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  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
  • C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
  • C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
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  • C08J7/0427—Coating with only one layer of a composition containing a polymer binder
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  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
  • C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
  • C08J7/04—Coating
  • C08J7/043—Improving the adhesiveness of the coatings per se, e.g. forming primers
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  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
  • C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
  • C08J7/04—Coating
  • C08J7/046—Forming abrasion-resistant coatings; Forming surface-hardening coatings
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
  • C12M35/06—Magnetic means
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
  • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
  • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
  • C12N11/082—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds
  • C12N11/087—Acrylic polymers
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
  • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
  • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
  • C12N11/089—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
  • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
  • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
  • C12N11/089—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
  • C12N11/093—Polyurethanes
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
  • C08J2300/00—Characterised by the use of unspecified polymers
  • C08J2300/14—Water soluble or water swellable polymers, e.g. aqueous gels
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
  • C08J2300/00—Characterised by the use of unspecified polymers
  • C08J2300/26—Elastomers
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
  • C08J2333/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides, or nitriles thereof; Derivatives of such polymers
  • C08J2333/24—Homopolymers or copolymers of amides or imides
  • C08J2333/26—Homopolymers or copolymers of acrylamide or methacrylamide
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
  • C08J2383/00—Characterised by the use of macromolecular compounds obtained by reactions forming in the main chain of the macromolecule a linkage containing silicon with or without sulfur, nitrogen, oxygen, or carbon only; Derivatives of such polymers
  • C08J2383/04—Polysiloxanes
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
  • C08J2489/00—Characterised by the use of proteins; Derivatives thereof
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
  • C08J2489/00—Characterised by the use of proteins; Derivatives thereof
  • C08J2489/04—Products derived from waste materials, e.g. horn, hoof or hair
  • C08J2489/06—Products derived from waste materials, e.g. horn, hoof or hair derived from leather or skin

Definitions

• Magnetoactive or electroactive composite material its use and method for influencing biological cells deposited on the magnetoactive or electroactive composite material
• the invention relates to biocompatible, soft, magneto-active polymers (MAP) whose elastomechanical properties can be adjusted by a magnetic field. Moreover, the invention relates to special magnetic field systems for the local control of the MAP and the movement, separation, control and influence of biological cells on the surface of the MAP material.
• MAP biocompatible, soft, magneto-active polymers
• Magnetorheological elastomers are composites consisting of an elastomer matrix and magnetizable particles contained therein.
• the application of a magnetic field gives rise to attractive interactions between the particles, which hardening of the composite material lead.
• another effect additionally occurs, which consists of stretching in an inhomogeneous magnetic field.
• Both processes are reversible and steplessly controllable.
• the reversible strain manifests itself in the relatively weakly networked MAP materials in a magnetic field-controlled shape memory effect.
• the magnetomechanical properties of the MRE can be adjusted over a wide range of composition. The main influencing parameters are the
• Iron powder which originate from various manufacturing processes, has hitherto predominantly been used as particulate material
• MAP magneto-active polymers
• ERE electrorheological elastomers
• the matrix material of the ERE can also be used silicone elastomer, while the polarizable particles of a polymer or inorganic materials such. B. zeolites can exist.
• Soft ERE will be electroactive in the following
• biomechanical stimuli can be exerted very advantageously by MAP materials if they are biocompatible.
• MAP materials are biocompatible.
• Elastomers fall on silicone, which is adjustable in its mechanical properties on the degree of chemical crosslinking in a wide range.
• the silicone surface is coated with a cell-compatible coating, e.g. modified from collagen or gelatin.
• the magnetic field generation system is to be a varia- allow for adjustable magnetic field strength in MAP. Due to the magnetic field strength, the hardness of the MAP material can be adjusted, which influences cell growth. In a specific embodiment, the magnetic field generating system can generate locally varying magnetic fields. Due to the resulting local differences in hardness on the MAP surface, cell migration can be triggered because different cells prefer different degrees of hardness of the substrate.
• Another specific embodiment of the magnetic induction generating system is capable of generating time-varying inhomogeneous magnetic fields. This uses the actoric effect of MAP materials so that the material follows and moves with the variable magnetic field.
• EAPs electroactive polymers
• the proliferation of different cell types can be influenced by a hardness of a MAP or EAP set by a static magnetic field or a static electric field.
• An actuatory stimulation of the cells can be produced by a temporally and / or locally variable hardness of the MAPs / EAPs.
• the cellular protein expression influenced by temporally and / or locally variable hardness of the MAPs / EAPs.
• a magnetoactive and / or electroactive composite material which contains magnetically and / or electrically polarisable particles in at least one polymer as matrix.
• the composite material is a biocompatible composite material or a biocompatible coated or surface-biocompatible modified
• both a biocompatible composite material or polymer and a non-biocompatible composite material or polymer may be biocompatible modified or coated on the surface.
• biocompatible it is preferred that the biocompatible
• the polymer is an elastomer, in particular a silicone elastomer, a polyurethane elastomer or a thermoplastic
• Elastomer or a gel, in particular a polyacrylamide gel.
• the polymer has a biocompatible surface modification which inhibits the binding of proteins, e.g. Collagen, laminin, fibronectin or gelatin.
• proteins e.g. Collagen, laminin, fibronectin or gelatin.
• the composite material according to the invention preferably has an E modulus 500 500 kPa, preferably 100 100 kPa, and more preferably 20 20 kPa.
• the magnetically polarisable particles are ferromagnetic and preferably selected from the group consisting of iron, iron alloys, such as iron / cobalt or iron / nickel, iron oxides, ferrites and / or mixtures thereof.
• the electrically polarizable particles are selected from the group consisting of polymers or doped polymers, preferably polyurethane, or of inorganic materials, preferably zeolites.
• the particles in this case preferably have an average particle size -S 100 ⁇ m, preferably 10 ⁇ m and particularly preferably 2 ⁇ m.
• the particles in the polymer matrix may have an anisotrope distribution.
• a method for influencing the invention magneto-active and / or electroactive composite material attached biological cells wherein the properties of the magneto-active and / or electroactive composite material can be changed by an applied magnetic field or electric field.
• the cells are stimulated by adjusting the mechanical hardness and / or the modulus of elasticity of the composite material.
• the cells can migrate due to an adjustment of a locally variable hardness and / or a locally variable modulus of elasticity of the composite material.
• the temporal and / or local change in the hardness and / or the modulus of elasticity of the composite material is used to control the cellular protein expression of the cells.
• the control according to the invention of the biocompatible polymer takes place with a magnetization system.
• a magnetization system consists of single or multiple elementary magnetic circuits in the various forms.
• Each elementary magnetic circuit consists of one or more magnetic field sources, magnetic flux carrying components, and the composite material that terminates the magnetic circuit.
• magnetic field sources permanent magnets and / or coils can be used.
• the magnetic flux conducting components are preferably made of a ferromagnetic Voll L, laminated material.
• the system can preferably also be equipped with a holding system and / or centering device. Furthermore, one or more coils may flow around the magnetic flux- be arranged the components.
• the permanent magnetic material may be disposed within or in the vicinity of the magnetic circuit components.
• the elementary circles may be arranged per sample as a single system or as an array of magnetic elementary circles in series, as a circle, or in another form.
• the magnetization system is below or to the side of one or more containers or trays containing the samples
• the magnetically flux-conducting components and magnetic heads / needles form a magnetic inference with the air gap between the container and the samples.
• the holding device connects the magnetization system with the containers used.
• the samples or containers are aligned with the magnetization system.
• the magnetic field generating coils can be embedded in the polymer.
• a further alternative according to the invention provides that an active mechanical stimulation of the cells by means of an optionally pulsative and / or
• the hardness of the MAP material can be individually and reversibly adjusted to the requirements of the cell culture by means of a magnetic field.
• a special magnetic field generation system can be used to set locally different magnetic fields and corresponding hardness ranges of the MAP material, which can trigger cell migrations. Furthermore, a corresponding movement and thus an active mechanical stimulation of the cell culture can be triggered by a time-varying inhomogeneous magnetic field in a soft AP material.
• the protein expression of the cells can be influenced by a temporally and / or locally variable hardness of the MAP material.
• Materials are additionally functionalized with special chemical groups, whereby the adhesion of cells and thus the effect of mechanical stimulation is improved.
• Magnetoactive or electroactive polymers of tissue-like elasticity are suitable as substrates for the growth, multiplication and differentiation of
• Nervous tissue can be of great importance.
• the invention likewise provides a system for influencing biological cells, which contains at least one unit for magnetic and / or electrical activation, the at least one permanent magnet and / or at least one electrical coil or at least one voltage supply and the magneto-active and / or electroactive
• the magnetic flux density generated at the surface of the composite material is the magnetic flux density generated at the surface of the composite material.
• B mT preferably B ⁇ 10 mT, particularly preferably B> 10 mT and B ⁇ 500 m.
• the gradient of the magnetic flux density and / or the electric field strength at the surface of the composite material is generated and can be changed over time.
• FIG. 1 shows a system according to the invention for influencing biological cells with a unit for magnetic activation. example
• Silicone composites is modified with a collagen coating.
• the magnetization system consists of several elementary magnetization systems (FIG. 1).
• Each elementary magnetization system consists of one or more permanent magnets 7, current-carrying coils 5 for controlling the magnetic field and yokes 3, 8 for conducting the magnetic flux within the magnetic circuit and the magnetic heads for conducting the magnetic flux to the MAP.
• Each elementary magnetic circuit is closed by the magnetizable MAP 10, 11.
• the MAP is thus used as magnetic inference.
• the elementary magnetic circuits are magnetically decoupled by non-magnetic connecting pieces 4 and can thus be controlled independently.
• the MAP filled Petri dish is attached by a holding device with a Zentri réelles Anlagenkeit and is di- right above the magnetization heads. All magnet-carrying components can be laminated to ensure fast time-dependent control options.
• the support rods 13 serve the centering and attachment of the individual elementary
• the plate 14 serves for the mechanical or actuatoric control of the permanent magnets. It serves to attenuate or switch off the permanent magnets by magnetically shorting the lower part of the magnetic circuit.
• the device can be operated in three modes. Depending on the required field strength and available power supply, electromagnetic operation via coils, operation via permanent magnets and hybrid operation via coils and PM is possible. For example, By changing the magnitude and direction of the currents through the coils, the magnetizing effect of the
• Permanent magnets strengthened or weakened.
• the purpose of the magnetization system is to generate both a spatial and a temporal change of the magnetic field within the MAP and thus to influence both the mechanical and temporal properties of the MAP and / or to generate a movement of the MAP surface. From the literature it is known that when egg ⁇ nes

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