CN103356692A - Composite antibacterial gel and preparation method thereof - Google Patents
Composite antibacterial gel and preparation method thereof Download PDFInfo
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- CN103356692A CN103356692A CN2013103135650A CN201310313565A CN103356692A CN 103356692 A CN103356692 A CN 103356692A CN 2013103135650 A CN2013103135650 A CN 2013103135650A CN 201310313565 A CN201310313565 A CN 201310313565A CN 103356692 A CN103356692 A CN 103356692A
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Abstract
The present invention discloses a composite antibacterial gel and a preparation method thereof. The composite antibacterial gel which has highly antibacterial performances is prepared by compositing gelatin, chitosan, amino acid and dimethyl octadecyl[3-(trimethoxysilyl)propyl]ammonium chloride. The composite antibacterial gel can stop bleeding, promote wound healing, has highly antibacterial properties, has a certain effect on treatment of vaginitis (bacterial vaginosis, candida vaginitis, mixed vaginitis, vulvitis) and cervical erosion and other gynecological diseases, and also has the effect of cleaning and nursing female privates, stopping bleeding, killing pain, diminishing inflammation, killing bacteria and promoting wound healing.
Description
Technical field
The invention belongs to the medical dressing formulation art, especially a kind of antibacterial gel and preparation method thereof.
Background technology
Hydrogel medical dressing is a kind of novel medical dressing that development in recent years is got up.It is the tridimensional network macromolecule swelling body that contains certain moisture, have good water absorption and biocompatibility, can closely fit and can not stick together with irregular wound surface, reduce the chance of bacteria breed and be easy to and changed, and can participate in the healing of various ingredients and somatomedin promotion wound.Compare with traditional dressing, hydrogel can better promote wound healing, alleviate patient's pain, and it can improve the microenvironment of wound surface, the growth of anti-bacteria, is specially adapted to common body surface wound, such as various skin injurys such as scratch, scuffing, decubital ulcers.The thermal capacity of aerogel dressing hydrogel itself is large, and wound is had gentle cooling effect, can alleviate the pain of wound.Therefore, Chinese scholars conducts extensive research and has obtained some achievements to aerogel dressing.At present, aerogel dressing accounts for the major part in global dressing and binder market.
General anti-bacterial hydrogel adopts chitosan as the body material of hydrogel more, but when infection was serious, there was significant limitation in the bacteriostasis of the subjective antisepsis hydrogels such as chitosan, therefore must the suitable antibacterial of load.
Aminoacid contains amido, also has certain antibiotic effect, and origin of amino acid is extensive, and is cheap.But the report that at present aminoacid is applied to anti-biotic material is less, existing bibliographical information adopts the mode of graft copolymerization that aminoacid is incorporated in the chitosan, but the percent grafting of the prepared product of the method is lower, and in preparation process, easily produce organic solvent, thereby cost and environment have all been caused certain pressure.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of gel with high-efficiency antimicrobial function and preparation method thereof is provided.The preparation of pluralgel agent of the present invention mainly is to have the plurality of raw materials of antibacterial functions, comprises that chitosan, aminoacid and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride and gelatin carry out composite according to a certain percentage.Result's demonstration, prepared pluralgel agent can promote wound healing, has the high-efficiency antimicrobial function, is specially adapted to infect more serious wound.Simultaneously gel safety gentleness, stable performance, preparation technology is simple, with low cost, environmental protection, with short production cycle, is fit to commercial production in enormous quantities.
The present invention solves the problems of the technologies described above with following technical scheme.
At first, invention provides a kind of complex antimicrobials, contains chitosan and aminoacid, and described aminoacid is the aminoacid of surface band positive charge.Described aminoacid is preferably one or more in lysine, arginine and the histidine.
Preferably, described chitosan and amino acid whose mass fraction are respectively 0.2 ~ 6% and 0.2 ~ 13%; Perhaps described chitosan and amino acid whose mass ratio are 1:1 ~ 1:2.5.
Preferably, described composite antibiosis gel contains dimethyl stearyl [3-(trimethoxy is silica-based) propyl group].
The mass fraction of described dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] is 0.1 ~ 10%; Perhaps described dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] is 1:1 ~ 1:2 with described amino acid whose mass ratio.
Except this, complex antimicrobials (gel) preferably further contains gelatin, and the mass fraction of described gelatin is 75 ~ 98%.
Preferably, the pH value of described gel is 6 ~ 7.
The present invention provides a kind of preparation method of antibacterial gel simultaneously, and it may further comprise the steps:
S1. getting a certain amount of chitosan, to add mass concentration be in the 3-8% acetic acid solution, stirs under the room temperature to chitosan to dissolve fully;
S2. under stirring condition, add the aminoacid of certain mass in the chitosan acetic acid solution, stir, it is dissolved fully;
S3. under stirring condition, add dimethyl stearyl [3-(trimethoxy the is silica-based) propyl group] ammonium chloride of certain mass in the above-mentioned mixed solution, stir, make its dissolving evenly;
S4. a certain amount of gelatin is added in the pure water heated and stirred dissolving under 30 ~ 40 ℃ of conditions;
S5. with in the gelatin solution after the mixed solution adding dissolving of chitosan, aminoacid and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, stir, then leave standstill and be cooled to room temperature; Regulating pH with lactic acid solution is 6 ~ 7;
S6. product sterilization encapsulation.
The present invention, the mass fraction of described chitosan in acetic acid solution is 0.2 ~ 0.9%;
The present invention, described amino acid whose kind comprises the aminoacid of the surface band positive charges such as lysine, arginine, histidine;
The present invention, described amino acid whose mass concentration is 0.2 ~ 2%;
The present invention, the mass fraction of described dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is 0.1 ~ 1.5%;
The present invention, the mass fraction of described gelatin in whole compound system is 15 ~ 50%;
The present invention, described gelatin is the derivant of collagen, contains the functional groups such as a large amount of amino, hydroxyl, carboxyl on the strand, has a large amount of hydrogen bonds in the strand and between strand, does not have immunogenicity.
The main sterilization mechanism of gel of the present invention is: (1) chitosan molecule enters in the bacterial cell, and destruction DNA transcribes to RNA's, causes bacterial reproduction to stop; (2) a large amount of amidos is contained on chitosan and aminoacid surface, the hydrion of amido in wound is combined, generate the amido of positively charged, absorption is with the antibacterial of negative charge, make the negative charge skewness on bacteria cell wall and the cell membrane, synthesizing of interference cell wall, Cell wall synthesis and dissolution equilibrium under naturalness have been broken, make cell wall trend towards dissolving, cell membrane is not because bearing the osmotic pressure deformation fracture, content such as water, the protein etc. of cell ooze out, and cause bacterolysis and death.(3) dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride has antibacterial, the bactericidal action of wide spectrum.
Gel of the present invention has the mechanism that promotes wound healing: aminoacid and chitosan can produce synergism, the activation aged cells, make it to be converted into healthy cell, can also promote the effect of weak cell or immunologic cellular activity simultaneously, thereby play the healing that promotes wound.
Amino acid whose kind can affect effect and the antibacterial action that concertedness between itself and the chitosan promotes cell viability, amino acid whose kind comprises nonpolar amino acid (alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, methionine), polarity neutral aminoacid (glycine, serine, threonine, cysteine, tyrosine, agedoite, glutamine), polarity zone positive charge aminoacid (lysine, arginine, histidine), polarity zone negative charge aminoacid (aspartic acid, glutamic acid).Wherein, because nonpolar amino acid is water insoluble, therefore be not suitable for the used aminoacid of the present invention, because the pH value of most of antibacterial isoelectric point, IPs is 3～4, and the character of bacterial cell surface charge is controlled by pH value in the water, be the pH value of water when being lower than the antibacterial isoelectric point, IP, bacterial cell surface band positive charge, on the contrary then electronegative.The prepared antimicrobial fluid of the present invention is near neutral water, therefore bacterial cell surface band negative charge during test.Therefore the aminoacid of polarity neutral aminoacid and polarity zone negative charge all can not be fully and electronegative antibacterial react, therefore only have the effective absorption of aminoacid ability of polarity zone positive charge with the antibacterial of negative charge, thereby play the effect of bacteriostasis and sterilization.
Simultaneously, whether amino acid whose kind also can affect between itself and dimethyl stearyl [3-(trimethoxy the is silica-based) propyl group] ammonium chloride can produce synergitic antibacterial effect, charging neutrality easily with between dimethyl stearyl [3-(trimethoxy the is silica-based) propyl group] ammonium chloride occurs and reacts in the carboxyl in polarity neutral and the electronegative aminoacid, therefore can affect the antibacterial effect of dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, and polarity zone positive charge aminoacid itself is positively charged, can not affect antibacterial effect separately after dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is combined.
Therefore, in the preferred version of the present invention, the aminoacid kind is polarity zone positive charge aminoacid, and most preferably, aminoacid is one or more in lysine, arginine and the histidine.
Compared with prior art, the present invention has following beneficial effect:
(1) not only can stop blooding, promoted wound healing but also have high-efficiency antimicrobial, bacteriostasis property, bio-safety is nontoxic, stable performance, the gynaecopathias such as vaginitis (bacterial vaginosis, monilial vaginitis, Combination vaginitis, vulvitis) and cervical erosion are had therapeutical effect, but and cleaning-nursing female private part, hemostasis and pain-relieving, antiinflammatory is antibacterial and the wound healing effect;
(2) simple, with low cost, the environmental protection, with short production cycle of technique is fit to commercial production in enormous quantities, has great application prospect.
The specific embodiment
The present invention will be further described below in conjunction with specific embodiment, but therefore do not limit the invention in the described example ranges.Employed test method is conventional method if no special instructions among the following embodiment; Employed raw material, reagent etc., if no special instructions, being can be from raw material and the reagent that commercial sources obtains such as routine is commercial.
Embodiment 1
Antibacterial effect contrast during different antibiotic raw material independent role
The 1g chitosan is dissolved in the acetic acid solution of 100mL 8%, 1g dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, 1g lysine, 1g arginine, 1g histidine, 1g aspartic acid, 1g glutamic acid are dissolved in respectively in the 100 mL sterilized water, do blank with sterile distilled water simultaneously, with the agar diffusion paper disk method escherichia coli and Candida albicans are done bacteriostatic experiment.
Antibiotic rate experimental result to escherichia coli and Candida albicans is as follows: chitosan 70 and 75%, dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride 72% and 78%, lysine 65% and 72%, arginine 62% and 67%, histidine 68% and 69%, aspartic acid 50% and 42%, glutamic acid 39% and 36%.
As can be seen from the above data, above-mentioned raw materials all has certain antibacterial effect to escherichia coli and Candida albicans, but antibiotic rate is all less than 80%.Simultaneously, can find out from independent amino acid whose data, the antibacterial effect of surface band positive charge amino acid lysine, arginine, histidine obviously is better than electronegative aminoacid aspartic acid, the antibacterial effect of glutamic acid.In addition, nonpolar amino acid is not here considered owing to being insoluble in water.
0.9g dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is dissolved in the 100 mL sterilized water respectively with after 0.1g lysine, 0.1g arginine, 0.1g histidine, 0.1g aspartic acid, 0.1g glutamic acid mix, do blank with sterile distilled water simultaneously, with the agar diffusion paper disk method escherichia coli and Candida albicans are done bacteriostatic experiment.Wherein, the antibacterial effect of each component and consumption are proportional.
Dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride and all kinds of amino acid whose mixture are as follows to the antibiotic rate experimental result of escherichia coli and Candida albicans:
The mixture 82% and 85% of lysine;
Arginic mixture 79% and 84%;
The mixture 78% and 82% of histidine;
The mixture 71% and 75% of aspartic acid;
The mixture 70% and 72% of glutamic acid.
Can be found out by above data, dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is with after positively charged amino acid lysine, arginine, histidine mixes, its antibacterial effect had collaborative increasing, and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is with after electronegative aminoacid aspartic acid and glutamic acid mix, and its antibacterial effect reduces on the contrary.
Embodiment 2:
It is in 3% the acetic acid solution that the 0.2g chitosan is added the 100mL mass concentration, stirs under the room temperature to chitosan to dissolve fully; Under stirring condition, add the lysine of 0.2g in the chitosan acetic acid solution, stirring dissolves it fully; Under stirring condition, in above-mentioned mixed solution, add 0.1g dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride equally, stir, make its dissolving evenly.The 15g gelatin is dissolved in the 100mL pure water, and heated and stirred is even to dissolving under 30 ℃ of conditions.In the gelatin solution after the mixed solution adding dissolving of chitosan, lysine and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, stir, then leave standstill and be cooled to room temperature, obtain product, regulating pH with lactic acid solution is 6.5, and with product sterilization encapsulation.
Anti-microbial property is carried out in the pluralgel agent for preparing under the above-mentioned condition detect, assay proves: this pluralgel agent reaches respectively 90% and 95% to the antibiotic rate of escherichia coli and Candida albicans.
The gel that meets for preparing under the above-mentioned condition is carried out repair in trauma test, and the testing procedure referenced patent application number is 200910248822.0 " a kind of preparation methoies of chitosan collagen gel ":
1. material
Chitosan composite antibiotic gel, sterile gauze
2. animal
Healthy guinea pig, body weight 200g
3. repair in trauma effect experiment
Get four of Cavia porcelluss, be divided into A group and B and organize two groups, 2 every group, with the guinea pig back cropping, alcohol disinfecting, at the holostrome skin of back excision 2 * 2cm, the A group with plural gel dressing be applied on the wound surface, sterile gauze wraps up; The B group is directly wrapped up wound surface with sterile gauze.Observed gel dressing in 1 day, 2 days, 4 days, 6 days, 12 days to the reparation situation of wound surface in postoperative, record wound healing situation, as shown in table 1.
Table 1 wound healing experimental result
| Group | The animal number of elements | Postoperative 1 day | Postoperative 2 days | Postoperative 4 days | Postoperative 6 days |
| The A group | 2 | Obviously rubescent | Slightly rubescent | Slightly rubescent | Normally |
| ? | ? | Obvious tumefaction | Slight swelling | Without swelling | Without swelling |
| ? | ? | Not bonding | Not bonding | Slightly bonding | Bonding |
| ? | ? | Without oozing of blood | Without oozing of blood | Without oozing of blood | Without oozing of blood |
| The B group | 2 | Obviously rubescent | Obviously rubescent | Slightly rubescent | Slightly rubescent |
| ? | ? | Obvious tumefaction | Slight swelling | Slight swelling | Slight swelling |
| ? | ? | Not bonding | Not bonding | Not bonding | Slightly bonding |
| ? | ? | Oozing of blood | Slight oozing of blood | Without oozing of blood | Without oozing of blood |
Postoperative was formed a mouthful all healed in 12 days two, without the swelling phenomenon.A group skin color is normal, and slight cicatrix is arranged.B group skin color is light rubescent, and larger cicatrix is arranged.
Experiment conclusion: A forms mouthful healing state and obviously is better than the B group, and A group sample (pluralgel) has the promotion wound healing, reduces the effect of cicatrix.And compare with application number 200910248822.0 used chitosan collagen gels, the prepared pluralgel of the present invention is more excellent to the repairing effect of wound.
Embodiment 3:
It is in 5% the acetic acid solution that the 0.9g chitosan is added the 100mL mass concentration, stirs under the room temperature to chitosan to dissolve fully; Under stirring condition, in the chitosan acetic acid solution, add the 2g arginine, stirring dissolves it fully; Under stirring condition, in above-mentioned mixed solution, add 1.5g dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride equally, stir, make its dissolving evenly.The 15g gelatin is dissolved in the 100mL pure water, and heated and stirred is even to dissolving under 40 ℃ of conditions.In the gelatin solution after the mixed solution adding dissolving of chitosan, arginine and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, stir, then leave standstill and be cooled to room temperature, regulating pH with lactic acid solution is 6, obtain product, and with product sterilization encapsulation.
Anti-microbial property is carried out in the pluralgel agent for preparing under the above-mentioned condition detect, assay proves: this pluralgel agent reaches respectively 96% and 98% to the antibiotic rate of escherichia coli and Candida albicans.
Haemostatic effect test is carried out in the pluralgel agent for preparing under the above-mentioned condition, and the testing procedure referenced patent application number is 200910248822.0 " a kind of preparation methoies of chitosan collagen gel ":
1. material
Composite antibiosis gel, sterile gauze (matched group)
2. animal
Healthy rabbits, body weight 2.0kg～2.5 kg
3. rabbit ear wound hemostasis experiment
Get 8 of rabbit, be divided at random two groups (A group and B groups), 4 every group, left and right sides ear is labeled as respectively the A group: A1, A2, B group: B1, B2.To it, in ear edge 1/3rd places, crosscut is the long wound of 1cm approximately with rabbit left and right sides ear, and wound is from ear edge crosscut rabbit ear, and all ears are done same treatment of wounds.A group: smear chitosan collagen gel dressing in 5 seconds, cover sterile gauze and pressed 1 minute; The B group was pressed one minute with gauze.Began later on to observe the bleeding of wound in one minute, and the beginning timing to oozing of blood no longer, hemorrhage be bleeding stopping period, record the result as shown in table 2.
The haemostatic effect test of table 2 wound
| Group | The animal number of elements | Bleeding stopping period |
| The A group | 4 | 1.90±0.10 |
| The B group | 4 | 3.10±0.10 |
Experiment conclusion: the result shows that A group, B group all have anastalsis in various degree, and wherein A group successful is better than the B group.This explanation composite antibiosis gel has preferably anthemorrhagic performance, and to be that bleeding stopping period A group 2.10 ± 0.10, the B group 3.35 ± 0.10 of 200910248822.0 chitosan collagen gel is compared shorter with application number simultaneously, and anthemorrhagic performance is more excellent.
Embodiment 4:
It is in 5% the acetic acid solution that the 0.5g chitosan is added the 100mL mass concentration, stirs under the room temperature to chitosan to dissolve fully; Under stirring condition, in the chitosan acetic acid solution, add the 1g histidine, stirring dissolves it fully; Under stirring condition, in above-mentioned mixed solution, add 1g dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride equally, stir, make its dissolving evenly.The 30g gelatin is dissolved in the 100mL pure water, and heated and stirred is even to dissolving under 40 ℃ of conditions.In the gelatin solution after the mixed solution adding dissolving of chitosan, histidine and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, stir, then leave standstill and be cooled to room temperature, regulating pH with lactic acid solution is 6.5, obtain product, and with product sterilization encapsulation.
Anti-microbial property is carried out in the pluralgel agent for preparing under the above-mentioned condition detect, assay proves: this pluralgel agent reaches respectively 92% and 95% to the antibiotic rate of escherichia coli and Candida albicans.
Claims (10)
1. a complex antimicrobials is characterized in that, contains chitosan and aminoacid, and described aminoacid is the aminoacid of surface band positive charge.
2. described a kind of complex antimicrobials according to claim 1 is characterized in that, described aminoacid is one or more in lysine, arginine and the histidine.
3. described a kind of complex antimicrobials according to claim 2 is characterized in that, described chitosan and amino acid whose mass fraction are respectively 0.2 ~ 6% and 0.2 ~ 13%; Perhaps described chitosan and amino acid whose mass ratio are 1:1 ~ 1:2.5.
4. described a kind of complex antimicrobials according to claim 1 is characterized in that, described composite antibiosis gel contains dimethyl stearyl [3-(trimethoxy is silica-based) propyl group].
5. described a kind of complex antimicrobials according to claim 4 is characterized in that, the mass fraction of described dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] is 0.1 ~ 10%; Perhaps described dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] is 1:1 ~ 1:2 with described amino acid whose mass ratio.
6. described a kind of complex antimicrobials according to claim 1 is characterized in that, described composite antibiosis gel contains gelatin, and the mass fraction of described gelatin is 75 ~ 98%.
7. described a kind of complex antimicrobials according to claim 1 is characterized in that, the pH value of described gel is 6 ~ 7.
8. the preparation method of a composite antibiosis gel is characterized in that, comprises following processing step:
S1. taking by weighing chitosan, to add mass concentration be in 3 ~ 8% the acetic acid solution, stirs under the room temperature to chitosan to dissolve fully;
S2. under stirring condition, in the chitosan acetic acid solution, add aminoacid, stir, it is dissolved fully;
S3. under stirring condition, in above-mentioned mixed solution, add dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, be stirred to dissolving evenly;
S4. take by weighing gelatin and add in the pure water, heated and stirred dissolving under 30 ~ 40 ℃ of conditions;
S5. the mixed solution of S3 step is added in the gelatin solution after the dissolving, stir, regulate 6 ~ 7 with lactic acid solution, leave standstill and be cooled to sterilization encapsulation after the room temperature.
9. the preparation method of described a kind of composite antibiosis gel according to claim 8, it is characterized in that, the addition of described chitosan is 0.2 ~ 6% mass fraction, described amino acid whose addition is 0.2 ~ 13% mass fraction, the addition of described dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is 0.1 ~ 10% mass fraction, and the addition of described gelatin is 75 ~ 98% mass fractions.
10. the preparation method of described a kind of composite antibiosis gel according to claim 8 is characterized in that, described aminoacid is the aminoacid of surface band positive charge.
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