CN109705369A - A kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel and the preparation method and application thereof - Google Patents
A kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel and the preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogels and the preparation method and application thereof.It is realized by following scheme: dopamine (DA) being introduced into neurotransmitter communication cell transmission chemical substance by chemical crosslinking grafted sodium alginate (SA) and reaches promotion wound healing effect, improve physical adhesion ability and improves Biocompatibility, the good and cheap and easily-available compound raising hydrogel physical property of material pharmaceutical grade Polyethylene alcohol (PVA) with biocompatibility again, to prepare a kind of higher hydrogel of mechanical strength.Hydrogel made from the application has good biocompatibility and excellent physical property, and reaction raw materials are cheap and easily-available, reaction condition is mild and operating procedure is easy.It has a wide range of applications in decubitus wound nursing and biomedicine.
Description
Technical field
The invention belongs to bioabsorbable polymer material fields, and in particular to a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel
Glue and the preparation method and application thereof.
Background technique
Bedsore is also known as pressure sore as clinical common complication.Causing the most important transient cause of pressure sore is pressure factor point
Not are as follows: pressure at right angle (local organization is by duration pressure at right angle such as body bone tuberosity protrusion), (skin rubs frictional force
Wipe), shearing (active force cause after imposing on object generate a parallel opposite direction planar slide, as lie down raise the head of a bed
When body glide, there is parallel frictional force with bed in skin, in addition the gravity of skin vertical direction, so as to cause the production of shearing
It is raw, cause local skin blood circulation disorder and pressure sore occurs).Clinically nursing is divided into two kinds at present: first is that conventional aseptic yarn
Cloth nursing；Second is that new pattern compress wound care, the nursing of conventional aseptic gauze has the barrier action of gauze poor, bacterial invasion
Possibility is higher, the surface of a wound granulation tissue is easy to grow in the mesh of gauze, and when dressing can damage cambium and cause to ache
Bitterly, to wound healing without be obviously promoted effect, sticky wound, for the damaged surface of a wound replace when not only pain may also result in it is secondary
The distinct disadvantages such as wound.
Researchers attempt to prepare novel bearing hydrocolloid dressing with natural and synthesis high molecular material to solve the problems, such as.However it is existing
There is bearing hydrocolloid dressing only to consider biocompatibility and mechanical property problem in design mostly, not introducing a kind of can promote wound
Healing substances, such dressing have obvious disadvantage: as preparation cost is excessively high, preparation process is many and diverse and is not easy to clean, reacts
The problems such as condition is excessively high and promotes wound healing effect bad, can not be than more comprehensively solving pressure sore later period nursing problem.Cause
This, there is an urgent need to further study with multi-functional and the excellent material of biocompatibility, more than realization can be solved disposably
Problem is applied to clinical diagnosis as early as possible and studies.
Summary of the invention
To solve the shortcomings and deficiencies of the prior art, the primary purpose of the present invention is that providing a kind of sodium alginate-
Dopamine/polyvinyl alcohol hydrogel.
Another object of the present invention is to provide a kind of above-mentioned sodium alginate-dopamine/polyvinyl alcohol hydrogel preparations
Method.
A further object of the present invention is to provide a kind of above-mentioned sodium alginate-dopamine/polyvinyl alcohol hydrogels in biology
The application in field.
The object of the invention is achieved through the following technical solutions:
A kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel preparation method, comprising the following steps:
(1) N- hydroxysuccinimidyl is added into sodium alginate (SA) aqueous solution that concentration is 1~2% (i.e. 1~2g/100ml)
Dopamine is added after mixing evenly in acid imide (NHS) and 1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride
(DA), 12~36h of reaction is stirred at room temperature, obtains sodium alginate-dopamine solution；
(2) sodium alginate-DOPA of step (1) is added into polyvinyl alcohol (PVA) aqueous solution that concentration is 40~60g/L
Amine aqueous solution stirs evenly, freeze-thaw, and dialysis obtains SA-DA/PVA hydrogel；
The molar ratio of sodium alginate and dopamine in sodium alginate aqueous solution described in step (1) is 1:1~2；
N-hydroxysuccinimide described in step (1) and 1- (3- dimethylamino-propyl) -3- ethyl carbodiimide hydrochloride
The molar ratio of salt is 1:1；Concentration in the reaction system of step (1) is 30~50mmol/L；
Sodium alginate-dopamine solution described in step (2) and the volume ratio of polyvinyl alcohol water solution are 1:1~2.
Preferably, the molar ratio of the sodium alginate in sodium alginate aqueous solution described in step (1) and dopamine is 1:1.
Preferably, sodium alginate aqueous solution described in step (1) the preparation method comprises the following steps: distilled water is added in sodium alginate
In, 10~15h is then stirred at 500~800rpm, is made it dissolve.
Preferably, speed of agitator described in step (1) is 500~800rpm.
Preferably, the reaction time described in step (1) is for 24 hours.
Preferably, sodium alginate-dopamine solution described in step (1) is also needed after the completion of reaction with distilled water dialysis 1
It is to remove extra NHS and EDC.
Preferably, sodium alginate-dopamine solution described in step (1) is sealed in 2-8 DEG C, the alginic acid
Sodium-dopamine is to react to connect in sodium alginate with the carboxyl in sodium alginate by chemical cross-linking agent using the amino in dopamine
On main chain.
Preferably, speed of agitator described in step (2) is 800~1000rpm, and the mixing time is 12~36h.
It is highly preferred that mixing time described in step (2) is for 24 hours.
Preferably, freeze-thaw method described in step (2) are as follows: freeze-thaw 3 is carried out between -20 DEG C and 4~8 DEG C
~5 times.
Preferably, dialysis process described in step (2) are as follows: distilled water is dialysed 3~4 days under room temperature.
A kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel made from the above method.
A kind of application of the above-mentioned sodium alginate-dopamine/polyvinyl alcohol hydrogel in biological field.
Compared with prior art, the present invention has the following advantages and beneficial effects:
1, sodium alginate is grafted dopamine, breaks tradition and introduces nerve conduction object communication promotion cytokine-expressing to promote
Into wound healing, improves physical adhesion ability and improve Biocompatibility
2, with synthesis macromolecule pharmaceutical grade Polyethylene alcohol composite frozen thawing technique plastic, natural macromolecular material can be improved
Mechanical property.
3, SA-DA/PVA hydrogel prepared by the present invention has good antibiotic property, promotees blood coagulation, has extensive biology doctor
Learn application prospect.
4, the present invention is grafted to neurotransmitter dopamine in sodium alginate by mild reaction conditions, passes through induction
Cell transmission signal promotes cytokine-expressing to reach and promote wound healing effect, and the hydrogel prepared can reach screen
Barrier effect, lower, gel contacts wound good water absorption and comfortable a possibility that bacterial invasion, when dressing, will not cause pain, together
The mechanical property of Shi Youliang is able to bear external various pressure factors and does not easily cause secondary insult.
Detailed description of the invention
Fig. 1 is sodium alginate-dopamine/polyvinyl alcohol hydrogel infrared spectrogram made from embodiment 1.
Fig. 2 is that sample is lyophilized under 1000 times in sodium alginate-dopamine made from embodiment 1/polyvinyl alcohol hydrogel
Scanning electron microscope diagram.
Fig. 3 is the stress-strain diagram of blank control group polyvinyl alcohol hydrogel made from embodiment 1.
Fig. 4 is sodium alginate-dopamine/polyvinyl alcohol hydrogel stress-strain diagram made from embodiment 1.
Fig. 5 is sodium alginate-dopamine/polyvinyl alcohol hydrogel and sodium alginate-Dopamine hydrochloride/polyvinyl alcohol hydrogel
The opposite appreciation rate (RGR) of gained l cell 3T3 after glue is handled 1 day.
Fig. 6 is sodium alginate-dopamine/polyvinyl alcohol hydrogel and sodium alginate-Dopamine hydrochloride/polyvinyl alcohol hydrogel
The opposite appreciation rate (RGR) of gained l cell 3T3 after glue is handled 2 days.
Fig. 7 is sodium alginate-dopamine/polyvinyl alcohol hydrogel and sodium alginate-Dopamine hydrochloride/polyvinyl alcohol hydrogel
The opposite appreciation rate (RGR) of gained l cell 3T3 after glue is handled 3 days.
Specific embodiment
Below with reference to embodiment and attached drawing, the present invention is described in further detail, but embodiments of the present invention are unlimited
In this.
Embodiment 1
(1) raw material, instrument sterilization treatment are tested
Experiment raw material are placed in super-clean bench ultraviolet irradiation 1h and carry out raw material sterilizing, glass apparatus is placed in 121 DEG C of high pressures
Under the conditions of sterilize 20 minutes.
(2) sodium alginate-dopamine preparation
2g sodium alginate (SA) is taken to be added in 100ml distilled water, 800rpm stirring and dissolving 12h is made into SA solution for later use.It is past
0.5755gN- HOSu NHS (NHS) and 0.9585g 1- (3- dimethylamino-propyl) -3- ethyl carbon two is added in SA solution
Inferior amine salt hydrochlorate is added according to molar ratio 1:1 amount and 800rpm stirs 20min, and 1.5473g dopamine (DA) room temperature is then added
And 800rpm is stirred to react 24 hours.Reaction terminates, and sodium alginate-dopamine (SA-DA) solution obtained is protected in 2-8 DEG C of sealing
It deposits.
(3) sodium alginate-dopamine/polyvinyl alcohol hydrogel preparation
It takes 30mlSA-DA solution to stir evenly for use, takes 5g polyvinyl alcohol (PVA) stirring to be dissolved in 100ml distilled water and be made
PVA solution mixes SA-DA solution and PVA solution according to volume 1:1, and room temperature, 800rpm are stirred 24 hours, are poured into and are ready to
Molding die freeze-thaw is carried out between -20 DEG C and 4 DEG C three times at hydrogel.Shaped hydrogel is taken out in room temperature condition
It is lower using distilled water dialyse 3 days sodium alginate-dopamine/polyvinyl alcohol (SA-DA/PVA) hydrogel.
Sodium alginate-dopamine/polyvinyl alcohol hydrogel infrared spectrogram obtained is shown in that attached drawing 1, freeze-drying sample exist
Scanning electron microscope diagram under 1000 times is shown in attached drawing 2, and SA-DA/PVA hydrogel is successfully made by attached Fig. 1 and 2 explanation.
Sodium alginate made from the present embodiment-dopamine/polyvinyl alcohol hydrogel mechanical property tests:
(A) sodium alginate-dopamine/polyvinyl alcohol obtained in 1 step of embodiment (3) without freeze-thaw is molten
The poly-vinyl alcohol solution (as blank control group) of liquid and 5g/L are poured into respectively with groove depth 10mm, the cylinder water that diameter is 12mm
Freeze-thaw is carried out in gel forming mold between -20 DEG C and 4 DEG C three times at hydrogel, is made test sample, experimental group and
Blank control group respectively prepares three same samples.
(B) compression test, design parameter are carried out to each sample with electronic universal tester (model ELF3220) are as follows:
1, compression speed 2mm/min；2, compression ratio is 60% length (i.e. each sample compression 6mm).
(C) access is according to stress-strain diagram is made, and as a result as shown in Figures 3 and 4, finding out slope is the sample elastic modulus.
The elasticity modulus of blank group polyvinyl alcohol hydrogel is 0.3KPa as can be drawn from Figure 3, and as can be drawn from Figure 4
Sodium alginate-dopamine/polyvinyl alcohol hydrogel elasticity modulus made from the application is 17.2KPa, hence it is evident that is better than polyethylene
Alcohol hydrogel, so sodium alginate-dopamine/polyvinyl alcohol hydrogel good mechanical performance made from the application and being able to satisfy wound
The requirement of mouth dressing mechanical property.
Comparative example 1
(1) raw material, instrument sterilization treatment are tested
With embodiment 1.
(2) sodium alginate-Dopamine hydrochloride preparation
2g sodium alginate (SA) is taken to be added in 100ml distilled water, 800rpm stirring and dissolving 12h is made into SA solution for later use.It is past
0.5755gN- HOSu NHS (NHS) and 0.9585g1- (3- dimethylamino-propyl) -3- ethyl carbon two is added in SA solution
Inferior amine salt hydrochlorate is added according to molar ratio 1:1 amount and 800rpm stirs 20min, and 1.9156g Dopamine hydrochloride room temperature is then added
And 800rpm is stirred to react 24 hours.Reaction terminates, and sodium alginate-Dopamine hydrochloride obtained is sealed in 2-8 DEG C.
(3) sodium alginate-Dopamine hydrochloride/polyvinyl alcohol hydrogel preparation
The step of referring to embodiment 1 (1)~(3), in addition to dopamine is replaced with Dopamine hydrochloride, other experimental raws and
Technological parameter condition is same as Example 1, and sodium alginate-Dopamine hydrochloride/polyvinyl alcohol hydrogel is made.
Embodiment 2
Sodium alginate-dopamine/polyvinyl alcohol hydrogel In vitro cell experiment
Co60 gamma-rays spoke is carried out to freeze-drying hydrogel sample made from embodiment 1 before carrying out vitro cytotoxicity experiment
Penetrate sterilization treatment.
(1) NIH 3T3 cell (purchase is in Wuhan Pu Nuosai Life Science Co., Ltd) is taken out in liquid nitrogen to be cultivated with DMEM
Liquid carries out recovery culture.
(2) carry out secondary culture for several times with DMEM culture solution, the cell of logarithmic growth phase and with cell counting board to NIH
3T3 cell is counted, and being adjusted to cell density is 5 × 104A/mL.
(3) take 96 orifice plates and thereto 30 holes be separately added into 100 μ L steps (2) containing cell density be 5 × 104A/mL
DMEM culture solution as experimental group；It in addition takes 3 holes to be arranged to blank control group simultaneously and is only separately added into 100 μ L DMEM
Culture solution (is free of cell), is placed in 37 DEG C, 5%CO2It is cultivated 24 hours in cell incubator.
Cell is adherent after (4) 24 hours, takes out the DMEM culture in 30 holes of 96 orifice plates and gettering step (3) experimental group
Liquid, then 15 holes thereto be separately added into 100 μ L concentration be 25mg/ml, 50mg/ml, 100mg/ml, 150mg/ml,
The SA-DA/PVA hydrogel leaching body fluid solution (the DMEM culture medium of leaching SA-DA/PVA hydrogel) of 200mg/ml, each concentration is set
3 holes are set, as experimental group；Negative control group is set by other 15 holes simultaneously, being separately added into 100 μ L concentration is 25mg/
The sodium alginate of ml, 50mg/ml, 100mg/ml, 150mg/ml, 200mg/ml/polyvinyl alcohol hydrogel soak body fluid solution, each
3 holes are arranged in concentration；Then 96 orifice plates are placed in 37 DEG C, 5%CO2It is cultivated respectively in cell incubator 24,48,72 hours；
Sodium alginate/the polyvinyl alcohol hydrogel is the preparation method comprises the following steps: the seaweed for being 2% (2g/100ml) by concentration
The polyvinyl alcohol water solution that acid sodium aqueous solution and concentration are 50g/L is mixed according to volume ratio 1:1, and room temperature, 800rpm stirring 24 are small
When, it pours into ready molding die and carries out freeze-thaw between -20 DEG C and 4 DEG C three times into hydrogel, take out molding water-setting
Glue uses distilled water to dialyse 3 days up to sodium alginate/polyvinyl alcohol hydrogel at room temperature.
(5) take out 96 orifice plates and absorb the SA-DA/PVA hydrogel in 15 holes of experimental group leaching body fluid solution and feminine gender it is right
Body fluid solution is soaked according to sodium alginate/polyvinyl alcohol hydrogel in 15 holes of group, then is separately added into 10 μ L into this 30 holes
Then 96 orifice plates are placed in 37 DEG C, 5%CO by DMEM culture solution and 10 μ L CCK-8 solution2It is cultivated 4 hours in cell incubator.
The absorbance that 96 orifice plates measure each hole using microplate reader in λ=450nm is taken out after (6) 4 hours, and according to following
The opposite appreciation rate (RGR) of formula calculating cell:
RGR%=(A experimental group-A blank group)/(A0 negative control group-A blank group) × 100%.
(7) access is according to cell processed with respect to appreciation rate figure.
Comparative example 2
Sodium alginate-Dopamine hydrochloride/polyvinyl alcohol hydrogel In vitro cell experiment
Co60 gamma-rays spoke is carried out to freeze-drying hydrogel sample made from comparative example 1 before carrying out vitro cytotoxicity experiment
Penetrate sterilization treatment.
Referring to 2 step of embodiment (1)~(3) recovery culture NIH 3T3 cell.
Cell is adherent after (4) 24 hours, takes out the DMEM culture in 30 holes of 96 orifice plates and gettering step (3) experimental group
Liquid, it is respectively 25mg/ml, 50mg/ml, 100mg/ml, 150mg/ that then 15 holes thereto, which are separately added into 100 μ L concentration,
Sodium alginate-Dopamine hydrochloride of ml, 200mg/ml/polyvinyl alcohol hydrogel leaching body fluid solution (leaching sodium alginate-hydrochloric acid DOPA
Amine/polyvinyl alcohol hydrogel DMEM culture medium) 3 holes of each concentration setting, as experimental group；Other 15 holes are set simultaneously
It is set to negative control group, being separately added into 100 μ L concentration is 25mg/ml, 50mg/ml, 100mg/ml, 150mg/ml, 200mg/ml
Sodium alginate/polyvinyl alcohol hydrogel soak body fluid solution, 3 holes are arranged in each concentration；Then by 96 orifice plates be placed in 37 DEG C,
5%CO2It is cultivated respectively in cell incubator 24,48,72 hours；
Sodium alginate/polyvinyl alcohol hydrogel preparation method is the same as embodiment 2.
Data are obtained referring to 2 step of embodiment (5)~(7) and cell processed is with respect to appreciation rate figure.
Fig. 5,6,7 are respectively sodium alginate-dopamine/polyvinyl alcohol hydrogel and sodium alginate-Dopamine hydrochloride/poly- second
The opposite appreciation rate (RGR) of gained l cell 3T3 after enol hydrogel is handled 1,2,3 day, as can be seen from Figure 5 the
Sodium alginate-Dopamine hydrochloride/polyvinyl alcohol hydrogel group is apparently without sodium alginate-dopamine/polyvinyl alcohol hydrogel within one day
The cell Proliferation of glue group it is fast.And it is can analyze out in Fig. 6 and Fig. 7 in sodium alginate-Dopamine hydrochloride/polyvinyl alcohol hydrogel
Do not have cytotoxicity substantially when soaking body fluid low concentration, but cell Proliferation will not be promoted, but with regard to body when concentration increases
Reveal with certain cytotoxicity, cell proliferation rate is lower than 100%.And sodium alginate-dopamine made from the application/poly-
Polyvinyl alcohol hydrogel shows as low concentration in the 2nd, 3 day has the function of certain promotion cell Proliferation；Promote when high concentration
Cell Proliferation effect is small, but does not have cytotoxicity, illustrates hydrogel biocompatibility made from the application and promotes cell
Cultivation effect is prominent.Pass through the NIH 3T3 cell proliferation experiment of the application, it can be deduced that hydrogel made from the application can be very
Good promotion mouse skin fibroblastic growth, so that promoting wound healing to reach a faster step repairs wound effect.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (10)
1. a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel preparation method, which comprises the following steps:
(1) n-hydroxysuccinimide and 1- (3- dimethylamino third is added in the sodium alginate aqueous solution for being 1~2% to concentration
Base) -3- ethyl-carbodiimide hydrochloride, after mixing evenly, addition dopamine is stirred at room temperature 12~36h of reaction, obtains alginic acid
Sodium-dopamine solution；
(2) it to polyvinyl alcohol sodium alginate-dopamine solution soluble in water that step (1) is added that concentration is 40~60g/L, stirs
It mixes uniformly, freeze-thaw, dialysis obtains SA-DA/PVA hydrogel；
The molar ratio of sodium alginate and dopamine in sodium alginate aqueous solution described in step (1) is 1:1~2；
N-hydroxysuccinimide described in step (1) and 1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride
Molar ratio is 1:1, and the concentration in the reaction system of step (1) is 30~50mmol/L；
Sodium alginate-dopamine solution described in step (2) and the volume ratio of polyvinyl alcohol water solution are 1:1~2.
2. a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel preparation method according to claim 1, feature
It is, the molar ratio of sodium alginate and dopamine in sodium alginate aqueous solution described in step (1) is 1:1.
3. according to right want 1 described in a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel preparation method, feature exists
In speed of agitator described in step (1) is 500~800rpm.
4. according to right want 1 or 3 described in a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel preparation method, feature
It is, the reaction time described in step (1) is for 24 hours.
5. according to right want 4 described in a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel preparation method, feature exists
In speed of agitator described in step (2) is 800~1000rpm；The mixing time is 12~36h.
6. according to right want 5 described in a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel preparation method, feature exists
In mixing time described in step (2) is for 24 hours.
7. according to right want 4 described in a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel preparation method, feature exists
In freeze-thaw method described in step (2) are as follows: carried out freeze-thaw 3~5 times between -20 DEG C and 4~8 DEG C.
8. according to right want 4 described in a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel preparation method, feature exists
In sodium alginate-dopamine solution described in step (1) also needs to be dialysed 1 day with distilled water after the completion of reaction；
Dialysis process described in step (2) are as follows: distilled water is dialysed 3~4 days under room temperature.
9. a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel made from preparation method according to any one of claims 1 to 8
Glue.
10. a kind of sodium alginate-dopamine/polyvinyl alcohol hydrogel as claimed in claim 9 is in the application of biological field.
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