CN113797229A - 一种用于抗炎和促组织再生的巨噬细胞来源外泌体制剂及其制备方法和应用 - Google Patents
一种用于抗炎和促组织再生的巨噬细胞来源外泌体制剂及其制备方法和应用 Download PDFInfo
- Publication number
- CN113797229A CN113797229A CN202111197930.7A CN202111197930A CN113797229A CN 113797229 A CN113797229 A CN 113797229A CN 202111197930 A CN202111197930 A CN 202111197930A CN 113797229 A CN113797229 A CN 113797229A
- Authority
- CN
- China
- Prior art keywords
- macrophage
- preparation
- derived
- exosome
- derived exosome
- 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.)
- Pending
Links
- 210000001808 exosome Anatomy 0.000 title claims abstract description 65
- 210000002540 macrophage Anatomy 0.000 title claims abstract description 63
- 238000002360 preparation method Methods 0.000 title claims abstract description 36
- 206010061218 Inflammation Diseases 0.000 title abstract description 16
- 230000004054 inflammatory process Effects 0.000 title abstract description 16
- 230000017423 tissue regeneration Effects 0.000 title abstract description 9
- 230000001737 promoting effect Effects 0.000 title abstract description 7
- 210000004322 M2 macrophage Anatomy 0.000 claims abstract description 20
- 239000000203 mixture Substances 0.000 claims abstract description 9
- 239000003814 drug Substances 0.000 claims abstract description 8
- 235000015872 dietary supplement Nutrition 0.000 claims abstract description 7
- 208000015122 neurodegenerative disease Diseases 0.000 claims abstract description 7
- 239000000047 product Substances 0.000 claims abstract description 6
- 208000027866 inflammatory disease Diseases 0.000 claims abstract description 4
- 238000009472 formulation Methods 0.000 claims abstract 8
- 238000000034 method Methods 0.000 claims description 18
- 210000004027 cell Anatomy 0.000 claims description 15
- 239000012528 membrane Substances 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 5
- 230000000451 tissue damage Effects 0.000 claims description 5
- 231100000827 tissue damage Toxicity 0.000 claims description 5
- 241001465754 Metazoa Species 0.000 claims description 4
- 241000699666 Mus <mouse, genus> Species 0.000 claims description 4
- 239000003937 drug carrier Substances 0.000 claims description 4
- 230000036541 health Effects 0.000 claims description 4
- 239000000017 hydrogel Substances 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 239000002244 precipitate Substances 0.000 claims description 4
- 108090000978 Interleukin-4 Proteins 0.000 claims description 3
- 238000001085 differential centrifugation Methods 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 239000000243 solution Substances 0.000 claims description 3
- 241000282693 Cercopithecidae Species 0.000 claims description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 2
- 241001494479 Pecora Species 0.000 claims description 2
- 241000282898 Sus scrofa Species 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 239000002674 ointment Substances 0.000 claims description 2
- 239000008055 phosphate buffer solution Substances 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims 1
- 239000002417 nutraceutical Substances 0.000 claims 1
- 235000021436 nutraceutical agent Nutrition 0.000 claims 1
- 230000029663 wound healing Effects 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 6
- 229940079593 drug Drugs 0.000 abstract description 4
- 239000000463 material Substances 0.000 abstract description 4
- 238000002659 cell therapy Methods 0.000 abstract description 2
- 230000008105 immune reaction Effects 0.000 abstract description 2
- 210000003690 classically activated macrophage Anatomy 0.000 description 22
- 238000011529 RT qPCR Methods 0.000 description 17
- 108090000623 proteins and genes Proteins 0.000 description 17
- 238000001262 western blot Methods 0.000 description 12
- 206010052428 Wound Diseases 0.000 description 11
- 208000027418 Wounds and injury Diseases 0.000 description 11
- 238000001514 detection method Methods 0.000 description 9
- 102000004169 proteins and genes Human genes 0.000 description 8
- 239000003550 marker Substances 0.000 description 7
- 101710089543 Nitric oxide synthase, inducible Proteins 0.000 description 6
- 230000007547 defect Effects 0.000 description 6
- 230000002757 inflammatory effect Effects 0.000 description 6
- 230000008672 reprogramming Effects 0.000 description 6
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000003556 assay Methods 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 238000003125 immunofluorescent labeling Methods 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 230000000770 proinflammatory effect Effects 0.000 description 3
- 238000010254 subcutaneous injection Methods 0.000 description 3
- 239000007929 subcutaneous injection Substances 0.000 description 3
- 238000010200 validation analysis Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 102000004452 Arginase Human genes 0.000 description 2
- 108700024123 Arginases Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 239000002033 PVDF binder Substances 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 239000006180 TBST buffer Substances 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 238000001574 biopsy Methods 0.000 description 2
- 230000007910 cell fusion Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 230000003111 delayed effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 230000035876 healing Effects 0.000 description 2
- 230000023597 hemostasis Effects 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 210000004969 inflammatory cell Anatomy 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000005199 ultracentrifugation Methods 0.000 description 2
- 230000037314 wound repair Effects 0.000 description 2
- 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 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 102100025222 CD63 antigen Human genes 0.000 description 1
- 101100298998 Caenorhabditis elegans pbs-3 gene Proteins 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 101000934368 Homo sapiens CD63 antigen Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 102100037850 Interferon gamma Human genes 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- 102000003777 Interleukin-1 beta Human genes 0.000 description 1
- 108090000193 Interleukin-1 beta Proteins 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 229940122907 Phosphatase inhibitor Drugs 0.000 description 1
- 238000010802 RNA extraction kit Methods 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 206010039710 Scleroderma Diseases 0.000 description 1
- 206010072170 Skin wound Diseases 0.000 description 1
- 206010047139 Vasoconstriction Diseases 0.000 description 1
- ABUBSBSOTTXVPV-UHFFFAOYSA-H [U+6].CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O Chemical compound [U+6].CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O ABUBSBSOTTXVPV-UHFFFAOYSA-H 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 238000000137 annealing Methods 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
- 238000009835 boiling Methods 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000010804 cDNA synthesis Methods 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000002296 dynamic light scattering Methods 0.000 description 1
- 230000007608 epigenetic mechanism Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 238000003205 genotyping method Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 208000037891 myocardial injury Diseases 0.000 description 1
- 201000008383 nephritis Diseases 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 201000008482 osteoarthritis Diseases 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- -1 poly (methyl vinyl acetate) -carbon Chemical compound 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 239000012474 protein marker Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 208000037816 tissue injury Diseases 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 230000025033 vasoconstriction Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Images
Classifications
-
- 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
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- 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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
-
- 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
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/04—Drugs for skeletal disorders for non-specific disorders of the connective tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biomedical Technology (AREA)
- Physical Education & Sports Medicine (AREA)
- Developmental Biology & Embryology (AREA)
- Epidemiology (AREA)
- Rheumatology (AREA)
- Immunology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Dermatology (AREA)
- Wood Science & Technology (AREA)
- Neurology (AREA)
- Genetics & Genomics (AREA)
- Urology & Nephrology (AREA)
- Food Science & Technology (AREA)
- Virology (AREA)
- Mycology (AREA)
- Polymers & Plastics (AREA)
- Pain & Pain Management (AREA)
- Heart & Thoracic Surgery (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Hematology (AREA)
- Cardiology (AREA)
- Nutrition Science (AREA)
Abstract
本发明公开了一种用于抗炎和促组织再生的巨噬细胞来源外泌体制剂及其制备方法和应用。所述制剂包括M2型巨噬细胞来源外泌体。本发明的巨噬细胞来源外泌体制剂可用于制备治疗炎性疾病、组织修复、退行性病变的药物、保健品或营养补充剂。本发明M2巨噬细胞外泌体制剂制备简单、材料来源广泛、储存方便，相对于细胞疗法，外泌体副作用小且不易引起免疫反应，应用后能调节炎症平衡、促进伤口愈合、减缓退变性疾病进展，可以多种形式应用，具有很好的临床应用价值。
Description
技术领域
本发明涉及生物医药技术领域，特别涉及一种用于抗炎和促组织再生的巨噬细胞来源外泌体制剂及其制备方法和应用。
背景技术
维持皮肤完整性对于机体至关重要，而组织损伤和修复是人体中极其复杂的过程，涉及各种分子机制及遗传表观学机制。到目前为止，已有研究证实了伤口愈合是通过四个精确且高度程序化的阶段实现的：止血、炎症、增殖和重塑。若这些阶段受到干扰，未能及时、适当地发生则会导致伤口延迟愈合或病理性伤口愈合。其中，炎症在生物学上被定义为"身体的免疫系统对某种刺激的反应"，主要是由各种病原体感染、或组织损伤引起的，被认为是机体的保护性反应，在组织修复中起着重要作用。因此，针对炎症的研究在改善和治疗慢性病领域具有良好的前景。
止血在组织损伤后会立即开始，伴随着血管收缩和纤维蛋白凝块的形成。一旦出血得以控制，即进入炎症阶段。该阶段在急性伤口中一般持续2-5天，而受种种因素影响，炎症期亦可持续数周或甚至数月，此时伤口转变为慢性伤口。早期伤口处的炎症可以起到保护伤口的作用，是对伤口愈合有利的，但一旦炎症迁延成慢性炎症，会使伤口转为慢性伤口，不利于伤口愈合，因此，调节控制促炎-抗炎平衡对伤口愈合很重要。
巨噬细胞是先天免疫细胞，在生理和病理过程中都扮演着极其重要的角色。在伤口修复的早期，巨噬细胞导致炎性细胞因子的产生以及病原体和碎片的清除。在伤口愈合炎症阶段的后期，巨噬细胞有助于炎症的消退并促进组织修复。巨噬细胞主要分为经典激活的促炎M1型巨噬细胞和交替激活的抗炎M2型巨噬细胞。M1巨噬细胞可分泌促炎因子如IL-1β、IL-6等促进炎症发展，而M2巨噬细胞则主要分泌IL-10、Arg-1等抑制炎症发展。因此，巨噬细胞表型的可塑性是皮肤伤口修复过程中重要的调控靶点，进而发明人认为，提供一种通过调节促炎-抗炎平衡从而治疗多种炎症相关疾病的制剂是非常有必要的。
发明内容
本发明为了解决上述技术问题，提供了一种用于抗炎和促组织再生的巨噬细胞来源外泌体制剂及其制备方法和应用。
第一方面，本申请提供了一种巨噬细胞来源外泌体制剂，是通过以下技术方案得以实现的。
一种巨噬细胞来源外泌体制剂，包括M2型巨噬细胞来源外泌体。
进一步的，制剂中M2型巨噬细胞来源外泌体的有效浓度大于等于50μg/mL。
进一步的，制剂总体积为200μL，外泌体含量大于等于10μg。
进一步的，M2型巨噬细胞来源外泌体的直径为30-150nm。优选的，M2型巨噬细胞来源外泌体的直径为50-90nm。
进一步的，所述M2型巨噬细胞来源于人或动物，所述动物选自鼠、兔、猪、羊或猴。
进一步的，所述制剂还包括药学上可接受的载体。
进一步的，所述药学上可接受的载体选自磷酸盐缓冲液和/或生理盐水，或具备缓释功能的水凝胶。所述具备缓释功能的水凝胶选自海藻酸盐水凝胶、壳聚糖、透明质酸、PEG中的一种或多种。
进一步的，所述制剂为注射剂、胶囊剂、片剂、粉末剂、软膏剂或喷雾剂。
进一步的，所述M2型巨噬细胞来源外泌体应用基因工程方法进行改造，优选为，负载具有特殊功能的mRNA、LcRNA的M2型巨噬细胞来源外泌体，得到具有特定功能的外泌体制剂。
第二方面，本申请提供了一种巨噬细胞来源外泌体制剂的制备方法，是通过以下技术方案得以实现的。
一种上述巨噬细胞来源外泌体制剂的制备方法，包括以下步骤：
S1.将M0巨噬细胞以1×105-5×105个的密度接进行接种，细胞贴壁后，用20-25ng/mL IL-4处理M0巨噬细胞48-60h，得到M2巨噬细胞；
S2.将M2巨噬细胞在无血清条件下孵育2-3d，收集上清，通过差速离心方法收集外泌体：4℃300-400g离心8-10min以移除细胞；2000-2500g离心8-10min以移除死细胞；130000-150000g离心90-100min以获取外泌体球；130000-150000g离心90-100min得到外泌体沉淀；用PBS溶液重悬外泌体沉淀，滤膜过滤后得到M2型巨噬细胞来源外泌体。
第三方面，本申请提供了巨噬细胞来源外泌体制剂的临床应用，是通过以下技术方案得以实现的。
一种上述巨噬细胞来源外泌体制剂在制备治疗炎症性疾病药物、保健品或营养补充剂中的应用。所述炎症性疾病包括类风湿性关节炎、肾炎、SLE或硬皮病。
一种上述巨噬细胞来源外泌体制剂在制备修复组织损伤药物、保健品或营养补充剂中的应用。所述组织损伤包括心梗后损伤，皮肤缺损、骨缺损、软骨损伤、肌肉损伤、肌腱损伤或神经缺损。
一种上述巨噬细胞来源外泌体制剂在制备治疗退变性疾病药物、保健品或营养补充剂中的应用。所述退变性疾病包括骨关节炎、脊柱退行性疾病。
本申请具有以下有益效果。
本发明采用M2型巨噬细胞外泌体引导的细胞重编程技术可实现从M1型巨噬细胞到M2型巨噬细胞的转变，从而使伤口部位通过促进血管生成、再上皮化和胶原沉积加速愈合。本发明M2巨噬细胞外泌体制剂制备简单、材料来源广泛、储存方便，相对于细胞疗法，外泌体副作用小且不易引起免疫反应，应用后能调节炎症平衡、促进伤口愈合、减缓退变性疾病进展，可以多种形式应用，具有很好的临床应用价值。
附图说明
图1是本发明RAW264.7细胞STR位点的基因分型结果图；
图2是本发明M2巨噬细胞的表型验证结果图(其中，A.Western Blot检测结果图；B.qRT-PCR检测结果图，**p<0.01)；
图3是本发明M2-Exo的表征结果图(其中，A.M2-Exo的TEM图；B.M2-Exo粒径的检测结果图；C.流式细胞分析术检测结果图；D.Western blot检测结果图)；
图4是本发明M1巨噬细胞的表型验证结果图(其中，A.Western Blot检测结果图；B.qRT-PCR检测结果图，**p<0.01)；
图5是本发明细胞摄取实验结果图；
图6是本发明M2-Exo重编程有效浓度的检测结果图(其中，A.不同浓度外泌体与M1巨噬细胞共培养后Western Blot检测结果图；B.不同浓度外泌体与M1巨噬细胞共培养后qRT-PCR检测结果图，**p<0.01；C.不同浓度外泌体与M1巨噬细胞共培养后免疫荧光染色图)；
图7是本发明M2-Exo重编程效应验证结果图(**p<0.01)；
图8是本发明M2-Exo处理促小鼠皮肤伤口愈合效果图；
图9是本发明皮下注射M2-Exo 7天后伤口的H&E染色图。
具体实施方式
以下结合附图和实施例对本发明进行进一步的说明。需要说明的是，下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
一、M2-Exo的制备
1、从武汉普诺赛生物公司购买小鼠单核巨噬细胞白血病细胞(RAW 264.7)，并进行相关鉴定(鉴定结果参见图1)，以确定为M0巨噬细胞。将细胞以1×105个的密度接种于75cm3培养瓶中，加入适量完全培养基，置于5％CO2，饱和湿度的37℃培养箱中常规培养，细胞融合度约为80％-90％时即可传代。
2、M2巨噬细胞的激活：将M0巨噬细胞以1×105个的密度接种于75cm3培养瓶中，细胞贴壁后，应用20ng/mL IL-4(R&D)处理M0巨噬细胞48h。
3、M2巨噬细胞的鉴定
3.1、qRT-PCR验证M2巨噬细胞相关基因的表达水平(Arginase)
使用RNA提取试剂盒提取总RNA，Nanodrop检测所提RNA浓度及纯度。按照试剂盒说明加入反转录酶，进行反转录及合成cDNA，-20℃保存。按照2×qPCR
Green Master的试剂盒说明配制反应体系，并上机检测。反应体系为：qPCR SYBR Green Master Mix10ul，上下游引物(参见表1，10uM)各0.4ul，cDNA 2ul，ddH2O 7.2ul。反应条件为：预变性95℃5min；变性95℃10s，退火56℃20s，延伸72℃20s，共50个循环；熔解曲线95℃15s,60℃1min,95℃1s。每组均进行3个重复，以GADPH为内参基因，使用2-△△Ct法测定相对mRNA表达水平。
表1 qRT-PCR引物序列信息
3.2、Western blot技术鉴定M2巨噬细胞特异性标志物(Arginase、CD206)
使用含有PMSF和磷酸酶抑制剂的RIPA裂解液，收集样品后置于冰上裂解1h，12000rpm 4℃离心20min，收集上清，并用BCA试剂盒进行蛋白定量。在上清中加入适量5xSDS蛋白上样缓冲液，95℃金属浴煮沸蛋白10min。配制PAGE凝胶，在浓缩胶中上样蛋白30μg，80V电压30min，120V电压60min进行蛋白电泳，待溴酚蓝跑出分离胶底部即可停止电泳。将PAGE凝胶转移到PVDF膜中，转膜220mA，90min。5％脱脂奶粉室温封闭1h后，按蛋白marker裁剪PVDF膜条带，在Arg-1、CD206一抗中4℃孵育过夜。TBST洗膜10min三次，辣根过氧化物酶标记的二抗室温孵育1h，TBST再次洗膜10min三次。加入ECL发光液，于化学发光显影系统中观察。
qRT-PCR和Western blot检测结果参见图2，从图2中可以看出，本申请制备得到的M2巨噬细胞能够表达M2表型相关基因Arg-1，且能够表达Arg-1、CD206特征性蛋白。
4、M2-Exo的分离：M2巨噬细胞在无血清条件下孵育2天。孵育2天后，收集上清，通过差速离心方法收集外泌体：4℃300g离心10min，移除细胞；2000g离心10min以移除死细胞；150000g离心90min以获取外泌体球。150000g离心90min得到外泌体沉淀。用1ml PBS溶液重悬外泌体沉淀，使用0.22μm滤膜过滤，用于后续实验。
5、M2-Exo的鉴定
直径：应用动态光散射技术(DLS,Zetasizer Nano,Malvern Instruments,UK)或Nanosight技术。
结构：应用2％多聚甲醛固定外泌体过夜，150000g离心30min后在无水乙醇中重悬。随后取2μL的悬液置于聚醋酸甲基乙烯脂-碳涂层的电镜格，电镜格预先已应用醋酸铀处理1min，最后进行透射电镜(Tecnai F20 G2)观察。
特异性标记物：采用流式细胞分析术验证外泌体特异性标记物的阳性表达率(CD9、CD63、CD81)。
标志物：Western blot技术鉴定外泌体表面标志物(CD9、Alix)：步骤如3.2所述。
实验结果参见图3，TEM显示M2-Exo为双层囊泡结构；NTA结果显示M2-Exo主要粒径为120nm左右；流式细胞分析验证了M2-Exo特异性标记物的阳性表达；Western blot验证了M2-Exo能够表达CD9、Alix外泌体特异性蛋白。
二、M2-Exo重编程M1巨噬细胞向M2巨噬细胞转变以实现促炎作用向抗炎作用的转变
1、M0巨噬细胞的提取与鉴定、M2巨噬细胞外泌体制备同上述方法。
2、M1巨噬细胞的激活：将M0巨噬细胞以1×105个的密度接种于75cm3培养瓶中，细胞贴壁后，应用20ng/mL IFN-γ(R&D)处理M0巨噬细胞48h。
3、M1巨噬细胞的鉴定：
3.1、qRT-PCR验证M1巨噬细胞相关基因的表达水平(iNOS)：方法同M2巨噬细胞的鉴定，qRT-PCR引物序列信息参见表2。
表2 qRT-PCR引物序列信息
3.2、Western blot技术鉴定M1巨噬细胞特异性标志物(iNOS、CD86)：方法同M2巨噬细胞的鉴定。
qRT-PCR和Western blot检测结果参见图4，从图4中可以看出，本申请制备得到的M1巨噬细胞能够表达M1表型相关基因iNOS，且能够表达iNOS、CD86特征性蛋白。
4、M1巨噬细胞摄取实验：用膜标记染料PKH67标记M2-Exo，120000g，90min超速离心去除未标记上的荧光染料，用PBS缓冲液重悬超速离心沉淀获得已标记外泌体。将外泌体悬液用0.22μm滤器过滤细菌后，加入铺有1×103个/孔M1巨噬细胞的6孔板中，于敷箱中孵育12h。4％多聚甲醛固定液室温下固定20min，PBS清洗3次，共聚焦显微镜下观察巨噬细胞摄取M2-Exo。
实验结果参见图5，将M2-Exo与M1巨噬细胞共培养，M2-Exo可被M1巨噬细胞摄取。
5、M2-Exo重编程M1巨噬细胞：
(1)当M1巨噬细胞融合率为70％-80％时，将不同浓度外泌体(10,25,50μg/mL)与M1巨噬细胞在无血清培养基中共同孵育24h，通过qRT-PCR验证M2巨噬细胞相关基因的表达水平(Arg-1)，Western blot技术鉴定M2巨噬细胞特异性标志物(Arg-1)，以及免疫荧光染色检测M1、M2巨噬细胞相关标记物(iNOS、CD206)的表达水平，以确定起效所需最低浓度。
实验结果参见图6，Western Blot显示50ug/mL M2-Exo可有效转换M1/M2型巨噬细胞表面标记物表达；qRT-PCR结果证明了50ug/mL M2-Exo处理M1巨噬细胞后M2型相关基因表达水平显著升高，M1型相关基因表达水平显著降低；免疫荧光染色进一步证实，随着M2-Exo处理浓度的增高，M1型标记物iNOS表达降低，M2型标记物CD206表达升高。综上所述，M2-Exo对M1巨噬细胞重编程作用的最低有效浓度为50μg/mL。
(2)以最低有效浓度50μg/mL M2-Exo处理M1细胞获得重编程后的巨噬细胞(RM2)，并利用qRT-PCR进行检测RM2炎症基因的表达水平，以进一步确定M2-Exo重编程M1巨噬细胞的作用，qRT-PCR引物序列信息参见表3。
表3 qRT-PCR引物序列信息
实验结果参见图7，应用50μg/mL M2-Exo处理M1巨噬细胞后获得重编程后的巨噬细胞(RM2)，可见RM2相关炎症基因表达水平显著降低。
三、M2-Exo促进伤口愈合
1、材料：5周龄雄性C57/b小鼠。
2、分组：根据处理不同分为PBS组，M2-Exo组。
3、手术处理：首先麻醉，然后电动剃须刀剃除背部毛发，70％酒精消毒后待干燥，使用8mm无菌活检针穿孔，推动活检针至背部皮肤中部，并左右扭转以穿透背部皮肤。术后分别在不同组伤口边缘皮下注射200ul PBS、M2-Exo(100ug溶于200ul PBS)处理。
4、术后4d、7d拍照确定伤口闭合情况。
实验结果如图8、图9所示，从图8中可以看出，与PBS组相比，M2-Exo处理组可显著促进皮肤组织愈合；从图9可以看出，皮下注射PBS、M2-Exo 7天后，PBS组可见大量的炎性细胞浸润，新生血管较少，而M2-Exo无炎性细胞浸润，新生血管较多。
本具体实施方式的实施例均为本发明的较佳实施例，并非依此限制本发明的保护范围，故：凡依本发明的结构、形状、原理所做的等效变化，均应涵盖于本发明的保护范围之内。
Claims (10)
1.一种巨噬细胞来源外泌体制剂，其特征在于：包括M2型巨噬细胞来源外泌体。
2.根据权利要求1所述的一种巨噬细胞来源外泌体制剂，其特征在于：制剂中M2型巨噬细胞来源外泌体的有效浓度大于等于50μg/mL。
3.根据权利要求1所述的一种巨噬细胞来源外泌体制剂，其特征在于：所述M2型巨噬细胞来源于人或动物，所述动物选自鼠、兔、猪、羊或猴。
4.根据权利要求1所述的一种巨噬细胞来源外泌体制剂，其特征在于：所述制剂还包括药学上可接受的载体。
5.根据权利要求4所述的一种巨噬细胞来源外泌体制剂，其特征在于：所述药学上可接受的载体选自磷酸盐缓冲液和/或生理盐水，或具备缓释功能的水凝胶。
6.根据权利要求1所述的一种巨噬细胞来源外泌体制剂，其特征在于：所述制剂为注射剂、胶囊剂、片剂、粉末剂、软膏剂或喷雾剂。
7.一种权利要求1-6任一所述巨噬细胞来源外泌体制剂的制备方法，其特征在于：包括以下步骤：
S1.将M0巨噬细胞以1×105-5×105个的密度接进行接种，细胞贴壁后，用20-25ng/mLIL-4处理M0巨噬细胞48-60h，得到M2巨噬细胞；
S2.将M2巨噬细胞在无血清条件下孵育2-3d，收集上清，通过差速离心方法收集外泌体：4℃300-400g离心8-10min以移除细胞；2000-2500g离心8-10min以移除死细胞；130000-150000g离心90-100min以获取外泌体球；130000-150000g离心90-100min得到外泌体沉淀；用PBS溶液重悬外泌体沉淀，滤膜过滤后得到M2型巨噬细胞来源外泌体。
8.一种权利要求1-6任一所述巨噬细胞来源外泌体制剂在制备治疗炎症性疾病药物、保健品或营养补充剂中的应用。
9.一种权利要求1-6任一所述巨噬细胞来源外泌体制剂在制备修复组织损伤药物、保健品或营养补充剂中的应用。
10.一种权利要求1-6任一所述巨噬细胞来源外泌体制剂在制备治疗退变性疾病药物、保健品或营养补充剂中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111197930.7A CN113797229A (zh) | 2021-10-14 | 2021-10-14 | 一种用于抗炎和促组织再生的巨噬细胞来源外泌体制剂及其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111197930.7A CN113797229A (zh) | 2021-10-14 | 2021-10-14 | 一种用于抗炎和促组织再生的巨噬细胞来源外泌体制剂及其制备方法和应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113797229A true CN113797229A (zh) | 2021-12-17 |
Family
ID=78937574
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111197930.7A Pending CN113797229A (zh) | 2021-10-14 | 2021-10-14 | 一种用于抗炎和促组织再生的巨噬细胞来源外泌体制剂及其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113797229A (zh) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114891728A (zh) * | 2022-04-07 | 2022-08-12 | 广东医科大学附属医院 | 聚电解质膜、巨噬细胞外泌体及其在促进BMSCs分化中的应用 |
| CN114949224A (zh) * | 2022-06-06 | 2022-08-30 | 山东大学第二医院 | Nrp2激动剂在制备复发性流产治疗药物中的应用 |
| CN115109750A (zh) * | 2022-08-10 | 2022-09-27 | 山西农业大学 | 一种巨噬细胞外泌体及其提取方法与应用 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2019528789A (ja) * | 2016-09-23 | 2019-10-17 | クスセルル メディカル ソルティオンス， エッセ．エッレ．Ｘｃｅｌｌ Ｍｅｄｉｃａｌ Ｓｏｌｕｔｉｏｎｓ， Ｓ．Ｌ． | 極性化マクロファージを用いた組織再生の細胞治療 |
| CN113046316A (zh) * | 2021-04-07 | 2021-06-29 | 中南大学湘雅医院 | M2型骨髓巨噬细胞外泌体及其应用、脊髓损伤治疗制剂 |
-
2021
- 2021-10-14 CN CN202111197930.7A patent/CN113797229A/zh active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2019528789A (ja) * | 2016-09-23 | 2019-10-17 | クスセルル メディカル ソルティオンス， エッセ．エッレ．Ｘｃｅｌｌ Ｍｅｄｉｃａｌ Ｓｏｌｕｔｉｏｎｓ， Ｓ．Ｌ． | 極性化マクロファージを用いた組織再生の細胞治療 |
| CN113046316A (zh) * | 2021-04-07 | 2021-06-29 | 中南大学湘雅医院 | M2型骨髓巨噬细胞外泌体及其应用、脊髓损伤治疗制剂 |
Non-Patent Citations (4)
| Title |
|---|
| 尹学红等: "脂肪间充质干细胞促进M1型巨噬细胞向M2型巨噬细胞转化", 《细胞与分子免疫学杂志》 * |
| 谢侃等: "巨噬细胞与脊髓损伤的研究进展", 《泸州医学院学报》 * |
| 郑文龙等: "巨噬细胞在血管再生和组织工程中的调控作用", 《国际口腔医学杂志》 * |
| 闻君侠等: "M2 型巨噬细胞来源的外泌体对类风湿关节炎小鼠的作用及其机制研究", 《中国现代医学杂志》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114891728A (zh) * | 2022-04-07 | 2022-08-12 | 广东医科大学附属医院 | 聚电解质膜、巨噬细胞外泌体及其在促进BMSCs分化中的应用 |
| CN114891728B (zh) * | 2022-04-07 | 2023-01-03 | 广东医科大学附属医院 | 聚电解质膜、巨噬细胞外泌体及其在促进BMSCs分化中的应用 |
| CN114949224A (zh) * | 2022-06-06 | 2022-08-30 | 山东大学第二医院 | Nrp2激动剂在制备复发性流产治疗药物中的应用 |
| CN114949224B (zh) * | 2022-06-06 | 2023-03-14 | 山东大学第二医院 | Nrp2激动剂在制备复发性流产治疗药物中的应用 |
| CN115109750A (zh) * | 2022-08-10 | 2022-09-27 | 山西农业大学 | 一种巨噬细胞外泌体及其提取方法与应用 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113797229A (zh) | 一种用于抗炎和促组织再生的巨噬细胞来源外泌体制剂及其制备方法和应用 | |
| JP6633056B2 (ja) | 移植片対宿主病（ｇｖｈｄ）または表皮水疱症（ｅｂ）のエキソソームによる治療方法 | |
| CN111437431A (zh) | 一种外泌体水凝胶伤口敷料及其制备方法 | |
| KR20150020113A (ko) | mTOR/STAT3 신호억제제 처리된 면역조절능을 갖는 간엽줄기세포 및 이를 포함하는 면역질환의 예방 또는 치료용 세포치료제 조성물 | |
| US10874114B2 (en) | Supplementation of milk formulas with microvesicles isolated from milk | |
| WO2017157131A1 (zh) | 绿原酸在制备治疗以lag-3为靶点的疾病的药物中的用途 | |
| CN112587720A (zh) | 一种cgf和脐带间充质干细胞外泌体混合物、制备及应用 | |
| CN110903348B (zh) | 一种促进伤口愈合的小肽及其应用 | |
| CN114107205A (zh) | 一种刺激细胞快速分泌外泌体的方法及其应用 | |
| CN110772483B (zh) | 硫化氢修饰的间充质干细胞外囊泡作为miRNA递送载体在缺氧缺血性脑损伤中的应用 | |
| CN113234811A (zh) | CircRNA000338的应用及其药物 | |
| Chiba et al. | Human Milk Exosomes Induce ZO-1 Expression via Inhibition of REDD1 Expression in Human Intestinal Epithelial Cells | |
| JP7389507B2 (ja) | クローナル幹細胞を含む移植片対宿主病の治療用薬学的組成物 | |
| CN114099545A (zh) | 外泌体在胸主动脉夹层/动脉瘤中的作用 | |
| CN107308178B (zh) | 一种用于肠外营养相关性肝病药物 | |
| TW202039834A (zh) | 包含克隆幹細胞的用於預防或治療特應性皮炎的藥學組合物 | |
| Liu et al. | Mesenchymal Stem Cell-Derived Exosomes as Drug Carriers for Delivering miRNA-29b to Ameliorate Inflammation in Corneal Injury Via Activating Autophagy | |
| CN110452860A (zh) | 一种唾液链球菌及其在治疗炎症性肠病药物中的应用 | |
| CN117159581B (zh) | 一种piRNA反义核苷酸药物组合物作为主动脉瓣钙化防治药的应用 | |
| CN108359671A (zh) | 肿瘤细胞糖摄取抑制剂及其应用 | |
| CN112458095B (zh) | 调控ADSCs成骨分化及组织再生LMO3基因及其应用 | |
| WO2019037644A1 (zh) | 一种人体免疫复合因子的制备方法及应用 | |
| CN115612670B (zh) | 一种微波制备肿瘤细胞微颗粒的方法 | |
| RU2766707C1 (ru) | Средство для лечения фиброза тканей на основе компонентов секретома мезенхимных стромальных клеток, способ получения и применения средства | |
| Lee et al. | Stem cell-derived exosomes as a biomaterial source for immune modulating therapy |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211217 |
|
| RJ01 | Rejection of invention patent application after publication |