CN116159182A - Nerve organoid-hydrogel system for treating spinal cord injury and preparation method thereof - Google Patents
Nerve organoid-hydrogel system for treating spinal cord injury and preparation method thereof Download PDFInfo
- Publication number
- CN116159182A CN116159182A CN202310076451.2A CN202310076451A CN116159182A CN 116159182 A CN116159182 A CN 116159182A CN 202310076451 A CN202310076451 A CN 202310076451A CN 116159182 A CN116159182 A CN 116159182A
- Authority
- CN
- China
- Prior art keywords
- hydrogel
- nerve
- organoid
- spinal cord
- cord injury
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000000017 hydrogel Substances 0.000 title claims abstract description 93
- 210000005036 nerve Anatomy 0.000 title claims abstract description 55
- 208000020431 spinal cord injury Diseases 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 210000002220 organoid Anatomy 0.000 claims abstract description 46
- 239000002243 precursor Substances 0.000 claims abstract description 25
- 239000007788 liquid Substances 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 15
- 238000004132 cross linking Methods 0.000 claims abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims abstract description 8
- 230000008569 process Effects 0.000 claims abstract description 7
- 108010010803 Gelatin Proteins 0.000 claims description 47
- 239000008273 gelatin Substances 0.000 claims description 47
- 229920000159 gelatin Polymers 0.000 claims description 47
- 235000019322 gelatine Nutrition 0.000 claims description 47
- 235000011852 gelatine desserts Nutrition 0.000 claims description 47
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 19
- 229920002674 hyaluronan Polymers 0.000 claims description 19
- 229960003160 hyaluronic acid Drugs 0.000 claims description 19
- 238000012258 culturing Methods 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 239000003999 initiator Substances 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 5
- 210000004263 induced pluripotent stem cell Anatomy 0.000 claims description 5
- 238000005286 illumination Methods 0.000 claims description 4
- 210000001671 embryonic stem cell Anatomy 0.000 claims description 3
- 229920001661 Chitosan Polymers 0.000 claims description 2
- 102000008186 Collagen Human genes 0.000 claims description 2
- 108010035532 Collagen Proteins 0.000 claims description 2
- 210000004504 adult stem cell Anatomy 0.000 claims description 2
- 229920001436 collagen Polymers 0.000 claims description 2
- 229920000747 poly(lactic acid) Polymers 0.000 claims description 2
- 239000004626 polylactic acid Substances 0.000 claims description 2
- 230000008929 regeneration Effects 0.000 abstract description 14
- 238000011069 regeneration method Methods 0.000 abstract description 14
- 230000006378 damage Effects 0.000 abstract description 11
- 210000001519 tissue Anatomy 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 8
- 230000004083 survival effect Effects 0.000 abstract description 7
- 210000003169 central nervous system Anatomy 0.000 abstract description 6
- 230000010261 cell growth Effects 0.000 abstract description 4
- 239000002131 composite material Substances 0.000 abstract description 4
- 238000000338 in vitro Methods 0.000 abstract description 4
- 230000014759 maintenance of location Effects 0.000 abstract description 4
- 238000013528 artificial neural network Methods 0.000 abstract description 3
- 210000000653 nervous system Anatomy 0.000 abstract description 3
- 230000012292 cell migration Effects 0.000 abstract description 2
- 238000003860 storage Methods 0.000 abstract description 2
- 230000001537 neural effect Effects 0.000 description 20
- 210000000278 spinal cord Anatomy 0.000 description 14
- 210000004027 cell Anatomy 0.000 description 12
- 239000002609 medium Substances 0.000 description 10
- 238000011084 recovery Methods 0.000 description 9
- 238000002054 transplantation Methods 0.000 description 9
- 208000027418 Wounds and injury Diseases 0.000 description 8
- 208000014674 injury Diseases 0.000 description 8
- 210000001178 neural stem cell Anatomy 0.000 description 8
- 210000002569 neuron Anatomy 0.000 description 8
- 210000000056 organ Anatomy 0.000 description 8
- 210000000130 stem cell Anatomy 0.000 description 8
- 241000700159 Rattus Species 0.000 description 7
- 230000004069 differentiation Effects 0.000 description 7
- 238000013508 migration Methods 0.000 description 5
- 230000003902 lesion Effects 0.000 description 4
- 230000005012 migration Effects 0.000 description 4
- 230000007659 motor function Effects 0.000 description 4
- 210000004126 nerve fiber Anatomy 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 3
- 230000007547 defect Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 210000003061 neural cell Anatomy 0.000 description 3
- 210000001778 pluripotent stem cell Anatomy 0.000 description 3
- 230000008439 repair process Effects 0.000 description 3
- 238000011476 stem cell transplantation Methods 0.000 description 3
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 2
- 102100039289 Glial fibrillary acidic protein Human genes 0.000 description 2
- 101710193519 Glial fibrillary acidic protein Proteins 0.000 description 2
- 101001092197 Homo sapiens RNA binding protein fox-1 homolog 3 Proteins 0.000 description 2
- 101150009249 MAP2 gene Proteins 0.000 description 2
- 102100035530 RNA binding protein fox-1 homolog 3 Human genes 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 238000002659 cell therapy Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 210000005046 glial fibrillary acidic protein Anatomy 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- -1 methacryloyl groups Chemical group 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 230000009689 neuronal regeneration Effects 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 230000000250 revascularization Effects 0.000 description 2
- 102100022928 DNA repair protein RAD51 homolog 1 Human genes 0.000 description 1
- 208000012661 Dyskinesia Diseases 0.000 description 1
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 101000620735 Homo sapiens DNA repair protein RAD51 homolog 1 Proteins 0.000 description 1
- 101000979001 Homo sapiens Methionine aminopeptidase 2 Proteins 0.000 description 1
- 101000969087 Homo sapiens Microtubule-associated protein 2 Proteins 0.000 description 1
- 102100021118 Microtubule-associated protein 2 Human genes 0.000 description 1
- QJJXYPPXXYFBGM-LFZNUXCKSA-N Tacrolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1\C=C(/C)[C@@H]1[C@H](C)[C@@H](O)CC(=O)[C@H](CC=C)/C=C(C)/C[C@H](C)C[C@H](OC)[C@H]([C@H](C[C@H]2C)OC)O[C@@]2(O)C(=O)C(=O)N2CCCC[C@H]2C(=O)O1 QJJXYPPXXYFBGM-LFZNUXCKSA-N 0.000 description 1
- ANCHPZQGZBCDBK-UHFFFAOYSA-N [PH2](O)=O.C1(=CC=CC=C1)C=1C(=C(C(=O)[Li])C(=CC1C)C)C Chemical compound [PH2](O)=O.C1(=CC=CC=C1)C=1C(=C(C(=O)[Li])C(=CC1C)C)C ANCHPZQGZBCDBK-UHFFFAOYSA-N 0.000 description 1
- 230000002164 acetylcholinergic effect Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 210000001130 astrocyte Anatomy 0.000 description 1
- 230000028600 axonogenesis Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000004641 brain development Effects 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000002932 cholinergic neuron Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000008611 intercellular interaction Effects 0.000 description 1
- 238000002684 laminectomy Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 210000001259 mesencephalon Anatomy 0.000 description 1
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 1
- DCUFMVPCXCSVNP-UHFFFAOYSA-N methacrylic anhydride Chemical compound CC(=C)C(=O)OC(=O)C(C)=C DCUFMVPCXCSVNP-UHFFFAOYSA-N 0.000 description 1
- 230000027939 micturition Effects 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 210000001428 peripheral nervous system Anatomy 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 210000004129 prosencephalon Anatomy 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 230000000862 serotonergic effect Effects 0.000 description 1
- 229940076279 serotonin Drugs 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 229960001967 tacrolimus Drugs 0.000 description 1
- QJJXYPPXXYFBGM-SHYZHZOCSA-N tacrolimus Natural products CO[C@H]1C[C@H](CC[C@@H]1O)C=C(C)[C@H]2OC(=O)[C@H]3CCCCN3C(=O)C(=O)[C@@]4(O)O[C@@H]([C@H](C[C@H]4C)OC)[C@@H](C[C@H](C)CC(=C[C@@H](CC=C)C(=O)C[C@H](O)[C@H]2C)C)OC QJJXYPPXXYFBGM-SHYZHZOCSA-N 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 208000037816 tissue injury Diseases 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 230000000472 traumatic effect Effects 0.000 description 1
Images
Classifications
-
- 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
- A61L27/3604—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 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/30—Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- 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
- A61L27/20—Polysaccharides
-
- 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
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/222—Gelatin
-
- 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
- A61L27/3641—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 characterised by the site of application in the body
- A61L27/3675—Nerve tissue, e.g. brain, spinal cord, nerves, dura mater
-
- 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
- A61L27/3683—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 subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—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 subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- 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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/32—Materials or treatment for tissue regeneration for nerve reconstruction
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Dermatology (AREA)
- Transplantation (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Neurosurgery (AREA)
- Molecular Biology (AREA)
- Neurology (AREA)
- Botany (AREA)
- Inorganic Chemistry (AREA)
- Zoology (AREA)
- Cell Biology (AREA)
- Developmental Biology & Embryology (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Virology (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Ophthalmology & Optometry (AREA)
- Dispersion Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Vascular Medicine (AREA)
- Urology & Nephrology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a nerve organ-hydrogel system for treating spinal cord injury and a preparation method thereof. The preparation method comprises the following steps: and uniformly mixing one or more nerve organoids with the precursor liquid of the hydrogel, and then performing a crosslinking reaction to form the nerve organoid-hydrogel system with a three-dimensional structure. The invention constructs the multipotent central nervous system organoid in vitro and composites the multipotent central nervous system organoid with hydrogel, can better ensure the simulated tissue structure, morphology and functional characteristics of the nervous system organoid in the storage and transportation processes, can form a nerve functional unit with a three-dimensional space structure and a larger size, can effectively prolong the retention time of the transplanted nerve organoid in a damaged area, improves the survival rate of the transplanted nerve organoid, supports cell migration and growth, improves the damage microenvironment, promotes nerve regeneration and neural network reconstruction, improves the exercise function and improves the treatment effect.
Description
Technical Field
The invention particularly relates to a nerve organ-hydrogel system for treating spinal cord injury, a preparation method and application thereof, and belongs to the technical field of medical treatment.
Background
The spinal cord is the central point of transmission of motor and sensory information between the brain and the peripheral nervous system, and is severely disabled after injury. The number of the existing traumatic spinal cord injury patients in China exceeds 200 ten thousand, and 10-14 ten thousand people are newly increased each year, so that a heavy burden is caused to families and society. Spinal cord injury repair is the most challenging regenerative medicine problem, and current clinical treatment protocols remain to reduce secondary injury and rehabilitation training.
A number of preclinical studies have shown that transplantation of stem cells (e.g., embryonic spinal cord-derived neural stem cells, embryonic brain-derived neural precursor cells, embryonic stem cells or neural precursor cells that induce differentiation of pluripotent stem cells, mesenchymal stem cells, etc.) can promote nerve regeneration and functional recovery after spinal cord injury (ACS biomatter. Sci. Eng.2020,6, 1671). Spinal cord formation is a highly ordered process in which stem cells proliferate, migrate, differentiate to produce multiple neural cells and establish complex neural circuits, however, single cell therapy has difficulty in mimicking multiple cell types and intercellular interactions in tissues, resulting in the difficulty of traditional stem cell transplantation therapy in mimicking the three-dimensional assembly characteristics of neural cells within spinal cord tissue in vivo. Therefore, how to realize nerve cell substitution and neural network reconstruction in the true sense of spinal cord tissue injury is the key of spinal cord injury cell treatment.
Organoid technology holds great promise for regenerative medicine. In recent years, several research teams internationally obtained brain-like organs that mimic the development of the cortex, as well as brain-region-specific brain-like organs such as forebrain, midbrain and hypothalamic-like organs, by differentiating pluripotent stem cells (pluripotent stem cells, PSC) by three-dimensional culture in vitro (nature.2013, 501:373;Cell Stem Cell.2016, 19:248; development.2019, 146:dev 166074). The brain-like organ provides a new research platform for researching brain development and disease processes and tissue regeneration, and the nerve-like organ comprises various nerve cell types and intercellular biological communication, provides a more suitable microenvironment for nerve regeneration, and can be better used for simulating the spinal cord generation process to promote injury repair.
On the other hand, the effect of simply transplanting stem cells to the spinal cord injury part is limited, mainly because the transplanted stem cells have short retention time in the injury area and are easily influenced by the injury microenvironment, thereby influencing the regeneration and repair effects. Transplanting stem cells and scaffold complexes is expected to improve therapeutic outcome. However, the superiority of central nervous organoids in treating diseases of spinal cord injury and lesions leading to dyskinesia, namely whether nerve regeneration and functional recovery after injury can be promoted has not been reported yet; meanwhile, in the existing research, stem cells and biological scaffolds are transplanted after being compounded, and considering that the central nervous organoids and the stem cells have obvious differences in some aspects, such as simulating three-dimensional multicellular composition of human central nervous tissues and providing a proper microenvironment, the central nervous organoids can achieve ideal treatment effects only by being compounded with the biological scaffolds, and the method is still the object of important research in the industry and is also the problem to be solved in the field.
Disclosure of Invention
The invention mainly aims to provide a nerve organ-hydrogel system for treating spinal cord injury, a preparation method and application thereof, so as to overcome the defects in the prior art.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
in one aspect, the invention provides a method of preparing a neuroorganoid-hydrogel system for treating spinal cord injury, comprising: and uniformly mixing one or more nerve organoids with the precursor liquid of the hydrogel, and then performing a crosslinking reaction to form the nerve organoid-hydrogel system with a three-dimensional structure.
In another aspect, the present invention provides a neuroorganoid-hydrogel system for treating spinal cord injury, which is prepared by the method.
In a further aspect the invention provides the use of the neuroorganoid-hydrogel system for the treatment of spinal cord injury.
Compared with the prior art, the invention has at least the following advantages:
(1) By constructing the multipotent central nervous system organoid in vitro and compounding the multipotent central nervous system organoid with hydrogel, on one hand, the simulated tissue structure, morphology and functional characteristics of the nervous system organoid can be better maintained, compared with cell transplantation, the nerve regeneration and nerve network reconstruction can be better realized, fusion with host tissues and vascularization are promoted, and on the other hand, a nerve functional unit with a three-dimensional space structure, which has larger size, meets the transplantation condition and fills the defect area, can be formed.
(2) The nerve organoid and the hydrogel are compounded to form a nerve organoid-hydrogel system, so that the retention time of the transplanted nerve organoid in a damaged area can be effectively prolonged, the survival rate of the transplanted nerve organoid is improved, the migration and the growth of cells are supported, the damage microenvironment is improved, the nerve regeneration and the nerve network reconstruction are promoted, the motor function is improved, and the treatment effect is improved.
Drawings
FIG. 1 shows the survival and migration levels of transplanted cells in a damaged area after different neural organoids have been cultured in accordance with one embodiment of the present invention.
Fig. 2a shows the level of promotion of nerve regeneration by different ratios of gelatin/hyaluronic acid hydrogel in an embodiment of the invention, wherein the neuronal markers: neuN and MAP2, nuclei: DAPI.
Fig. 2b shows the level of gelatin/hyaluronic acid hydrogel in different ratios to promote nerve fiber regeneration in an embodiment of the invention, wherein NF: neurofibrillary markers, GFAP: astrocyte markers, DAPI: and (3) cell nucleus.
Figure 2c shows that different ratios of gelatin/hyaluronic acid hydrogel increased the level of NeuN positive neuronal regeneration in an embodiment of the invention.
FIG. 2d shows that different ratios of gelatin/hyaluronic acid hydrogel increase Map2 positive neuronal regeneration levels in an embodiment of the invention.
FIG. 2e shows the increase in the number of NeuN and Map2 biscationic neurons with different ratios of gelatin/hyaluronic acid hydrogel in an embodiment of the invention.
FIG. 2f shows different ratios of gelatin/hyaluronic acid hydrogel to enhance NF+ nerve fiber growth in an embodiment of the invention.
FIG. 3 shows the survival of GFP-labeled neural stem cells and GFP-labeled neural organoids in the damaged area and integration with host spinal cord tissue in the control group (SCI), the hydrogel-grafted Group (GL), the hydrogel-entrapped neural stem cell-grafted group (hi-NSC-sc/GL), and the hydrogel-entrapped neural organoids-grafted group (hi-NSC-organoid/GL) after 10 days and 60 days of treatment in an embodiment of the invention.
FIG. 4 shows Tuj1 positive neurons regenerated in the lesion area in a control group (SCI), a hydrogel-grafted Group (GL), a hydrogel-entrapped neural stem cell-grafted group (hi-NSC-sc/GL), and a hydrogel-entrapped neural organoid-grafted group (hi-NSC-organoid/GL) 60 days after treatment in an embodiment of the invention.
FIG. 5 shows NF positive nerve fibers regenerated in spinal cord injury areas in a control group (SCI), a hydrogel-grafted Group (GL), a hydrogel-entrapped neural stem cell-grafted group (hi-NSC-sc/GL), and a hydrogel-entrapped neural organoid-grafted group (hi-NSC-organoid/GL) 60 days after treatment in an embodiment of the invention.
FIGS. 6 a-6 b show serotonin positive neurons and cholinergic neurons regenerated in the spinal cord injury zone in a control group (SCI), a hydrogel-grafted Group (GL), a hydrogel-entrapped neural stem cell grafted group (hi-NSC-sc/GL), and a hydrogel-entrapped neural organoid grafted group (hi-NSC-organoid/GL), respectively, 60 days after treatment in an embodiment of the invention.
FIG. 7 shows the angiogenesis in a control group (SCI), a hydrogel-grafted Group (GL), a hydrogel-entrapped neural stem cell-grafted group (hi-NSC-sc/GL), and a hydrogel-entrapped neural organoid-grafted group (hi-NSC-organoid/GL) after 60 days of treatment in an embodiment of the invention.
Fig. 8 a-8 b show the motor function recovery of rats in the control group (SCI), the hydrogel-grafted Group (GL), the hydrogel-entrapped neural stem cell-grafted group (hi-NSC-sc/GL), and the hydrogel-entrapped neural organoid-grafted group, respectively, after 60 days of treatment in an embodiment of the present invention, fig. 8a shows BBB scoring, and fig. 8b shows a swash plate experiment.
Detailed Description
Some embodiments of the present invention provide a method of preparing a neuroorganoid-hydrogel system for treating spinal cord injury comprising: and uniformly mixing one or more nerve organoids with the precursor liquid of the hydrogel, and then performing a crosslinking reaction to form the nerve organoid-hydrogel system with a three-dimensional structure.
In one embodiment, the preparation method comprises the following steps: the neuroorganoid is obtained by at least culturing one or more of embryonic stem cells, induced pluripotent stem cells, and adult stem cells.
In one embodiment, the preparation method specifically includes: culturing the induced pluripotent stem cells for 7-10 days to form a nerve organoid, uniformly mixing the nerve organoid with the precursor liquid of the hydrogel, and carrying out the crosslinking reaction.
In one embodiment, the hydrogel includes any one or more of gelatin hydrogel, collagen hydrogel, hyaluronic acid hydrogel, chitosan hydrogel, polylactic acid hydrogel, and the like, and is not limited thereto.
In one embodiment, the hydrogel comprises gelatin hydrogel and hyaluronic acid hydrogel in a mass ratio of 9:1 to 19:1.
In one embodiment, the precursor liquid of the hydrogel comprises methacryloylated gelatin and methacryloylated hyaluronic acid.
In one embodiment, the hydrogel precursor solution is present at a concentration of 5-10w/v% in a range that facilitates cell growth and migration and mass exchange.
In one embodiment, the precursor solution of the hydrogel further comprises 0.01-0.2w/v% of an initiator, which in this concentration range facilitates rapid crosslinking of the hydrogel in situ at the lesion and has a high biosafety. The initiator may be a photoinitiator, a thermal initiator, etc., which are common in the art, preferably a visible light initiator, such as phenyl-2, 4, 6-trimethylbenzoyl-lithium phosphinate (LAP), etc., and is not limited thereto.
In one embodiment, the preparation method specifically includes: the crosslinking reaction is stimulated by illumination.
In one embodiment, the density of the neuroorganoid in the precursor fluid of the hydrogel is 1 x 10 6 -1*10 7 This density range is more favorable for survival and differentiation of central nervous organoids in the damaged area and for participation in neural network remodeling.
In one embodiment, the neuroorganoid-hydrogel system is on the order of millimeters to centimeters in size to better meet the clinical treatment of acute and chronic spinal cord injury implant needs.
Some embodiments of the invention also provide a neuroorganoid-hydrogel system for treating spinal cord injury, which is made by the methods described herein.
Some embodiments of the invention also provide for the use of the neuroorganoid-hydrogel system for treating spinal cord injury in the manufacture of a medicament for treating spinal cord injury.
In one embodiment, the medicament may further include a pharmaceutically acceptable carrier, other bioactive factors, or pharmaceutical compounds, etc., and is not limited thereto.
The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The reagents and starting materials used in the following examples were all commercially available; the test methods in which specific conditions are not specified are generally conducted under conventional conditions or under conditions recommended by each manufacturer.
In the following examples, the process of culturing the neuroorganoid includes: single cell suspensions of human induced pluripotent stem cells were seeded at a density of 9000 cells/well into 96 well ultra low adhesion plates and 10. Mu.MY-27632 EB in the medium. The EB-forming medium was changed to one without Y-27632 every 2 days. After 5 days, EBs were transferred to 24-well ultra-low adhesion plates containing induction medium for differentiation. Organoids after 2 days of differentiation can be used for transplantation.
In the following examples, the preparation process of the precursor liquid of the hydrogel specifically includes: methacryloylated gelatin (mGL) and methacryloylated hyaluronic acid (mHA) were prepared by introducing methacryloyl groups onto the molecular chains of Gelatin (GL) and Hyaluronic Acid (HA), respectively, according to the methods described in the literature. mGL and mHA of different mass ratios were dissolved in PBS and small amounts of LAP were added, respectively, to prepare gelatin/hyaluronic acid precursor solutions (containing 0.02% w/v initiator) at a concentration of 6% w/v (total concentration of mGL and mHA), i.e. precursor solutions of hydrogels. Under photoinitiation condition, mGL and mHA can be crosslinked through a methacryloyl group to form a biomaterial scaffold, the biomaterial scaffold has good biocompatibility and gel forming performance, and the crosslinking degree and mechanical property can be regulated and controlled according to the grafting rate of methacrylic anhydride.
In the following examples, the preparation of the neuroorganoid-hydrogel system includes: the prepared multiple nerve organs were mixed with the hydrogel precursor solution uniformly, and a culture medium containing 10. Mu.MY-27632 EB was added to an ultra-low adhesion culture plate. The EB-forming medium was changed to one without Y-27632 every 2 days. After 5 days, EBs were transferred to 24-well ultra-low adhesion plates containing induction medium for differentiation. After differentiating for 2 days, the organoids are then cross-linked by illumination with a 5W LED lamp with an emission wavelength of 395nm for 90 seconds at room temperature to form three-dimensional neuroorganoid-hydrogel systems with a size ranging from millimeter to centimeter, which can be used for implantation to fill a large segment of injury.
Example 1 in this example, when culturing neuroorganoids, culture was performed in ultra-low adhesion plates in 10 mu M Y-27632 EB medium for 2 days, followed by 3 days in EB forming medium without Y-27632, and then EBs were transferred to 24-well ultra-low adhesion plates containing induction medium for 2 and 9 days. CollectingThe mass ratio of mGL to mHA contained in the precursor liquid of the hydrogel was 9:1. And, the precursor liquid of the nerve organ and the hydrogel is up to 2X 10 according to the cell concentration 6 The two neuroorganoid-hydrogel systems were prepared by mixing in the ratio of individual/mL followed by the light illumination described.
Thereafter, treatment of spinal cord injury using these neuroorganoid-hydrogel systems, specific procedures include: midline incisions were made in the backs of rats to expose the T8-T10 vertebrae. Laminectomy was performed at the T8-T9 vertebral level using a #11 surgical blade. The T8-T9 spinal cord was excised and resected, forming a 3mm full defect, and the bleeding at the spinal cord injury site was controlled with gelatin sponge. After 10 days of treatment, recovery from the spinal cord injury site was observed and the results are shown in FIG. 1. Wherein green fluorescent GFP corresponds to the transplanted organoids; white GFAP corresponds to host spinal cord tissue cells.
It can be seen that transplantation after culturing the nerve organoid for 7 days is better in survival and migration level of transplanted cells in damaged areas, which is helpful for functional recovery, compared to transplantation after culturing the nerve organoid for a long period of time.
In fact, similar experiments show that the treatment effect is obviously improved when the nerve organoid is transplanted after being cultured for 6-8 days compared with the nerve organoid after being cultured for 10-30 days.
Example 2 in this example, the precursor solution for the hydrogels used contained mGL and mHA in mass ratios of 10/0, 9/1, 8/2 and 7/3, respectively. The neuroorganoids were cultured for a total of 7 days in the manner described in reference to example 1. The preparation of the neuroorganoid-hydrogel system was also similar to example 1. Treatment of spinal cord injury using these hydrogel systems was performed in the same manner as in example 1.
After 60 days of treatment, recovery from the spinal cord injury site was observed and the results are shown in FIGS. 2 a-2 f. It can be seen that the neural differentiation is better promoted when the mass ratio of mGL and mHA is 9/1, thereby more effectively improving motor function recovery in spinal cord injured rats.
Example 3
In this embodiment, the nerve organoids are embedded in the cultureThe organ remained cultured in the expansion medium for 4 days, and at day 5, organoids were transferred to the differentiation medium and continued to be cultured until day 7, respectively. The mass ratio of mGL to mHA contained in the precursor liquid of the hydrogel is 9/1. And the precursor fluid of the nerve organoid and hydrogel is according to 2 x 10 6 Individual cells/mL were mixed and then subjected to the light described to produce a neuroorganoid-hydrogel system.
Thereafter, spinal cord injured rats were produced in the same manner as in example 1. And all spinal cord injured rats were randomly divided into four groups: control group (SCI), hydrogel-grafted Group (GL), hydrogel-entrapped neural stem cell grafted group (hi-NSC-sc/GL), and hydrogel-entrapped neural organoid grafted group (hi-NSC-organoid/GL). Control group: PBS was added to the spinal cord injury area; hydrogel graft group: transplanting gelatin/hyaluronic acid composite hydrogel in the spinal cord injury area; hydrogel-entrapped neural stem cell transplantation group: transplanting gelatin/hyaluronic acid composite hydrogel loaded with neural stem cells in the spinal cord injury area; hydrogel-entrapped nerve organoid graft group: the spinal cord injury area is transplanted with gelatin/hyaluronic acid composite hydrogel which is used for carrying nerve organoids. Finally, the muscle and skin layers are sutured. Tacrolimus and assisted urination are injected daily after surgery.
After 10 and 60 days of treatment of spinal cord injury by the transplantation of the neuroid-hydrogel system, more cells survived the transplantation of the neuroid-hydrogel system in the injured area and integrated with the host spinal cord tissue compared to the transplantation of the neural stem cell-hydrogel system (fig. 3). More surviving cells in the neuroorganoid-hydrogel system will effectively enhance the efficacy of cell therapy.
After 60 days of spinal cord injury treatment with the transplanted neuroid-hydrogel system, more Tuj positive neural cell regeneration and NF+ nerve fiber growth were observed in the hydrogel-entrapped neuroid-transplanted group (hi-NSC-organoid/GL) compared to the control group (SCI), the hydrogel-transplanted Group (GL), and the hydrogel-entrapped neural stem cell transplanted group (hi-NSC-sc/GL) (FIGS. 4 and 5). This suggests that the transplanted nerve organoid-hydrogel system may be more effective in promoting nerve regeneration and axon growth following injury. More importantly, the transplanted neuroid-hydrogel system can better promote spinal cord functional neurons such as serotonergic neurons (5-HT, see FIG. 6 a) and acetylcholinergic neurons (Chat, see FIG. 6 b).
In addition, the transplanted neuroorganoid-hydrogel system can better promote the regeneration of blood vessels in the spinal cord injury area. The hydrogel-entrapped neural stem cell transplantation group (hi-NSC-sc/GL) showed increased endothelial cells and vessel detail immunostained with the central specific marker RECA for spinal cord injury lesions (fig. 7), indicating that it promotes revascularization. Increased revascularization of the damaged area will be more beneficial to nerve regeneration.
The motor function recovery effect of transplanted neuroorganoid-hydrogel systems after full transection injury of spinal cord was assessed by measuring Basso-Beattie-Bresnahan (BBB) scores and a swash plate experiment (fig. 8 a-8 b). The BBB score and the swash plate angle of the hydrogel-entrapped nerve-like organ-transplanted rats (hi-NSC-organoid/GL) were both greatly increased compared to the control group (SCI), the hydrogel-transplanted Group (GL) and the hydrogel-entrapped nerve stem cell-transplanted group (hi-NSC-sc/GL), with an increase in the BBB average score of about 9.5, indicating that some rats occasionally walk, and the swash plate average angle increased to 33 °, indicating an increase in muscle strength. The transplanted nerve organoid-hydrogel system can better promote functional recovery after spinal cord injury.
The embodiment of the invention constructs the multipotent central nervous system organoid in vitro and compounds the multipotent central nervous system organoid with hydrogel, can better ensure the simulated tissue structure, morphology and functional characteristics of the nervous system organoid in the storage and transportation processes, can form a nerve functional unit with a three-dimensional space structure and a larger size, can effectively prolong the retention time of the transplanted nerve organoid in a damaged area, improve the survival rate of the transplanted nerve organoid, support the migration and growth of cells, improve the damage microenvironment, promote the regeneration of nerves and the reconstruction of a nerve network, improve the exercise function and improve the treatment effect.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (10)
1. A method of preparing a neuroorganoid-hydrogel system for treating spinal cord injury, comprising: and uniformly mixing one or more nerve organoids with the precursor liquid of the hydrogel, and then performing a crosslinking reaction to form the nerve organoid-hydrogel system with a three-dimensional structure.
2. The preparation method according to claim 1, characterized by comprising: the neuroorganoid is obtained by at least culturing one or more of embryonic stem cells, induced pluripotent stem cells, and adult stem cells.
3. The preparation method according to claim 2, characterized by comprising: culturing the induced pluripotent stem cells for 7-10 days to form a nerve organoid, uniformly mixing the nerve organoid with the precursor liquid of the hydrogel, and carrying out the crosslinking reaction.
4. A production method according to any one of claims 1 to 3, characterized in that: the hydrogel comprises any one or more of gelatin hydrogel, collagen hydrogel, hyaluronic acid hydrogel, chitosan hydrogel and polylactic acid hydrogel.
5. The method of manufacturing according to claim 4, wherein: the hydrogel comprises gelatin hydrogel and hyaluronic acid hydrogel with the mass ratio of 9:1-19:1.
6. The method of manufacturing according to claim 5, wherein: the precursor liquid of the hydrogel comprises methacryloylated gelatin and methacryloylated hyaluronic acid; and/or the concentration of the precursor liquid of the hydrogel is 5-10w/v%.
7. The production method according to any one of claims 1 to 3, 5 to 6, characterized in that: the precursor liquid of the hydrogel also comprises 0.01-0.2w/v% of an initiator; the preparation method specifically comprises the following steps: the crosslinking reaction is stimulated by illumination.
8. The production method according to any one of claims 1 to 3, 5 to 6, characterized in that: the density of the nerve organoid in the precursor liquid of the hydrogel is 1 x 10 6 -1*10 7 individual/mL; and/or the neuroorganoid-hydrogel system is on the order of millimeters to centimeters in size.
9. A neuroorganoid-hydrogel system for treating spinal cord injury, characterized by: which is obtainable by the process according to any one of claims 1 to 8.
10. Use of a neuroorganoid-hydrogel system for treating spinal cord injury according to claim 9 in the manufacture of a medicament for treating spinal cord injury.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310076451.2A CN116159182B (en) | 2023-02-02 | 2023-02-02 | Nerve organoid-hydrogel system for treating spinal cord injury and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310076451.2A CN116159182B (en) | 2023-02-02 | 2023-02-02 | Nerve organoid-hydrogel system for treating spinal cord injury and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116159182A true CN116159182A (en) | 2023-05-26 |
| CN116159182B CN116159182B (en) | 2024-09-24 |
Family
ID=86415935
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310076451.2A Active CN116159182B (en) | 2023-02-02 | 2023-02-02 | Nerve organoid-hydrogel system for treating spinal cord injury and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116159182B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105378062A (en) * | 2013-05-08 | 2016-03-02 | 再生医学Tx有限责任公司 | Organoids comprising isolated renal cells and uses thereof |
| CN110741078A (en) * | 2017-06-13 | 2020-01-31 | 乔治-奥古斯特-哥廷根大学公共利益基金会 | Methods for producing bioengineered neuronal organoids (BENO) and uses thereof |
| US20200078493A1 (en) * | 2017-03-09 | 2020-03-12 | Georgia Tech Research Corporation | Synthetic hydrogel carriers for cellular structures, generation of organoids, and treatment of tissue injury |
| US20220389375A1 (en) * | 2019-11-06 | 2022-12-08 | UCL Business Ltd. | Extracellular Matrix Gels, and Organoid Cultures Comprising the Same |
-
2023
- 2023-02-02 CN CN202310076451.2A patent/CN116159182B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105378062A (en) * | 2013-05-08 | 2016-03-02 | 再生医学Tx有限责任公司 | Organoids comprising isolated renal cells and uses thereof |
| US20200078493A1 (en) * | 2017-03-09 | 2020-03-12 | Georgia Tech Research Corporation | Synthetic hydrogel carriers for cellular structures, generation of organoids, and treatment of tissue injury |
| CN110741078A (en) * | 2017-06-13 | 2020-01-31 | 乔治-奥古斯特-哥廷根大学公共利益基金会 | Methods for producing bioengineered neuronal organoids (BENO) and uses thereof |
| US20200208105A1 (en) * | 2017-06-13 | 2020-07-02 | Georg-August-Universität Göttingen | Methods of producing bioengineered neuronal organoids (benos) and uses thereof |
| US20220389375A1 (en) * | 2019-11-06 | 2022-12-08 | UCL Business Ltd. | Extracellular Matrix Gels, and Organoid Cultures Comprising the Same |
Non-Patent Citations (3)
| Title |
|---|
| HE SHEN ET AL.: "Advances in Biomaterial-Based Spinal Cord Injury Repair", 《ADVANCED FUNCTIONAL MATERIALS》, vol. 32, no. 13, pages 10 * |
| XINHAO ZHAO ET AL.: "Optimized, visible light-induced crosslinkable hybrid gelatin/hyaluronic acid scaffold promotes complete spinal cord injury repair", 《BIOMEDICAL MATERIALS》, vol. 17, pages 1 - 2 * |
| 卢新培: "《大气压非平衡等离子体射流 II 生物医学应用》", vol. 2021, 30 November 2021, 华中科技大学出版社 , pages: 133 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116159182B (en) | 2024-09-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Choi et al. | A 3D cell printed muscle construct with tissue-derived bioink for the treatment of volumetric muscle loss | |
| Liu et al. | Current methods for skeletal muscle tissue repair and regeneration | |
| US7601525B2 (en) | Alginate gel scaffold having a plurality of continuous parallel microtubular copper capillaries | |
| JP6677813B2 (en) | Novel tissue-engineered neural construction via microRNA genes and its use in repairing neural defects | |
| Jabbari et al. | The matrix reloaded: the evolution of regenerative hydrogels | |
| Mukhatyar et al. | Tissue engineering strategies designed to realize the endogenous regenerative potential of peripheral nerves | |
| CA2845516C (en) | Muscle tissue regeneration using muscle fiber fragments | |
| WO2018028224A1 (en) | Silk fibroin with different degradation velocity and use thereof | |
| Wang et al. | Functional acellular matrix for tissue repair | |
| Wang et al. | The spatial arrangement of cells in a 3D-printed biomimetic spinal cord promotes directional differentiation and repairs the motor function after spinal cord injury | |
| Tang et al. | China’s landscape in regenerative medicine | |
| Li et al. | Collagen-based bioinks for regenerative medicine: Fabrication, application and prospective | |
| US7560275B2 (en) | Compositions and methods for generating skin | |
| Hashemi et al. | Bilayer scaffolds for interface tissue engineering and regenerative medicine: a systematic reviews | |
| CN115501393A (en) | Hydrogel for repairing nerve defects and preparation method and application thereof | |
| EP1838240B1 (en) | Hyaluronic acid derivative and neural stem cells for sci and pnt regeneration | |
| CN111450119A (en) | Perinatal tissue-derived extracellular matrix hydrogel preparation for promoting organ injury repair | |
| CN116159182B (en) | Nerve organoid-hydrogel system for treating spinal cord injury and preparation method thereof | |
| CN112656816A (en) | Exosome-loaded fibrin glue and application thereof | |
| CN112494727A (en) | VEGF secretion promoting hydrogel/fibroin nanofiber/neural stem cell integrated transplant and preparation method and application thereof | |
| He et al. | Human menstrual blood-derived stem cells combined with a new 3D bioprinted composite scaffold for spinal cord injury treatment | |
| Zavan et al. | New 3D hyaluronan-based scaffold for in vitro reconstruction of the rat sciatic nerve | |
| US20220378844A1 (en) | Rotator cuff therapy using muscle fiber fragments | |
| JP7510651B2 (en) | Composite product for bone and joint regeneration of cartilage lesions | |
| Wang et al. | Dual‐Crosslinked Bioactive Hydrogel Scaffold for Accelerated Repair of Genital Tract Defect |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |