CN108635376A - Purposes of the Cord blood regenerated particle and combinations thereof in treating skin wound healing - Google Patents
Purposes of the Cord blood regenerated particle and combinations thereof in treating skin wound healing Download PDFInfo
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- CN108635376A CN108635376A CN201810312745.XA CN201810312745A CN108635376A CN 108635376 A CN108635376 A CN 108635376A CN 201810312745 A CN201810312745 A CN 201810312745A CN 108635376 A CN108635376 A CN 108635376A
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- cord blood
- regenerated particle
- blood regenerated
- particle
- skin
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- 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
- A61K35/48—Reproductive organs
- A61K35/51—Umbilical cord; Umbilical cord blood; Umbilical stem cells
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- 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
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a kind of purposes of the Cord blood regenerated particle and combinations thereof in treating skin wound healing, the Cord blood regenerated particle has following feature：Ranging from 15 μm of the diameter of the Cord blood regenerated particle；The Cord blood regenerated particle contains minim DNA ingredient；The Cord blood regenerated particle expresses Oct3/4, Nanog, Sox2 albumen.The study found that therapeutic agent prepared by Cord blood regenerated particle of the present invention and combinations thereof can significantly promote the healing of skin wound, and the rate of wound healing is significantly accelerated, and haemostatic effect is good, while also can effectively prevent the formation of scar.
Description
Technical field
The present invention relates to the fields such as biomedical sector, especially histocytology, regenerative medicine, disease treatment, especially
It is related to purposes of the Cord blood regenerated particle and combinations thereof in treating skin wound healing.
Background technology
Wound healing refers to the natural process that damaged tissues are repaired by body.Most of slight wounds can be effective by body itself
Ground cures, but some more serious wounds may need suture needle or the help of other precautionary measures.The process of healing is one multiple
Miscellaneous process, although can not possibly what externally visibly be had occurred.When wound occurs first, body starts tightening in damage blood
Pipe, this contributes to limit blood to flow, and excessive bleeding will not thus occur.Most of minor cut or wounds, this is that there is no problem,
But big wound still bleed because body cannot pressor speed it is not fast enough, to prevent Hemostatic Oral Liquid be lost in.It may need additional
Emergency procedure, such as around region be that a piece of cloth flows to region to cut off blood.Once blood flow slows down, it is small to form blood
Plate is combined together to form grumeleuse in the opening of wound.Clot is added to prevent it from moving or detaching in additional substance.It is covered
Lid wound prevents additional bleeding, prevents foreign matter from entering wound.The next step of wound healing helps to prevent wound infection.In people
In class, a kind of solution of antibacterial can be added to help this process, but body also has natural mechanism to help to prevent to feel
Dye.Because wound has been formed a scab and closed, blood vessel, which reopens, allows more red blood cells and leucocyte to enter this area.Then white
Cell can find and kill any bacterium possibly into wound.
Finally, wound healing is related to the reconstruction of tissue and skin.The skin of the lower wound both sides of incrustation finally flexes outward,
It meets at the center of wound.This sometimes results in visible scar, this depends on the severity of wound.With pushing away for time
It moves, the tissue bound up a wound becomes stronger, and final scar tissue can fall off or reuptake in body.Some situations may need
Additional wound healing is wanted to help.Very serious injury is such as stabbed into body depths, it may be necessary to medical assistance, to prevent
It only infects, closes wound, prevent bleeding.In addition to the normal healing process of body, antibiotic, suture and blood-clotting agent may need
It manages.
Wound is the rupture of skin outer layer, referred to as epidermis.Wound is usually caused by incised wound or scratch.Different types of wound
Mouth, which can be distinguished, to be treated, this depends on their occurring mode and severity.Other wounds include puncture wound, wound
(reduction), bedsore, anal fissure, extravasation (drug can cause outside the vein of tissue damage accidentally), cause incontinence skin injury (bladder control
System is insufficient), skin-grafting healing (thickness), diabetic ulcer, Skin graft ischemic (blood flow lacks operation connection skin).
It is the reaction made to the injury of sequence of events to treat more.In addition to bone, institute has scar outside some to be cured in a organized way
It closes.The purpose correctly nursed is to try to reduce the possibility of infection and cicatrization.
Substantially there are four the agglutinations in stage：Inflammatory phase, proliferative phase and remodeling phase, epithelialization stage.
Inflammation phase starts from damage itself.In inflammation phase, bleeding, blood vessel is narrow immediately, and clot is formed, and release is various
Chemical substance enters wound, starts agglutination.Special cell (macrophage) removes the wound of fragment in several days.
Followed by proliferative phase, matrix or gitter cell are formed.On this matrix, new Skin Cell and blood vessel can shapes
At.It is new thin vessels (being known as capillary), is cicatrized a wound, pinkiness or aubergine.These new blood vessels will be
Reconstituted cell provides oxygen and nutrition, to maintain the growth of neoblast, and supports the generation of protein (mainly collagen).
Frame of the collagen as new tissue construction.Collagen is the main matter in final scar.
The remodeling phase starts after 2-3 weeks.Skeleton (collagen) becomes more organized, keeps tissue strongr.Vessel density becomes
Small, wound starts to lose pink.At past six months, increases in the area of intensity, be finally reached the intensity of strong side skin
70%.
Epithelium is laid with during new skin or epithelial cell, cell.Skin is formed between external environment and body
One of barrier.Its main purpose is to prevent excessive water loss and bacterium.This layer rebuilds damage in the several hours started
Wound and completion are being cleaned in 24-48 hours, are sewed up a wound (seam).Open wound may need 7-10 days, because of inflammation mistake
Journey can extend, so as to cause cicatrization.Scar betides damage and (enters skin corium, i.e. second layer skin beyond deep skin
Skin).
Invention content
Technical problems based on background technology can be with the present invention provides a kind of safe therapeutic agent or composition
It is effectively used for the treatment of skin wound healing, to realize the shape for substantially speeding up and effectivelying prevent scar of wound healing time
At.
Technical scheme is as follows：
The stem cell precursor active material that Cord blood regenerated particle is found as current research, can be followed by blood of human body
Loop system reaches the different zones of skin wound, by region microenvironment stimulation and influence, it is concurrent by assembling, merging
Educate and be divided into skin progenitor cell, fibroblast, basal layer cell etc., so be significantly reduced wound healing time and effectively
Ground prevents the formation of scar.
The Cord blood regenerated particle and combinations thereof of the present invention can be significantly reduced the time of skin wound healing.
The Cord blood regenerated particle and combinations thereof that the present invention obtains can be effectively prevented during skin wound healing
The formation of scar.
The Cord blood regenerated particle and combinations thereof of the present invention is specifically for use among the treatment of skin burn.
The Cord blood regenerated particle and combinations thereof of the present invention is also particularly applied to skin and scratches (machinery or on-mechanical)
Among treatment.
The Cord blood regenerated particle and combinations thereof of the present invention is also particularly applied among the treatment of wound healing.
The Cord blood regenerated particle and combinations thereof of the present invention is also particularly applied among the treatment of amputation wounds healing.
It is cured for skin wound in preparation as therapeutic agent for this purpose, first aspect present invention provides Cord blood regenerated particle
Close the purposes in the therapeutic agent of class disease.
Therapeutic agent according to a first aspect of the present invention, wherein the Cord blood regenerated particle, there is following feature：
The Cord blood regenerated particle is derived from the regenerated particle of Cord blood.
Ranging from 1-5 μm of the diameter of the Cord blood regenerated particle.
The Cord blood regenerated particle contains minim DNA ingredient.
The Cord blood regenerated particle expresses Oct3/4, the albumen such as Nanog, Sox2.
The Cord blood regenerated particle is for mammal, such as people, the dosage of the Cord blood regenerated particle
Be per kilogram patient's weight dosage be (1-10) x109A Cord blood regenerated particle.Such as per kilogram patient's weight dosage is 1x109
A Cord blood regenerated particle, per kilogram patient's weight dosage are 5x109A Cord blood regenerated particle, per kilogram patient's weight dosage
For 10x109A Cord blood regenerated particle.
Further, second aspect of the present invention provides a kind of therapeutic agent for skin wound healing class disease, is in
The form of composition, the composition include mainly Cord blood regenerated particle liquid and injection buffer solution.
Composition according to a second aspect of the present invention, wherein the Cord blood regenerated particle liquid, there is following feature：
The Cord blood regenerated particle liquid comes from Cord blood.
Ranging from 1-5 μm of the diameter of Cord blood regenerated particle in the Cord blood regenerated particle liquid.
Cord blood regenerated particle in the Cord blood regenerated particle liquid contains minim DNA ingredient.
Cord blood regenerated particle in the Cord blood regenerated particle liquid expresses Oct3/4, the albumen such as Nanog, Sox2.
The Cord blood regenerated particle liquid is for mammal, such as people, the use of the Cord blood regenerated particle liquid
Dosage is that per kilogram patient's weight dosage is (1-10) x109A Cord blood regenerated particle.Such as per kilogram patient's weight dosage is
1x109A Cord blood regenerated particle, per kilogram patient's weight dosage are 5x109A Cord blood regenerated particle, per kilogram patient's weight
Dosage is 10x109A Cord blood regenerated particle.
Therapeutic agent according to a second aspect of the present invention, wherein the injection buffer solution is when for mammal, example
Such as people, the injection buffer solution in the therapeutic agent is 0.9% sodium chloride injection.
According to a first aspect of the present invention and the purposes of second aspect, ability may be used in the Cord blood regenerated particle used in
It is prepared by method known to domain.Such as the Cord blood regenerated particle obtained by Cord blood, it is referred to Chinese Patent Application No.
Method in CN201610871433.3 obtains.
The invention has the beneficial effects that：
The present invention provides a kind of Cord blood regenerated particles and combinations thereof to prepare for skin wound healing therapeutic agent
In purposes.The study found that therapeutic agent of the present invention can be significant promotion skin wound healing, and the rate of wound healing
Significantly accelerated, haemostatic effect is good, while also can effectively prevent the formation of scar.
Description of the drawings
Fig. 1：The form that Cord blood regenerated particle is presented when cultivating in the medium；
Fig. 2：The regenerated particle of culture detects expression Oct3/4, Nanog, Sox2 by Immunofluorescence test technology；
Fig. 3：At 12 days, regenerated particle group and physiological saline group, the actual conditions of wounds in mice healing；
Fig. 4：At 24 days, regenerated particle group and physiological saline group, the actual conditions of wounds in mice wound healing；
Fig. 5：Surface of a wound area data statistical analysis in wound；Surface of a wound area is measured each day or each alternate day,
Calculate data.
Specific implementation mode
Embodiment 1
The preparation of Cord blood regenerated particle
In the present embodiment, the preparation of Cord blood regenerated particle is illustrated.
Blood source：Umbilical cord blood collection to the blood bag containing anti-coagulants in, anti-coagulants is original sodium citrate in blood bag, is passed through
Cord blood is transported to and prepares laboratory by cold chain transportation mode (temperature is maintained at 2 to 8 DEG C).
Condition：The processing procedure for obtaining Cord blood regenerated particle needs whole sterile working.
It is as follows：
(1) Cord blood is taken to carry out 200g centrifugations, centrifugation time is 10 minutes, is divided into two parts after centrifugation, upper solution A and
Lower sediment A.
(2) a layer precipitate A is removed, is by volume 1:2-5 is added PBS and slightly washs, and carries out 200g centrifugations, and centrifugation time is
10 minutes, two parts, upper solution B and lower sediment B are divided into after centrifugation.
(3) a layer precipitate B is removed, is by volume 1:5-10 is added erythrocyte cracked liquid and is placed in horizontal shaker, shakes 20 points
Clock carries out 2000g centrifugations later, and centrifugation time is 10 minutes, and two parts, upper solution C and lower sediment C are divided into after centrifugation,
Upper solution C is discarded.
(4) a layer precipitate C is removed, is by volume 1:5-10 be added PBS acutely shake, jitter time be 2 minutes, it is laggard
Row 200g centrifugations, centrifugation time is 10 minutes, two parts, upper solution D and lower sediment D is divided into after centrifugation, lower sediment D is abandoned
Fall.
(5) upper solution A, upper solution B, upper solution D are taken respectively, carries out 5000g centrifugations, and centrifugation time is 10 points
Clock after centrifugation, collects all precipitations.
(6) culture medium (the mixing serum of umbilical cord blood for containing 20%) that the precipitation being collected into is placed in MEM is cultivated, and is cultivated
Condition is 37 DEG C, and in 5%CO2 incubators, it is primary that culture medium was replaced per 2-3 days.
(7) culture medium by culture 10 days is collected, and 5000g centrifugations is carried out, to be enriched with Cord blood regenerated particle (regenerated particle
Form is shown in Fig. 1).
Embodiment 2
Regenerated particle expresses embryo's albuminoid label
It is cultivated on the slide that collagen covers by the regenerated particle (GFP labels) for obtaining embodiment 1, after fixed
Detect their protein expression, coloration result shows that sperm cell expresses Oct3/4, Nanog and Sox2 (see Fig. 2).
Embodiment 3
Regenerated particle repairs the experiment of diabetic mice skin wound
Mouse used is diabetic mice in experiment, 6 in total, every diabetic mice, is made at skin of back
The skin wound of one 1.5cmx1.5cm.It is randomly divided into 2 groups, every group 3 according to weight.Regenerated particle group is noted by tail vein
The regenerated particle (GFP labels) for penetrating the acquisition of embodiment 1 arrives experimental diabetes mouse, and physiological saline group injects the physiology of equal volume
Brine is to experimental diabetes mouse.
At 12 days, the skin wound of the diabetic mice of all regenerated particle groups be less than physiological saline group (see figure
3), this shows that the wound repair speed for receiving the mouse of regenerated particle treatment is accelerated.
Wound repair is completed, and has a hair newly to grow out at the wounds in mice position of injection regenerated particle group, and physiology
Brine group is without this phenomenon (see Fig. 4).In addition, the rear scar area of regenerated particle group is again smaller than physiological saline group.
By the analysis of the surface of a wound, the diabetic mice for receiving regenerated particle injection is found at 10 days after the surface of a wound is formed,
The rate of reparation starts to substantially speed up (see Fig. 5).This implies that regenerated particle has more compared to physiological saline, clear-cutting forestland process
Advantage, and the rear scar smaller of the surface of a wound, can incidentally grow new hair.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Any one 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 Cord blood regenerated particle treated in skin wound healing, which is characterized in that the Cord blood regenerated particle
Ranging from 1-5 μm of diameter；The Cord blood regenerated particle contains minim DNA ingredient；The Cord blood regenerated particle expression
Oct3/4, Nanog, Sox2 albumen.
2. Cord blood regenerated particle as described in claim 1, which is characterized in that the promotion skin wound that it can be significant
Healing.
3. Cord blood regenerated particle as described in claim 1, which is characterized in that it can be effectively prevented skin wound healing
The formation of scar in the process.
4. Cord blood regenerated particle as described in claim 1, which is characterized in that its can be applied to the treatment of skin burn it
In.
5. Cord blood regenerated particle as described in claim 1, which is characterized in that its can be applied to skin scratch treatment it
In.
6. Cord blood regenerated particle as described in claim 1, which is characterized in that it can be applied to controlling for wound healing
Among treatment.
7. Cord blood regenerated particle as described in claim 1, which is characterized in that it can be applied to controlling for amputation wounds healing
Among treatment.
8. Cord blood regenerated particle as described in claim 1, which is characterized in that it can be used for preparing treatment skin wound and is cured
Close the therapeutic agent or composition of class disease.
9. Cord blood regenerated particle as claimed in claim 8, which is characterized in that the therapeutic agent mainly include Cord blood again
Raw particle liquid and injection buffer solution.
10. Cord blood regenerated particle as claimed in claim 9, which is characterized in that the injection buffer solution is 0.9%
Sodium chloride injection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810312745.XA CN108635376A (en) | 2018-04-09 | 2018-04-09 | Purposes of the Cord blood regenerated particle and combinations thereof in treating skin wound healing |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810312745.XA CN108635376A (en) | 2018-04-09 | 2018-04-09 | Purposes of the Cord blood regenerated particle and combinations thereof in treating skin wound healing |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108635376A true CN108635376A (en) | 2018-10-12 |
Family
ID=63745541
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810312745.XA Pending CN108635376A (en) | 2018-04-09 | 2018-04-09 | Purposes of the Cord blood regenerated particle and combinations thereof in treating skin wound healing |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101031639A (en) * | 2004-07-01 | 2007-09-05 | 马克罗珀尔生物外科公司 | Methods of using regenerative cells to promote wound healing |
| CN106377547A (en) * | 2016-09-30 | 2017-02-08 | 孔五 | Extraction method and purpose of renewable particles of umbilical cord blood |
| CN106459918A (en) * | 2014-04-24 | 2017-02-22 | 得克萨斯州大学系统董事会 | Application of induced pluripotent stem cells to generate adoptive cell therapy products |
| CN107709544A (en) * | 2015-06-12 | 2018-02-16 | 隆萨沃克斯维尔股份有限公司 | Use the nuclear reprogramming method of synthesis transcription factor |
-
2018
- 2018-04-09 CN CN201810312745.XA patent/CN108635376A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101031639A (en) * | 2004-07-01 | 2007-09-05 | 马克罗珀尔生物外科公司 | Methods of using regenerative cells to promote wound healing |
| CN106459918A (en) * | 2014-04-24 | 2017-02-22 | 得克萨斯州大学系统董事会 | Application of induced pluripotent stem cells to generate adoptive cell therapy products |
| CN107709544A (en) * | 2015-06-12 | 2018-02-16 | 隆萨沃克斯维尔股份有限公司 | Use the nuclear reprogramming method of synthesis transcription factor |
| CN106377547A (en) * | 2016-09-30 | 2017-02-08 | 孔五 | Extraction method and purpose of renewable particles of umbilical cord blood |
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Application publication date: 20181012 |
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