WO2015192832A1 - Two-photon polymerization initiators for use in organic solvents - Google Patents

Two-photon polymerization initiators for use in organic solvents Download PDF

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WO2015192832A1
WO2015192832A1 PCT/DE2015/100239 DE2015100239W WO2015192832A1 WO 2015192832 A1 WO2015192832 A1 WO 2015192832A1 DE 2015100239 W DE2015100239 W DE 2015100239W WO 2015192832 A1 WO2015192832 A1 WO 2015192832A1
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photoinitiator
photon polymerization
photon
initiators
polymerization initiators
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German (de)
French (fr)
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Michael Gottschaldt
Cornelia Bader
Leander Poocza
Felix Siedenbiedel
Gerhard Hildebrand
Klaus Liefeith
Ullrich S. SCHUBERT
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Friedrich-Schiller-Universität Jena
Institut für Bioprozess- und Analysenmesstechnik e.V.
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C219/00—Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton
    • C07C219/02—Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton having esterified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C219/04—Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton having esterified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C219/06—Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton having esterified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having the hydroxy groups esterified by carboxylic acids having the esterifying carboxyl groups bound to hydrogen atoms or to acyclic carbon atoms of an acyclic saturated carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F136/00—Homopolymers of compounds having one or more unsaturated aliphatic radicals, at least one having two or more carbon-to-carbon double bonds
    • C08F136/02—Homopolymers of compounds having one or more unsaturated aliphatic radicals, at least one having two or more carbon-to-carbon double bonds the radical having only two carbon-to-carbon double bonds
    • C08F136/20—Homopolymers of compounds having one or more unsaturated aliphatic radicals, at least one having two or more carbon-to-carbon double bonds the radical having only two carbon-to-carbon double bonds unconjugated
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F2/00—Processes of polymerisation
    • C08F2/46—Polymerisation initiated by wave energy or particle radiation
    • C08F2/48—Polymerisation initiated by wave energy or particle radiation by ultraviolet or visible light
    • C08F2/50—Polymerisation initiated by wave energy or particle radiation by ultraviolet or visible light with sensitising agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/06—Systems containing only non-condensed rings with a five-membered ring
    • C07C2601/08—Systems containing only non-condensed rings with a five-membered ring the ring being saturated

Definitions

  • the invention relates to new initiators for two-photon polymerization, which are suitable for use in organic solvents.
  • the two-photon polymerization (2PP) of various monomer and macro- monomer mixtures is increasingly used in the realization of high data density optical data storage devices or in the fabrication of three-dimensional objects (KS Lee, DY Yang, SH Park, RH Kim: "Recent developments in the Advance Technol. 2006, 7, 72-82).
  • initiators which can be excited by means of two-photon absorption, leads to the applicability of larger wavelengths (about 800 nm) and thus to a greater penetration depth of the light into the medium.
  • the polymerization can be controlled so that it takes place only at one point (a defined volume in a three-dimensional space) (BH Cumpston, SP Ananthavel, S. Barlow, DL Dyer, JE Ehrlich, LL Erskine, AA Heikai, SM Kuebler, IYS Lee, D. McCord-Maughon, JQ Qin, H. Rockel, M. Rumi, XL Wu, SR Marder, JW Perry : "Two-photon polymerization initiators for three-dimensional optical data storage and microfabrication", Nature 1999, 398, 51-54).
  • UV initiators were initially used (eg Irgacure 369 TM), but these have low photosensitivity and low initiation rates in combination with NIR ultrashort pulse lasers (KJ Schafer, JM Haies, M. Balu, KD Belfield, EW Van Stryland, DJ Hagan: “Two-photon absorption cross-sections of common photoinitiators", Photochem Photobiol A Chem., 2004, 162, 497-502).
  • Organoluble known photoinitiators based on 2,5-dibenzylidenecyclopentanone are easy to prepare and have high absorption cross sections and good solubilities, but are not suitable for this use due to their lacking initiation properties.
  • Z. Li N. Pucher, K. Cicha, J. Torgersen, SC Ligon, A. Ajami, W. Husinsky, A. Rosspeintner, E. Vauthey, S. Naumov, T. Scherzer, J. Stampfl, R. Liska: "A straightforward synthesis and structure-activity relationship of highly efficient initiators for two-photon polymerization", Macromolecules 2013, 46, 352-361):
  • two-photon polymerization initiators which consist of a basic structure of the general formula IX are proposed for achieving this object:
  • R 1, R 2, R 3, R 4 independently of one another are alkyl or aryl radicals.
  • 2PP initiators have the advantage of high 2-photon absorption cross sections, as they have per se known 2,5-Dibenzylidencyclopentanon initiators, however simultaneously combined with organo-solubilizing radicals and polar groups in the side chains.
  • the reactivity and solubility can be controlled application-specific via the incorporation of different radicals and polar coupling groups.
  • the new 2PP initiators even if only added in low concentrations of ⁇ 0.1%, give surprisingly high resolutions for the two-photon polymerization in organic solvents (minimally writable voxels with a diameter of 250 nm and writing speeds> 5 mm / s).
  • organosoluble 2PP initiators according to the invention is possible by well-established and well-known in the art synthesis methods, as they are known for the preparation of other 2PP initiators.
  • the invention will be explained in more detail below with reference to the said photoinitiator in formula X.
  • the purification is carried out by column chromatography in tert-butyl methyl ether over silica gel. Due to the poor solubility of the 2PP initiator in tert-butyl methyl ether this is dissolved in a little chloroform, added to the column and then eluted with tert-butyl methyl ether (R f - value 0.21). It may be necessary to carry out further column chromatography in ethyl acetate (R f 0.67). After concentration, a dark red oil is obtained in 35% yield, which dissolves very well in acetone and chloroform.
  • Laser wavelength 780 to 800 nm
  • laser power 100 to 350 mW
  • writing speed 0.1 to 5 mm / s
  • lens 63x (NA: 0.75) or 100x (NA: 1.4 oil immersion)
  • SD structures Hatch: up to 13 ⁇ .
  • the structure development was carried out by washing or rinsing in acetone.
  • the UDMA (urethane dimethacrylate) could be written / structured with the known Irgacure 369 only from a minimum concentration of 2 wt.% With the aforementioned laser parameters. However, writing defects occurred regularly, especially at the edges of the structure. At a concentration of 0.1% by weight, it was not possible to detect any structure formation with Irgacure 369 in UDMA.
  • the aim of these investigations was to carry out general suitability tests of the photoinitiator according to the invention for producing complex 3D structures by means of two-photon polymerization.
  • Geometrically complex 3D structures were created using the 2PP.
  • the templates for the 3D structures to be created were various STL files (Surface Tessellation Language for describing 3D surfaces), which were programmed into the corresponding control software of the 2PP system and converted by the latter into 3D polymer structures.
  • the precursor used was a copolymer consisting of polycaprolactone and polylactide in which 0.1% by weight of the photoinitiator according to the invention was admixed.
  • the 3D model was each represented by a mathematical minimal surface (gyroid), which was used as the unit cell for the 2PP production of a 3D array.
  • the height of the 3D array here was about 600 ⁇ .
  • the structure was implemented according to the STL file in detail and to scale.
  • the determination of a possible cytotoxic effect of the photoinitiator was carried out by XTT assay based on the guidelines of ISO / EN 10993 part 5 and using the following cell types: L929 - mouse fibroblasts, HepG2 - human hepatocytes, HEK - human embryonic kidney cells and bovChz - bovine chondrocytes ).
  • the cells were seeded in 96-well microtiter plates at a density of 1 x 10 4 cells / well and cultured for 24 h to form a semiconfluent monolayer cell lawn.
  • the organosoluble photoinitiator was first dissolved in DMSO (10% by weight or 100 mg / ml).
  • DMSO content in the final photoinitiator test solutions 10 and 1%, respectively.
  • the DMSO control solutions used were appropriately prepared cell culture medium with a DMSO content of 10 and 1%, respectively.
  • Per well of 96-well plate were by aspirating the medium from the 24-h pre-cultured cells, respectively 100 ⁇ ⁇ the photoinitiator - where test solutions or the DMSO control solutions to the cells and for 24 h at 37 ° C and 5% C0 2 , After incubation, the cells were washed once with PBS and added with 150 ⁇ ⁇ XTT-PMS medium solution per well to be tested and incubated for an additional 3 h to 5 h at 37 ° C and 5% C0 2 .
  • the OD values of the control solutions correspond to a viability of 100% and 0% reduction / inhibition of metabolic cell activity respectively, with a decrease in viability values to less than 70% of the control to a significant cytotoxic potential the test compound indicates.
  • the vitality and intact morphology of the cells after contact with photoinitiator Living-dead staining and fluorescence microscopy evaluation confirmed.
  • the cells were seeded analogously to the XTT test in 96-well microtiter plates and treated with different photoinitiator concentrations or b.) Prepared photoinitiator films and then populated with cells.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Polymerisation Methods In General (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

The aim of the invention was to produce non-toxic two-photon polymerization initiators which are effective in organic solvents, are suitable for applications with NIR ultrashort pulse lasers, have a high absorbing effect, and are able to quickly form radicals for radially initiated polymerization or cross-linking reactions. Said aim was achieved by two-photon polymerization initiators consisting of a basic structure of general formula I.

Description

Beschreibung der Erfindung  Description of the invention
Zwei-Photonenpolymerisations-Initiatoren für die Anwendung in organischen Lösungsmitteln Two-photon polymerization initiators for use in organic solvents
Die Erfindung betrifft neue Initiatoren für Zwei-Photonenpolymerisation, welche für Anwendung in organischen Lösungsmitteln geeignet sind. The invention relates to new initiators for two-photon polymerization, which are suitable for use in organic solvents.
Die Zwei-Photonenpolymerisation (2PP) unterschiedlichster Monomer- und Makro- monomergemische findet zunehmend Anwendung bei der Realisierung von optischen Datenspeichern mit hoher Informationsdichte oder bei der Herstellung dreidimensionaler Objekte (K. S. Lee, D. Y. Yang, S. H. Park, R. H. Kim: "Recent developments in the use of two-photon polymerization in precise 2D and 3D microfabrications", Polym. Adv. Technol. 2006, 7, 72-82).  The two-photon polymerization (2PP) of various monomer and macro- monomer mixtures is increasingly used in the realization of high data density optical data storage devices or in the fabrication of three-dimensional objects (KS Lee, DY Yang, SH Park, RH Kim: "Recent developments in the Advance Technol. 2006, 7, 72-82).
Anwendungsmöglichkeiten sind u.a. die Herstellung dreidimensional mikro- und nanostrukturierter polymerer Formkörper auf der Grundlage von Computermodellen (Nano-Rapid Prototyping) sowie die Herstellung dreidimensional mikro- und nanostrukturierter polymerer funktionaler Beschichtungen (S. C. Singh, H. Zeng, "Nanomaterials and nanopatterns based on laser processing: A brief review on current State of art", Sei. Adv. Mater. 2012, 4, 368-390). Applications are u.a. the production of three-dimensional microstructured and nanostructured polymeric moldings based on computer models (nano-rapid prototyping) and the production of three-dimensional microstructured and nanostructured polymeric functional coatings (SC Singh, H. Zeng, "Nanomaterials and nanopatterns based on laser processing: A letter Review of Current State of Art, p. Adv. Mater. 2012, 4, 368-390).
Es ist daher allgemein bekannt, dass bei der 2-Photonen-Polymerisation die Polymerisation von Mono-/Makromerengemischen im Fokus eines Laserstrahls ausgelöst werden kann (M. T. Raimondi, S. M. Eaton, M. M. Nava, M. Lagana, G. Cerullo, R. Osellame: "Two- photon laser polymerization: From fundamentals to biomedical application in tissue engineering and regenerative medicine", J. Appl. Biomater. Funt. Mater. 2012, 10, 55-65). Grundvoraussetzung ist dabei die Anwesenheit eines geeigneten Photoinitiators. It is therefore generally known that in the 2-photon polymerization the polymerization of mono- / macromer mixtures in the focus of a laser beam can be triggered (MT Raimondi, SM Eaton, MM Nava, M. Lagana, G. Cerullo, R. Osellame: "Two-photon laser polymerization: From fundamental to biomedical application in tissue engineering and regenerative medicine", J. Appl. Biomater., Funt. Mater., 2012, 10, 55-65). The basic requirement is the presence of a suitable photoinitiator.
Die Verwendung von Initiatoren, welche mittels Zwei-Photonenabsorption angeregt werden können, führt zur Anwendbarkeit größerer Wellenlängen (ca. 800 nm) und damit zu einer größeren Eindringtiefe des Lichtes in das Medium. Außerdem kann die Polymerisation dadurch so gesteuert werden, dass sie nur an einer Stelle (einem definierten Volumen in einem dreidimensionalen Raum) stattfindet (B. H. Cumpston, S. P. Ananthavel, S. Barlow, D. L. Dyer, J. E. Ehrlich, L. L. Erskine, A. A. Heikai, S. M. Kuebler, I. Y. S. Lee, D. McCord-Maughon, J. Q. Qin, H. Rockel, M. Rumi, X. L. Wu, S. R. Marder, J. W. Perry: "Two-photon polymerization Initiators for three- dimensional optical data storage and microfabrication", Nature 1999, 398, 51-54). The use of initiators, which can be excited by means of two-photon absorption, leads to the applicability of larger wavelengths (about 800 nm) and thus to a greater penetration depth of the light into the medium. In addition, the polymerization can be controlled so that it takes place only at one point (a defined volume in a three-dimensional space) (BH Cumpston, SP Ananthavel, S. Barlow, DL Dyer, JE Ehrlich, LL Erskine, AA Heikai, SM Kuebler, IYS Lee, D. McCord-Maughon, JQ Qin, H. Rockel, M. Rumi, XL Wu, SR Marder, JW Perry : "Two-photon polymerization initiators for three-dimensional optical data storage and microfabrication", Nature 1999, 398, 51-54).
Weitere entscheidende Parameter sind die zu realisierende Schreibgeschwindigkeit sowie die Effizienz der Polymerisation und die dafür benötigte Laserintensität. Alle diese Parameter hängen unmittelbar von der Art des eingesetzten Photoinitiators, dessen photophysikalischen Eigenschaften, seiner Löslichkeit, der Wechselwirkungen des Initiators mit den Komponenten des Mediums sowie seiner Stabilität ab. Further decisive parameters are the writing speed to be realized as well as the efficiency of the polymerization and the laser intensity required for this. All these parameters depend directly on the type of photoinitiator used, its photophysical properties, its solubility, the interactions of the initiator with the components of the medium and its stability.
Dabei fanden zunächst kommerzielle UV-Initiatoren Anwendung (z. B. Irgacure 369™), diese weisen jedoch eine geringe Photosensitivität sowie zu geringe Initiationsgeschwindigkeiten in Verbindung mit NIR-Ultrakurzpulslasern auf (K. J. Schafer, J. M. Haies, M. Balu, K. D. Belfield, E. W. Van Stryland, D. J. Hagan: "Two-photon absorption cross-sections of common photoinitiators", . Photochem. Photobiol. A Chem. 2004, 162, 497-502). Commercial UV initiators were initially used (eg Irgacure 369 ™), but these have low photosensitivity and low initiation rates in combination with NIR ultrashort pulse lasers (KJ Schafer, JM Haies, M. Balu, KD Belfield, EW Van Stryland, DJ Hagan: "Two-photon absorption cross-sections of common photoinitiators", Photochem Photobiol A Chem., 2004, 162, 497-502).
Figure imgf000003_0001
Figure imgf000003_0001
Als besser geeignete Initiatoren für die Zwei-Photonenpolymerisation können verschiedene andere Farbstoffe mit unterschiedlichen chromophoren Gruppen eingesetzt werden. Beispiele hierfür sind: aromatische Ketone: As more suitable initiators for two-photon polymerization, various other dyes having different chromophore groups can be used. Examples include: aromatic ketones:
Figure imgf000003_0002
(wie Z. Li, M. Siklos, N. Pucher, K. Cicha, A. Ajami, W. Husinsky, A. Rosspeintner, E. Vauthey, G. Gescheidt, J. Stampfl, R. Liska: "Synthesis and structure-activity relationship of several aromatic ketone-based two-photon Initiators", J. Polym. Sei., PartA: Polym. Chem. 2011, 49, 3688-3699,1-2),
Figure imgf000003_0002
(See, for example, Z. Li, M. Siklos, N. Pucher, K. Cicha, A. Ajami, W. Husinsky, A. Rosspeintner, E. Vauthey, G. Gescheidt, J. Stampfl, R. Liska: "Synthesis and structure -activity relationship of several aromatic ketone-based two-photon initiators ", J. Polym. Sci., PartA: Polym. Chem. 2011, 49, 3688-3699, 1-2),
Fluorene: fluorenes:
Figure imgf000004_0001
Figure imgf000004_0001
(wie A. R. Morales, K. D. Belfield, J. M. Haies, E. W. Van Stryland, D. J. Hagan: "Synthesis of two-photon absorbing unsymmetrical fluorenyl-based chromophores", Chem. Mater. 2006, 18, 4972-4980; M. Jin, J.-P. Malval, D.-L. Versace, F. Morlet-Savary, H. Chaumeil, A. Defoin, X. Allonas, J.-P. Fouassier: "Two-photon absorption and polymerization ability of intramolecular energy transfer based photoinitiating Systems", Chem. Commun. 2008, 6540-6542.) oder E-stilbene:  (as AR Morales, KD Belfield, JM Haies, EW Van Stryland, DJ Hagan: "Synthesis of two-photonically absorbing unsymmetrical fluorenyl-based chromophores", Chem. Mater., 2006, 18, 4972-4980, M. Jin, J. Malval, D.L. Versace, F. Morlet-Savary, H. Chaumeil, A. Defoin, X. Allonas, J.-P. Fouassier: "Two-photon absorption and polymerization ability of intramolecular energy transfer based photoinitiating Systems ", Chem. Commun. 2008, 6540-6542.) or E-style level:
Figure imgf000004_0002
Figure imgf000004_0002
(wie B. H. Cumpston, S. P. Ananthavel, S. Barlow, D. L. Dyer, J. E. Ehrlich, L. L. Erskine, A. A. Heikai, S. M.Kuebler, I. Y. S. Lee, D. McCord-Maughon, J. Q. Qin, H. Rockel, M. Rumi, X. L. Wu, S. R. Marder, J. W. Perry: "Two-photon polymerization Initiators for three-dimensional optical data storage and microfabrication", Nature 1999, 398, 51-54). (see BH Cumpston, SP Ananthavel, S. Barlow, DL Dyer, JE Ehrlich, LL Erskine, AA Heikai, SM Kuebel, IYS Lee, D. McCord-Maughon, JQ Qin, H. Rockel, M. Rumi, XL Wu, SR Marder, JW Perry: "Two-photon polymerization initiators for three-dimensional optical data storage and microfabrication", Nature 1999, 398, 51-54).
Bekannt ist auch, 2,5-Dibenzylidencyclo-alkanon-basierte Farbstoffe als Initiatoren für die Zwei-Photonenpolymerisation einzusetzen (US 5,112,721 A): It is also known to use 2,5-dibenzylidenecyclo-alkanone-based dyes as initiators for two-photon polymerization (US 5,112,721 A):
Figure imgf000004_0003
Sowohl die Ringgröße und Substitution des zentralen Alkanons als auch die Substituenten in der Peripherie der chemischen Struktur zeigen einen Einfluss auf die Wirksamkeit des 2PP-Photoinitiators (Z. Li, N. Pucher, K. Cicha, J. Torgersen, S. C. Ligon, A. Ajami, W. Husinsky, A. Rosspeintner, E. Vauthey, S. Naumov, T. Scherzer, J. Stampfl, R. Liska: "A straightforward synthesis and structure-activity relationship of highly efficient Initiators for two-photon polymerization", Macromolecules 2013, 46, 352-361; Q. Zou, Y. Zhao, N. S. Makarov, J. Campo, H. Yuan, D.-C. Fang, J. W. Perry, F. Wu: "Effect of alicyclic ring size on the photophysical and photochemical properties of bis(arylidene)cycloalkanone Compounds", Phys. Chem. Chem. Phys. 2012, 14, 11743- 11752).
Figure imgf000004_0003
Both the ring size and substitution of the central alkanone and the substituents in the periphery of the chemical structure have an influence on the effectiveness of the 2PP photoinitiator (Z. Li, N. Pucher, K. Cicha, J. Torgersen, SC Ligon, A. Ajami, W. Husinsky, A. Rosspeintner, E. Vauthey, S. Naumov, T. Scherzer, J. Stampfl, R. Liska: "A straightforward synthesis and structure-activity relationship of highly efficient initiators for two-photon polymerization", Macromolecules 2013, 46, 352-361; Q. Zou, Y. Zhao, NS Makarov, J. Campo, H. Yuan, D.-C. Fang, JW Perry, F. Wu: "Effect of alicyclic ring size on the photophysical and photochemical properties of bis (arylidene) cycloalkanone compounds ", Phys. Chem. Chem. Phys. 2012, 14, 11743-11752).
Organolösliche bekannte Photoinitiatoren auf der Basis von 2,5-Dibenzyliden- cyclopentanon (u. a. die kommerziell erhältlichen Dimethylamino- und Diethylamino- Derivate) sind einfach herzustellen und besitzen hohe Absorptionsquerschnitte sowie gute Löslichkeiten, sind aber aufgrund ihrer mangelnden Initiationseigenschaften nicht für diesen Einsatz geeignet. (Z. Li, N. Pucher, K. Cicha, J. Torgersen, S. C. Ligon, A. Ajami, W. Husinsky, A. Rosspeintner, E. Vauthey, S. Naumov, T. Scherzer, J. Stampfl, R. Liska: " A straightforward synthesis and structure-activity relationship of highly efficient Initiators for two-photon polymerization", Macromolecules 2013, 46, 352-361): Organoluble known photoinitiators based on 2,5-dibenzylidenecyclopentanone (inter alia the commercially available dimethylamino and diethylamino derivatives) are easy to prepare and have high absorption cross sections and good solubilities, but are not suitable for this use due to their lacking initiation properties. (Z. Li, N. Pucher, K. Cicha, J. Torgersen, SC Ligon, A. Ajami, W. Husinsky, A. Rosspeintner, E. Vauthey, S. Naumov, T. Scherzer, J. Stampfl, R. Liska: "A straightforward synthesis and structure-activity relationship of highly efficient initiators for two-photon polymerization", Macromolecules 2013, 46, 352-361):
Figure imgf000005_0001
Figure imgf000005_0001
Speziell für diese 2,5-Dibenzylidencyclopentanon-Initiatoren wurde daher der Einfluss der Substituenten an den peripheren Phenylringen untersucht, wobei auch unsymmetrische Molekülstrukturen beschrieben wurden (C. A. Zoto, R. E. Connors: "Photophysical properties of an asymmetrical 2,5-diarylidene-cyclopentanone dye possessing electron donor and acceptor substituents", J. Mol. Struct. 2010, 982, 121-126; J. Xue, Y. Zhao, H. Wu, F. Wu: "Novel benzylidene cyclopentanone dyes for two-photon photopolymeriza- tion", J. Photochem. Photobiol. A Chem. 2008, 195, 261-266): The influence of the substituents on the peripheral phenyl rings was therefore investigated specifically for these 2,5-dibenzylidenecyclopentanone initiators, with asymmetric molecular structures also being described (CA Zoto, RE Connors: "Photophysical properties of an asymmetrical 2,5-diarylidenes-cyclopentanone dye possessing electron donor and acceptor substituents, J. Mol. Struct. 2010, 982, 121-126, J. Xue, Y. Zhao, H. Wu, F.Wu: "Novel benzylidene cyclopentanone dyes for two-photon photopolymerization", J. Photochem. Photobiol. A Chem. 2008, 195, 261-266):
Figure imgf000006_0001
Figure imgf000006_0001
Auch diese Derivate erscheinen wegen ihrer schlechten Löslichkeit, ihrer aufwendigen Herstellung oder geringen Initiationseigenschaften als nicht geeignet. These derivatives also do not appear to be suitable because of their poor solubility, their elaborate preparation or their low initiation properties.
Eine Steigerung der Schreibgeschwindigkeit unter gleichzeitiger Verringerung der Initiatorkonzentration wird bei der Verwendung eines PIs mit zentraler 4-Methyl- cyclohexanone-Struktureinheit beschrieben (Z. Li, N. Pucher, K. Cicha, J. Torgersen, S. C. Ligon, A. Ajami, W. Husinsky, A. Rosspeintner, E. Vauthey, S. Naumov, T. Scherzer, J. Stampfl, R. Liska: "A straightforward synthesis and structure-activity relationship of highly efficient Initiators for two-photon polymerization", Macromolecules 2013, 46, 352-361). Allerdings wurde auch hier eine PI- Konzentration von > 2% verwendet. An increase in writing speed with concomitant reduction in initiator concentration is described using a PI having a central 4-methylcyclohexanone moiety (Z. Li, N. Pucher, K. Cicha, J. Torgersen, SC Ligon, A. Ajami, W Husinsky, A. Rosspeintner, E. Vauthey, S. Naumov, T. Scherzer, J. Stampfl, R. Liska: "A straightforward synthesis and structure-activity relationship of highly efficient initiators for two-photon polymerization", Macromolecules 2013, 46, 352-361). However, a PI concentration of> 2% was also used here.
Zusammenfassend lässt sich feststellen, dass der Fachwelt zwar hinreichend Photoinitiatoren für die Zwei-Photonenpolymerisation mit entsprechender Effizienz bekannt sind, nicht aber Photoinitiatoren, welche für die Reaktion Organolöslichkeit erfordern und dennoch hohe Empfindlichkeit und Schreibgeschwindigkeit aufweisen sowie nicht toxisch sind. In summary it can be stated that although the art is well known photoinitiators for the two-photon polymerization with corresponding efficiency, but not photoinitiators, which require the reaction for organosolubility and yet high sensitivity and writing speed and are not toxic.
Der Erfindung liegt daher die Aufgabe zugrunde, in organischen Lösungsmitteln wirksame, nicht toxische und für Anwendungen mit NIR-Ultrakurzpulslasern geeignete Zwei-Photonenpolymerisations-Initiatoren mit hohem Absorptionseffekt und schnellem Radikalbildungsvermögen für radikalisch ausgelöste Polymerisations- bzw. Vernetzungsreaktionen zu schaffen. It is therefore an object of the present invention to provide two-photon polymerization initiators which are effective in organic solvents and which are suitable for applications with NIR ultrashort pulse lasers and have a high absorption effect and a fast radical-forming capacity for free-radical polymerization or crosslinking reactions.
Erfindungsgemäß werden zur Lösung dieser Aufgabe Zwei-Photonenpolymerisations- Initiatoren vorgeschlagen, die aus einer Grundstruktur der allgemeinen Formel IX bestehen: According to the invention, two-photon polymerization initiators which consist of a basic structure of the general formula IX are proposed for achieving this object:
Figure imgf000007_0001
mit
Figure imgf000007_0001
With
W, X, Y. Z: unabhängig voneinander NH, O  W, X, Y. Z: independently of one another NH, O
R 1 , R2 , R 3 , R 4 : unabhängig voneinander Alkyl- oder Arylrest.  R 1, R 2, R 3, R 4: independently of one another are alkyl or aryl radicals.
Als vorteilhaft hat sich dabei ein Photoinitiator gemäß nachstehender Formel X gezeigt: A photoinitiator according to the following formula X has proved to be advantageous:
Figure imgf000007_0002
Figure imgf000007_0002
Diese 2PP-Initiatoren besitzen den Vorteil hoher 2-Photonen-Absorptionsquerschnitte, wie sie an sich bekannte 2,5-Dibenzylidencyclopentanon-Initiatoren aufweisen, jedoch gleichzeitig vereint mit organolöslichkeitsvermittelnden Resten sowie polaren Gruppen in den Seitenketten. Die Reaktivität und Löslichkeit können über den Einbau unterschiedlicher Reste und polarer Kopplungsgruppen anwendungsspezifisch gesteuert werden. In Verbindung mit NIR-Ultrakurzpulslasern liefern die neuen 2PP-Initiatoren, selbst wenn auch nur in geringer Konzentration < 0.1 % zugesetzt, für die Zwei- Photonenpolymerisation in organischen Lösungsmitteln überraschend hohe Auflösungen (minimal schreibbarer Voxel mit einem Durchmesser von 250 nm und Schreibgeschwindigkeiten > 5 mm/s). These 2PP initiators have the advantage of high 2-photon absorption cross sections, as they have per se known 2,5-Dibenzylidencyclopentanon initiators, however simultaneously combined with organo-solubilizing radicals and polar groups in the side chains. The reactivity and solubility can be controlled application-specific via the incorporation of different radicals and polar coupling groups. In combination with NIR ultrashort pulse lasers, the new 2PP initiators, even if only added in low concentrations of <0.1%, give surprisingly high resolutions for the two-photon polymerization in organic solvents (minimally writable voxels with a diameter of 250 nm and writing speeds> 5 mm / s).
Zelltests belegen zudem, dass die neuen PIs im relevanten Konzentrationsbereich keine zytotoxischen Eigenschaften aufweisen.  Cell tests also show that the new PIs have no cytotoxic properties in the relevant concentration range.
Die Herstellung der erfindungsgemäßen organolöslichen 2PP-Initiatoren ist durch etablierte und der Fachwelt hinreichend bekannte Syntheseverfahren, wie sie für die Herstellung anderer 2PP-Initiatoren bekannt sind, möglich. Die Erfindung soll nachstehend anhand des besagten Photoinitiators in Formel X näher erläutert werden.  The preparation of the organosoluble 2PP initiators according to the invention is possible by well-established and well-known in the art synthesis methods, as they are known for the preparation of other 2PP initiators. The invention will be explained in more detail below with reference to the said photoinitiator in formula X.
Zur Synthese dieses Photoinitiators werden in einem 100 mL Zweihalskolben 4-[Bis[2-(- acetyloxy)ethyl]amino]benzaldehyd (4 g, 13.6 mmol), Cyclopentanon (576 mg, 6.8 mmol) und Cyanurylchlorid (252 mg, 1.37 mmol) vorgelegt. Man gibt einen Tropfen Wasser zur Mischung und erwärmt diese unter Rühren auf 75 bis 80 °C. Nach wenigen Minuten erhält man eine dunkelgrüne homogene Masse. Man lässt die Mischung noch ca. eine Stunde rühren. Nach dem Abkühlen wird das Rohprodukt in Chloroform gelöst, mit Wasser ausgeschüttelt und über Natriumsulfat getrocknet. Die Reinigung erfolgt durch Säulenchromatographie in tert.-Butylmethylether über Kieselgel. Auf Grund der schlechten Löslichkeit des 2PP-Initiators in tert.-Butylmethylether wird dieser in wenig Chloroform gelöst, auf die Säule gegeben und anschließend mit tert.-Butylmethylether eluiert (Rf- Wert 0.21). Eventuell ist eine weitere Säulenchromatographie in Essigester notwendig (Rf 0.67). Nach dem Einengen erhält man ein dunkelrotes Öl in 35%iger Ausbeute, welches sich sehr gut in Aceton und Chloroform löst. To synthesize this photoinitiator, in a 100 mL two-necked flask, add 4- [bis [2- (acetyloxy) ethyl] amino] benzaldehyde (4 g, 13.6 mmol), cyclopentanone (576 mg, 6.8 mmol) and cyanuryl chloride (252 mg, 1.37 mmol ) submitted. A drop of water is added to the mixture and heated with stirring to 75 to 80 ° C. After a few minutes, a dark green homogeneous mass is obtained. The mixture is allowed to stir for about an hour. After cooling, the crude product is dissolved in chloroform, shaken out with water and dried over sodium sulfate. The purification is carried out by column chromatography in tert-butyl methyl ether over silica gel. Due to the poor solubility of the 2PP initiator in tert-butyl methyl ether this is dissolved in a little chloroform, added to the column and then eluted with tert-butyl methyl ether (R f - value 0.21). It may be necessary to carry out further column chromatography in ethyl acetate (R f 0.67). After concentration, a dark red oil is obtained in 35% yield, which dissolves very well in acetone and chloroform.
C35H4209N2 (M = 634.73 g/mol). Elementar Analyse berechnet: C 66.23; H 6.67; N 4.41; gefunden: C 64.53; H 6.50; N 4.26. Zur Untersuchung der Polymerisationsinitiierung wurde eine an sich bekannte Zweiphotonen-Polymerisationsanlage (2PP-Anlage) M3DL (Laserzentrum Hannover) inklusive ABL 101000 luftgelagertes Achssystem für eine nm-aufgelöste Probenbewegung (Fa. Aerotech) verwendet. Als Laserquelle dient ein Ti:Sa Laser (Vision II, Fa. Coherent). Wesentliche Parameter sind: C 35 H 42 O 9 N 2 (M = 634.73 g / mol). Elemental analysis calculated: C 66.23; H 6.67; N 4.41; found: C 64.53; H 6.50; N 4.26. To investigate the polymerization initiation, a known two-photon polymerization plant (2PP plant) M3DL (Laserzentrum Hannover) including ABL 101000 air-supported axial system for an nm-resolved sample movement (Aerotech) was used. The laser source is a Ti: Sa laser (Vision II, Coherent). Essential parameters are:
Laserwellenlänge: 780 bis 800 nm, Laserleistung: 100 bis 350 mW, Schreibgeschwindigkeit: 0,1 bis 5 mm/s, Objektiv: 63x (NA: 0,75) oder lOOx (NA: 1,4 Ölimmersion), SD- Strukturen: Hatch: bis zu 13 μιη.  Laser wavelength: 780 to 800 nm, laser power: 100 to 350 mW, writing speed: 0.1 to 5 mm / s, lens: 63x (NA: 0.75) or 100x (NA: 1.4 oil immersion), SD structures: Hatch: up to 13 μιη.
Die Strukturentwicklung erfolgte durch Waschen bzw. Spülen in Aceton.  The structure development was carried out by washing or rinsing in acetone.
Bestimmung der maximalen Auflösung und optimaler Laserintensität: Determination of the maximum resolution and optimal laser intensity:
Um die Eigenschaften des erfindungsgemäßen Photoinitiators zu untersuchen, wurde die Anwendung desselben in der vorgenannten 2PP- Anlage zum Vergleich mit der adäquaten Verwendung des bekannten und kommerziell erhältlichen organolöslichen Photoinitiators Irgacure 369 in derselben Anlage mit den ausgewiesenen Prozessparametern getestet und diesem gegenübergestellt. To study the properties of the photoinitiator of the present invention, its use in the aforementioned 2PP plant was tested and compared to the appropriate process parameters for comparison with the adequate use of the known and commercially available organosoluble photoinitiator Irgacure 369 in the same facility.
In diesen Untersuchungen an UDMA (Urethandimethacrylat) zeigte sich, dass mit beiden gegenübergestellten Photoinitiatoren ein minimal schreibbarer Voxel mit einem Durchmesser von 250 nm realisiert werden kann. Dies entspricht einem minimal schreibbaren Volumen von 0,008 fl. Beide getesteten Photoinitiatoren ermöglichen demnach eine minimale Auflösung von 250 nm. Im Ergebnis dieser Untersuchungen konnte festgestellt werden, dass Laserleistungen von ca. 200 mW für beide getesteten und gegenübergestellten Photoinitiatoren zu den besten Strukturierungsergebnissen führten. Bestimmung der minimal notwendige Photoinitiatorkonzentration: In these investigations on UDMA (urethane dimethacrylate) it was shown that a minimally writable voxel with a diameter of 250 nm can be realized with both opposing photoinitiators. This corresponds to a minimally writable volume of 0.008 fl. Both photoinitiators tested therefore allow a minimum resolution of 250 nm. As a result of these investigations, it was found that laser powers of about 200 mW led to the best structuring results for both tested and compared photoinitiators. Determination of the minimum necessary photoinitiator concentration:
Zur Untersuchung der Effizienz des besagten erfindungsgemäßen Photoinitiators wurden in der besagten 2PP-Anlage weiterhin Vergleichstests der gegenübergestellten Photo- initatoren mit jeweils zwei unterschiedlichen Photoinitiatorkonzentrationen durchgeführt (2% und 0.1%). To test the efficiency of the said photoinitiator according to the invention, comparison tests of the opposite photoinitiators, each with two different photoinitiator concentrations (2% and 0.1%), were also carried out in said 2PP system.
Zwecks besserer Vergleichbarkeit wurden Laserleistung (145 mW) und Schreib geschwin- digkeit (5 mm/s) jeweils konstant gehalten. Das UDMA (Urethandimethacrylat) ließ sich mit dem bekannten Irgacure 369 erst ab einer minimalen Konzentration von 2 Gew.% mit den vorgenannten Laserparametern schreiben/strukturieren. Allerdings traten regelmäßig, besonders an den Rändern der Struktur, Schreibdefekte auf. Bei einer Konzentration von 0.1 Gew.% konnte mit Irgacure 369 im UDMA keine Strukturbildung nachgewiesen werden. For better comparability, laser power (145 mW) and write speed (5 mm / s) were kept constant. The UDMA (urethane dimethacrylate) could be written / structured with the known Irgacure 369 only from a minimum concentration of 2 wt.% With the aforementioned laser parameters. However, writing defects occurred regularly, especially at the edges of the structure. At a concentration of 0.1% by weight, it was not possible to detect any structure formation with Irgacure 369 in UDMA.
Im Gegensatz hierzu war es bereits mit 0.1 Gew.% möglich, mit dem erfindungsgemäßen Photoinitiator den entstehenden Precursor vollständig und fehlerfrei zu strukturieren. Sogar eine Konzentration dieses Photoinitiators von 0,01Gew.% zeigte sich als ausreichend, um einen UDMA Precursor erfolgreich zu strukturieren. Hierzu bedurfte es allerdings der Reduzierung der Schreibgeschwindigkeit auf 1 mm/s.  In contrast, it was already possible with 0.1% by weight, with the photoinitiator according to the invention, to structure the resulting precursor completely and without errors. Even a 0.01% by weight concentration of this photoinitiator was found to be sufficient to successfully pattern a UDMA precursor. However, this required the reduction of the write speed to 1 mm / s.
Strukturierung komplexer 3D-Objekte: Structuring complex 3D objects:
Ziel dieser Untersuchungen war die Durchführung von generellen Eignungstestungen des erfindungsgemäßen Photoinitiators zur Herstellung komplexer 3D-Strukturen mittels der Zwei-Photonenpolymerisation. Es wurden geometrisch komplexe 3D-Strukturen mittels der 2PP erstellt. Vorlage für die zu erstellenden 3D-Strukturen waren verschiedene STL- Files (Surface Tesselation Language zur Beschreibung von 3D-Oberflächen), welche in die entsprechende Steuerungssoftware der 2PP-Anlage programmiert und von dieser in 3D-Polymerstrukturen umgesetzt wurden. The aim of these investigations was to carry out general suitability tests of the photoinitiator according to the invention for producing complex 3D structures by means of two-photon polymerization. Geometrically complex 3D structures were created using the 2PP. The templates for the 3D structures to be created were various STL files (Surface Tessellation Language for describing 3D surfaces), which were programmed into the corresponding control software of the 2PP system and converted by the latter into 3D polymer structures.
Als Precursor diente ein Copolymer, bestehend aus Polycaprolacton und Polylaktid, in welches 0,1 Gew.% des erfindungsgemäßen Photoinitiators beigemischt wurden. The precursor used was a copolymer consisting of polycaprolactone and polylactide in which 0.1% by weight of the photoinitiator according to the invention was admixed.
Das 3D-Modell wurde jeweils durch eine mathematische Minimalfläche (Gyroid) verkörpert, die als Elementarzelle für die 2PP-Herstellung eines 3D-Arrays benutzt wurde. Die Höhe des 3D-Arrays betrug hierbei ca. 600 μιη. Die Struktur wurde jeweils gemäß dem STL-File detail- und maßstabsgerecht umgesetzt. The 3D model was each represented by a mathematical minimal surface (gyroid), which was used as the unit cell for the 2PP production of a 3D array. The height of the 3D array here was about 600 μιη. The structure was implemented according to the STL file in detail and to scale.
Darüber hinaus wurde ein 3D-Scaffold mit den Abmaßen 5x4x2 mm generiert, dessen Elementarzelle eine Schwarz-P Minimalfläche darstellt. Die Entwicklung der 2PP- geschriebenen Scaffoldstruktur erfolgte in Aceton. Der verwendete Precursor, inklusive des erfindungsgemäßen Photoinitiators, konnte somit durch photoinitiierte Zwei- Photonenpolymerisation hergestellt werden. Die Prozesszeit betrug ca. 12 Stunden. Prüfungen auf in vitro Zytotoxizität des Photoinitiators: In addition, a 3D scaffold with the dimensions 5x4x2 mm was generated, whose unit cell represents a black-P minimal area. The development of the 2PP-written scaffold structure was carried out in acetone. The precursor used, including the photoinitiator according to the invention, could thus be prepared by photoinitiated two-photon polymerization. The process time was about 12 hours. Tests for in vitro cytotoxicity of the photoinitiator:
Die Bestimmung einer möglichen zytotoxischen Wirkung des Photoinitiators erfolgte mittels XTT-Assay auf Grundlage der Richtlinien nach ISO/EN 10993 Teil 5 und unter Verwendung folgender Zelltypen: L929 - Mausfibroblasten, HepG2 - menschliche Hepatozyten, HEK - menschliche embryonale Nierenzellen sowie bovChz - Rinder- Chondrozyten). Die Zellen wurden in 96 Loch-Mikrotiterplatten mit einer Dichte von 1 x 104 Zellen/Loch ausgesät und für 24 h kultiviert, um einen semikonfluenten einschichtigen Zellrasen zu bilden. Für die zellbiologische Testung wurde zunächst der organolösliche Photoinitiator in DMSO gelöst (10 Gew.% bzw. 100 mg/mL). Durch die Zugabe von Zellkulturmedium erfolgte anschließend die Herstellung der finalen Testlösungen mit einer Konzentration von 1 bzw. 0,1% (DMSO Gehalt in den finalen Photoinitiator - Testlösungen: 10 bzw. 1%). Als DMSO-Kontrolllösungen diente entsprechend präpariertes Zellkulturmedium mit einem DMSO Gehalt von 10 bzw. 1%. The determination of a possible cytotoxic effect of the photoinitiator was carried out by XTT assay based on the guidelines of ISO / EN 10993 part 5 and using the following cell types: L929 - mouse fibroblasts, HepG2 - human hepatocytes, HEK - human embryonic kidney cells and bovChz - bovine chondrocytes ). The cells were seeded in 96-well microtiter plates at a density of 1 x 10 4 cells / well and cultured for 24 h to form a semiconfluent monolayer cell lawn. For cell biological testing, the organosoluble photoinitiator was first dissolved in DMSO (10% by weight or 100 mg / ml). The addition of cell culture medium then resulted in the preparation of the final test solutions at a concentration of 1 or 0.1% (DMSO content in the final photoinitiator test solutions: 10 and 1%, respectively). The DMSO control solutions used were appropriately prepared cell culture medium with a DMSO content of 10 and 1%, respectively.
Je Vertiefung der 96-Lochplatte wurden nach Absaugen des Mediums von den für 24 h vorkultivierten Zellen jeweils 100 μΐ^ der Photoinitiator - Testlösungen bzw. der DMSO- Kontrolllösungen zu den Zellen gegeben und für 24 h bei 37 °C und 5% C02 kultiviert. Nach der Inkubation wurden die Zellen einmal mit PBS gewaschen und mit 150 μΐ^ XTT- PMS-Mediumlösung pro zu prüfender Vertiefung versetzt und für weitere 3 h bis 5 h bei 37 °C und 5% C02 bebrütet. Anschließend wurden die Platten vorsichtig geschwenkt, ein aliquoter Anteil von 100 μΐ^ aus jeder Vertiefung in die entsprechende Vertiefung einer neuen Platte überführt und die Absorption bei 450-nm (Referenz -Wellenlänge 630 nm) in einem Mikrotiterplatten-Lesegerät gemessen. Per well of 96-well plate were by aspirating the medium from the 24-h pre-cultured cells, respectively 100 μΐ ^ the photoinitiator - where test solutions or the DMSO control solutions to the cells and for 24 h at 37 ° C and 5% C0 2 , After incubation, the cells were washed once with PBS and added with 150 μΐ ^ XTT-PMS medium solution per well to be tested and incubated for an additional 3 h to 5 h at 37 ° C and 5% C0 2 . Subsequently, the plates were gently swirled, an aliquot of 100 μΐ ^ transferred from each well into the appropriate well of a new plate and the absorbance at 450 nm (reference wavelength 630 nm) measured in a microtiter plate reader.
Bei der Berechnung der Verringerung der Lebensfähigkeit im Vergleich zur Kontrollprobe entsprechen die ermittelten OD Werte der Kontrolllösungen einer Viabilität von 100% bzw. 0% Reduktion/Inhibition der metabolischen Zellaktivität, wobei eine Abnahme der Viabilitätswerte auf unter 70% der Kontrollprobe auf ein deutliches zytotoxisches Potenzial der Testverbindung hinweist.  When calculating the viability reduction compared to the control, the OD values of the control solutions correspond to a viability of 100% and 0% reduction / inhibition of metabolic cell activity respectively, with a decrease in viability values to less than 70% of the control to a significant cytotoxic potential the test compound indicates.
Im beschriebenen XTT Test wurde bei Photoinitiators - Konzentrationen von 0.1 Gew.% (1.58 μΜ) bzw. bei 1% (15.8 μΜ) keine zytotoxische oder wachstumshemmende Wirkung im Vergleich zur DMSO Kontrolle beobachtet.  In the described XTT test, no cytotoxic or growth inhibitory effect was observed at photoinitiator concentrations of 0.1% by weight (1.58 μΜ) or at 1% (15.8 μΜ) compared to the DMSO control.
Zusätzlich zur Bestimmung der metabolischen Aktivität mittels XTT Test wurde die Vitalität und intakte Morphologie der Zellen nach Kontakt mit Photoinitiator mittels Lebend-Totfärbung und fluoreszenzmikroskopischer Auswertung bestätigt. Hierfür wurden entweder a.) die Zellen analog zum XTT Test in 96-Loch Mikrotiterplatten ausgesät und mit verschiedenen Photoinitiator Konzentrationen behandelt bzw. b.) Photoinitiator Filme präpariert und anschließend mit Zellen besiedelt. In beiden Fällen erfolgte nach 24 h Photoinitiator Exposition der Zellen ein einmaliges Waschen mit PBS und anschliessendes 10 minütiges Färben mit Fluoresceindiacetat (1 μg/mL) / Propidiumiodid (1 μg/mL) / Hoechst 33342 (1 μg/mL). Nach erneutem Waschen mit PBS wurden anschließend Vitalität sowie eine intakte Morphologie der mit Photoinitiator exponierten Zellen (sowohl in Lösung als auch in Form eines Filmes) mittels Hellfeld- und Fluoreszenzmikroskopie untersucht und bestätigt. In addition to the determination of the metabolic activity by XTT test, the vitality and intact morphology of the cells after contact with photoinitiator Living-dead staining and fluorescence microscopy evaluation confirmed. For this either a.) The cells were seeded analogously to the XTT test in 96-well microtiter plates and treated with different photoinitiator concentrations or b.) Prepared photoinitiator films and then populated with cells. In both cases, after 24 h of photoinitiator exposure of the cells, a single wash with PBS followed by dyeing with fluorescein diacetate (1 μg / ml) / propidium iodide (1 μg / ml) / Hoechst 33342 (1 μg / ml) for 10 minutes. After washing again with PBS, vitality and an intact morphology of the photoinitiator-exposed cells (both in solution and in the form of a film) were then examined and confirmed by bright field and fluorescence microscopy.

Claims

Patentansprüche claims
1. Zwei-Photonenpolymerisations-Initiatoren, bestehend aus einer Grundstruktur allgemeinen Formel IX: 1. Two-photon polymerization initiators consisting of a basic structure of general formula IX:
Figure imgf000013_0001
mit
Figure imgf000013_0001
With
W, X, Y. Z: unabhängig voneinander NH, O  W, X, Y. Z: independently of one another NH, O
R 1 , R2 , R 3 , R 4 : unabhängig voneinander Alkyl- oder Alkylrest R 1, R 2, R 3, R 4 independently of one another are alkyl or alkyl radicals
2. Zwei-Photonenpolymerisations-Initiator gemäß Anspruch 1, gekennzeichnet durch einen Photoinitiator der nachfolgenden Formel X: 2. Two-photopolymerization initiator according to claim 1, characterized by a photoinitiator of the following formula X:
Figure imgf000013_0002
Figure imgf000013_0002
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5112721A (en) 1990-01-29 1992-05-12 E. I. Du Pont De Nemours And Company Photopolymerizable compositions containing sensitizer mixtures

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5112721A (en) 1990-01-29 1992-05-12 E. I. Du Pont De Nemours And Company Photopolymerizable compositions containing sensitizer mixtures

Non-Patent Citations (16)

* Cited by examiner, † Cited by third party
Title
"Novel benzylidene cyclopentanone dyes for two-photon photopolymerization", J. PHOTOCHEM. PHOTOBIOL. A CHEM., vol. 195, 2008, pages 261 - 266
A. R. MORALES; K. D. BELFIELD; J. M. HALES; E. W. VAN STRYLAND; D. J. HAGAN: "Synthesis of two-photon absorbing unsymmetrical fluorenyl-based chromophores", CHEM. MATER., vol. 18, 2006, pages 4972 - 4980
B. H. CUMPSTON; S. P. ANANTHAVEL; S. BARLOW; D. L. DYER; J. E. EHRLICH; L. L. ERSKINE; A. A. HEIKAL; S. M. KUEBLER; I. Y. S. LEE;: "Two-photon polymerization initiators for three-dimensional optical data storage and microfabrication", NATURE, vol. 398, 1999, pages 51 - 54
B. H. CUMPSTON; S. P. ANANTHAVEL; S. BARLOW; D. L. DYER; J. E. EHRLICH; L. L. ERSKINE; A. A. HEIKAL; S. M.KUEBLER; I. Y. S. LEE; D: "Two-photon polymerization initiators for three-dimensional optical data storage and microfabrication", NATURE, vol. 398, 1999, pages 51 - 54
C. A. ZOTO; R. E. CONNORS: "Photophysical properties of an asymmetrical 2,5-diarylidene-cyclopentanone dye possessing electron donor and acceptor substituents", J. MOL. STRUCT., vol. 982, 2010, pages 121 - 126
K. J. SCHAFER; J. M. HALES; M. BALU; K. D. BELFIELD; E. W. VAN STRYLAND; D. J. HAGAN: "Two-photon absorption cross-sections of common photoinitiators", J. PHOTOCHEM. PHOTOBIOL. A CHEM., vol. 162, 2004, pages 497 - 502
K. S. LEE; D. Y. YANG; S. H. PARK; R. H. KIM: "Recent developments in the use of two-photon polymerization in precise 2D and 3D microfabrications", POLYM. ADV. TECHNOL, vol. 17, 2006, pages 72 - 82
M. JIN; J.-P. MALVAL; D.-L. VERSACE; F. MORLET-SAVARY; H. CHAUMEIL; A. DEFOIN; X. ALLONAS; J.-P. FOUASSIER: "Two-photon absorption and polymerization ability of intramolecular energy transfer based photoinitiating systems", CHEM. COMMUN., 2008, pages 6540 - 6542
M. T. RAIMONDI; S. M. EATON; M. M. NAVA; M. LAGANA; G.CERULLO; R. OSELLAME: "Two-photon laser polymerization: From fundamentals to biomedical application in tissue engineering and regenerative medicine", J. APPL. BIOMATER. FUNT. MATER., vol. 10, 2012, pages 55 - 65
Q. ZOU; Y. ZHAO; N. S. MAKAROV; J. CAMPO; H. YUAN; D.-C. FANG; J. W. PERRY; F. WU: "Effect of alicyclic ring size on the photophysical and photochemical properties of bis(arylidene)cycloalkanone compounds", PHYS. CHEM. CHEM. PHYS., vol. 14, 2012, pages 11743 - 11752
S. C. SINGH; H. ZENG: "Nanomaterials and nanopatterns based' on laser processing: A brief review on current state of art", SCI. ADV. MATER., vol. 4, 2012, pages 368 - 390
XUE ET AL: "Novel benzylidene cyclopentanone dyes for two-photon photopolymerization", JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY, A: CHEMISTRY, ELSEVIER SEQUOIA, LAUSANNE, CH, vol. 195, no. 2-3, 22 October 2007 (2007-10-22), pages 261 - 266, XP022512167, ISSN: 1010-6030, DOI: 10.1016/J.JPHOTOCHEM.2007.10.012 *
Z. LI; M. SIKLOS; N. PUCHER; K. CICHA; A. AJAMI; W. HUSINSKY; A. ROSSPEINTNER; E. VAUTHEY; G. GESCHEIDT; J. STAMPFL: "Synthesis and structure-activity relationship of several aromatic ketone-based two-photon initiators", J. POLYM. SCI., PART A: POLYM. CHEM., vol. 49, 2011, pages 3688 - 3699,1-2
Z. LI; N. P,UCHE; K. CICHA; J. TORGERSEN; S. C. LIGON; A. AJAMI; W. HUSINSKY; A. ROSSPEINTNER; E. VAUTHEY; S. NAUMOV: "A straightforward synthesis and structure-activity relationship of highly efficient iriitiators for two-photon polymerization", MACROMOLECULES, vol. 46, 2013, pages 352 - 361
Z. LI; N. PUCHER; K. CICHA; J. TORGERSEN; S. C. LIGON; A. AJAMI; W. HUSINSKY; A. ROSSPEINTNER; E. VAUTHEY; S. NAUMOV: "A straightforward synthesis and structure-activity relationship of highly efficient Initiators for two-photon polymerization", MACROMOLECULES, vol. 46, 2013, pages 352 - 361
ZOTO C A ET AL: "Photophysical properties of an asymmetrical 2,5-diarylidene-cyclopentanone dye possessing electron donor and acceptor substituents", JOURNAL OF MOLECULAR STRUCTURE, ELSEVIER, AMSTERDAM, NL, vol. 982, no. 1-3, 8 October 2010 (2010-10-08), pages 121 - 126, XP027290010, ISSN: 0022-2860, [retrieved on 20100814] *

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