CN106581758A - Antibacterial acellular dermal dressing with vascularization inducing capability and preparation method thereof - Google Patents
Antibacterial acellular dermal dressing with vascularization inducing capability and preparation method thereof Download PDFInfo
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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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
- A61L27/3633—Extracellular matrix [ECM]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/60—Materials for use in artificial skin
-
- 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
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
-
- 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
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- 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
Abstract
The invention belongs to the technical field of biomedical engineering, in particular relates to the technical field of skin wound dressing preparation, and discloses an antibacterial acellular dermal dressing with vascularization inducing capability and a preparation method thereof. The antibacterial acellular dermal matrix dressing comprises porcine acellular dermal matrix, poly dopamine, vascular endothelial growth factor (VEGF) composition and antibacterial peptide LL37. The antibacterial acellular dermal dressing with the vascularization inducing capability has the advantages of low immunogenicity, good biocompatibility and antibacterial ability, and has obvious wound healing and vascularization promoting effect.
Description
Technical field
The invention belongs to biomedical engineering technology field, particularly skin wound dressing preparing technical field, disclose
A kind of antibacterial acellular dermal dressing of tool induced vascularization ability and preparation method thereof.
Background technology
With the progress to skin injury Therapy study, organization engineering skin is the focus of Recent study, has wide
Potential applicability in clinical practice.Skin is badly in need of obtaining when facing large-area burns or damaging substantial amounts of epidermis covering to promote skin group
The regeneration knitted and reparation.Acellular dermal matrix has many advantages, such as preferable skin regeneration material：（1）Price is just
Preferably, wide material sources；（2）There is good histocompatibility, nontoxic, antigenicity itself is low；（3）Degradability, material were being degraded
Gradually absorbed in journey and kept stable volume；（4）Growth cellular morphology thereon is maintained, promotes cell adhesion and growth；
（5）With certain porosity；（6）With certain mechanical strength and toughness, it is adapted to working cut.But due to acellular dermal
Substrate is a kind of inactive timbering material, and the low particularly Xenogenic acellular dermal matrix of vascularization ability, is facing in use
In bed application, another problem is how to solve the source of nutrition that bulk tissue or organ implant, it is obvious that
To realize in the case of without blood supply that the survival in vivo of a large amount of living cells is impossible, how to solve acellular dermal matrix
The problem of vascularization scarce capacity, is to break through the key that acellular dermal matrix promotes bottleneck in clinical practice in the application.This
Outward, acellular dermal matrix does not have antibacterial ability, easily receives bacterium infection and causes traumatic infection, have when using as dressing again
There is the danger of cross infection, also limit its application as dressing clinically.
VEGF (VEGF) can promote vascular endothelial cell division and propagation, increase venule, venule
Permeability, induce the expression of serine protease and interstitial collagenase, assemble Cytoplasm calcium, induction of vascular is generated, in wound
Recover from injury and play an important role in closing.The degraded Human Umbilical Vein Endothelial Cells migration of basement membrane is very necessary, and is pathologic vessels hypertrophy
The key link of startup.VEGF can induce the expression of MMP, MMP that necessary effect has been migrated to VEGF inducing endothelial cells.This
Outer VEGF can promote the release of plasminogen activator and its proteolytic enzyme, so as to capillary basement membrane of degrading, have
Beneficial to proliferating endothelial cells migration and chemotactic by its stimulation, lure that the capillary endothelium of convergence invades fiber gel into,
Form pipe spline structure or already present blood vessel and new vesselses are formed with form of germinateing.Additionally, VEGF can promote endothelial cell proliferation
Signal path and Endothelial Cell Survival effect and the signal path of Anti-G value.
Antimicrobial peptide LL-37 is the C-terminal cleaved products of cathelicidin peptide Hcap-18, can directly kill antibacterial, funguses
And virus.Antimicrobial peptide LL-37 broad-spectrum antiseptic, has inhibitory action to antibacterial, funguses, virus, mycoplasma.Wound can especially be suppressed
The common infection bacteria species of wound, such as Strep A (Group A Streptococcus), staphylococcus epidermidiss
(Staphylococcus epidermidis), staphylococcus aureuses (Staphylococcus aureus), bacillus pyocyaneus
(Pseudomonas aeruginosa), escherichia coli (Escherichia coli), Klebsiella Pneumoniae (Klebsiella
Pneumoniae), its minimal inhibitory concentration (MIC) be respectively 16,1. 5,6. 5,3. 2,6. 5,0. 8 μm of ol/L.
Except direct bactericidal action, antimicrobial peptide LL-37 also suppresses bacteriogenic endotoxin and CD14 positive cells to combine, and contributes to
Improve pyemia, reduce the death that infection is caused.Additionally, antimicrobial peptide LL-37 is also promoted by chemotactic immunocyte, regulation inflammatory
Enter the secretion of the factor/inhibitive factor, coordinate the function such as the natural immunity and acquired immunity, play immunoregulation effect.Antibacterial peptide
LL-37 may also act to vascular endothelial cell and epithelial cell, stimulates angiogenesis and promotes injury repairing.
The content of the invention
Present invention aims to the vascularization ability of existing acellular dermal matrix dressing is low, energy is repaired in induction
Power is poor, and does not possess anti-microbial property, there is provided a kind of antibacterial acellular dermal dressing of tool induced vascularization ability and its preparation side
Method.The present invention is by with the property acellular dermal dressing of pig source as substrate, being given birth to blood vessel endothelium after poly-dopamine surface modification
The long factor（VEGF）Its surface is fixed on antimicrobial peptide LL-37, and to prepare a kind of de- cell of antibacterial of tool induced vascularization ability true
Skin dressing.The acellular dermal dressing immunogenicity is low, and material source enriches, with good biocompatibility and antibiotic property
Can, and with good vascularization ability.
For achieving the above object, the technical scheme is that a kind of antibacterial acellular dermal of tool induced vascularization ability
Dressing is by acellular dermal matrix, VEGF（VEGF）With antibacterial peptide LL37.
Further, described acellular dermal dressing is counted by weight by 10 parts of acellular dermal matrix, poly- DOPA
Amine 0.25-0.5 parts, VEGF 0.01-0.05 parts, antibacterial peptide LL37 0.1-0.5 compositions.
Preferably, described VEGF（VEGF）, antibacterial peptide LL37 is purchased from U.S. Thermo Fisher
Scientifi companies.
For achieving the above object, another technical scheme of the invention is：A kind of antibacterial of tool induced vascularization ability is de- thin
The preparation method of born of the same parents' corium dressing, described preparation method are realized by following steps：（1）Acellular dermal matrix poly-dopamine
Surface modification,（2）VEGF（VEGF）Fixation,（3）The fixation of antibacterial peptide LL37.
Further, the acellular dermal matrix poly-dopamine surface modification is concretely comprised the following steps：
（1）Dopamine Hydrochloride is dissolved in trishydroxymethylaminomethane-hydrochloric acid buffer solution that pH is 8.5, concentration is configured to
For the Dopamine Hydrochloride solution of 10mg/mL；（2）Then acellular porcine dermal matrix is soaked in Dopamine Hydrochloride solution and is stirred
Reaction 24 hours, then deionized water ultrasonic cleaning 30 minutes, is placed in lyophilization in -80 DEG C of vacuum freeze driers.
Further, the VEGF（VEGF）Fixation it is specific as follows：
（1）By VEGF（VEGF）It is dissolved in trishydroxymethylaminomethane-hydrochloric acid buffer solution that pH is 8.5
In, gentle agitation 1 hour is configured to the VEGF solution that concentration is 1 ~ 5 μ g/mL, is then placed into standby under oxygen-free environment；（2）
The acellular porcine dermal matrix that table Jing poly-dopamines are modified, is infiltrated with sterilized deionized water, is then soaked in VEGF molten
It is placed in liquid in 37 DEG C of CO2 gas incubator and is incubated 24 hours, then with the deionized water rinsing of sterilizing, is placed in -80 DEG C of vacuum
Lyophilization in freezer dryer.
Further, the fixation of the antibacterial peptide LL37 is specific as follows：
（1）Antibacterial peptide LL37 is dissolved in the phosphate buffered solution that pH is 7.4, gentle agitation 1 hour, being configured to concentration is
The antibacterial peptide LL37 solution of 10 ~ 50 μ g/mL, is then placed into standby under gnotobasiss；
（2）By intravascular endothelial cell growth factor (ECGF)（VEGF）Fixing process acellular dermal matrix, be soaked in antibacterial peptide LL37
It is placed in solution in 37 DEG C of CO2 gas incubator and is incubated 12 hours, then with the deionized water rinsing of sterilizing, is placed in -80 DEG C
Lyophilization in vacuum freeze drier.Obtain final product the antibacterial acellular dermal matrix dressing of tool induction repair ability.
Further, described acellular porcine dermal matrix is application reference number for disclosed in 200410022506.9
Obtained by the preparation method of acellular dermal matrix material.
Beneficial effects of the present invention are：
（1）The present invention is by the method for poly-dopamine surface modification so that itself do not possess the acellular dermal of reaction site
The active reaction site of substrate, improving the cell adhesion ability of acellular dermal matrix strengthens its biocompatibility.
（2）The present invention passes through VEGF（VEGF）The acellular dermal base being modified with Jing poly-dopamines surface
The reaction of the catechol and amino isoreactivity group on matter surface, by VEGF（VEGF）It is fixed on acellular dermal
In substrate, acellular dermal matrix vascularization ability is being given.And by this kind of fixing meanss energy effectively solving blood vessel endothelium life
The long factor（VEGF）The short and prominent problem released of Half-life in vivo.
（3）The present invention is that antimicrobial component passes through primary amino radical and acellular dermal matrix on its peptide chain with antimicrobial peptide LL-37
The quinoid structure of upper poly-dopamine reacts, and antimicrobial peptide LL-37 is fixed in acellular dermal dressing, gives de- cell true
The excellent antibacterial ability of skin dressing.In addition, antibacterial peptide LL37 can recruit endothelial precursor cell to injury repairing site, and in activating
Chrotoplast, and VEGF（VEGF）Endothelial cell proliferation can be promoted, can be carried significantly under both synergism
The ability of the vascularization of high acellular dermal dressing.Additionally, by the fixing meanss of antimicrobial peptide LL-37 disclosed in this invention,
Can overcome antimicrobial peptide LL-37 because dash forward release caused by caused excessive concentration to the toxic effect of host cell, occur haemolysis
The deficiencies such as phenomenon.
Description of the drawings
Fig. 1 is the antibacterial ability evaluation experimental comparative result figure of embodiment 1～3 and comparative example；
Fig. 2 is Cytotoxic evaluation Comparative result after embodiment 1～3 and comparative example are co-cultured 1 day and 7 days with vascular endothelial cell
Figure；
Fig. 3 is the VEGF-ELISA testing result comparison diagrams of embodiment 1～3 and comparative example.
Specific embodiment
Technical scheme is described further with reference to embodiment.
Embodiment 1
A kind of antibacterial acellular dermal dressing of tool induced vascularization ability is by acellular dermal matrix, poly-dopamine, Ink vessel transfusing
Skin growth factor（VEGF）Constitute with antibacterial peptide LL37；Counted by 10 parts of acellular dermal matrix, poly-dopamine by weight
0.25 part, 0.01 part of VEGF, antibacterial peptide LL37 0.1 constitute；Described VEGF
（VEGF）, antibacterial peptide LL37 is purchased from Thermo Fisher Scientifi companies of the U.S..
Embodiment 2
A kind of antibacterial acellular dermal dressing of tool induced vascularization ability is by acellular dermal matrix, poly-dopamine, Ink vessel transfusing
Skin growth factor（VEGF）Constitute with antibacterial peptide LL37；Counted by 10 parts of acellular dermal matrix, poly-dopamine 0.3 by weight
Part, 0.025 part of VEGF, antibacterial peptide LL37 0.3 are constituted；Described VEGF（VEGF）, resist
Bacterium peptide LL37 is purchased from Thermo Fisher Scientifi companies of the U.S..
Embodiment 3
A kind of antibacterial acellular dermal dressing of tool induced vascularization ability is by acellular dermal matrix, poly-dopamine, Ink vessel transfusing
Skin growth factor（VEGF）Constitute with antibacterial peptide LL37；Counted by 10 parts of acellular dermal matrix, poly-dopamine 0.5 by weight
Part, 0.05 part of VEGF, antibacterial peptide LL370.5 composition；Described VEGF（VEGF）, antibacterial
Peptide LL37 is purchased from Thermo Fisher Scientifi companies of the U.S..
Embodiment 4
A kind of antibacterial acellular dermal dressing of tool induced vascularization ability of 1～embodiment of embodiment, 3 arbitrary example, its preparation method
Comprise the following steps that：
1. acellular dermal matrix poly-dopamine surface modification：
（1）Dopamine Hydrochloride is dissolved in trishydroxymethylaminomethane-hydrochloric acid buffer solution that pH is 8.5, concentration is configured to
For the Dopamine Hydrochloride solution of 10mg/mL；
（2）Then acellular porcine dermal matrix is soaked in into stirring reaction 24 hours in Dopamine Hydrochloride solution, then spend from
Sub- water is cleaned by ultrasonic 30 minutes, is placed in lyophilization in -80 DEG C of vacuum freeze driers.
2. VEGF（VEGF）Fixation：
（1）By VEGF（VEGF）It is dissolved in trishydroxymethylaminomethane-hydrochloric acid buffer solution that pH is 8.5
In, gentle agitation 1 hour is configured to the VEGF solution that concentration is 1 ~ 5 μ g/mL, is then placed into standby under oxygen-free environment；
（2）The acellular porcine dermal matrix that table Jing poly-dopamines are modified, is infiltrated with sterilized deionized water, is then soaked in
It is placed in VEGF solution in 37 DEG C of CO2 gas incubator and is incubated 24 hours, then with the deionized water rinsing of sterilizing, is placed in -80
Lyophilization in DEG C vacuum freeze drier.
3. the fixation of antibacterial peptide LL37：
（1）Antibacterial peptide LL37 is dissolved in the phosphate buffered solution that pH is 7.4, gentle agitation 1 hour, being configured to concentration is
The antibacterial peptide LL37 solution of 10 ~ 50 μ g/mL, is then placed into standby under gnotobasiss；
（2）By intravascular endothelial cell growth factor (ECGF)（VEGF）Fixing process acellular dermal matrix, be soaked in antibacterial peptide LL37
It is placed in solution in 37 DEG C of CO2 gas incubator and is incubated 12 hours, then with the deionized water rinsing of sterilizing, is placed in -80 DEG C
Lyophilization in vacuum freeze drier.Obtain final product the antibacterial acellular dermal matrix dressing of tool induction repair ability.
Embodiment 5
Comparative example：For single acellular porcine dermal matrix（Application reference number is the de- cell disclosed in 200410022506.9
Obtained by the preparation method of dermal matrix material）.
Experimental group 1 ~ 3：A kind of antibacterial acellular dermal dressing of the tool induced vascularization ability obtained by embodiment 1～3,
It is prepared from using the method for embodiment 4.
By a kind of antibacterial acellular dermal dressing of the tool induced vascularization ability prepared by above-described embodiment 1～3 with it is right
Ratio carries out antibacterial ability evaluation experimental, and comparative example 1 ~ 3 and comparative example contact 24 and to escherichia coli, staphylococcus aureus
Fungistatic effect after 72 hours.Experimental result such as Fig. 1 shows.
Understand embodiment 1 ~ 3 to contact 24 with escherichia coli and staphylococcus aureus little from anti-microbial property evaluation test result
When after bacteriostasis rate can reach more than 90%, after contact 72 hours, bacteriostasis rate can remain to be maintained at more than 90%, illustrate by this
1 ~ 3 pair of suppression having with escherichia coli and staphylococcus aureus rapidly and efficiently of embodiment obtained by bright disclosed method
Effect, and comparative example is antibacterial equal less than 10% after contacting 24 and 72 hours with escherichia coli and staphylococcus aureus, hence it is evident that do not have
There are bacteriostasis.
Embodiment 6
Comparative example：For single acellular porcine dermal matrix（Application reference number is the de- cell disclosed in 200410022506.9
Obtained by the preparation method of dermal matrix material）.
Experimental group 1 ~ 3：The antibacterial acellular dermal dressing of the tool induced vascularization ability obtained by embodiment 1～3, adopts
The method of embodiment 4 is prepared from.
By the dressing of antibacterial acellular dermal and comparative example of the tool induced vascularization ability prepared by above-described embodiment 1～3
Carry out Cytotoxic evaluation experiment（Tested by GB GB/T 16886.5-2003）, comparative example 1 ~ 3 and comparative example
It is right.Experimental result such as Fig. 2 shows.
Cytotoxicity testing result shows embodiment 1 ~ 3, and after co-culturing 1 day and 7 days with vascular endothelial cell, which is corresponding
With respect to the rate of increase more than 95%, cytotoxicity is rated 0 grade to cell, it was demonstrated which has good describing property of cell, and contrasts
Example co-culture 1 day and 7 days with vascular endothelial cell after the relative rate of increase of its corresponding cell 80% or so, cytotoxicity is graded
For 2 grades, with slight cytotoxicity.Additionally, the relative of the prolongation embodiment 1 ~ 3 of the time of co-cultivation is compared the rate of increase and has bright
Aobvious to improve, its cell is high with respect to more negative group of the rate of increase after 7 days for embodiment 1 ~ 3（It is above 130%）, it was demonstrated that using this
Acellular dermal matrix dressing obtained by the disclosed preparation method of invention has significant promotion vascular endothelial cell growth
Effect, is conducive to the formation of undamaged portion fresh blood tubing.
Embodiment 7
Comparative example：For single acellular porcine dermal matrix（Application reference number is the de- cell disclosed in 200410022506.9
Obtained by the preparation method of dermal matrix material）.
Experimental group 1 ~ 3：The antibacterial acellular dermal dressing of the tool induced vascularization ability obtained by embodiment 1～3, adopts
The method of embodiment 4 is prepared from.
By the dressing of antibacterial acellular dermal and comparative example of the tool induced vascularization ability prepared by above-described embodiment 1～3
VEGF contents are detected using VEGF-ELISA Kit methods, by the content of VEGF how much evaluating its vascularization ability
Height（The content of VEGF and vascularization ability positive correlation）.As a result as Fig. 3 shows.
VEGF-ELISA testing results find that the VEGF contents of embodiment 1 ~ 3 want containing for much higher VEGF relative to comparative example
Amount is higher, and which is more susceptible to the extracellular region specific binding of receptor.VEGF and its receptor are in extracellular specific binding
Afterwards, by tyrosine residue autophosphorylation in the allosteric inducing cell of space, and in active cell signal transduction and play biology
Effect is learned, is promoted vascular endothelial cell division and propagation, is increased venule, venular permeability, induce serine protease
With the expression of interstitial collagenase, assemble Cytoplasm calcium, induction of vascular is generated.
Embodiment 8
Comparative example：For single acellular porcine dermal matrix（Application reference number is the de- cell disclosed in 200410022506.9
Obtained by the preparation method of dermal matrix material）.
Experimental group 1 ~ 3：The antibacterial acellular dermal dressing of the tool induced vascularization ability obtained by embodiment 1～3, adopts
The method of embodiment 4 is prepared from.
By the dressing of antibacterial acellular dermal and comparative example of the tool induced vascularization ability prepared by above-described embodiment 1～3
Carry out animal wound repair experiment, contrast experiment's group and repairing effect of the comparative example to wound surface.Experimental result is as shown in table 1：
1 wound repair experimental result of table
| Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example | |
| The healing conjunction time | 7 | 7 | 5 | 10 |
| Healing state | NIP in wound healing process, the shape such as suppurate Condition is produced without cicatrix after there is healing. | NIP in wound healing process, suppurates Situation is produced without cicatrix after there is healing. | NIP in wound healing process, the situation such as suppurate are sent out Produce without cicatrix after raw healing. | With inflammation, the situation such as suppurate, wound heal wound There is obvious cicatrix to produce after conjunction. |
As seen from the above table, the antibacterial acellular dermal dressing of the tool induced vascularization ability prepared by embodiment 1 ~ 3 is in wound
Effectively can prevent because of bacterium infection in agglutination, and can effectively shorten the time of wound healing.And comparative example is in application
During with obvious bacterium infection situation.
Obviously, the above embodiment of the present invention is only intended to clearly illustrate example of the present invention, and is not right
The restriction of embodiments of the present invention；For those of ordinary skill in the field, may be used also on the basis of the above description
To make other changes in different forms, there is no need to be exhaustive to all of embodiment；It is all this
Any modification, equivalent and improvement made within the spirit and principle of invention etc., should be included in the claims in the present invention
Protection domain within.
Claims (7)
1. it is a kind of tool induced vascularization ability antibacterial acellular dermal dressing, it is characterised in that by following parts by weight into
It is grouped into：10 parts of acellular dermal matrix, poly-dopamine 0.25-0.5 part, VEGF 0.01-0.05 parts, antibacterial
Peptide LL37 0.1-0.5.
2. the antibacterial acellular dermal dressing of tool induced vascularization ability according to claim 1, it is characterised in that described
Acellular dermal matrix be pig source property acellular dermal matrix.
3. the preparation method of the antibacterial acellular dermal dressing of a kind of tool induced vascularization ability described in claim 1, which is special
Levy and be, described preparation method is realized by following steps：Acellular dermal matrix poly-dopamine surface modification, blood vessel endothelium
The fixation of somatomedin, the fixation of antibacterial peptide LL37.
4. it is according to claim 3 it is a kind of tool induced vascularization ability antibacterial acellular dermal dressing preparation method,
Characterized in that, the acellular dermal matrix poly-dopamine surface modification is concretely comprised the following steps：
（1）Dopamine Hydrochloride is dissolved in trishydroxymethylaminomethane-hydrochloric acid buffer solution that pH is 8.5, quality is configured to
Dopamine Hydrochloride solution of the concentration for 10mg/mL；
（2）Acellular dermal matrix is soaked in into stirring reaction 24 hours in the Dopamine Hydrochloride solution, deionization is then used
Water is cleaned by ultrasonic 30 minutes, is placed in lyophilization in -80 DEG C of vacuum freeze driers.
5. it is according to claim 3 it is a kind of tool induced vascularization ability antibacterial acellular dermal dressing preparation method,
Characterized in that, the fixation of the VEGF is concretely comprised the following steps：
（1）VEGF is dissolved in trishydroxymethylaminomethane-hydrochloric acid buffer solution that pH is 8.5, softly
Stirring 1 hour, is configured to the VEGF solution that mass concentration is 1 ~ 5 μ g/mL, is then placed into standby under oxygen-free environment；
（2）The acellular porcine dermal matrix that Jing poly-dopamines are modified, is infiltrated with sterilized deionized water, is then soaked in
It is placed in VEGF solution in 37 DEG C of CO2 gas incubator and is incubated 24 hours, then with the deionized water rinsing of sterilizing, is placed in -80
Lyophilization in DEG C vacuum freeze drier.
6. it is according to claim 3 it is a kind of tool induced vascularization ability antibacterial acellular dermal dressing preparation method,
Characterized in that, the fixation of the antibacterial peptide LL37 is concretely comprised the following steps：
（1）Antibacterial peptide LL37 is dissolved in the phosphate buffered solution that pH is 7.4, gentle agitation 1 hour is configured to quality dense
The antibacterial peptide LL37 solution for 10 ~ 50 μ g/mL is spent, is then placed into standby under gnotobasiss；
（2）By the acellular dermal matrix of the fixing process of intravascular endothelial cell growth factor (ECGF), the antibacterial peptide LL37 is soaked in molten
It is placed in liquid in 37 DEG C of CO2 gas incubator and is incubated 12 hours, then with the deionized water rinsing of sterilizing, is placed in -80 DEG C very
Lyophilization in vacuum freecing-dry machine, obtains final product the antibacterial acellular dermal dressing of the tool induced vascularization ability.
7. the preparation side according to the antibacterial acellular dermal dressing of the arbitrary described tool induced vascularization ability of claim 3 to 6
Method, it is characterised in that the acellular dermal matrix is pig source property acellular dermal matrix.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710021832.5A CN106581758A (en) | 2017-01-12 | 2017-01-12 | Antibacterial acellular dermal dressing with vascularization inducing capability and preparation method thereof |
Applications Claiming Priority (1)
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| CN109260518A (en) * | 2018-11-28 | 2019-01-25 | 广州聚明生物科技有限公司 | A kind of Oral Defects repair membrane and preparation method thereof |
| CN109498840A (en) * | 2018-11-28 | 2019-03-22 | 广州聚明生物科技有限公司 | A kind of Ocular surface healing film and preparation method thereof |
| CN109771691A (en) * | 2019-01-24 | 2019-05-21 | 广州创赛生物医用材料有限公司 | A kind of conductive hydrogel material and preparation method thereof |
| CN111035806A (en) * | 2018-10-12 | 2020-04-21 | 上海市静安区闸北中心医院 | Anti-infection biological material and preparation method thereof |
| CN111097067A (en) * | 2020-01-10 | 2020-05-05 | 温州医科大学 | Antibacterial medical dressing for promoting wound to heal rapidly and preparation method thereof |
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| CN111744051A (en) * | 2020-07-15 | 2020-10-09 | 中国人民解放军西部战区总医院 | Preparation method and wound healing method of graphene oxide-lysozyme/alkaline fibroblast growth factor composite dressing |
| CN111744050A (en) * | 2020-07-15 | 2020-10-09 | 中国人民解放军西部战区总医院 | Preparation method and wound healing method of graphene oxide-daptomycin/epidermal growth factor composite dressing |
| CN111921000A (en) * | 2020-07-15 | 2020-11-13 | 中国人民解放军西部战区总医院 | Preparation method of graphene oxide-nano silver/insulin-like growth factor-1 composite dressing and wound healing method |
| CN114870034A (en) * | 2022-02-17 | 2022-08-09 | 上海交通大学医学院附属仁济医院 | Gene transfection nano material with efficient anti-infection capacity and preparation thereof |
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