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WO2019179978A1 - Procédé pour l'amélioration et/ou le contrôle de l'adhésion cellulaire et implant médical, système de culture cellulaire ou essai cellulaire présentant une adhésion cellulaire améliorée - Google Patents

Procédé pour l'amélioration et/ou le contrôle de l'adhésion cellulaire et implant médical, système de culture cellulaire ou essai cellulaire présentant une adhésion cellulaire améliorée Download PDF

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WO2019179978A1
WO2019179978A1 PCT/EP2019/056769 EP2019056769W WO2019179978A1 WO 2019179978 A1 WO2019179978 A1 WO 2019179978A1 EP 2019056769 W EP2019056769 W EP 2019056769W WO 2019179978 A1 WO2019179978 A1 WO 2019179978A1
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cell
areas
partial areas
cell adhesion
cells
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German (de)
English (en)
Inventor
Andreas Radeloff
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Carl Von Ossietzky Universitaet Oldenburg
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Carl Von Ossietzky Universitaet Oldenburg
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/28—Materials for coating prostheses
    • A61L27/34—Macromolecular materials
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • A61L27/3886—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells comprising two or more cell types
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
    • C09D183/00—Coating compositions based on 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; Coating compositions based on derivatives of such polymers
    • C09D183/04—Polysiloxanes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/18—Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment

Definitions

  • the present invention relates to a method for improving and / or controlling cell adhesion on a surface to be coagulated or on a surface area to be coagulated, as well as a medical implant, a cell culture system or a cell assay with a surface produced in this way or with a surface area produced in this way ,
  • Cochlear implants are electrical hearing prostheses that are used in deaf patients and electrically stimulate the processes of the auditory nerve.
  • an electrode carrier made of silicone rubber is introduced into the cochlea, on which electrode contacts are located. The connection of the electrode contacts to the Hömervenfort accounts has so far not been optimally successful, since the electrode carriers of cochlear implants are difficult to colonize due to the very high hydrophobicity of the silicone surface of cells, especially of the nerve cells.
  • the object is achieved by a method for improving and / or controlling cell adhesion, comprising a) providing a surface to be covered or a surface area, b) defining partial areas A on the surface or on the surface area on which the cell adhesion is to take place, and of subareas B on which cell adhesion should not occur,
  • the contrasting structuring takes place on the nanometer and / or micrometer scale and the partial areas A are hydrophilized and the partial areas B are made hydrophobic.
  • the partial areas A on the surface or on the surface area are first defined, on which a good cell adhesion is to be promoted, and the subareas B, on which no promotion of cell adhesion is desired, are conceptually differentiated.
  • portions A may be selected on the surfaces of the electrode contacts in which an attraction of the auditory nerve fibers to the cochlear implant is to be achieved, and conversely those portions B in which no attraction of nerve fibers is to be achieved. In this way it can be achieved that a control of the growth direction of the nerve fibers on the cochlear implant is made possible.
  • specifying portions A and B may be directed to achieving application of cells for artificial target innervation.
  • neurotrophin-producing cells such as stem cells from adipose tissue or genetically modified cells on implants for neuro / sensory prostheses, because a very precise positioning of the applied cells and avoidance of uncontrolled migration is in these as well displayed in related application areas.
  • partial regions A and partial regions B on cell culture systems.
  • two or more cell types can be cocultured next to one another in a culture system, which can each be applied to partial areas A separated from partial areas B in one and the same culture system.
  • partial regions A and partial regions B may be the production of very differentiated chemotaxis assays or neuritogenesis assays.
  • central partial areas A with comb-like channels or tracks for applying the cells to be examined can be provided, and at the same time separate partial areas A separated by partial areas B, in which different cytokine sources or similar substances, cells or negative controls can be arranged.
  • step c) of the method according to the invention a contrast is produced between the partial areas A and the partial areas B by means of structuring.
  • the term "structuring” is understood here and below to mean that an arrangement of the subregions relative to one another, and thus an articulated structure, as it were an outline, of the surface or of the surface region is undertaken.
  • a surface texture is created There are subareas that mutually alternate with each other or form contexts and a self-contained whole.
  • structuring not that a relief-like design surface is made with elevations or depressions.
  • the term contrast is understood here and below as meaning that there is a difference in a property of the subregions.
  • the predominant property to be defined is a difference in the hydrophilicity or in the hydrophobicity of the surface of the partial areas.
  • nanoscale and / or micrometer rod is understood to mean that the extent of the partial regions A and / or the partial regions B on the surface or surface region to be structured is approximately between 2 nanometers and 5 millimeters (5,000 Micrometer), for example, the partial areas A or the partial areas B may have a length of 0.05 mm to 3 mm and a width of 5-300 micrometers, or the partial areas A or the partial areas B may have an extension between 10 nanometers and 250 Microns in one direction.
  • the extent of the partial areas A or the partial areas B is determined and determined on the basis of the application.
  • a portion A for an application in the application of cells for artificial target innervation or for controlling the growth direction of the nerve fibers to the cochlear implant on a corresponding electrode contact will be made on a nano- or small micrometer scale, while the extension of a portion A in an application as a defined range in a cell culture rather on a millimeter scale. All of these orders of magnitude are intended to be encompassed within the meaning of the present invention by the term nano and / or micrometer scale.
  • the partial areas A are hydrophilized, while the partial areas B are made hydrophobic.
  • Hydrophilic means "water-loving" and indicates that a substance or surface preferably interacts with water or other polar species.
  • hydrophilicity is the solvation tendency of a molecule in water.
  • hydrophilicity is not synonymous with water solubility. Hydrophilicity therefore refers only to the interaction with water or other polar groups, and neither solubility nor the ability to attract and bind water.
  • the opposite of hydrophilicity is hydrophobicity and refers to the tendency of a molecule or surface to attract or interact with nonpolar substances such as fats. Hydrophobic surfaces are characterized by the fact that they repel water and have a large contact angle to water.
  • the size of the contact angle Q between water as a liquid and the solid of the surface can be measured.
  • a surface can be assumed to be hydrophilic.
  • a surface may be considered to be more hydrophobic.
  • one often speaks of superhydrophobic surfaces about 100 ° to about 170 °).
  • the contact angle Q can be measured with a contact angle goniometer.
  • the step of hydrophilizing the portions A may be performed in any manner known to those skilled in the art.
  • a plasma treatment a treatment with a polydop amine solution or a poly-D-lysine solution, application of small protein sequences, a coating with phosphorylcholine, a CVD (Chemical Vapor Deposition) coating, a Metallization or a glaze can be called.
  • Typical methods for coating with liquid agents are, for example, the dip coating (dip coating), the stamp printing coating, the spin coating and the roll coating (roll coating).
  • the hydrophilic portions A are not formed, or not alone, by plasma treatment.
  • not solely by plasma treatment is meant, for example, that it is conceivable that a method is used in which first a masking is made, then a plasma treatment of the unmasked areas is made, and then the masking is washed off, after which a further treatment for the hydrophilization of the partial areas A can follow.
  • portions B are inventively hydrophobic.
  • portions B are those portions of the originally provided surface that are not treated by hydrophilization. Due to the mostly hydrophobic surface of the respective preferred materials for the selected application, such as, for example, silicone or PTFE surfaces of the implants and metallic surfaces of the electrode contacts, the partial regions B are often designed to be hydrophobic without further treatment. It is alternatively or additionally equally possible to bring about the hydrophobic formation of the portions B by a treatment step or reinforce. All methods known to those skilled in the art for hydrophobizing a surface are suitable for this purpose. As examples may be mentioned in this regard the treatment or coating with silicones, PTFE, or waxes.
  • the procedure of the method according to the invention makes it possible to provide new applications, such as the use of a cell culture with two of the more different cell types in coculture, or the application of the Zellas says, the targeted channels or lanes for migration to be examined For example, allow cells to access different cytokine sources.
  • the method according to the invention is a widely applicable new form of directed Controlling cell adhesion in preselected subregions and also providing directional prevention of cell adhesion in adjacent subregions of the surface.
  • the applications mentioned here and below are only a small part of the many possibilities offered by the new process.
  • silicone elastomers and / or silicone oils can be used for the hydrophobicization of the partial regions B.
  • Silicone elastomers and / or silicone oils are widely used in biological or medical applications.
  • Silicone rubbers are materials that can be converted into the rubber-elastic state and contain poly (organo) siloxanes.
  • Silicone rubbers contain reinforcing substances and fillers whose type and quantity significantly influence the mechanical and chemical behavior of the silicone elastomers resulting from the cross-linking. Silicones are non-toxic to the human body and very resistant. They also withstand extreme mechanical demands and also have a high degree of purity in clinical applications, resulting in very good biocompatibility.
  • a polydopamine solution can be used to hydrophilize the partial areas A.
  • Polydopamine (PDA) has been shown in recent years to be an easily accessible synthetic analogue of naturally occurring melanin. The structure of the polymer is dependent on various synthesis conditions and plays a central role in the adsorption of molecules to ensure the versatile functionality of a layer, for example optical absorption and fluorescence, adhesive properties, biocompatibility or biodegradation.
  • Hydrophilization of the partial areas A can be carried out by wet-chemical methods, for example by dip-coating methods. Stamp printing processes or soft lithography processes are also possibilities for hydrophilizing the areas A.
  • a PDA (polydopamine) / Tris-HCl buffer solution (2 mg / ml) can be used at a pH of 8.5, but other concentrations and pH ranges of the solution are also useful.
  • an amount of 5 mg dopamine in 50 mM Tris buffer at pH 8.8 was used. The polymerization takes place only on the surface. Also a solution in ethanol is possible.
  • a coating which adheres only to these partial areas can be applied to the partial areas A.
  • an additional coating can be applied in such a way that it adheres only in the predetermined subregions. This can be achieved, for example, by a corresponding pretreatment of the partial areas A.
  • Such pretreatment is generally known to those skilled in the art.
  • a plasma treatment with various gases, such as argon, carbon dioxide or oxygen and mixtures thereof, or other agents may be mentioned.
  • gases such as argon, carbon dioxide or oxygen and mixtures thereof, or other agents.
  • Other types of pretreatment are already mentioned above as hydrophilization procedures.
  • coatings may be mentioned which consist of glycoproteins of the extracellular matrix such as laminin or amino acid polymers such as poly-lysine, receptors or their ligands, cell adhesion molecules (CAMs), amino acid sequences such as the RGD sequence or consist of or include sequences of RNA or DNA.
  • laminin amino acid polymers
  • CAMs cell adhesion molecules
  • amino acid sequences such as the RGD sequence or consist of or include sequences of RNA or DNA.
  • Such a coating can be prepared, for example, by wetting with a poly-lysine solution (typically 10-150 ⁇ g / ml poly-lysine in PBS), which typically remains on the surface for 8-24 hours at 4 ° C.
  • a poly-lysine solution typically 10-150 ⁇ g / ml poly-lysine in PBS
  • a laminin solution typically 10 pg / ml laminin in PBS
  • concentrations of 100 pg / ml poly-D (or L-) lysine are also used.
  • a solution of 0.5 mg / ml RGD peptide in carbonate buffer which remains on the surface for 3 hours, is suitable.
  • siRNA solutions eg 200 nM siRNA in ultrapure water
  • siRNA solutions can be used which remain on the surfaces to be treated for a few hours.
  • the contrasting structuring can be carried out by means of stamp printing method or soft lithography method. These methods are characterized by the fact that they are easy to perform at desired dimensional accuracy and do not make great demands on the equipment used.
  • an increase of cells on the subareas A is carried out as step d).
  • those cells and cell types can be selectively applied only in the subareas A, while the subareas B remain uncovered or the adhesion of the vegetation with cells in the subregions B is so low that the cells can be washed out.
  • the structured application of cells can also take place via the detour of a coating which adheres only to the hydrophilic areas.
  • Suitable coatings typically consist of or include proteins such as laminin or poly-D-lysine, receptors or their ligands or suitable amino acid sequences. These coatings regularly delaminate from hydrophobic substrates, making them unsuitable for microstructuring as a cell pad on silicone implants alone.
  • the said hydrophilic / hydrophobic microstructuring can also be used to provide surfaces of implants and cell culture systems in step d) with coatings which are applied with the aid of aqueous solutions. For this purpose, a hydrophilic / hydrophobic structuring takes place in a first step.
  • an aqueous solution with bioactive ingredients such as, for example, amino acids, proteins, DNA / RNA, pharmaceutically active substances, is then applied in a planar manner. This then forms a coating in the hydrophilic areas A, while the hydrophobic areas B are recessed, optionally after suitable rinsing operations.
  • bioactive ingredients such as, for example, amino acids, proteins, DNA / RNA, pharmaceutically active substances
  • step d) may be a nerve cell nerve destination, and in particular the cells in step d) may be horseshoe cells.
  • a further subject of the present invention is a medical implant comprising a surface to be covered with cells or a surface area on which or areas A are provided which are intended for cell adhesion and on which or areas B are not fixed Cell adhesion are provided, characterized in that for the improvement and / or control of cell adhesion, a contrasting structuring on the nanometer or micrometer scale is applied, which forms the partial areas A hydrophilic and the partial areas B hydrophobic forms.
  • an implant of the present invention it can be achieved in a very targeted manner that cells, such as nerve cells, are directed along the predefined subregions A, which are designed, for example, as channels or pathways are able to grow and thus allow a connection of damaged nerve endings or other sensory cells.
  • cells such as nerve cells
  • the application of narrow hydrophilic structures (about 5 pm) in corresponding subareas A can be used to direct nerve fibers along these structures.
  • the nerve fibers of the Hömerv grow transversely to the electrode carrier and not along the electrode carrier, since the latter would lead to a less selective stimulation. It therefore makes sense to apply structures perpendicular to the longitudinal axis of the electrode carrier.
  • the partial areas A can be made hydrophilic by polydopamine.
  • the application of a polydopamine layer per se has already been described above and known to the person skilled in the literature, for example in "Collagen-Polydopamine Coating and Colonization of Cochlear Implants with Adipose-Derived Stem Cells", German Society for Otolaryngology. Medical science, head and neck surgery. 88th Annual Meeting of the German Society for Otorhinolaryngology, Head and Neck Surgery. Erfurt, May 24-27, 2017 or in by Schendzierlorz et al. described.
  • the implants according to the invention are made possible for the first time with the implants according to the invention.
  • mammary implants on the surface of which a contrasting structuring according to the present invention is applied.
  • the Kapselkon Wick rate or the tendency to inflammation can be significantly reduced while suppressing unwanted adhesions or be avoided.
  • the surface of silicone implants, such as in Mammarekonstrutation be chosen so that the resulting connective tissue reaction arises in a functional or aesthetically favorable manner, it is possible to produce directed fibers instead of flat capsules.
  • suitable cells on implants can be important in many areas of medicine in the future. Typically it will be These are cells that naturally or after biotechnological interventions produce substances that attract or repel other cells or cell processes.
  • This precise application of cells is possible with the inventive concept of hydrophilic / hydrophobic micro structuring, ie the contrasting structuring.
  • An example of an application already briefly cited above is the support of nerve fiber target innervation where a target, for example an implant, is colonized at precisely defined locations, for example electrode contacts, with cells attracting nerve fibers, while other areas remain uninhabited so do not create attraction. This is always important when nerve cells or their extensions connect with implants, see electrode-nerve interfaces: sensory prostheses such as cochlear implant, retinal implant, future implants of the spinal cord after paraplegia, can take advantage of this, the fiction, contemporary implant offers.
  • Artificial target innervation of implants such as cochlear implants or other neuro / sensory prostheses can be achieved by applying neurotrophin-producing cells to the implants.
  • Suitable for this are, for example, stem cells from adipose tissue (so-called adipose-derived stromal cells, ASCs) or genetically modified cells.
  • ASCs adipose-derived stromal cells
  • ASCs adipose-derived stromal cells
  • a hydrophilic / hydrophobic micro structuring in the sense of the present invention. This is designed so that hydrophilic areas A are applied where cells are supposed to adhere, while other areas B are rendered hydrophobic.
  • Correspondingly designed surfaces are introduced into a cell suspension.
  • the cells in suspension then adhere only to hydrophilic areas A, while the adherence to hydrophobic areas B takes place very weakly or not at all.
  • cells which loosely adhere to hydrophobic regions B may be detached by a washing step.
  • the hydrophilic / hydrophobic structuring can also be used to modify the response of the body to passive implants such as endoprostheses in orthopedics, dental implants, or implants for augmentation of the breast, etc.
  • the surrounding connective tissue or the surrounding bone can be stimulated to a structured growth. This can be used, for example, to control the growth of bones on endoprostheses according to the force vectors that occur.
  • Another object of the present invention is the use of a method according to the invention for the production of cell culture systems or cell assays in which two or more cell types in co-culture occurrence, without the cell types come into contact with each other, as well as a cell culture system or a thus prepared assay.
  • a method of the invention can be advantageously used to prepare experimental cell culture systems for research and diagnosis.
  • the hydrophilic-hydrophobic contrasting structure can be used to ensure a spatially defined growth of cells. Separated regions can be created with the use of the method according to the invention, in which two or more cell types are cocultured without these cells coming into direct contact with each other (see FIG. 1). Cytokines are then released or absorbed via the overlying medium.
  • already known coculture systems work with an insert that is hung in a culture dish. The cells in this insert deliver the cytokines to the medium through a membrane at the bottom of the insert. The second cell type is cultured in the culture dish. Delivery of the substances from the insert through the membrane into the culture medium is uncertain because the exchange area is small.
  • chemo taxisassays can be prepared by means of the use according to the invention of the present method, wherein the cells to be examined are located in a central region, while in separate areas two or more competing cytokine sources or similar substances are produced (see FIG. 2).
  • assays for neuritogenesis can be displayed.
  • neurons are located in a central area, from which longitudinal narrow structures in the form of tracks or channels lead to the periphery. Due to their narrow configuration, the latter are only covered by nerve processes, not by cell bodies. Competing neuroattractive substances, cells or the like are located in the periphery in subareas A. The neuritogenesis and attraction can be easily read from the filled lanes (see Figure 3).
  • hydrophilic / hydrophobic microstructuring can also be used to provide surfaces of implants and cell culture systems with coatings that are applied with the aid of aqueous solutions.
  • a hydrophilic / hydrophobic structuring takes place in a first step.
  • an aqueous solution with bioactive ingredients such as, for example, amino acids, proteins, DNA / RNA, pharmaceutically active substances, is then applied in a planar manner. This then forms a coating in the hydrophilic areas A, while the hydrophobic areas B are recessed, optionally after suitable rinsing operations.
  • a cell culture system or assay produced by a method according to the invention is claimed in which the partial regions A are formed as separated regions and one, two, three or more identical or different cell types, neurons or cytokine sources can be applied to the partial regions A ,
  • the present invention will be further explained by the figures 1 to 3 below.
  • the figures show illustrative examples of various applications of the invention which, however, do not represent all the applications described and are therefore presented as exemplary for a better understanding of the invention.
  • Figures 1A and 1B show two cell culture systems according to the present invention for coculture of different cells in a schematic plan view
  • FIG. 3 shows a neuritogenesis / neuroattraction assay according to the present invention.
  • Figs. 1A and 1B examples are shown for each of a cell culture system 10 having two (Fig. 1A) or four (Fig. 1B) fields A1 and A2, and Al, A2, A3 and A4 for coculture of the same or different cells, respectively.
  • the hydrophilic portions A according to the invention are shown as Al and A2 or Al, A2, A3 and A4 and allow cell adhesion, while the portions B are arranged around the partial areas A surrounding and are left white in the figure.
  • This offset representation of the partial regions A (hydrophilic, medium gray) and the partial regions B (hydrophobic, white) is also used uniformly in the following figures for the partial regions A and B according to the invention.
  • FIG. 2 shows an example of a chemotaxis assay 10 according to the present invention.
  • Gray areas A1, A2 and A3 are hydrophilic as partial areas A and enable cell adhesion.
  • cytokine sources or the like are applied in the area Al and A2 co-cultured cells.
  • the cells to be examined 12 are then seeded in the central region A3 and are shown here dark gray as dots.
  • the light gray area around the area A3 is initially covered and prevents the primary adherence of cells in this area. He is then revealed.
  • the cells 12 now travel along the gray partial area A-paths in the direction of the separate partial areas A separated therefrom, which in the figure as Al and A2 are designated.
  • the chemotactic effect of the cells / substances in the regions A or A2 belonging to the subregions A can then be easily quantified.
  • An example of a neuritogenesis / neuroattraction assay 10 is shown in FIG.
  • Neurons or neuronal stem cells 12 are shown in dark gray as dots and seeded in the middle hydrophilic portion A region A3.
  • the likewise hydrophilic paths, which lead from the central portion A comb-like in the periphery, are so narrow that they can be grown only by nerve processes, but not by cell bodies.
  • Cells, cytokine sources, negative controls or the like are present in the further hydrophilic subarea A-representing regions A1 and A2. Nerve sets now grow towards Al or A2. The growth can be easily quantified and exemplified by dark gray areas in the paths.

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  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

La présente invention concerne un procédé pour l'amélioration et/ou le contrôle de l'adhésion cellulaire, comprenant les étapes consistant à a) fournir une surface à cultiver ou une zone de surface, b) déterminer les zones partielles A sur la surface ou la zone de surface, sur lesquelles l'adhésion cellulaire doit s'effectuer, et les zones partielles B, sur lesquelles l'adhésion cellulaire ne doit pas s'effectuer, c) créer un contraste entre les zones partielles A et les zones partielles B au moyen d'une structuration, la structuration contrastante s'effectuant à l'échelle du nanomètre et/ou du micromètre et les zones partielles A étant rendues hydrophiles et les zones partielles B hydrophobes. L'invention concerne également un implant médical, un système de culture cellulaire et un essai qui présentent une structuration contrastante précitée de la surface ou d'une zone de la surface.
PCT/EP2019/056769 2018-03-20 2019-03-19 Procédé pour l'amélioration et/ou le contrôle de l'adhésion cellulaire et implant médical, système de culture cellulaire ou essai cellulaire présentant une adhésion cellulaire améliorée Ceased WO2019179978A1 (fr)

Applications Claiming Priority (2)

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DE102018204203.9A DE102018204203A1 (de) 2018-03-20 2018-03-20 Verfahren zur Verbesserung und/oder Steuerung der Zelladhäsion und medizinisches Implantat, Zellkultursystem oder Zellassay mit verbesserter Zelladhäsion
DE102018204203.9 2018-03-20

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WO2019179978A1 true WO2019179978A1 (fr) 2019-09-26

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DE (1) DE102018204203A1 (fr)
WO (1) WO2019179978A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102005054941A1 (de) 2005-11-17 2007-05-31 Gelita Ag Nervenleitschiene
WO2010075933A1 (fr) * 2008-12-08 2010-07-08 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Substrats permettant de sélectionner et d'influencer spécifiquement le fonctionnement de cellules
EP2662054A1 (fr) * 2012-05-07 2013-11-13 SNU R&DB Foundation Iris artificiel à régulation automatique et son procédé de fabrication

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102005054941A1 (de) 2005-11-17 2007-05-31 Gelita Ag Nervenleitschiene
WO2010075933A1 (fr) * 2008-12-08 2010-07-08 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Substrats permettant de sélectionner et d'influencer spécifiquement le fonctionnement de cellules
EP2662054A1 (fr) * 2012-05-07 2013-11-13 SNU R&DB Foundation Iris artificiel à régulation automatique et son procédé de fabrication

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
PHILIPP SCHENDZIELORZ ET AL: "A polydopamine peptide coating enables adipose-derived stem cell growth on the silicone surface of cochlear implant electrode arrays : A POLYDOPAMIN PEPTIDE COATING ENABLES ASC ADHESION", JOURNAL OF BIOMEDICAL MATERIALS RESEARCH. PART B: APPLIED BIOMATERIALS, vol. 106, no. 4, 22 July 2017 (2017-07-22), US, pages 1431 - 1438, XP055595896, ISSN: 1552-4973, DOI: 10.1002/jbm.b.33947 *
SCHENDZIERLORZ: "Kollagen-Polydopamin-Beschichtung und Besiedelung von Cochlea Implantaten mit adipose-derived stem cells", DEUTSCHE GESELLSCHAFT FÜR HALS-NASEN-OHREN-HEILKUNDE, KOPF- UND HALS-CHIRURGIE. 88. JAHRESVERSAMMLUNG DER DEUTSCHEN GESELLSCHAFT FÜR HALS-NASEN-OHREN-HEILKUNDE, KOPF-UND HALS-CHIRURGIE, 24 May 2017 (2017-05-24)

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