CN102940903B - Method for preparing medical dressing of polysaccharide cavernous body - Google Patents
Method for preparing medical dressing of polysaccharide cavernous body Download PDFInfo
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- CN102940903B CN102940903B CN201210473901.3A CN201210473901A CN102940903B CN 102940903 B CN102940903 B CN 102940903B CN 201210473901 A CN201210473901 A CN 201210473901A CN 102940903 B CN102940903 B CN 102940903B
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Abstract
The invention relates to the preparation technique of tissue engineering scaffold materials and provides a crosslinking method for preparing medical dressing of a polysaccharide cavernous body. Chitosan and sodium alga acid serve as the base material, and by means of crosslinking of a couple reaction and the freezing drying technique, water absorption capability of the medical dressing of the polysaccharide cavernous body is improved, and effective protection on skin wound is achieved. When used, the medical dressing of the polysaccharide cavernous body is directly compressed on the wound surface, has excellent adsorption effect, and is particularly suitable for skin diseases of burning, pus, wound drainage, ulcer and the like. The method for preparing medical dressing of the polysaccharide cavernous body is simple in process, low in cost, low in crosslinking temperature, fast in curing, and suitable for large-scale industrial production. By means of the method, water adsorption capability of the dressing is obviously improved, residue of toxic chemical cross-linked agents is avoided simultaneously, chemical stimulation on the skin caused by materials is reduced, and biocompatibility and safety of the dressing are improved.
Description
Technical field
The invention belongs to the crosslinking technological of tissue engineering bracket material, particularly a kind of preparation method of natural polysaccharide spongy body medical dressing.
Background technology
Dressing is a kind of medical material that temporary covering wound uses that is used for, and its topmost function is to control transudate and the protection wound of wound, to avoid the pollution of antibacterial and grit, to provide being beneficial to the environment of wound healing.Select suitable dressing to cover on wound, can assist Bleeding control, protect from infection and absorb secretions, thereby promote wound healing.Now dressing plays protection wound surface, prevent that body fluid and protein run off, prevent that antibacterial from invading the effect causing inflammation, and proliferative cell is provided support.
The wound of skin is clinical common disease, is difficult in the situation of healing when large defect or short time appear in skin, of crucial importance in the dressing of wound surface coverage.For a comparatively long period of time, using the most general surgical wound dressings is medical absorbent cotton and gauze.It can play certain facilitation to the treatment of wound, but has very large deficiency.For example, when it uses, wound surface granulation tissue causes adhesion to growth in dressing; When releasing, easily cause secondary insult; Easily cause that due to wound surface hydrops antibacterial infects.
Along with the progress in epoch and scientific and technological development, progressively developed into the wound dressing of the high-tech component content of today by traditional gauze, in the nearly more than ten years, there is breakthrough variation in the composition of dressing and kind.The clinical research and development of dressing of need to giving have proposed requirements at the higher level, also become the power that carries out new pattern compress exploitation.Therefore, develop a kind of new pattern compress, make it can not only flap coverage, can also help wound healing, prevent bacteria attack, reduce superelevation metabolism and the malnutrition of wound area, alleviate wound pain, accelerating wound, have become scientific research personnel's main direction.Therefore, extremely important to improve its quality and water absorbing properties to the crosslinking Treatment of dressing.
The matrix material that is generally used for dressing has two classes: natural macromolecular material and synthesized polymer material.Natural macromolecular material mainly contains collagen, gelatin, chitosan, chondroitin sulfate, alginic acid, cellulose etc.Conventional artificial macromolecular material comprises Polyethylene Glycol (PEG), polyurethane (PU), polyvinylpyrrolidone (PVP) etc.In general, natural polymer has excellent biocompatibility and degradability, and synthetic macromolecule has good physical and mechanical properties, processing characteristics and chemical stability.These material controllabilitys are good, and practicality is high, have advantage and disadvantage separately, therefore obtained application widely as biomaterial in organizational project and regenerative medicine field.
Preparing at present the method that dressing is conventional is chemical reagent cross-linking method and uv cross-linking method.Chemical reagent cross-linking method is by adding chemical cross-linking agent (as paraformaldehyde, glutaraldehyde and water-soluble carbodiimide etc.) as condensing agent in matrix, and its effect is that the active group in substrate molecule (as amino or carboxyl etc.) is reached to crosslinked object through condensation reaction.Uv cross-linking method is first matrix to be passed through to chemical modification, makes it with unsaturated double-bond, then adds initiator, causes the adduction of unsaturated double-bond by ultraviolet light irradiation, thereby it is crosslinked that matrix is occurred.Adopt the crosslinked material of these methods more stable in chemical constitution, can meet the technological requirement that material is made, but owing to producing unavoidably the residual of poisonous chemical reagent or initiator, therefore material has stimulation in various degree to human body, and safety can not be protected.In addition, these chemical cross-linking agents are micromolecule conventionally, and the intermolecular distance of crosslinked rear material is less, causes water absorbing properties lower, directly affects protection effect and the result of use of dressing.Therefore, avoiding using poisonous micromolecule chemical cross-linking agent, is the effective way that reduces dressing toxicity and improve material swelling behavior.But, in prior art, there is not the method that adopts nontoxic chemical reagent or initiator crosslinked poly sugar medical dressing.
Natural polysaccharide molecule (as chitosan, alginic acid etc.) is the ideal material of medical dressing, they have good biocompatibility on the one hand, on its strand, there is on the other hand the multiple active function groups reacting, can carry out chemical modification by modified methods such as grafting easily, can effectively improve the Performance and quality of material.At present a lot of spongy body dressing, all based on this class material, if avoid using poisonous chemical reagent to be cross-linked, such as adopting bimolecular conjugation reaction cross-linking method, is expected to obtain the polysaccharide spongy body medical dressing of high swelling ratio and good biocompatibility.
Summary of the invention
The object of the present invention is to provide the preparation method of a kind of conjugation reaction crosslinked poly easy and simple to handle sugar dressing.
The technical solution that realizes the object of the invention is: a kind of preparation method of polysaccharide spongy body medical dressing, adopt Freeze Drying Technique, and taking chitosan and sodium alginate as base material, form by Diels-Alder conjugation reaction is crosslinked, specifically comprise the following steps:
Under step 1, room temperature, chitosan is dissolved in to lactic acid solution, adds succinic anhydrides to react with it, after dialysis lyophilizing, obtain N-succinyl-chitosan;
Under step 2, room temperature, N-succinyl-chitosan is dissolved in buffer solution, drips furfuryl amine-2-furylamine and react with it, after dialysis lyophilizing, obtain furan chitosan;
Under step 3, room temperature, sodium alginate is soluble in water, under lucifuge condition, drip Potassium metaperiodate. solution, after reaction a period of time, dialysis lyophilizing obtain aldehyde radical sodium alginate;
Step 4, by soluble in water aldehyde radical sodium alginate, add 4-(4-N-maleimide phenyl) butyrate reaction, after dialysis lyophilizing, obtain maleimide amination sodium alginate;
Step 5, prepare certain density furan chitosan and maleimide amination sodium alginate respectively, under room temperature, fully mix in proportion, after lyophilization, obtain spongy body dressing.
The reagent source that the present invention is used: chitosan, sodium alginate, Potassium metaperiodate., furfuryl amine-2-furylamine, water-soluble carbodiimide (EDAC), bovine serum albumin (BSA), 3-(4,5-dimethylthiazole)-2,5-diphenyl tetrazole bromine salt (MTT), is purchased from Sigma company; Succinic anhydrides, analytical pure, Chinese Medicine group; Lactic acid, analytical pure, Solution on Chemical Reagents in Shanghai company limited; 4-(4-N-maleimide phenyl) butyrate (MPBH), is purchased from Thermo Fisher company.Sodium chloride, potassium chloride, analytical pure, Shanghai reagent three factories; Sodium hydrogen phosphate, potassium dihydrogen phosphate, analytical pure, Hangzhou chemical reagent company limited; Dimethyl sulfoxide, dodecyl sodium sulfate, analytical pure, Solution on Chemical Reagents in Shanghai factory.Protein determination kit, is purchased from green skies biotechnology research institute.DMEM culture medium, Giboco company; Calf serum, Hangzhou Ilex purpurea Hassk.[I.chinensis Sims biomaterial Graduate School of Engineering; Penicillin, streptomycin, Huabei Pharmaceutic Co., Ltd.
The instrument that the present invention is used: scanning electron microscope (JSM-6330F, JEOL), microplate reader (Biorad, Model 550).
The present invention compared with prior art, is characterized in that: 1) based on conjugation reaction principle of crosslinking, the present invention avoids having used poisonous chemical cross-linking agent, therefore in dressing, does not have the residual of toxic reagent, has ensured the safety of dressing; 2) because conjugation reaction related in the present invention results between macromole-macromole, protect space structure and distance between macromole, therefore improve the water absorbing properties of dressing, realize the effective protection to wound; 3) in the present invention, matrix used material is natural polysaccharide material, demonstrates biocompatibility admirably, and body sense is good; 4) the crosslinked spongy body dressing water absorption of the present invention is strong, high with the bond strength of wound surface, has avoided coming off in materials'use, and has had good permeability; 5) the technology of the present invention has the advantages such as crosslinking temperature is low, curing rate fast, the crosslinking Treatment cycle is short; 6) simple, the Yi Hang of technology and equipment of the present invention, handling safety, without toxic reagent, does not produce pollution to environment, and the cost of raw material of use is cheap, is applicable to large-scale industrial production.
Further illustrate the present invention below by embodiment.
Brief description of the drawings
Fig. 1 adopts the crosslinked schematic diagram of preparing chitin-sodium alginate spongy body medical dressing of Diels-Alder conjugation reaction.
Fig. 2 is the stereoscan photograph of chitin-sodium alginate spongy body medical dressing internal structure.
The water absorption rate (chitosan/sodium alginate soln volume ratio is respectively 1/2,1/1 and 2/1) of the chitin-sodium alginate spongy body medical dressing of Fig. 3 different proportion.
The moisture evaporation rate (chitosan/sodium alginate soln volume ratio is respectively 1/2,1/1 and 2/1) of the chitin-sodium alginate spongy body medical dressing of Fig. 4 different proportion.
The albumen adhesiveness of the chitin-sodium alginate spongy body medical dressing of Fig. 5 different proportion.
The cell adhesion of the chitin-sodium alginate spongy body medical dressing of Fig. 6 different proportion.
Detailed description of the invention
Lyophilization is a kind of common technology of preparing polymer tissue engineering scaffold material, and the preparation method of a kind of polysaccharide spongy body medical dressing of the present invention is taking chitosan and sodium alginate as base material, forms by Diels-Alder conjugation reaction crosslinking curing.Chitosan and sodium alginate are carried out respectively to modification, make them respectively carry the active group that can react, can automatically cross-linked molding under temperate condition after making it to mix, without adding poisonous chemical cross-linking agent, ensure the safety of material.Specifically comprise the following steps:
Under step 1, room temperature, in the lactic acid aqueous solution of 5% (v/v), add a gram chitosan, stir 3 hours, form homogeneous solution, add subsequently succinic anhydrides, under room temperature, stir 12 ~ 24 hours, then by reaction solution dialysis 3 ~ 5 days, last lyophilizing obtains N-succinyl-chitosan, wherein lactic acid aqueous solution quality is chitosan 4 times, succinic anhydrides quality is chitosan 1 ~ 4 times;
Under step 2, room temperature, N-succinyl-chitosan is dissolved in phosphate buffered solution, then drip furfuryl amine-2-furylamine of 100 ~ 500 microlitres, add after water-soluble carbodiimide and stir 12 ~ 24 hours under room temperature, the lyophilizing after 3 ~ 5 days of dialysing obtains furan chitosan, wherein the quality of N-succinyl-chitosan is 1 ~ 5 times of furfuryl amine-2-furylamine, the quality of phosphate buffer is 80 times of N-succinyl-chitosan, and water-soluble carbodiimide quality is 0.5 times of N-succinyl-chitosan;
Under step 3, room temperature, sodium alginate is dissolved in the water, then drip the sodium periodate solution that concentration is 0.5M, under lucifuge condition, stir 1 ~ 4 hour, the lyophilizing after 3 ~ 5 days of dialysing obtains aldehyde radical sodium alginate, wherein the mass ratio of sodium alginate and water is 1:100, the quality of sodium alginate is sodium metaperiodate 200 ~ 500 times;
Under step 4, room temperature, aldehyde radical sodium alginate is dissolved in the water, then adding concentration is 0.5% 4-(4-N-maleimide phenyl) butyrate solution, under room temperature, react 0.5 ~ 3 hour, the lyophilizing after 3 ~ 5 days of dialysing obtains maleimide amination sodium alginate, wherein the mass ratio of aldehyde radical sodium alginate and water is 1:100, and the quality of 4-(4-N-maleimide phenyl) butyrate is 0.015 ~ 0.06 times of aldehyde radical sodium alginate;
Step 5, compound concentration is furan chitosan and the maleimide amination sodium alginate aqueous solution of 0.5% ~ 2.0% (w/v) respectively, and under room temperature, by volume ratio 1:2/1:1/2:1 fully mixes, 37
oc transfers and sets to 0 .5 ~ 2 hour, is then placed on-20
ounder C freezing 2 hours, finally-50
ounder C, lyophilization obtains spongy body dressing for 24 hours.
Wherein, the preparation of phosphate buffered solution (PBS): take 8 grams, analytical pure sodium chloride, 0.2 gram, potassium chloride, 2.9 grams of sodium hydrogen phosphates, 0.2 gram of potassium dihydrogen phosphate, be dissolved in 1000 ml distilled waters, regulating pH is 7.4.
The constructed observation of dressing: through-50
oc lyophilization (FD1A50, Beijing rich doctor health) is after 24 hours, by dressing metal spraying (Cressington 108 Auto), then in the upper internal microstructure of observing of scanning electron microscope (JSM-6330F, JEOL).
The water absorption of dressing detects: (W first weighs dressing
d), 25
ounder C, in phosphate buffered solution, soak 6 hours fully swelling to ensure.When test, take out dressing from solution, suck the water on dressing surface with filter paper, (W at once weighs
w), each sample parallel testing 5 times.Calculate dressing water absorbing properties P
aformula be P
a=(W
w– W
d)/W
d.
The rate of perviousness of dressing detects: the quality (unit: mgcm of the aqueous vapor that what rate of perviousness represented is passes through on unit interval area
-2h
-1).(capacity is 5 milliliters to the moisture-inhibiting bottle of the fixing dressing of mensuration, includes 4 milliliters of distilled water, and aperture area is 1.18cm
2) quality (W
0), then moisture-inhibiting bottle is placed in to the exsiccator of constant temperature and humidity.With reference to general moisture-inhibiting method of testing ASTM method E96-90 standard, in exsiccator, place a large amount of silica gel, keep humidity in 40RH left and right.The quality (Wt) of measuring moisture-inhibiting bottle with the testing time, obtains quality curve over time.Calculate the rate of perviousness of dressing according to slope of a curve, computing formula is rate of perviousness=L/S
0.Wherein L is moisture-inhibiting slope of a curve, and the representation unit time is by the quality (unit: mgh of aqueous vapor
-1); S
0for the effective area of penetrating aqueous vapor, i.e. moisture-inhibiting bottle aperture area, is 1.18cm
2).
The protein adsorption of dressing detects: the protein solution of preparation 0.2wt%, 25
ounder C, dressing is dipped in this solution, after 24 hours, takes out dressing, use a large amount of distilled water flushings, then dressing is placed in to the sodium dodecyl sulfate solution of 0.1wt%, 37
oshaking table 24 hours under C, being adsorbed in the albumen stripping of dressing.Protein adsorption quantity measuring method is carried out according to protein determination kit Standard Operating Procedure.
The cell adhesion of dressing detects: dressing scissors are circular, and size and 24 well culture plate (Nunc
tM, Denmark) and aperture is consistent, by dressing confluent cultures plate bottom.Before use first by the sterilization in 2 hours of 75% soak with ethanol for dressing, then with phosphate buffered solution repeatedly rinsing remove ethanol.Be 4 × 10 in dressing surface seeding quantity
4human body fibroblast, and add 2 milliliters of culture fluid (DMEM culture medium+10% calf serum+100 units per ml penicillin/streptomycin), be put in 37
ounder C, hatch after 4 hours and take out, repeatedly rinse to remove the cell not adhering to by phosphate buffered solution, then add the phosphate buffered solution of 20 microlitre 0.5wt%MTT, be placed in 37
ounder C, hatch after 4 hours and add 200 microlitre dimethyl sulfoxide, after vibration evenly, adopt microplate reader (Biorad, Model 550) to measure the absorbance of purple material in 570 nanometers.
The present invention adopts ANOVA method of analysis of variance, significant difference value
pbe made as≤0.05.
The prepared dressing of the present invention is sponge loose structure, and average pore size is 200 microns of left and right and mutually runs through, and is conducive to the absorption of ventilative and moisture.
Below in conjunction with embodiment, the present invention is done to further detailed description:
embodiment 1:
Concrete operation step is:
(1) in the lactic acid solution of 40 milliliter 5% (v/v), add 0.5 gram of chitosan, under room temperature, stir 3 hours, form homogeneous solution, add subsequently 2.0 grams of succinic anhydrides, under room temperature, stir 24 hours, then by reaction solution dialysis 3 days, last lyophilizing obtains N-succinyl-chitosan;
(2) under room temperature, 0.5 gram of N-succinyl-chitosan is dissolved in 40 milliliters of buffer solution, then drip furfuryl amine-2-furylamine of 100 microlitres, add after 0.2 gram of water-soluble carbodiimide and stir 18 hours under room temperature, the lyophilizing after 3 days of dialysing obtains furan chitosan;
(3) under room temperature, 1.0 grams of sodium alginates are dissolved in 100 ml waters, then drip the sodium periodate solution that 5 ml concns are 0.5M, stir 2 hours under lucifuge condition, the lyophilizing after 3 days of dialysing obtains aldehyde radical sodium alginate;
(4) under room temperature, 1.0 grams of aldehyde radical sodium alginates are dissolved in 100 ml waters, then adding 6 ml concns is 0.5% 4-(4-N-maleimide phenyl) butyrate solution, under room temperature, react 0.5 hour, the lyophilizing after 3 days of dialysing obtains maleimide amination sodium alginate;
(5) compound concentration is furan chitosan and the maleimide amination sodium alginate aqueous solution of 0.5% (w/v) respectively, and under room temperature, by volume ratio 1:1 fully mixes, 37
ounder C, place 1 hour, be then placed on-20
ounder C freezing 2 hours, finally-50
ounder C, lyophilization obtains spongy body dressing for 24 hours.
embodiment 2:
Concrete operation step is:
(1) in the lactic acid solution of 40 milliliter 5% (v/v), add 0.5 gram of chitosan, under room temperature, stir 3 hours, form homogeneous solution, add subsequently 1.8 grams of succinic anhydrides, under room temperature, stir 20 hours, then by reaction solution dialysis 3 days, last lyophilizing obtains N-succinyl-chitosan;
(2) under room temperature, 0.5 gram of N-succinyl-chitosan is dissolved in 40 milliliters of buffer solution, then drip furfuryl amine-2-furylamine of 200 microlitres, add after 0.2 gram of water-soluble carbodiimide and stir 16 hours under room temperature, the lyophilizing after 3 days of dialysing obtains furan chitosan;
(3) under room temperature, 1.0 grams of sodium alginates are dissolved in 100 ml waters, then drip the sodium periodate solution that 4 ml concns are 0.5M, stir 4 hours under lucifuge condition, the lyophilizing after 3 days of dialysing obtains aldehyde radical sodium alginate;
(4) under room temperature, 1.0 grams of aldehyde radical sodium alginates are dissolved in 100 ml waters, then adding 8 ml concns is 0.5% 4-(4-N-maleimide phenyl) butyrate solution, under room temperature, react 1 hour, the lyophilizing after 3 days of dialysing obtains maleimide amination sodium alginate;
(5) compound concentration is furan chitosan and the maleimide amination sodium alginate aqueous solution of 0.5% (w/v) respectively, and under room temperature, by volume ratio 1:1 fully mixes, 37
ounder C, place 1.5 hours, be then placed on-20
ounder C freezing 2 hours, finally-50
ounder C, lyophilization obtains spongy body dressing for 24 hours.
embodiment 3:
Concrete operation step is:
(1) in the lactic acid solution of 40 milliliter 5% (v/v), add 0.5 gram of chitosan, under room temperature, stir 3 hours, form homogeneous solution, add subsequently 1.5 grams of succinic anhydrides, under room temperature, stir 18 hours, then by reaction solution dialysis 4 days, last lyophilizing obtains N-succinyl-chitosan;
(2) under room temperature, 0.5 gram of N-succinyl-chitosan is dissolved in 40 milliliters of buffer solution, then drip furfuryl amine-2-furylamine of 250 microlitres, add after 0.2 gram of water-soluble carbodiimide and stir 24 hours under room temperature, the lyophilizing after 4 days of dialysing obtains furan chitosan;
(3) under room temperature, 1.0 grams of sodium alginates are dissolved in 100 ml waters, then drip the sodium periodate solution that 5 ml concns are 0.5M, stir 3 hours under lucifuge condition, the lyophilizing after 4 days of dialysing obtains aldehyde radical sodium alginate;
(4) under room temperature, 1.0 grams of aldehyde radical sodium alginates are dissolved in 100 ml waters, then adding 10 ml concns is 0.5% 4-(4-N-maleimide phenyl) butyrate solution, under room temperature, react 1.5 hours, the lyophilizing after 4 days of dialysing obtains maleimide amination sodium alginate;
(5) compound concentration is 1.0% respectively furan chitosan and maleimide amination sodium alginate aqueous solution, under room temperature, by volume ratio 1:2 fully mixes, and 37
ounder C, place 2 hours, be then placed on-20
ounder C freezing 2 hours, finally-50
ounder C, lyophilization obtains spongy body dressing for 24 hours.
embodiment 4:
Concrete operation step is:
(1) in the lactic acid solution of 40 milliliter 5% (v/v), add 0.5 gram of chitosan, under room temperature, stir 3 hours, form homogeneous solution, add subsequently 1.2 grams of succinic anhydrides, under room temperature, stir 16 hours, then by reaction solution dialysis 4 days, last lyophilizing obtains N-succinyl-chitosan;
(2) under room temperature, 0.5 gram of N-succinyl-chitosan is dissolved in 40 milliliters of buffer solution, then drip furfuryl amine-2-furylamine of 300 microlitres, add after 0.2 gram of water-soluble carbodiimide and stir 20 hours under room temperature, the lyophilizing after 4 days of dialysing obtains furan chitosan;
(3) under room temperature, 1.0 grams of sodium alginates are dissolved in 100 ml waters, then drip the sodium periodate solution that 3 ml concns are 0.5M, stir 2.5 hours under lucifuge condition, the lyophilizing after 4 days of dialysing obtains aldehyde radical sodium alginate;
(4) under room temperature, 1.0 grams of aldehyde radical sodium alginates are dissolved in 100 ml waters, then adding 3 ml concns is 0.5% 4-(4-N-maleimide phenyl) butyrate solution, under room temperature, react 2 hours, the lyophilizing after 4 days of dialysing obtains maleimide amination sodium alginate;
(5) compound concentration is furan chitosan and the maleimide amination sodium alginate aqueous solution of 1.0% (w/v) respectively, and under room temperature, by volume ratio 1:2 fully mixes, 37
oc transfers and sets to 0 .5 hour, is then placed on-20
ounder C freezing 2 hours, finally-50
ounder C, lyophilization obtains spongy body dressing for 24 hours.
embodiment 5:
Concrete operation step is:
(1) in the lactic acid solution of 40 milliliter 5% (v/v), add 0.5 gram of chitosan, under room temperature, stir 3 hours, form homogeneous solution, add subsequently 1.0 grams of succinic anhydrides, under room temperature, stir 24 hours, then by reaction solution dialysis 5 days, last lyophilizing obtains N-succinyl-chitosan;
(2) under room temperature, 0.5 gram of N-succinyl-chitosan is dissolved in 40 milliliters of buffer solution, then drip furfuryl amine-2-furylamine of 400 microlitres, add after 0.2 gram of water-soluble carbodiimide and stir 12 hours under room temperature, the lyophilizing after 5 days of dialysing obtains furan chitosan;
(3) under room temperature, 1.0 grams of sodium alginates are dissolved in 100 ml waters, then drip the sodium periodate solution that 4 ml concns are 0.5M, stir 2 hours under lucifuge condition, the lyophilizing after 5 days of dialysing obtains aldehyde radical sodium alginate;
(4) under room temperature, 1.0 grams of aldehyde radical sodium alginates are dissolved in 100 ml waters, then adding 8 ml concns is 0.5% 4-(4-N-maleimide phenyl) butyrate solution, under room temperature, react 2.5 hours, the lyophilizing after 5 days of dialysing obtains maleimide amination sodium alginate;
(5) compound concentration is furan chitosan and the maleimide amination sodium alginate aqueous solution of 0.8% (w/v) respectively, and under room temperature, by volume ratio 2:1 fully mixes, 37
ounder C, place 2 hours, be then placed on-20
ounder C freezing 2 hours, finally-50
ounder C, lyophilization obtains spongy body dressing for 24 hours.
embodiment 6:
Concrete operation step is:
(1) in the lactic acid solution of 40 milliliter 5% (v/v), add 0.5 gram of chitosan, under room temperature, stir 3 hours, form homogeneous solution, add subsequently 0.5 gram of succinic anhydrides, under room temperature, stir 12 hours, then by reaction solution dialysis 3 days, last lyophilizing obtains N-succinyl-chitosan;
(2) under room temperature, 0.5 gram of N-succinyl-chitosan is dissolved in 40 milliliters of buffer solution, then drip furfuryl amine-2-furylamine of 500 microlitres, add after 0.2 gram of water-soluble carbodiimide and stir 24 hours under room temperature, the lyophilizing after 5 days of dialysing obtains furan chitosan;
(3) under room temperature, 1.0 grams of sodium alginates are dissolved in 100 ml waters, then drip the sodium periodate solution that 2 ml concns are 0.5M, stir 1 hour under lucifuge condition, the lyophilizing after 5 days of dialysing obtains aldehyde radical sodium alginate;
(4) under room temperature, 1.0 grams of aldehyde radical sodium alginates are dissolved in 100 ml waters, then adding 12 ml concns is 0.5% 4-(4-N-maleimide phenyl) butyrate solution, under room temperature, react 3 hours, the lyophilizing after 5 days of dialysing obtains maleimide amination sodium alginate;
(5) compound concentration is furan chitosan and the maleimide amination sodium alginate aqueous solution of 0.8% (w/v) respectively, and under room temperature, by volume ratio 2:1 fully mixes, 37
ounder C, place 1.5 hours, be then placed on-20
ounder C freezing 2 hours, finally-50
ounder C, lyophilization obtains spongy body dressing for 24 hours.
Dressing prepared by the present invention is sponge loose structure, and average pore size is 200 microns of left and right and mutually runs through, and is conducive to the absorption (Fig. 1) of ventilative and moisture.The water absorbing capacity of this dressing is extremely strong, can absorb the liquid (Fig. 2) that is equivalent to 52 ~ 57 times of own wts.The water evaporation speed of this dressing is at 2100g/m
2left and right (Fig. 3), it is generally acknowledged and control moisture evaporation rate at 2000 ~ 2500g/m every day
2can meet preferably the requirement of wound surface to wound, therefore this dressing meets the requirement to water vapor transmittance completely.The protein adsorption ability of this dressing, significantly lower than conventional hospital gauze (Fig. 4), illustrates that this dressing hydrophilic is strong, thereby is not easy to cause the adhesion of cell.Cell adhesion is that dressing design needs a very important problem of considering.The dressing causing because material is incorrect adheres to granulation tissue, thereby causes the secondary damage bringing when dressing removes, and comprises the adhesion to granulation, and the epithelial cell to migration, fibroblastic infringement of hypertrophy are even more serious than not using dressing.This dressing superficial cell adhesion value is very low, significantly, lower than tissue culturing plate and hospital gauze (Fig. 5), this is consistent with the protein adsorption result of dressing, illustrates that the hydrophilic of dressing is high, water absorbing capacity is strong, thereby be unfavorable for the adhesion of cell, such dressing is unlikely causes tissue adhesion.
Above embodiment has been contained the most representative experimental data.
Claims (7)
1. a method of preparing polysaccharide spongy body medical dressing, is characterized in that, comprises the following steps:
Step 1, chitosan is dissolved in to lactic acid aqueous solution, stirs and form homogeneous solution, add succinic anhydrides, stirring reaction under room temperature, obtains N-succinyl-chitosan after dialysis lyophilizing;
Step 2, N-succinyl-chitosan is dissolved in phosphate buffered solution, drips furfuryl amine-2-furylamine, add after water-soluble carbodiimide stirring reaction under room temperature, after dialysis lyophilizing, obtain furan chitosan;
Step 3, sodium alginate is soluble in water, drips Potassium metaperiodate. solution, stirring reaction under lucifuge condition, and dialysis lyophilizing obtains aldehyde radical sodium alginate;
Step 4, by soluble in water aldehyde radical sodium alginate, add 4-(4-N-maleimide phenyl) butyrate to react under room temperature, dialysis obtains maleimide amination sodium alginate after lyophilizing;
Step 5, furan chitosan and maleimide amination sodium alginate aqueous solution are at room temperature fully mixed, room temperature place after freezing placement, after lyophilization, obtain spongy body dressing.
2. the method for preparing polysaccharide spongy body medical dressing according to claim 1, it is characterized in that, in step 1, lactic acid aqueous solution concentration is percent by volume 5%, described lactic acid aqueous solution quality is chitosan 4 times, succinic anhydrides quality is 1 ~ 4 times of chitosan, after chitosan is dissolved in to lactic acid aqueous solution, stir 3 hours, stir 12 ~ 24 hours after adding succinic anhydrides.
3. the method for preparing polysaccharide spongy body medical dressing according to claim 1, it is characterized in that, furfuryl amine-2-the furylamine dripping in step 2 is 100 ~ 500 microlitres, the quality of described N-succinyl-chitosan is 1 ~ 5 times of furfuryl amine-2-furylamine, the quality of buffer is 80 times of N-succinyl-chitosan, water-soluble carbodiimide quality is 0.5 times of N-succinyl-chitosan, stirs 12 ~ 24 hours.
4. the method for preparing polysaccharide spongy body medical dressing according to claim 1, it is characterized in that, the Potassium metaperiodate. solution dripping in step 3 is 2 ~ 5 milliliters, concentration is 0.5M, stir 1 ~ 4 hour, the mass ratio of described sodium alginate and water is 1:100, the quality of sodium alginate is Potassium metaperiodate. 200 ~ 500 times.
5. the method for preparing polysaccharide spongy body medical dressing according to claim 1, it is characterized in that, 4-(4-N-maleimide phenyl) the butyrate solution adding in step 4 is 3 ~ 12 milliliters, concentration is quality percent by volume 0.5%, react 0.5 ~ 3 hour, the mass ratio of described aldehyde radical sodium alginate and water is 1:100, and the quality of 4-(4-N-maleimide phenyl) butyrate is 0.015 ~ 0.06 times of aldehyde radical sodium alginate.
6. the method for preparing polysaccharide spongy body medical dressing according to claim 1, is characterized in that, in step 5, the reaction volume of furan chitosan and maleimide amination sodium alginate is than being 1:2,1:1 or 2:1; Wherein participating in the furan chitosan of reaction and the concentration of maleimide amination sodium alginate is respectively quality percent by volume 0.5% ~ 2.0%; Room temperature is placed 0.5 ~ 2 hour, freezing being placed as-20
ounder C freezing 2 hours.
7. the method for preparing polysaccharide spongy body medical dressing according to claim 1, is characterized in that, the time of described dialysis is 3 ~ 5 days.
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| CN104672484A (en) * | 2013-11-27 | 2015-06-03 | 南京理工大学 | Cross-linked polysaccharide tissue engineering porous scaffold preparation method |
| CN105327356B (en) * | 2014-08-11 | 2018-04-03 | 南京理工大学 | A kind of preparation method of magnetic polysaccharide nano-gel material |
| CN104174061A (en) * | 2014-08-22 | 2014-12-03 | 山东颐诺生物科技有限公司 | Chitosan-alginic acid compound antibacterial slow-release material |
| CN105770903B (en) * | 2014-12-25 | 2019-01-18 | 南京理工大学 | A kind of preparation method of temperature control drug-releasing polymer micro-sphere material |
| CN105169460B (en) * | 2015-10-26 | 2018-09-25 | 广东泰宝医疗科技股份有限公司 | A kind of magnetic therapy antibacterial anti hemorrhagic dressing and preparation method thereof |
| CN107432951B (en) * | 2017-07-14 | 2020-05-01 | 四川省中医药科学院 | Sodium alginate-chitosan dressing loaded with tetrahydrocurcumin nanoparticles and preparation method thereof |
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| US6200595B1 (en) * | 1998-04-24 | 2001-03-13 | Kuraray Co., Ltd. | Medical adhesive |
| CN102137684A (en) * | 2008-04-25 | 2011-07-27 | 医疗行业产品有限公司 | Haemostatic material |
| WO2012056465A1 (en) * | 2010-10-27 | 2012-05-03 | Institute Of Life Sciences | A process for preparing curcumin encapsulated chitosan alginate sponge useful for wound healing |
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| US6200595B1 (en) * | 1998-04-24 | 2001-03-13 | Kuraray Co., Ltd. | Medical adhesive |
| CN102137684A (en) * | 2008-04-25 | 2011-07-27 | 医疗行业产品有限公司 | Haemostatic material |
| WO2012056465A1 (en) * | 2010-10-27 | 2012-05-03 | Institute Of Life Sciences | A process for preparing curcumin encapsulated chitosan alginate sponge useful for wound healing |
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