CN105709270B - A kind of aerogel dressing for stem-cell therapy - Google Patents
A kind of aerogel dressing for stem-cell therapy Download PDFInfo
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- CN105709270B CN105709270B CN201610156699.XA CN201610156699A CN105709270B CN 105709270 B CN105709270 B CN 105709270B CN 201610156699 A CN201610156699 A CN 201610156699A CN 105709270 B CN105709270 B CN 105709270B
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- 239000004964 aerogel Substances 0.000 title claims abstract description 67
- 238000009168 stem cell therapy Methods 0.000 title claims abstract description 11
- 238000009580 stem-cell therapy Methods 0.000 title claims abstract description 11
- 210000000130 stem cell Anatomy 0.000 claims abstract description 89
- 239000000017 hydrogel Substances 0.000 claims abstract description 73
- 108010010803 Gelatin Proteins 0.000 claims abstract description 22
- 239000008273 gelatin Substances 0.000 claims abstract description 22
- 229920000159 gelatin Polymers 0.000 claims abstract description 22
- 235000019322 gelatine Nutrition 0.000 claims abstract description 22
- 235000011852 gelatine desserts Nutrition 0.000 claims abstract description 22
- 230000003287 optical effect Effects 0.000 claims abstract description 13
- 239000000463 material Substances 0.000 claims abstract description 8
- 238000004132 cross linking Methods 0.000 claims description 11
- 230000004048 modification Effects 0.000 claims description 10
- 238000012986 modification Methods 0.000 claims description 10
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 4
- 238000011534 incubation Methods 0.000 claims description 4
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 4
- JCBWQNLTYXTHBZ-UHFFFAOYSA-N 2-azidobenzoic acid Chemical compound OC(=O)C1=CC=CC=C1N=[N+]=[N-] JCBWQNLTYXTHBZ-UHFFFAOYSA-N 0.000 claims description 3
- 229920001661 Chitosan Polymers 0.000 claims description 3
- 239000000499 gel Substances 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 230000035755 proliferation Effects 0.000 abstract description 17
- 230000008439 repair process Effects 0.000 abstract description 5
- 230000012010 growth Effects 0.000 abstract description 4
- 230000001737 promoting effect Effects 0.000 abstract description 3
- 208000027418 Wounds and injury Diseases 0.000 description 30
- 206010052428 Wound Diseases 0.000 description 29
- 210000004027 cell Anatomy 0.000 description 29
- 230000000694 effects Effects 0.000 description 9
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- 230000000975 bioactive effect Effects 0.000 description 8
- 238000000034 method Methods 0.000 description 8
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- 238000012360 testing method Methods 0.000 description 8
- 239000003102 growth factor Substances 0.000 description 6
- 210000001626 skin fibroblast Anatomy 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
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- 230000006378 damage Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 230000036259 sexual stimuli Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 230000004069 differentiation Effects 0.000 description 3
- PQXPAFTXDVNANI-UHFFFAOYSA-N 4-azidobenzoic acid Chemical compound OC(=O)C1=CC=C(N=[N+]=[N-])C=C1 PQXPAFTXDVNANI-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
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- 210000001339 epidermal cell Anatomy 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
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- 239000007850 fluorescent dye Substances 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 239000003292 glue Substances 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 2
- 230000035699 permeability Effects 0.000 description 2
- 238000000016 photochemical curing Methods 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 230000029663 wound healing Effects 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 206010072170 Skin wound Diseases 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 1
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
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- 239000003814 drug Substances 0.000 description 1
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- 210000000630 fibrocyte Anatomy 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
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- 238000002955 isolation Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000001473 noxious effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000007420 reactivation Effects 0.000 description 1
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- 231100000397 ulcer Toxicity 0.000 description 1
Classifications
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- 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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/008—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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0014—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0023—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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0028—Polypeptides; Proteins; Degradation products thereof
- A61L26/0038—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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0057—Ingredients of undetermined constitution or reaction products thereof
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- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
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- General Health & Medical Sciences (AREA)
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Abstract
The present invention provides a kind of aerogel dressings for stem-cell therapy.Aerogel dressing provided by the invention includes hydrogel layer and hydrogel modified layer, and hydrogel modified layer material therefor is the gelatin containing optical active group.Subject hydrogel dressing is conducive to the adherency and proliferation of stem cell, and the active factors for promoting surface of a wound growth to repair can be discharged into the surface of a wound by stem cell with this configuration, while being avoided stem cell and being contacted with the direct of the surface of a wound, realizes the security application of stem cell.
Description
Technical field
The present invention relates to field of medicaments, in particular to a kind of aerogel dressing for stem-cell therapy.
Background technique
Skin is the maximum organ of human body, and total surface area is about 1.5-2.0m2, play the role of protection and support body.
Once skin is because damage and failure occur in burn, wound, ulcer etc., body will lose large quantity of moisture, electrolyte, protein
Deng the easy injury by bacterium and external environment at the same time.
In recent years, the stem cell of the potential with self-renewing, hyperproliferation and Multidirectional Differentiation has been increasingly becoming the surface of a wound and has been cured
Close the focus of repairing research.Stem cell (such as mesenchymal stem cell, fat mesenchymal stem cell etc.) is a kind of with self
It updates, the cell of Multidirectional Differentiation and high proliferative capacity, it can have effective feeling by surrounding microenvironment, and carry out standard according to microenvironment
Really break up and secrete corresponding regulatory factor, promotes the reparation and regeneration of tissue.Therefore, stem cell is repaired applied to skin wound
The enough effective healings for accelerating the surface of a wound of reactivation, reach better repairing effect.
Currently, common stem-cell therapy generallys use direct injection, carrier or biological support and implants, stem cell is straight
It connects and human contact and participates in process of tissue reparation.However, the Spain of " cancer research " report and Danish scientist discovery are aobvious
Show: in vitro in breeding, having part stem cell that canceration may occur, it means that stem cell directly contacts with body and is used for
Tissue repair has potential risk.The security application of stem cell gradually causes extensive concern and the attention of scientist.Such as
What can be safer while promoting skin repair using stem cell, and avoid brought by stem cell directly contacts with body
Potential danger becomes the hot spot of medical research.
Hydrogels Wound dressing has as a kind of ideal Wound dressing and maintains the enough humidity of the surface of a wound, and absorbs big
The advantages of measuring diffusate and noxious material.At the same time, the pore structure of hydrogel is conducive to the gas exchanges of the surface of a wound again, guarantees wound
The good gas permeability in face.Moreover, many Hydrogels dressing (such as PVA) are due to its smooth surface texture, wound tissue is not easy
In being adhered, the secondary damage of more change dressings is alleviated.In view of the good permeability of aerogel dressing and mass exchange performance,
Stem cell is inoculated in the surface of hydrogel, the growth factor of the promotion skin histology reparation that stem cell is discharged (such as VEGF,
BFGF) the isoreactivity factor penetrates into wound surface by the porous structure of hydrogel, and to cannot pass through hydrogel direct for stem cell
Contacted with the surface of a wound, thus using stem cell secretion active factors while, avoid stem cell and directly contacted with body, realize
Safer stem cell application.Therefore, Hydrogels Wound dressing plays a significant role in wound repair, it has application letter
Just, rapidly, the lower advantage of cost, be suitable for extensive wound reparation and first aid.However, hydrogel is unfavorable for cell adherence is
" double-edged sword ", it be both avoided that be adhered with the surface of a wound reduce more change dressings secondary damage, but be unfavorable for surface on it and connect
Kind stem cell, to promote wound repair using the active factors of stem cell secretion.
Currently, there is research that stem cell is mixed into hydrogel, directly spreads on the surface of a wound and carry out repairing and treating.The technology is due to water-setting
The limitation of glue inner space, while cell exists with free state, is neither able to maintain the good state of stem cell, is unfavorable for simultaneously
Stem cells hyperplasia differentiation, preferably to discharge the active factors for promoting wound repair.Meanwhile this method stem cell is inevitable
Meeting it is in direct contact with the skin, however it remains potential safety wind it is dangerous.Therefore it provides one kind is conducive to stem cell adherency
, safe aerogel dressing has important practical significance.
Summary of the invention
In view of this, the present invention provides a kind of aerogel dressings for stem-cell therapy.The aerogel dressing is advantageous
In the adherency and proliferation of stem cell, while it can realize the security application of stem cell.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides a kind of aerogel dressing for stem-cell therapy, the aerogel dressing include hydrogel layer and
Hydrogel modified layer, hydrogel modified layer material therefor are the gelatin containing optical active group.
Gelatin containing optical active group is fixed on the one side of aerogel dressing by UV crosslinking technology by the present invention, real
It is now modified to the surface of aerogel dressing, in favor of the adherency and proliferation of stem cell, stem cell is inoculated in the modification of hydrogel
Upper surface, completely cut off stem cell and the surface of a wound, the active factors of stem cell secretion can pass through aerogel dressing and penetrate into the surface of a wound, from
And while reaching wound healing effect, avoid stem cell contacts with the direct of the surface of a wound, realizes stem cell security application.
Preferably, the gelatin containing optical active group is the gelatin modified through 4- azidobenzoic acid.4- azidobenzoic acid
Under ultraviolet excitation, active nitrogen is generated, so that gelatin is grafted in hydrogel carbochain.
Preferably, aerogel dressing the preparation method comprises the following steps: hydrogel modified layer is fixed on by UV crosslinking technology
On hydrogel layer.
Preferably, the time of UV crosslinking is 10~60s.
In some embodiments provided by the invention, the time of UV crosslinking is 20s.
Preferably, hydrogel layer material therefor is hydrogel, hydrogel is polyvinyl alcohol and/or chitosan.But this hair
Bright to be not limited to this, those skilled in the art think that feasible hydrogel material is within the scope of the present invention.
Preferably, the concentration of the gelatin containing optical active group is 0.01~10mg/mL.
In some embodiments provided by the invention, the concentration of the gelatin containing optical active group is 1mg/mL.
Preferably, the surface being in contact on hydrogel layer with hydrogel modified layer is provided with sunk structure.This recess
On the one hand structure is conducive to stem cell and survive, grow and breed in sunk structure, formation cell colony, holding cell activity with
The expression and release of bioactive substance；On the other hand, it is inoculated in stem cell in sunk structure by hydrogel completely and skin
Skin isolation avoids stem cell and direct body contact and bring safety issue.Meanwhile the growth factor of stem cell secretion
Equal bioactive substances can be released to wound surface by the network structure of hydrogel, promote the reparation of wound.And stem cell
The formation of colony can preferably secrete various cell factors, improve skin since there are mutual contact sexual stimulus between cell
The efficiency of skin reparation.Hydrogel lower surface still maintains its characteristic for being unfavorable for cell adherence, and dressing will not stick together, be convenient for
The replacement of dressing.
In some embodiments provided by the invention, sunk structure is hole and/or slot.But present invention is not limited to this,
Those skilled in the art think that feasible sunk structure is within the scope of the present invention.
It In some embodiments provided by the invention, further include stem cell, stem cell is inoculated in hydrogel modified layer.
In some embodiments provided by the invention, the incubation time of stem cell is 1~14 day.
The present invention provides a kind of aerogel dressing for stem-cell therapy, the aerogel dressing include hydrogel layer and
Hydrogel modified layer, hydrogel modified layer material therefor are the gelatin containing optical active group.The present invention at least has following excellent
One of gesture:
1, the gelatin containing optical active group is fixed on the one side of aerogel dressing by UV crosslinking technology by the present invention,
It realizes and the surface of aerogel dressing is modified, in favor of the adherency and proliferation of stem cell；
2, stem cell is inoculated in the upper surface of the modification of hydrogel, completely cuts off stem cell and the surface of a wound, stem cell secretion
Active factors can pass through aerogel dressing and penetrate into the surface of a wound, to avoid dry thin while reaching wound healing effect
Born of the same parents directly contact with the surface of a wound, realize stem cell security application；
3, the surface being in contact on hydrogel layer with hydrogel modified layer is provided with sunk structure, this one side of sunk structure
Face is conducive to stem cell and survives, grows and breed in sunk structure, forms cell colony, keeps cell activity and bioactivity
The expression and release of substance；On the other hand, the stem cell being inoculated in sunk structure is isolated with skin completely by hydrogel, is kept away
Stem cell and direct body contact are exempted from and bring safety issue.And the formation of stem cell colonies, due to being deposited between cell
In mutual contact sexual stimulus, various cell factors can be preferably secreted, the efficiency of skin repair is improved；
4, aerogel dressing lower surface still maintains its characteristic for being unfavorable for cell adherence, and dressing will not stick together, just
In the replacement of dressing.
Detailed description of the invention
Fig. 1 shows pass aerogel dressing structural schematic diagram；Wherein, a is pass aerogel dressing top view, and 1 shows hydrogel
Layer, 2 show hydrogel modified layer；B is pass aerogel dressing sectional drawing, and 1 shows hydrogel layer, and 2 show hydrogel modified layer；
Fig. 2 shows the schematic diagram of stem cell inoculation aerogel dressing；
Fig. 3 shows that stem cell adheres to situation in the growth of dressing surface；Wherein, a shows that control group, b show test group；
Fig. 4 is shown as fibrocyte proliferation situation；
Fig. 5 shows groove profile aerogel dressing top view；1 shows hydrogel layer, and 2 show hydrogel modified layer；
Specific embodiment
The invention discloses a kind of aerogel dressing for stem-cell therapy, those skilled in the art can be used for reference herein
Content is suitably modified realization of process parameters.In particular, it should be pointed out that all similar substitutions and modifications are to those skilled in the art
It is it will be apparent that they are considered as being included in the present invention for member.Method and application of the invention has passed through preferably real
Example is applied to be described, related personnel obviously can not depart from the content of present invention, in spirit and scope to method described herein
It is modified or appropriate changes and combinations with application, carrys out implementation and application the technology of the present invention.
It is available on the market provided by the present invention for biomaterial used in the aerogel dressing of stem-cell therapy.
Below with reference to embodiment, the present invention is further explained:
Embodiment 1
The structural schematic diagram of aerogel dressing (chitosan) is shown in Fig. 1.Wherein, Fig. 1-a shows the vertical view of pass aerogel dressing
Figure, Fig. 1-b show the sectional drawing of pass aerogel dressing.Surface manufactures micropore on the hydrogel, and the bottom in hole still retains one layer of water
Gel completely cuts off it with the surface of a wound.In dressing before use, stem cell need to be only inoculated in the upper surface of hydrogel, that is, can be applied to
Wound repair.On the one hand the pass aerogel dressing is conducive to stem cell and survives, grows and breed in hole, form cell collection
It falls, keeps the expression and release of cell activity and bioactive substance；On the other hand, the stem cell being inoculated in hole passes through water-setting
Glue is isolated with skin completely, avoids stem cell and direct body contact and bring safety issue.
It is modified to carry out surface to aerogel dressing: (light is living for the optical active group that contains of configuration a certain concentration (1mg/mL)
Property group be four azidobenzoic acids by covalent bond mode in conjunction with the amino of gelatin after obtain) gelatin.Gained light is living
Property gelatin solution be laid in the surface of hydrogel, stand 12h.Hydrogel is placed in UV crosslinking instrument and carries out UV crosslinking 20s, is obtained
Obtain modified aerogel dressing.The present invention carries out photocuring to hydrogel upper surface by photolytic activity gelatin and is modified, can either
The hole of aerogel dressing upper surface is set to be conducive to the adherency proliferation of stem cell, while hydrogel lower surface still maintains it and is unfavorable for
The characteristic of cell adherence, dressing will not stick together, convenient for the replacement of dressing.
Be inoculated with stem cell: stem cell is inoculated in the modification of hydrogel by disinfection treatment by modified aerogel dressing
Face, the inoculum density of stem cell are 104A/cm2, it cultivates 10 days, the schematic diagram of the aerogel dressing after stem cell inoculation is shown in Fig. 2,
The bioactive substances such as the growth factor of stem cell secretion can be in contact by the network structure of hydrogel with epidermal cell；Together
When control group is set, control group is inoculated with stem cell by above-mentioned identical method using unmodified aerogel dressing, and carries out thin
Born of the same parents' culture.Growth by Fluorescent Staining Observation stem cell in different dressing surfaces adheres to situation (Fig. 3).
From the figure 3, it may be seen that aerogel dressing after modified, can promote the proliferation of stem cell.As it can be seen that the modification of aerogel dressing
Be conducive to the adherency and proliferation of cell, the bioactive substances such as growth factor of stem cell secretion can pass through the network of hydrogel
Structure release promotes the reparation of wound in wound surface.And the formation of stem cell colonies, due to being connect between cell there are mutual
Sexual stimulus is touched, various cell factors can be preferably secreted, improve the efficiency of skin repair.
External reparative experiment:
The test is the proliferation test of skin fibroblasts, specific test operation are as follows: be inoculated with skin fibroblasts
In culture plate, after cell is adherent, the culture that embodiment 1 is obtained has the aerogel dressing of stem cell to be placed on attached cell
Aerogel dressing is taken out after culture 3 days on surface.Meanwhile be arranged control group, control group using it is above-mentioned it is unmodified, through being inoculated with
Aerogel dressing after stem cell.Have to culture and MTT is added in fibroblastic hole, 37 DEG C of incubation 4h discard culture solution, add
Enter dimethyl sulfone (DMSO), 490nm detects absorbance value, analyzes different dressing and covers lower fibroblast proliferation situation.As a result
See Fig. 4.
As shown in Figure 4, the aerogel dressing of the embodiment of the present invention 1 can significantly promote the proliferation of skin fibroblasts.
Embodiment 2
The structure of aerogel dressing (polyvinyl alcohol) is groove profile aerogel dressing, and structural schematic diagram is shown in Fig. 5 (top view).?
Hydrogel upper surface manufactures microflute, and the bottom of slot still retains one layer of hydrogel, completely cuts off it with the surface of a wound.In dressing before use, only
Stem cell need to be inoculated in the upper surface of hydrogel, that is, can be applied to wound repair.The groove profile aerogel dressing is on the one hand advantageous
Survive, grow and breed in slot in stem cell, form cell colony, keep cell activity and bioactive substance expression and
Release；On the other hand, the stem cell being inoculated in slot is isolated with skin completely by hydrogel, and it is straight with human body to avoid stem cell
It contacts and bring safety issue.
It is modified to carry out surface to aerogel dressing: (light is living for the optical active group that contains of configuration a certain concentration (10mg/mL)
Property group be four azidobenzoic acids by covalent bond mode in conjunction with the amino of gelatin after obtain) gelatin.Gained light is living
Property gelatin solution be laid in the surface of hydrogel, stand for 24 hours.Hydrogel is placed in UV crosslinking instrument and carries out UV crosslinking 20s, is obtained
Obtain modified aerogel dressing.The present invention carries out photocuring to hydrogel upper surface by photolytic activity gelatin and is modified, can either
The hole of aerogel dressing upper surface is set to be conducive to the adherency proliferation of stem cell, while hydrogel lower surface still maintains it and is unfavorable for
The characteristic of cell adherence, dressing will not stick together, convenient for the replacement of dressing.
Be inoculated with stem cell: stem cell is inoculated in the modification of hydrogel by disinfection treatment by modified aerogel dressing
Face, the inoculum density of stem cell are 104A/cm2, cultivate 14 days, the bioactive substances such as growth factor of stem cell secretion can be with
It is in contact by the network structure of hydrogel with epidermal cell；Control group is set simultaneously, and control group uses unmodified hydrogel
Dressing is inoculated with stem cell by above-mentioned identical method, and carries out cell culture.By Fluorescent Staining Observation stem cell different
The growth of dressing surface adheres to situation (close with Fig. 3).
According to the experimental results, aerogel dressing after modified, can promote the proliferation of stem cell.As it can be seen that aerogel dressing
Modification be conducive to the adherency and proliferation of cell, the bioactive substances such as growth factor of stem cell secretion can pass through hydrogel
Network structure be released to wound surface, promote the reparation of wound.And the formation of stem cell colonies, since there are that between cell
This contact sexual stimulus can preferably secrete various cell factors, improve the efficiency of skin repair.
External reparative experiment:
The test is the proliferation test of skin fibroblasts, specific test operation are as follows: be inoculated with skin fibroblasts
In culture plate, after cell is adherent, the culture that embodiment 2 is obtained has the aerogel dressing of stem cell to be placed on attached cell
Aerogel dressing is taken out after culture 3 days on surface.Meanwhile be arranged control group, control group using it is above-mentioned it is unmodified, through being inoculated with
Aerogel dressing after stem cell.Have to culture and MTT is added in fibroblastic hole, 37 DEG C of incubation 4h discard culture solution, add
Enter dimethyl sulfone (DMSO), 490nm detects absorbance value, analyzes different dressing and covers lower fibroblast proliferation situation.As a result
It is close with Fig. 4.
By above-mentioned test result it is found that the aerogel dressing of the embodiment of the present invention 2 can significantly promote skin fibroblasts
Proliferation.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (8)
1. a kind of aerogel dressing for stem-cell therapy, which is characterized in that the aerogel dressing includes stem cell, water-setting
Glue-line and hydrogel modified layer, the hydrogel modified layer material therefor are the gelatin containing optical active group；
The surface being in contact on hydrogel layer with hydrogel modified layer is provided with sunk structure, and the stem cell is inoculated in the water
In gel modified layer.
2. aerogel dressing according to claim 1, which is characterized in that the gelatin containing optical active group is through 4-
The gelatin of azidobenzoic acid modification.
3. aerogel dressing according to claim 1, which is characterized in that the aerogel dressing the preparation method comprises the following steps: logical
It crosses UV crosslinking technology the hydrogel modified layer is fixed on hydrogel layer.
4. aerogel dressing according to claim 3, which is characterized in that the time of the UV crosslinking is 10~60s.
5. aerogel dressing according to claim 1, which is characterized in that the hydrogel layer material therefor is hydrogel,
The hydrogel is polyvinyl alcohol and/or chitosan.
6. aerogel dressing according to claim 5, which is characterized in that the concentration of the gelatin containing optical active group
For 0.01~10mg/mL.
7. aerogel dressing according to claim 1, which is characterized in that the sunk structure is hole and/or slot.
8. aerogel dressing according to claim 1, which is characterized in that the incubation time of the stem cell is 1~14 day.
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