WO2024084040A1 - Solution aqueuse de bioencre destinée à être utilisée dans des applications de bio-impression à base de lumière, et procédés d'utilisation de la solution aqueuse de bioencre - Google Patents

Solution aqueuse de bioencre destinée à être utilisée dans des applications de bio-impression à base de lumière, et procédés d'utilisation de la solution aqueuse de bioencre Download PDF

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Publication number
WO2024084040A1
WO2024084040A1 PCT/EP2023/079273 EP2023079273W WO2024084040A1 WO 2024084040 A1 WO2024084040 A1 WO 2024084040A1 EP 2023079273 W EP2023079273 W EP 2023079273W WO 2024084040 A1 WO2024084040 A1 WO 2024084040A1
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Prior art keywords
aqueous
cells
bioink
light
bioink solution
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PCT/EP2023/079273
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English (en)
Inventor
Wei Zhu
Original Assignee
Cellink Bioprinting Ab
Allegro 3D Inc
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Application filed by Cellink Bioprinting Ab, Allegro 3D Inc filed Critical Cellink Bioprinting Ab
Publication of WO2024084040A1 publication Critical patent/WO2024084040A1/fr

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    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03F—PHOTOMECHANICAL PRODUCTION OF TEXTURED OR PATTERNED SURFACES, e.g. FOR PRINTING, FOR PROCESSING OF SEMICONDUCTOR DEVICES; MATERIALS THEREFOR; ORIGINALS THEREFOR; APPARATUS SPECIALLY ADAPTED THEREFOR
    • G03F7/00—Photomechanical, e.g. photolithographic, production of textured or patterned surfaces, e.g. printing surfaces; Materials therefor, e.g. comprising photoresists; Apparatus specially adapted therefor
    • G03F7/0037—Production of three-dimensional images
    • 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
    • A61L27/16—Macromolecular materials obtained 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
    • 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
    • A61L27/52—Hydrogels or hydrocolloids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y70/00—Materials specially adapted for additive manufacturing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y80/00—Products made by additive manufacturing
    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03F—PHOTOMECHANICAL PRODUCTION OF TEXTURED OR PATTERNED SURFACES, e.g. FOR PRINTING, FOR PROCESSING OF SEMICONDUCTOR DEVICES; MATERIALS THEREFOR; ORIGINALS THEREFOR; APPARATUS SPECIALLY ADAPTED THEREFOR
    • G03F7/00—Photomechanical, e.g. photolithographic, production of textured or patterned surfaces, e.g. printing surfaces; Materials therefor, e.g. comprising photoresists; Apparatus specially adapted therefor
    • G03F7/004—Photosensitive materials
    • G03F7/027—Non-macromolecular photopolymerisable compounds having carbon-to-carbon double bonds, e.g. ethylenic compounds
    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03F—PHOTOMECHANICAL PRODUCTION OF TEXTURED OR PATTERNED SURFACES, e.g. FOR PRINTING, FOR PROCESSING OF SEMICONDUCTOR DEVICES; MATERIALS THEREFOR; ORIGINALS THEREFOR; APPARATUS SPECIALLY ADAPTED THEREFOR
    • G03F7/00—Photomechanical, e.g. photolithographic, production of textured or patterned surfaces, e.g. printing surfaces; Materials therefor, e.g. comprising photoresists; Apparatus specially adapted therefor
    • G03F7/004—Photosensitive materials
    • G03F7/027—Non-macromolecular photopolymerisable compounds having carbon-to-carbon double bonds, e.g. ethylenic compounds
    • G03F7/028—Non-macromolecular photopolymerisable compounds having carbon-to-carbon double bonds, e.g. ethylenic compounds with photosensitivity-increasing substances, e.g. photoinitiators
    • G03F7/029—Inorganic compounds; Onium compounds; Organic compounds having hetero atoms other than oxygen, nitrogen or sulfur

Definitions

  • the present disclosure relates to an aqueous bioink solution for use in light-based bioprinting applications, a method of using the aqueous bioink solution including live cells and a method of using the aqueous bioink solution without live cells (acellularly). More specifically, the disclosure relates to an aqueous bioink solution for use in light-based bioprinting applications, and methods of using the aqueous bioink solution as defined in the introductory parts of the independent claims.
  • 3D bioprinting technologies have become the forefront in innovative tools to create customizable scaffolds or tissues with varying degrees of complexity for applications in tissue engineering, regenerative medicine, microfluidics, drug discovery, etc.
  • 3D bioprinting platforms such as inkjet bioprinting, extrusion bioprinting, digital light processing (DLP) bioprinting, laser assisted bioprinting [1], Among them, DLP bioprinting stands out for its superior printing speed, resolution, scalability, and throughput in creating highly complex 3D tissues, biomimetic scaffolds as well as microfluidic chips.
  • DLP bioprinting employs a digital micromirror device (DMD), composed of millions of micromirrors, to modulate the light patterns projected onto the photopolymer solution.
  • the exposed area of the photopolymer solution will be photo-crosslinked.
  • a 3D structure can be printed either layer-by-layer or continuously [2- 6].
  • the design of the projected light patterns can be derived from computer aided design (CAD) models as well as medical imaging data such as magnetic resonance imaging (MRI) and computerized tomography (CT) scans, enabling the fabrication of highly complex biomedical devices (e.g., microfluidic chips) and patient-specific tissue models or implants.
  • CAD computer aided design
  • MRI magnetic resonance imaging
  • CT computerized tomography
  • light absorbers e.g., tartrazine, quinoline yellow
  • the photoinitiator Another key component of the prepolymer solution for light-based bioprinting (including DLP bioprinting and laser assisted bioprinting) is the photoinitiator. Because the prepolymer solution for bioprinting is typically aqueous to allow incorporation of live cells during or post the bioprinting process, the photoinitiator must be water soluble. More importantly, to bioprint functional tissues and scaffolds for in vitro and in vivo biomedical applications (e.g., tissue engineering, drug testing, transplantation, cell and tissue therapies, etc.), the photoinitiator must be biocompatible with minimal cytotoxicity and minimal adverse side effects to human health.
  • Lithium acylphosphinate (LAP), a type of lithium salt, has been used together with light absorbers (i.e., tartrazine, quinoline yellow) in the aqueous prepolymer solution for fabricating complex 3D hydrogels [6,8,9].
  • light absorbers i.e., tartrazine, quinoline yellow
  • lithium salts are known to cause kidney injury and adverse effects to the neurological and renal systems of the human body[10,ll].
  • Lithium ions are known to accumulate in the kidney collecting duct cells leading to kidney injury and disease, which is a common side effect of lithium treatment for psychiatric disorders [10,12,13].
  • the cause of such lithium cytotoxicity is that the influx sodium channels on the cell membranes have higher affinity for lithium ions than sodium ions, but the sodium efflux ATPase pumps have lower affinity for lithium than sodium, which leads to excess accumulation of lithium within the cells [10,12,13],
  • Metal-AP metal-acylphosphinate
  • physiologically relevant metal ions include, but are not limited to, sodium, magnesium, calcium, and potassium.
  • sodium and magnesium acylphosphinates have been synthesized in literature [9], to the inventor's knowledge, there have been no published studies that combine them with light absorber in a prepolymer solution to bioprint high- fidelity complex 3D acellular scaffolds or cell-laden tissue constructs using light based bioprinting platforms for tissue engineering and regenerative medicine applications. Also, there are no commercial products available for any other metal-acylphosphinates than those including lithium.
  • an aqueous bioink solution for use in light-based bioprinting applications, comprising: 0.5-95 w/v (%) water-soluble photocrosslinkable prepolymer; 0.001-5 w/v (%) biocompatible metal acylphosphinate photoinitiator; 0.001-10 w/v (%) biocompatible light absorber; and 5 to 99.5 w/v (%) solvent.
  • aqueous bioink solution according to the first aspect on light-based bioprinting platforms to create acellular tissues or hydrogel constructs or scaffolds, such as for in vivo implantation, in vitro cell seeding and therapeutic applications, wherein the aqueous bioink solution with desired composition is loaded directly on the light-based bioprinting platforms to create acellular 3D tissues, hydrogel constructs or scaffolds and thereafter using the 3D tissues, hydrogel constructs or scaffolds for in vivo implantation, in vitro cell seeding or therapeutic applications.
  • AP refers to acylphosphinate(s).
  • biocompatible material By a “biocompatible” material is meant that the material is not harmful or toxic to living tissue.
  • the biocompatibility of a material, such as a bioink or ingredients of a bioink may depend on e.g., the cell type, other components of the bioink and printing conditions.
  • tissues, hydrogel constructs or scaffolds is, in the context of this disclosure, meant any 3D-bioprinted product or structure, with or without cells, produced by using the aqueous bioink solution of the present disclosure.
  • Figure 1 shows Live/dead viability results of C2C12 cells bioprinted in Na-AP (left) and LAP (right) based bioinks.
  • Cells in Na-AP based bioink demonstrated healthier morphology than those in LAP based bioink at Day 1 post bioprinting.
  • Figure 2 shows printing results comparing Na-AP vs LAP based bioinks.
  • Figure 3 discloses devices and 3D structures printed with bioinks composed of two types of PEGDA, Na-AP and light absorber using a DLP bioprinter, (a-b) Microfluidic chips with horizontal channels. The hollow horizonal channels in the devices were perfused with red and blue dyes to demonstrate the high printing fidelity, (c) A free-standing lattice structure, (d) A free-standing mesh stent.
  • an improved aqueous bioink solution for use in light-based bioprinting applications comprising: 0.5-95 w/v (%) water-soluble photocrosslinkable prepolymer; 0.001-5 w/v (%) biocompatible metal acylphosphinate photoinitiator; 0.001-10 w/v (%) biocompatible light absorber; and 5 to 99.5 w/v (%) solvent.
  • the present disclosure relates to biocompatible aqueous bioink solutions with at least a water soluble photocrosslinkable prepolymer, a biocompatible metal acylphosphinate photoinitiator and a biocompatible light absorber.
  • the biocompatible metal acylphosphinate photoinitiator is chosen from the group comprising Na-AP, Mg-AP, Ca-AP and K-AP.
  • the biocompatible metal acylphosphinate photoinitiator is Na-AP.
  • Na-AP has shown promising results in relation to the previously known alternative LAP.
  • the biocompatible light absorber is chosen from the group comprising quinoline yellow, ponceau 4r, sunset yellow, yellow food dyes, micro/nanoparticles, of different sizes and types, riboflavin, phenol red, curcumin, saffron, turmeric, beta carotene, carbon black and tartrazine.
  • the biocompatible light absorber is quinoline yellow.
  • potential toxic effects as for other regularly used light absorbers such as e.g., tartrazine [14] does not appear to be known for quinoline yellow, an improved biocompatibility is obtained.
  • quinoline yellow may exhibit a higher absorbance at 405 nm per gram than other light absorbers, such as tartrazine.
  • quinoline yellow may be used at a lower concentration than e.g., tartrazine, resulting in less material to be removed from the construct for better imaging, and/or less negative effects (if any).
  • the biocompatible metal acylphosphinate photoinitiator is Na-AP and the biocompatible light absorber is quinoline yellow.
  • a bioink having an improved toxicity profile, while maintaining or improving the bioprinting properties in light-based bioprinting applications, is provided.
  • an improved biocompatibility behaviour is obtained.
  • Na-AP is used in a concentration of 0.001-5.0% wt, or 0.01-2% wt, or 0.1-l%wt, or 0.5-1.0% wt
  • quinoline yellow is used in a concentration of 0.001-10% wt, or 0.01-1% wt, or 0.05-0.2% wt, respectively.
  • the water-soluble prepolymer is chosen from monomers or oligomers with photocrosslinkable moieties such as vinyl, acrylic, arylamide, acrylate, thiol-ene and epoxide.
  • the water-soluble prepolymer is chosen from polyethylene glycol)diacrylate, gelatin methacrylate, methacrylated hyaluronic acid, methacrylated alginate, methacrylated silk, and mixtures of two or more of PEGDA, GelMA, HAMA, AlgMA and Silk- MA.
  • PEGDA is chosen from the group comprising 700, IK, 3.4K, 6K, 10K and 20K MW. Other PEGDA prepolymer molecular weights may also be used.
  • the solvent is chosen from the group comprising water, phosphate- buffered saline and cell culture medium.
  • Other solvents may also be used, depending on application and context.
  • cell culture media may comprise ingredients such as carbohydrates, amino acids, vitamins, minerals, and a pH buffer system. Other ingredients may also be included, e.g., depending on application and cell type.
  • Table 1 lists representative ink formulations for the disclosed biocompatible aqueous bioink solutions.
  • the solution further comprises living cells.
  • the aqueous bioink solutions can be used in applications requiring living cells, which can be added prior to or after bioprinting depending on application.
  • the living cells are chosen from the group comprising cells or cell lines of human and/or animal origin, being primary cells, immortalized and iPSC- or ESC- derived, such as keratinocytes, melanocytes, fibroblasts, sebocytes, dendritic cells, macrophages, stem cells, induced pluripotent stem cells, adipocytes, glandular cells or follicle cells, as well as myoblast cells, hepatic cells, human primary renal proximal tubule epithelial cells, collecting duct cells or adipose-derived mesenchymal stem cells. Any other cells and/or cell types may also be used.
  • the cells may for example be related to dermal, hepatic, cardiac, kidney, lung, muscle, cartilage or neural tissue. Any other tissue types may also be used.
  • the aqueous bioink solution may comprise one or more additional ingredients, such as cryoprotectants, DMEM media, serum, proteins, lipids, nucleic acids, carbohydrates, hormones, glycosaminoglycans, collagens, methacrylate collagen, gelatin, cellulose, na nofi brilla r cellulose, alginate, chitosan, acacia gum, tara gum, glucomannan, pectin, locust bean gum, guar gum, carrageenan or tragacanth, elastin, proteoglycans, aggrecans, isolated laminins, glycol-aminoglycans, such as hyaluronic acid and heparin, growth factors, thickeners, such as a polysaccharide-based substance, such as nanocellulose, glucomannan, xanthan gum, gellan gum
  • additional ingredients such as cryoprotectants, DMEM media, serum, proteins, lipids, nucleic acids
  • a typical bioink for the present disclosure is produced by mixing the selected components from the above table.
  • Some of the components e.g., prepolymers, light absorbers, solvents and any additional ingredients
  • metal acylphosphinate photoinitiators see e.g., [9], which is hereby incorporated as a reference.
  • light-based bioprinting refers to methods utilizing light and/or a light pattern to photo-crosslink 3D printed layers and structures, including digital light processing (DLP) bioprinting, laser assisted bioprinting, stereolithography (SLA) bioprinting and two photon polymerization (TPP) bioprinting.
  • DLP digital light processing
  • SLA stereolithography
  • TPP two photon polymerization
  • the disclosed biocompatible aqueous bioink solutions can be used on light-based bioprinting platforms to create hydrogel constructs for cell encapsulation or seeding for in vitro tissue engineering, disease modelling, drug discovery, etc.
  • the above disclosed biocompatible aqueous bioink solutions can also be used on light-based bioprinting platforms to create cell-laden tissues or acellular hydrogel constructs for in vivo implantation and therapeutic applications.
  • bioink solution with desired compositions can be loaded directly on the light-based bioprinting platforms to create 3D scaffolds, which can be subsequently used for in vivo implantation or in vitro cell seeding.
  • biocompatible aqueous bioink solutions can be used for fabrication of various 3D tissue models, such as cardiac tissues, kidney tissues, lung tissues, nerve tissues, muscle tissues, cartilage tissues, skin tissues, hepatic tissues etc.
  • tissue products can feature biomimetic 3D geometries as well as cellular compositions mimicking native tissues and in vivo environment, which cannot be achieved by conventional 2D cell cultures.
  • a method of using the aqueous bioink solution according to the first aspect on light-based bioprinting platforms for the creation of cell-laden tissues, hydrogel constructs or scaffolds, such as for cell encapsulation, seeding, in vitro tissue engineering, disease modelling or drug discovery wherein cells are mixed with the aqueous bioink solution to the desired concentration and then loaded to the bioprinting platform to create 3D tissues, hydrogel constructs or scaffolds containing cells, and thereafter using the 3D tissues, hydrogel constructs or scaffolds containing cells for cell encapsulation, seeding, in vitro tissue engineering, disease modelling or drug discovery.
  • a method of using the aqueous bioink solution according to the first aspect on light-based bioprinting platforms to create acellular tissues, hydrogel constructs or scaffolds, such as for in vivo implantation, in vitro cell seeding and therapeutic applications wherein the aqueous bioink solution with desired composition is loaded directly on the light-based bioprinting platforms to create acellular 3D tissues, hydrogel constructs or scaffolds and thereafter using the 3D tissues, hydrogel constructs or scaffolds for in vivo implantation, in vitro cell seeding or therapeutic applications.
  • a first novel aspect of the present disclosure is the use of Na-AP, Mg-AP, Ca-AP, and K- AP as photoinitiator in the disclosed bioinks, which are more biocompatible and physiologically relevant photoinitiators than LAP.
  • Na, Mg, Ca, and K are ions that naturally exist in human body fluid and are essential nutrients for human body. They are also essential components in cell culture medium. Therefore, using Na-AP, Mg-AP, Ca-AP, and K-AP as a photoinitiator supports better cell viability, proliferation, and differentiation than LAP.
  • the inventor used Na-AP as an example to compare its cytotoxicity with LAP.
  • the inventor formulated two bioinks to bioprint tissues encapsulated with myoblast cells (i.e., C2C12): (1) 5% GelMA + 0.25% Na-AP + 2 million/ml C2C12 cells (a type of myoblast cell); (2) 5% GelMA + 0.25% LAP + 2 million/ml C2C12 cells.
  • a DLP bioprinter (BIONOVA X, from CELLINK) was used to print tissue constructs with these two cellladen bioinks and compared cell viability on Day 1 post bioprinting.
  • a commercial cytotoxicity kit (LIVE/DEADTM Viability/Cytotoxicity Kit for mammalian cells, Catalog# L3224, ThermoFisher Scientific) was used to evaluate the cell viability. Staining and imaging were performed according to manufacturer's recommendations. As shown in Fig. 1, the cells encapsulated in the Na-AP based bioink demonstrated healthier cell morphology (cells more spread out) than the cells in the LAP based bioink (cells balled up).
  • the inventors have also planned using other cells to further compare and systematically investigate the cytotoxicity of LAP vs. Na-AP/Mg-AP/Ca-AP/K-AP.
  • Some representative cell types include but are not limited to, human primary renal proximal tubule epithelial cells and collecting duct cells, as lithium is known to target the renal system and kidney collecting duct cells. The cells will be exposed to following experimental conditions to evaluate their viability, morphology, proliferation, and metabolism:
  • Bioinks will be mixed in bioinks made with different photoinitiators (i.e., LAP vs Na-AP vs Mg-AP vs Ca-AP vs. K-AP) and immediately printed into tissues with light-based bioprinting (e.g., DLP bioprinting). Except for the photoinitiator, all the other components of the bioinks (i.e., prepolymer, light absorber, solvent, etc.) will be controlled to be the same. The printed tissues will be assayed after a certain time of in vitro culturing (e.g. 1, 4, 7 days after bioprinting).
  • photoinitiators i.e., LAP vs Na-AP vs Mg-AP vs Ca-AP vs. K-AP
  • light-based bioprinting e.g., DLP bioprinting
  • all the other components of the bioinks i.e., prepolymer, light absorber, solvent, etc.
  • the printed tissues
  • cell assay kits that can be used to evaluate the viability, morphology, proliferation, and metabolism include but are not limited to LIVE/DEADTM Viability/Cytotoxicity Kit, Alamar Blue assay, CyQuant Direct cell assay, Neutral Red Uptake cytotoxicity assay, MTT assay, Cell Painting assay, etc.
  • the inventor will also evaluate the impact of different photoinitiators on cell differentiation.
  • the inventor will use Adipose-Derived Mesenchymal Stem Cells (ADSCs, ATCC PCS-500-011) as the representative cell type for this study.
  • Cells will be mixed in bioinks made with different photoinitiators (i.e., LAP vs Na-AP vs Mg-AP vs Ca-AP vs. K- AP) and immediately printed into tissues with light-based bioprinting (e.g., DLP bioprinting). Except for the photoinitiator, all the other components of the bioinks (i.e., prepolymer, light absorber, solvent, etc.) will be controlled to be the same.
  • Adipocyte Differentiation Toolkit Adipocyte Differentiation Toolkit
  • ADSCs printed in bioinks made with Na-AP/Mg- AP/Ca-AP/K-AP are expected, as compared to those in LAP-based bioinks.
  • bioinks with different photoinitiators e.g., Na-AP vs. LAP
  • Na-AP/Mg-AP/Ca-AP/K-AP based bioinks without light absorber can be used for light-based printing, there is no control over the light penetration depth in axial direction, leading to very poor axial resolution (i.e., hundreds of microns to a few millimeters).
  • the low axial resolution limits the use of such bioinks without light absorber to only printing 2.5D structures with no or minimal design variation in the axial direction, such as the printing results shown in Figure 2.
  • complex 3D designs such as overhanging structures and horizontal hollow channels cannot be printed using light-based printing, especially DLP-based bioprinting.
  • biocompatible light absorber can be added to the bioink to reduce light penetration in the bioink solution and improve axial resolution.
  • biocompatible light absorbers include but are not limited to Quinoline yellow, ponceau 4R, sunset yellow, yellow food dyes, micro/nanoparticles, riboflavin, phenol red, curcumin, saffron, turmeric, beta carotene, carbon black, tartrazine etc.

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Abstract

La présente divulgation concerne une solution aqueuse de bioencre destinée à être utilisée dans des applications de bio-impression à base de lumière, comprenant : (a) 0,5 à 95 % en poids d'un prépolymère soluble dans l'eau ; (b) 0,001 à 5 % en poids d'un photoinitiateur à base d'acylphosphinate métallique biocompatible ; (c) 0,001 à 10 % en poids d'un absorbeur de lumière biocompatible ; et (d) 5 à 99,5 % en poids de solvant. La présente divulgation concerne en outre un procédé d'utilisation de la solution aqueuse de bioencre comprenant des cellules vivantes et un procédé d'utilisation de la solution aqueuse de bioencre sans cellules vivantes (de manière acellulaire).
PCT/EP2023/079273 2022-10-20 2023-10-20 Solution aqueuse de bioencre destinée à être utilisée dans des applications de bio-impression à base de lumière, et procédés d'utilisation de la solution aqueuse de bioencre WO2024084040A1 (fr)

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