion the outermost polymer layer, giving physical (magnetic) and biological (specific recognition) properties to the whole structure. These microcapsules were utilized to obtain a magnetic immunosensor able to specifically recognize and extract horseradish peroxidase (used as protein model) from a solution.

Zusammenfassung:
Diese kumulativ verfasste Doktorarbeit behandelt die Verwendung mehrerer Polymere für unterschiedliche Anwendungen der Nanotechnologie. Diesbezüglich ist die Synthese und Charakterisierung verschiedener Nano- sowie Mikrostrukturen beschrieben, welche mit Hilfe der Polymere oberflächenmodifiziert werden. Als Polymerisation wird der Vorgang bezeichnet wenn mehrere, gleiche molekulare Bausteine (Monomere) sich über chemische Bindungen zu einem großen Polymer zusammenfügen. Die Monomereinheiten sind nach einem spezifischen Muster aneinander gereiht, welches neben den molekularen Eigenschaften die Funktion des Polymers vorgibt. Man findet Polymere in einer Vielzahl von Anwendungen wie der Entwicklung neuer Materialien in Bereichen der Luftfahrtindustrie, Biotechnologie und Medizin. Die vorliegende Arbeit beschreibt erweiternde Anwendungen für den generellen Gebrauch von Polymeren in der Nanotechnologie, sowie die Einführung von amphiphil, zwitterionischen Polymeren und deren Charakterisierung über ihre Stabilität innerhalb biomedizinischer Anwendungen. Der Fokus dieser Arbeit liegt auf der Untersuchung verschiedener Polymere im Bezug auf drei Anwendungsgebiete der Nanotechnologie. Auf diesem Gebiet stellt die Auftragung (eng. coating) von amphiphilen Polymeren auf anorganische Nanopartikel (NPs), welche ursprünglich in organischem Lösungsmittel synthetisiert werden, eine der wichtigsten Anwendungen dar. Die mit Polymer ummantelten NPs können im Anschluss über funktionelle Gruppen an ihrer Oberfläche weiter modifiziert und somit funktionalisiert werden. Dieses Verfahren wird für NPs unterschiedlicher Größe (4 bis 29 nm) und Material (Gold und Eisenoxid) verwendet. Die Polymere können außerdem zur Stabilisierung von Wasser- und Sauerstoffsensitiven Metallcluster verwendet werden und diese somit vor Agglomeration in wässriger Lösung bewahren. Hierfür wurden 4 nm große Gold- Nanopartikel (Au NPs) als Trägermaterial verwendet und die Cluster zwischen der Oberfläche der Au NPs und der amphiphilen Polymerhülle angeordnet. Die kinetische Aktivität wurde in wässriger Lösung untersucht und innerhalb der ersten 24 h nach dem coating ermittelt. Zusätzlich wurden verschiedene amphiphil, zwitterionische Polymere synthetisiert und deren stabilisierende Wirkung auf Nanopartikel in wässriger Lösung optimiert. Der Einfluss von Parametern wie pH, Proteinkonzentration und der Stabilität gegen erhöhte Ionenkonzentration wurden untersucht, um so ein genaues Bild über die Stabilität dieser Partikel, ummantelt mit zwitterionischen Polymeren, zu bekommen und sie so mit NPs mit einfacher Oberflächenladung zu vergleichen (z.B. komplett positive oder negative Oberflächenladung). Eine dritte Anwendung beschreibt die Eigenassemblierung von abwechselnd positiv und negativ geladenen Polyelektrolyt-Schichten auf Kalziumkarbonat Kerne, was zu hohlen Mikrosphären aus Polymer mit physikalisch, biologischen Eigenschaften führt. Diese Eigenschaften werden zum einen über Einbettung von Eisenoxid NPs innerhalb der Polyelektrolyt-Schichten (Magnetisierung), sowie dem Anheften von spezifischen Antikörpern an die äußerste Lage der Sphären (biologische Molekülerkennung) gewehrleistet. Die Kapseln wurden als magnetischer Immunosensor benutzt, der in der Lage ist spezifisch Meerrettichperoxidase (Beispielprotein) in Lösung zu erkennen und zu extrahieren.

Bibliographie / References

  1. Torrano, A.A., et al., 2013. A fast analysis method to quantify nanoparticle uptake on a single cell level. Nanomedicine 8 (11), 1815-1828.
  2. Ghinea, N., Simionescu, N., 1985. Anionized and cationized hemeundecapeptides as probes for cell surface charge and permeability studies. J. Cell Biol. 100, 606-612.
  3. Semmling, M., et al., 2008. A novel flow-cytometry-based assay for cellular uptake studies of polyelectrolyte microcapsules. Small 4 (10), 1763-1768.
  4. Takeuchi, H., Omogo, B., Heyes, C.D., 2013. Are bidentate ligands really better than monodentate ligands for nanoparticles? Nano Lett.
  5. Meng, H., et al., 2011. Aspect ratio determines the quantity of mesoporous silica nanoparticle uptake by a small GTPase-dependent macropinocytosis mechanism. ACS Nano 5 (6), 4434-4447.
  6. Stern, S.T., Adiseshaiah, P.P., Crist, R.M., 2012. Autophagy and lysosomal dysfunction as emerging mechanisms of nanomaterial toxicity. Part. Fibre Toxicol. 9.
  7. Lai, P.Y., et al., 2015. Biomimetic stem cell membrane-camouflaged iron oxide nanoparticles for theranostic applications. RSC Adv. 5 (119), 98222-98230.
  8. Fadeel, B., et al., 2013. Bridge over troubled waters: understanding the synthetic and biological identities of engineered nanomaterials. Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol. 5 (2), 111-129.
  9. Russ, K.A., et al., 2016. C-60 fullerene localization and membrane interactions in RAW 264.7 immortalized mouse macrophages. Nanoscale 8 (7), 4134-4144.
  10. Shi, X., et al., 2011. Cell entry of one-dimensional nanomaterials occurs by tip recognition and rotation. Nat. Nanotechnol. 6, 714-719.
  11. Lin, J.Q., Alexander-Katz, A., 2013. Cell membranes open “doors” for cationic nanoparticles/biomolecules: insights into uptake kinetics. ACS Nano 7 (12), 10799-10808.
  12. Parak, W.J., et al., 2002. Cell motility and metastatic potential studies based on quantum dot imaging of phagokinetic tracks. Adv. Mater. 14 (12), 882-885.
  13. Wang, T., et al., 2012. Cellular uptake of nanoparticles by membrane penetration: a study combining confocal microscopy with FTIR spectroelectrochemistry. ACS Nano 6 (2), 1251-1259.
  14. Muñoz Javier, A., et al., 2006. Combined atomic force microscopy and optical microscopy measurements as a method to investigate particle uptake by cells. Small 2 (3), 394-400.
  15. Boldt, K., et al., 2006. Comparative examination of the stability of semiconductor quantum dots in various biochemical buffers. J. Phys. Chem. B 110 (5), 1959-1963.
  16. Delehanty, J.B., et al., 2010. Delivering quantum dot-peptide bioconjugates to the cellular cytosol: escaping from the endolysosomal system. Integr. Biol. 2 (5-6), 265-277.
  17. Chithrani, B.D., Ghazan, A.A., Chan, C.W., 2006. Determining the size and the shape dependence of gold nanoparticle uptake into mammalian cells. Nano Lett. 6 (4), 662-668.
  18. He, C., et al., 2010. Effects of particle size and surface charge on cellular uptake and biodistribution of polymeric nanoparticles. Biomaterials 31 (13), 3657-3666.
  19. Yoo, J.W., Doshi, N., Mitragotri, S., 2010. Endocytosis and intracellular distribution of plga particles in endothelial cells: effect of particle geometry. Macromol. Rapid Commun. 31 (2), 142-148.
  20. Iversen, T.G., Skotland, T., Sandvig, K., 2011. Endocytosis and intracellular transport of nanoparticles: present knowledge and need for future studies. Nano Today 6 (2), 176-185.
  21. Canton, I., Battaglia, G., 2012. Endocytosis at the nanoscale. Chem. Soc. Rev. 41 (7), 2718-2739.
  22. Sahay, G., Alakhova, D.Y., Kabanov, A.V., 2010. Endocytosis of nanomedicines. J. Control. Release 145 (3), 182-195.
  23. Braun, G.B., et al., 2014. Etchable plasmonic nanoparticle probes to image and quantify cellular internalization. Nat. Mater. 13 (9), 904-911.
  24. Huefner, A., et al., 2014. Gold nanoparticles explore cells: cellular uptake and their use as intracellular probes. Methods 68 (2), 354-363.
  25. Florez, L., et al., 2012. How shape influences uptake: interactions of anisotropic polymer nanoparticles and human mesenchymal stem cells. Small 8 (14), 2222-2230.
  26. Yameen, B., et al., 2014. Insight into nanoparticle cellular uptake and intracellular targeting. J. Control. Release 190, 485-499.
  27. Rivera_Gil, P., et al., 2009. Intracellular processing of proteins mediated by biodegradable polyelectrolyte capsules. Nano Lett. 9 (12), 4398-4402.
  28. Soenen, S.J., et al., 2015. (Intra)cellular stability of inorganic nanoparticles: effects on cytotoxicity, particle functionality, and biomedical applications. Chem. Rev. 115 (5), 2109-2135.
  29. Nazarenus, M., et al., 2014. In vitro interaction of colloidal nanoparticles with mammalian cells: what have we learned thus far? Beilstein J. Nanotechnol. 5, 1477-1490.
  30. Feliu, N., et al., 2016a. In vivo degradation and the fate of inorganic nanoparticles. Chem. Soc. Rev. 45, 2440-2457.
  31. Kreyling, W.G., et al., 2015. In vivo integrity of polymer-coated gold nanoparticles. Nat. Nanotechnol. 10 (7), 619-623.
  32. Mattoussi, H., Palui, G., Na, H.B., 2012. Luminescent quantum dots as platforms for probing in vitro and in vivo biological processes. Adv. Drug Deliv. Rev. 64 (2), 138-166.
  33. Zhang, L.W., Monteiro-Riviere, N.A., 2009. Mechanisms of quantum dot nanoparticle cellular uptake. Toxicol. Sci. 110, 138-155.
  34. Mani, T., et al., 2015. Microplastics profile along the Rhine River. Sci. Rep. 5, 17988.
  35. Kastl, L., et al., 2013. Multiple internalization pathways of polyelectrolyte multilayer capsules into mammalian cells. ACS Nano 7 (8), 6605-6618.
  36. Welsher, K., Yang, H., 2014. Multi-resolution 3D visualization of the early stages of cellular uptake of peptide-coated nanoparticles. Nat. Nanotechnol. 9 (3), 198-203.
  37. Feliu, N., et al., 2016b. Nanoparticle dosage-a nontrivial task of utmost importance for quantitative nanosafety research. Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol.
  38. Dalal, C., Saha, A., Jana, N.R., 2016. Nanoparticle multivalency directed shifting of cellular uptake mechanism. J. Phys. Chem. C 120 (12), 6778-6786.
  39. Lundqvist, M., et al., 2008. Nanoparticle size and surface properties determine the protein corona with possible implications for biological impacts. Proc. Natl. Acad. Sci. U. S. A. 105 (38), 14265-14270.
  40. Krug, H.F., 2014. Nanosafety research-are we on the right track? Angew. Chem. Int. Ed. 53 (46), 12304-12319.
  41. Oberdörster, G., Oberdörster, E., Oberdörster, J., 2005. Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ. Health Perspect. 113 (7), 823-839.
  42. Sharma, V.K., et al., 2015. Natural inorganic nanoparticles-formation, fate, and toxicity in the environment. Chem. Soc. Rev. 44 (23), 8410-8423.
  43. Bedard, M.F., et al., 2009. On the mechanical stability of polymeric microcontainers functionalized with nanoparticles. Soft Matter 5 (1), 148-155.
  44. Mayor, S., Pagano, R.E., 2007. Pathways of clathrin-independent endocytosis. Nat. Rev. Mol. Cell Biol. 8, 603-612.
  45. Goy-Lopez, S., et al., 2012. Physicochemical characteristics of protein-NP bioconjugates: the role of particle curvature and solution conditions on human serum albumin conformation and fibrillogenesis inhibition. Langmuir 28 (24), 9113-9126.
  46. Chanana, M., et al., 2013. Physicochemical properties of protein-coated gold nanoparticles in biological fluids and cells before and after proteolytic digestion. Angew. Chem. Int. Ed. 52 (15), 4179-4183.
  47. Hühn, D., et al., 2013. Polymer-coated nanoparticles interacting with proteins and cells: focusing on the sign of the net charge. ACS Nano 7 (4), 3253-3263.
  48. Lerch, S., et al., 2013. Polymeric nanoparticles of different sizes overcome the cell membrane barrier. Eur. J. Pharm. Biopharm. 84 (2), 265-274.
  49. Kreft, O., et al., 2007. Polymer microcapsules as mobile local pH-sensors. J. Mater. Chem. 17, 4471-4476.
  50. Forte, M., et al., 2016. Polystyrene nanoparticles internalization in human gastric adenocarcinoma cells. Toxicol. in Vitro 31, 126-136.
  51. Weidner, A., et al., 2015. Preparation of core-shell hybrid materials by producing a protein corona around magnetic nanoparticles. Nanoscale Res. Lett. 10, 1-11.
  52. Casals, E., et al., 2014. Programmed iron oxide nanoparticles disintegration in anaerobic digesters boosts biogas production. Small 10 (14), 2801-2808.
  53. Cheng, X.J., et al., 2015. Protein corona influences cellular uptake of gold nanoparticles by phagocytic and nonphagocytic cells in a size-dependent manner. ACS Appl. Mater. Interfaces 7 (37), 20568-20575.
  54. Schweiger, C., et al., 2012. Quantification of the internalization patterns of superparamagnetic iron oxide nanoparticles with opposite charge. J. Nanobiotechnol. 10 (1), 28.
  55. Medintz, I.L., et al., 2005. Quantum dot bioconjugates for imaging, labelling and sensing. Nat. Mater. 4 (6), 435-446.
  56. Taylor, U., et al., 2014. Rational design of gold nanoparticle toxicology assays: a question of exposure scenario, dose and experimental setup. Nanomedicine 9 (13), 1971-1989.
  57. Bastus, N.G., et al., 2008. Reactivity of engineered inorganic nanoparticles and carbon nanostructures in biological media. Nanotoxicology 2 (3), 99-112.
  58. Conner, S.D., Schmid, S.L., 2003. Regulated portals of entry into the cell. Nature 422 (6927), 37-44.
  59. Shimoni, O., et al., 2013. Shape-dependent cellular processing of polyelectrolyte capsules. ACS Nano 7 (1), 522-530.
  60. Gliga, A.R., et al., 2014. Size-dependent cytotoxicity of silver nanoparticles in human lung cells: the role of cellular uptake, agglomeration and Ag release. Part. Fibre Toxicol. 11.
  61. Jiang, X., et al., 2010. Specific effects of surface amines on polystyrene nanoparticles in their interactions with mesenchymal stem cells. Biomacromolecules 11 (3), 748-753.
  62. Hartmann, R., et al., 2015. Stiffness-dependent in vitro uptake and lysosomal acidification of colloidal particles. Angew. Chem. Int. Ed. 54 (4), 1365-1368.
  63. Feuser, P.E., et al., 2016. Superparamagnetic poly(methyl methacrylate) nanoparticles surface modified with folic acid presenting cell uptake mediated by endocytosis. J. Nanopart. Res. 18, 104.
  64. Mutsaers, S., Papadimitriou, J., 1988. Surface charge of macrophages and their interaction with charged particles. J. Leukoc. Biol. 44 (1), 17-20.
  65. Cesbron, Y., et al., 2015. TAT and HA2 facilitate cellular uptake of gold nanoparticles but do not lead to cytosolic localisation. PLoS One 10 (4).
  66. Mahmoudi, M., et al., 2013. Temperature: the “ignored” factor at the nanobio interface. ACS Nano 7 (8), 6555-6562.
  67. Westmeier, D., et al., 2016a. The bio-corona and its impact on nanomaterial toxicity. Eur. J. Nanomedicine 7, 153-168.
  68. Wang, F.J., et al., 2013. The biomolecular corona is retained during nanoparticle uptake and protects the cells from the damage induced by cationic nanoparticles until degraded in the lysosomes. Nanomed. Nanotechnol. Biol. Med. 9 (8), 1159-1168.
  69. Rivera Gil, P., et al., 2013. The challenge to relate the physicochemical properties of colloidal nanoparticles to their cytotoxicity. Acc. Chem. Res. 46 (3), 743-749.
  70. Westmeier, D., Stauber, R.H., Docter, D., 2016b. The concept of bio-corona in modulating the toxicity of engineered nanomaterials (ENM). Toxicol. Appl. Pharmacol. 299, 53-57.
  71. Bargheer, D., et al., 2015a. The distribution and degradation of radiolabelled SPIOs and quantum dots in mice. Beilstein J. Nanotechnol. 6, 111-123.
  72. Bargheer, D., et al., 2015b. The fate of a designed protein corona on nanoparticles in vitro and in vivo. Beilstein J. Nanotechnol. 6, 36-46.
  73. Parakhonskiy, B., et al., 2015. The influence of the size and aspect ratio of anisotropic, porous CaCO3 particles on their uptake by cells. J. Nanobiotechnol. 13 (1), 53.
  74. Kolosnjaj-Tabi, J., et al., 2015. The one year fate of iron oxide coated gold nanoparticles in mice. ACS Nano 9 (8), 7925-7939.
  75. Sun, H.L., et al., 2015. The role of capsule stiffness on cellular processing. Chem. Sci. 6 (6), 3505-3514.
  76. Casals, E., et al., 2010. Time evolution of the nanoparticle protein corona. ACS Nano 4 (7), 3623-3632.
  77. Kim, B., et al., 2010. Tuning payload delivery in tumour cylindroids using gold nanoparticles. Nat. Nanotechnol. 5 (6), 465-472.
  78. Beddoes, C.M., Case, C.P., Briscoe, W.H., 2015. Understanding nanoparticle cellular entry: a physicochemical perspective. Adv. Colloid Interf. Sci. 218, 48-68.
  79. Cedervall, T., et al., 2007. Understanding the nanoparticle-protein corona using methods to quantify exchange rates and affinities of proteins for nanoparticles. Proc. Natl. Acad. Sci. U. S. A. 104 (7), 2050-2055.
  80. Muñoz_Javier, A., et al., 2008. Uptake of colloidal polyelectrolyte coated particles and polyelectrolyte multilayer capsules by living cells. Adv. Mater. 20 (22), 4281-4287.


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