CN110496139A - A kind of biological agent for skin wounds reparation - Google Patents
A kind of biological agent for skin wounds reparation Download PDFInfo
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- CN110496139A CN110496139A CN201910880979.9A CN201910880979A CN110496139A CN 110496139 A CN110496139 A CN 110496139A CN 201910880979 A CN201910880979 A CN 201910880979A CN 110496139 A CN110496139 A CN 110496139A
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- 239000003124 biologic agent Substances 0.000 title claims abstract description 40
- 206010072170 Skin wound Diseases 0.000 title claims abstract description 36
- 210000002901 mesenchymal stem cell Anatomy 0.000 claims abstract description 30
- 150000004676 glycans Chemical class 0.000 claims abstract description 29
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 29
- 239000005017 polysaccharide Substances 0.000 claims abstract description 29
- HPUXDMUGCAWDFW-UHFFFAOYSA-N Osthole Natural products COc1ccc2CCC(=O)Oc2c1C=CC(=O)C HPUXDMUGCAWDFW-UHFFFAOYSA-N 0.000 claims abstract description 21
- MBRLOUHOWLUMFF-UHFFFAOYSA-N osthole Chemical compound C1=CC(=O)OC2=C(CC=C(C)C)C(OC)=CC=C21 MBRLOUHOWLUMFF-UHFFFAOYSA-N 0.000 claims abstract description 21
- 239000002904 solvent Substances 0.000 claims abstract description 15
- HSTZMXCBWJGKHG-UHFFFAOYSA-N (E)-piceid Natural products OC1C(O)C(O)C(CO)OC1OC1=CC(O)=CC(C=CC=2C=CC(O)=CC=2)=C1 HSTZMXCBWJGKHG-UHFFFAOYSA-N 0.000 claims abstract description 12
- 229960003764 polydatin Drugs 0.000 claims abstract description 12
- HSTZMXCBWJGKHG-CUYWLFDKSA-N trans-piceid Polymers O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=CC(\C=C\C=2C=CC(O)=CC=2)=C1 HSTZMXCBWJGKHG-CUYWLFDKSA-N 0.000 claims abstract description 12
- 239000000243 solution Substances 0.000 claims description 13
- -1 Veratryl alcohol glycosides Chemical class 0.000 claims description 12
- 210000004027 cell Anatomy 0.000 claims description 12
- 229930182470 glycoside Natural products 0.000 claims description 12
- 238000002360 preparation method Methods 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 10
- 102000004142 Trypsin Human genes 0.000 claims description 7
- 108090000631 Trypsin Proteins 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- 239000012679 serum free medium Substances 0.000 claims description 7
- 239000012588 trypsin Substances 0.000 claims description 7
- 230000007910 cell fusion Effects 0.000 claims description 5
- 239000006285 cell suspension Substances 0.000 claims description 5
- 238000011081 inoculation Methods 0.000 claims description 5
- 238000005119 centrifugation Methods 0.000 claims description 4
- 239000002504 physiological saline solution Substances 0.000 claims description 4
- 210000003716 mesoderm Anatomy 0.000 claims description 3
- 210000000130 stem cell Anatomy 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 210000001519 tissue Anatomy 0.000 claims description 2
- 210000000988 bone and bone Anatomy 0.000 claims 2
- 235000002710 Ilex cornuta Nutrition 0.000 claims 1
- 241001310146 Ilex cornuta Species 0.000 claims 1
- 235000010326 Osmanthus heterophyllus Nutrition 0.000 claims 1
- 229910017435 S2 In Inorganic materials 0.000 claims 1
- 210000004271 bone marrow stromal cell Anatomy 0.000 claims 1
- OEGPRYNGFWGMMV-UHFFFAOYSA-N veratryl alcohol Natural products COC1=CC=C(CO)C=C1OC OEGPRYNGFWGMMV-UHFFFAOYSA-N 0.000 claims 1
- 230000029663 wound healing Effects 0.000 abstract description 13
- 230000001900 immune effect Effects 0.000 abstract description 4
- 230000001225 therapeutic effect Effects 0.000 abstract description 4
- 206010052428 Wound Diseases 0.000 description 13
- 208000027418 Wounds and injury Diseases 0.000 description 13
- 230000000052 comparative effect Effects 0.000 description 9
- 239000007924 injection Substances 0.000 description 7
- 238000002347 injection Methods 0.000 description 7
- 230000000694 effects Effects 0.000 description 6
- QNVSXXGDAPORNA-UHFFFAOYSA-N Resveratrol Natural products OC1=CC=CC(C=CC=2C=C(O)C(O)=CC=2)=C1 QNVSXXGDAPORNA-UHFFFAOYSA-N 0.000 description 5
- 229940016667 resveratrol Drugs 0.000 description 5
- 235000021283 resveratrol Nutrition 0.000 description 5
- 241000219317 Amaranthaceae Species 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- 235000014676 Phragmites communis Nutrition 0.000 description 3
- 241000489523 Veratrum Species 0.000 description 3
- 210000001185 bone marrow Anatomy 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 230000035876 healing Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 239000004017 serum-free culture medium Substances 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 241000356446 Cnidium monnieri Species 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 206010063560 Excessive granulation tissue Diseases 0.000 description 1
- 102000003777 Interleukin-1 beta Human genes 0.000 description 1
- 108090000193 Interleukin-1 beta Proteins 0.000 description 1
- 235000019084 Selinum monnieri Nutrition 0.000 description 1
- 208000028990 Skin injury Diseases 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003851 biochemical process Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 210000001126 granulation tissue Anatomy 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000007420 reactivation Effects 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 210000002268 wool Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
- A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
- A61K31/37—Coumarins, e.g. psoralen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
- A61K36/815—Lycium (desert-thorn)
-
- 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
-
- 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
- C12N5/0663—Bone marrow mesenchymal stem cells (BM-MSC)
-
- 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
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
Abstract
The invention discloses a kind of biological agents for skin wounds reparation, including mesenchymal stem cell, Osthole, polydatin, polysaccharides and solvent；The mesenchymal stem cell is 0.5 × 106‑5×106A/mL, Osthole are 1-2 μ g/mL, and polydatin is 0.5-1 μ g/mL, and polysaccharides is 20-30 μ g/mL.Biological agent of the invention can be used for skin wounds reparation, has excellent histocompatbility, will not cause immunological rejection, and Wound healing speed is fast, therapeutic effect is good.
Description
Technical field
The present invention relates to biologic product technology field more particularly to a kind of biological agents for skin wounds reparation.
Background technique
Skin is body and extraneous mechanical barrier, is often easy to happen damage, is difficult to the wound to heal for a long time once being formed
Mouthful, people's lives and work quality will be seriously affected.Wound healing process can be divided into inflammatory reaction, granulation tissue increases
Reconstruction process after raw, surface of a wound re-epithelialization and wound healing, metabolism, gene promoter comprising intraor extracellular substance etc. are a series of
Biochemical processes.Currently, mainly having skin graft, skin substitutes and various biological agents for skin wounds reparation
Deng.Mesenchymal stem cell can not only participate in the reparation of skin histology, promote Wound healing, and will not cause immunological rejection
Reaction has good skin wounds reparative potential.But it is repaired using the skin of mesenchymal stem cell biological agent merely
Reactivation power is limited, it is difficult to reach expected therapeutic effect.
Summary of the invention
Technical problems based on background technology, the invention proposes a kind of biology systems for skin wounds reparation
Agent has excellent histocompatbility, will not cause immunological rejection, and Wound healing speed is fast, therapeutic effect is good.
A kind of biological agent for skin wounds reparation proposed by the present invention, including mesenchymal stem cell, cnidium monnieri
Sub- element, polydatin, polysaccharides and solvent.
Preferably, the mesenchymal stem cell is 0.5 × 106-5×106A/mL, Osthole are 1-2 μ g/mL,
Polydatin is 0.5-1 μ g/mL, and polysaccharides is 20-30 μ g/mL.
It is highly preferred that the mesenchymal stem cell is 3 × 106A/mL, Osthole are 1.8 μ g/mL, white black false hellebore
Alcohol glycosides is 0.6 μ g/mL, and polysaccharides is 25 μ g/mL.
Preferably, the mesenchymal stem cell is P3-P5 generation.
Preferably, the molecular weight of the polysaccharides is 1-3 ten thousand.
Preferably, the solvent is PBS solution or physiological saline.
Preferably, the biological agent for skin wounds reparation the preparation method is as follows: the marrow that will be prepared
Mescenchymal stem cell is resuspended to obtain cell suspension with solvent, and Osthole, polydatin and polysaccharides, mixing is then added
Uniformly, the biological agent for skin wounds reparation is obtained.
Preferably, the preparation method of the mesenchymal stem cell includes the following steps:
S1, it takes myeloid tissue to be digested with trypsin solution in 37 DEG C of conditions, then is terminated and digested with serum free medium, from
It is washed after the heart with D-Hanks liquid, is centrifuged again, cell is resuspended with D-Hanks liquid；
S2: the cell inoculation that step S1 is obtained is added serum free medium culture, more renews after 1-2d in culture bottle
Fresh culture medium, every 2-3d replaces fresh culture later, when cell fusion reaches 80-90%, passes on, obtains according to 1:3 ratio
To mesenchymal stem cell.
Preferably, in the step S1, the concentration of trypsin solution is 1.5-2.5%.
Preferably, in the step S2, condition of culture is 37 DEG C, 5%CO2。
Beneficial effects of the present invention are as follows:
The present invention is compounded mesenchymal stem cell, Osthole, polydatin, polysaccharides to obtain skin
The biological agent of wound reparation.Wherein, mesenchymal stem cell, Osthole, polydatin, polysaccharides cooperation, energy
Efficient activation Wnt signal path, so that the healing rate of skin wounds is increased substantially, Osthole, polydatin, fructus lycii
Polysaccharide can also improve the proliferation activity of mesenchymal stem cell by activation Wnt signal path, promote medulla mesenchyma dry thin
Intracrine more facilitates the cell factor of Wound healing, further promotes the healing of skin wounds；Polysaccharides, resveratrol
Glycosides cooperation, moreover it is possible to play the degeneration-resistant protective effect of cell, mitigate oxidative stress, improve mesenchymal stem cell in skin wounds
The activity of survival rate, proliferation activity and secrete cytokines, so that enhancing mesenchymal stem cell promotes wound healing
Effect；Polysaccharides can also inhibit proinflammatory factor TNF-α and IL-1 β to discharge, and reduce inflammatory factor level, inhibit the excessive of inflammation
Occur, to further increase the reparation speed of wound, while reducing the formation of scar, improves wound repairing effect.The present invention
Biological agent can overcome single mesenchymal stem cell biological agent for skin wounds reparation wound healing efficiency
Low defect not only has excellent histocompatbility, will not cause immunological rejection, and can increase substantially wound
Healing rate has good therapeutic effect.
Specific embodiment
In the following, technical solution of the present invention is described in detail by specific embodiment.
Embodiment 1
A kind of biological agent for skin wounds reparation, including mesenchymal stem cell, Osthole, resveratrol
Glycosides, polysaccharides and solvent, wherein mesenchymal stem cell is 0.5 × 106A/mL, Osthole are 1 μ g/mL, white black false hellebore
Alcohol glycosides is 0.5 μ g/mL, and polysaccharides is 20 μ g/mL.
Wherein, the molecular weight of polysaccharides is 10,000, and solvent is PBS solution.
Biological agent for skin wounds reparation the preparation method is as follows:
S1, it takes bone marrow of mice to be digested with the trypsin solution that concentration is 1.5% in 37 DEG C of conditions, then uses serum-free
Culture medium terminates digestion, is washed after centrifugation with D-Hanks liquid, is centrifuged again, and cell is resuspended with D-Hanks liquid；
In culture bottle serum free medium is added at 37 DEG C in S2, the cell inoculation for obtaining step S1,5%CO2Condition
Lower culture replaces fresh culture after 1d, and every 2d replaces fresh culture later, when cell fusion reaches 80%, according to 1:3
Ratio passage, obtains mesenchymal stem cell；
S3, P3 is resuspended to obtain cell suspension with solvent for mesenchymal stem cell, Osthole, white Chenopodiaceae is then added
Reed alcohol glycosides and polysaccharides are uniformly mixed, obtain the biological agent for skin wounds reparation.
Embodiment 2
A kind of biological agent for skin wounds reparation, including mesenchymal stem cell, Osthole, resveratrol
Glycosides, polysaccharides and solvent, wherein mesenchymal stem cell is 2 × 106A/mL, Osthole are 1.8 μ g/mL, white black false hellebore
Alcohol glycosides is 0.6 μ g/mL, and polysaccharides is 24 μ g/mL.
Wherein, the molecular weight of polysaccharides is 20,000, and solvent is PBS solution or physiological saline.
Biological agent for skin wounds reparation the preparation method is as follows:
S1, it takes bone marrow of mice to be digested with the trypsin solution that concentration is 2% in 37 DEG C of conditions, then is trained with serum-free
It supports base and terminates digestion, washed after centrifugation with D-Hanks liquid, be centrifuged again, cell is resuspended with D-Hanks liquid；
In culture bottle serum free medium is added at 37 DEG C in S2, the cell inoculation for obtaining step S1,5%CO2Condition
Lower culture replaces fresh culture after 1d, and every 2d replaces fresh culture later, when cell fusion reaches 80%, according to 1:3
Ratio passage, obtains mesenchymal stem cell；
S3, P3 is resuspended to obtain cell suspension with solvent for mesenchymal stem cell, Osthole, white Chenopodiaceae is then added
Reed alcohol glycosides and polysaccharides are uniformly mixed, obtain the biological agent for skin wounds reparation.
Embodiment 3
A kind of biological agent for skin wounds reparation, including mesenchymal stem cell, Osthole, resveratrol
Glycosides, polysaccharides and solvent, wherein mesenchymal stem cell is 5 × 106A/mL, Osthole are 2 μ g/mL, resveratrol
Glycosides is 1 μ g/mL, and polysaccharides is 30 μ g/mL.
Wherein, the molecular weight of polysaccharides is 30,000, and solvent is PBS solution or physiological saline.
Biological agent for skin wounds reparation the preparation method is as follows:
S1, it takes bone marrow of mice to be digested with the trypsin solution that concentration is 2.5% in 37 DEG C of conditions, then uses serum-free
Culture medium terminates digestion, is washed after centrifugation with D-Hanks liquid, is centrifuged again, and cell is resuspended with D-Hanks liquid；
In culture bottle serum free medium is added at 37 DEG C in S2, the cell inoculation for obtaining step S1,5%CO2Condition
Lower culture replaces fresh culture after 2d, and every 3d replaces fresh culture later, when cell fusion reaches 90%, according to 1:3
Ratio passage, obtains mesenchymal stem cell；
S3, P3 is resuspended to obtain cell suspension with solvent for mesenchymal stem cell, Osthole, white Chenopodiaceae is then added
Reed alcohol glycosides and polysaccharides are uniformly mixed, obtain the biological agent for skin wounds reparation.
Comparative example 1
The difference of comparative example 1 and embodiment 1 are as follows: be free of Osthole, remaining raw material composition is all the same with preparation method.
Comparative example 2
The difference of comparative example 2 and embodiment 1 are as follows: be free of polydatin, remaining raw material composition is all the same with preparation method.
Comparative example 3
The difference of comparative example 3 and embodiment 1 are as follows: be free of polysaccharides, remaining raw material composition is all the same with preparation method.
Test example mouse skin wound repairing test
Back wool will be removed after 72 mouse anesthesias, forms the skin injury wound that diameter is 1.5cm at back with sterile cut
Face.It is divided into 6 processing groups, each processing group 12 after modeling success.Wherein, 1 is handled: the biological agent of injection embodiment 1；Place
Reason 2: the biological agent of injection embodiment 2；Processing 3: the biological agent of injection embodiment 3；Processing 4: the biology of injection comparative example 1
Preparation；Processing 5: the biological agent of injection comparative example 2；Processing 6: the biological agent of injection comparative example 3.Injecting method is as follows: enclosing
Biological agent, every 100 μ l are injected around wound 5 injection methods of Zhou Caiyong.Mouse is put back in cage after end of operation and is raised, is freely drunk
Food, and be observed continuously.Respectively at the 3rd, 7,10 day, each processing group surface of a wound image, Image-Pro are acquired using digital camera
Plus 6.0 analyzes software and measures surface of a wound area.Wound healing rate: Wound healing rate={ (original wound is calculated using following formula
The current surface of a wound area of face area -)/original surface of a wound area } × 100%.Test result is as shown in table 1:
1 each group Wound healing rate of table
As can be seen from the above table, biological agent of the invention has excellent wound healing effect, can increase substantially wound
The Healing Rate in face.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (10)
1. a kind of biological agent for skin wounds reparation, which is characterized in that including mesenchymal stem cell, frutus cnidii
Element, polydatin, polysaccharides and solvent.
2. the biological agent according to claim 1 for skin wounds reparation, which is characterized in that the medulla mesenchyma
Stem cell is 0.5 × 106-5×106A/mL, Osthole are 1-2 μ g/mL, and polydatin is 0.5-1 μ g/mL, and fructus lycii is more
Sugar is 20-30 μ g/mL.
3. the biological agent according to claim 1 or 2 for skin wounds reparation, which is characterized in that between the marrow
Mesenchymal stem cells are 3 × 106A/mL, Osthole are 1.8 μ g/mL, and polydatin is 0.6 μ g/mL, and polysaccharides is 25 μ
g/mL。
4. the biological agent according to claim 1-3 for skin wounds reparation, which is characterized in that the bone
Bone marrow-drived mesenchymal stem is P3-P5 generation.
5. the biological agent according to claim 1-4 for skin wounds reparation, which is characterized in that the Chinese holly
The molecular weight of Qi polysaccharide is 1-3 ten thousand.
6. the biological agent according to claim 1-5 for skin wounds reparation, which is characterized in that described molten
Agent is PBS solution or physiological saline.
7. the biological agent according to claim 1-6 for skin wounds reparation, which is characterized in that preparation side
Method is as follows: the mesenchymal stem cell being prepared being resuspended to obtain cell suspension with solvent, Osthole, white is then added
Veratryl alcohol glycosides and polysaccharides are uniformly mixed, obtain the biological agent for skin wounds reparation.
8. the biological agent according to claim 7 for skin wounds reparation, which is characterized in that the medulla mesenchyma
The preparation method of stem cell includes the following steps:
S1, it takes myeloid tissue to be digested with trypsin solution in 37 DEG C of conditions, then is terminated and digested with serum free medium, after centrifugation
It is washed with D-Hanks liquid, is centrifuged again, cell is resuspended with D-Hanks liquid；
S2, the cell inoculation for obtaining step S1 are added serum free medium culture, replace fresh training after 1-2d in culture bottle
Base is supported, every 2-3d replaces fresh culture later, when cell fusion reaches 80-90%, passes on according to 1:3 ratio, obtains bone
Bone marrow-drived mesenchymal stem.
9. the biological agent according to claim 8 for skin wounds reparation, which is characterized in that in the step S1,
The concentration of trypsin solution is 1.5-2.5%.
10. the biological agent for skin wounds reparation according to claim 8 or claim 9, which is characterized in that the step S2
In, condition of culture is 37 DEG C, 5%CO2。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111012653A (en) * | 2019-12-27 | 2020-04-17 | 衡欣实业有限公司 | Skin restoration instrument based on yeast extraction cartridge bag |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107823632A (en) * | 2017-11-10 | 2018-03-23 | 南京九圣生物科技股份有限公司 | A kind of mesenchymal stem cells MSCs parenteral solution and preparation method |
-
2019
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107823632A (en) * | 2017-11-10 | 2018-03-23 | 南京九圣生物科技股份有限公司 | A kind of mesenchymal stem cells MSCs parenteral solution and preparation method |
Non-Patent Citations (2)
| Title |
|---|
| HUI ZHAO ET AL: "Chapter 5 Dermatologic Uses and Effects of Lycium Barbarum", 《LYCIUM BARBARUM AND HUMAN HEALTH》 * |
| 卞徽宁 等: "虎杖苷对成纤维细胞生物学特性的影响", 《中国组织工程研究》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111012653A (en) * | 2019-12-27 | 2020-04-17 | 衡欣实业有限公司 | Skin restoration instrument based on yeast extraction cartridge bag |
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