CN103356692B - A kind of composite antibacterial gel and preparation method thereof - Google Patents
A kind of composite antibacterial gel and preparation method thereof Download PDFInfo
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- CN103356692B CN103356692B CN201310313565.0A CN201310313565A CN103356692B CN 103356692 B CN103356692 B CN 103356692B CN 201310313565 A CN201310313565 A CN 201310313565A CN 103356692 B CN103356692 B CN 103356692B
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Abstract
The invention discloses a kind of pluralgel agent and preparation method thereof, mainly gelatin, chitosan, aminoacid and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride are carried out compound, prepare a kind of medical pluralgel agent with high-efficiency antimicrobial, bacteriostasis property.This pluralgel agent can stop blooding, promote wound healing, there is again high-efficiency antimicrobial, bacteriostasis property, bio-safety is nontoxic, to gynaecopathias such as treatment vaginitis (bacterial vaginosis, monilial vaginitis, Combination vaginitis, vulvitis) and cervical erosion, there is certain effect, in addition, the effects such as the antibacterial and wound healing of cleaning-nursing female private part, hemostasis and pain-relieving, antiinflammatory can also be played.
Description
Technical field
The invention belongs to 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 swelling body of tridimensional network macromolecule containing certain moisture, there is good water absorption and biocompatibility, can closely fit with irregular wound surface and can not stick together, decrease the chance of bacteria breed and be easy to change, and the healing of various ingredient and somatomedin promotion wound can be participated in.Compared with traditional dressing, hydrogel can better promote wound healing, alleviate the pain of patient, and it can improve the microenvironment of wound surface, the growth of anti-bacteria, is specially adapted to common body surface wound, as various skin injurys such as scratch, scuffing, decubital ulcers.The thermal capacity of aerogel dressing hydrogel itself is large, has gentle cooling effect, can alleviate the pain of wound to wound.Therefore, Chinese scholars conducts extensive research aerogel dressing and achieves some achievements.At present, aerogel dressing accounts for the major part in global dressing and binder market.
General anti-bacterial hydrogel many employings chitosan is as the body material of hydrogel, but in time infecting serious, the bacteriostasis of the subjective antisepsis hydrogels such as chitosan exists significant limitation, the antibacterial that therefore necessary load is suitable.
Aminoacid contains amido, also has certain antibiotic effect, and origin of amino acid is extensive, cheap.But the report at present aminoacid being applied to anti-biotic material is less, existing bibliographical information adopts the mode of graft copolymerization to be incorporated in chitosan by aminoacid, but the percent grafting of product is lower prepared by the method, and in preparation process, easily produce organic solvent, thus certain pressure is all caused to cost and environment.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, provide a kind of and there is gel of high-efficiency antimicrobial function and preparation method thereof.The preparation of pluralgel agent of the present invention mainly will have the plurality of raw materials of antibacterial functions, comprise chitosan, aminoacid and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride and gelatin carries out composite according to a certain percentage.Result shows, and prepared pluralgel agent can promote wound healing, has high-efficiency antimicrobial function, is specially adapted to infect more serious wound.Simultaneously gentle, the stable performance of gel safety, preparation technology is simple, with low cost, environmental protection, with short production cycle, is applicable to commercial production in enormous quantities.
The present invention solves the problems of the technologies described above with following technical scheme.
First, invention provides a kind of complex antimicrobials, and containing chitosan and aminoacid, described aminoacid is the aminoacid of surface band positive charge.Described aminoacid be preferably in lysine, arginine and histidine one or more.
Preferably, described chitosan and amino acid whose mass fraction are respectively 0.2 ~ 6% and 0.2 ~ 13%; Or described chitosan and amino acid whose mass ratio are 1:1 ~ 1:2.5.
Preferably, described composite antibacterial 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%; Or described dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] and described amino acid whose mass ratio are 1:1 ~ 1:2.
Except this, complex antimicrobials (gel) is preferably further containing gelatin, and the mass fraction of described gelatin is 75 ~ 98%.
Preferably, the pH value of described gel is 6 ~ 7.
Invention also provides a kind of preparation method of antibacterial gel, it comprises the following steps:
S1. getting a certain amount of chitosan, to add mass concentration be in 3-8% acetic acid solution, and stirred at ambient temperature evenly dissolves completely to chitosan;
S2. under agitation, in chitosan acetic acid solution, add the aminoacid of certain mass, stir, make it dissolve completely;
S3. under agitation, in above-mentioned mixed solution, add dimethyl stearyl [3-(trimethoxy the is silica-based) propyl group] ammonium chloride of certain mass, stir, make it be uniformly dissolved;
S4. add in pure water by a certain amount of gelatin, under 30 ~ 40 DEG C of conditions, heated and stirred is dissolved;
S5. the mixed solution of chitosan, aminoacid and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is added in the gelatin solution after dissolving, stir, then leave standstill and be cooled to room temperature; PH is regulated to be 6 ~ 7 with lactic acid solution;
S6. product sterilizing 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, containing a large amount of functional groups such as amino, hydroxyl, carboxyl on strand, there is a large amount of hydrogen bond, do not have immunogenicity in strand and between strand.
The major antibacterial mechanism of inventive gel agent is: (1) chitosan molecule enters in bacterial cell, destroys transcribing of DNA to RNA, causes bacterial reproduction to stop; (2) chitosan and amino acid surface contain a large amount of amidos, the hydrion of amido in wound is combined, generate the amido of positively charged, adsorb the antibacterial with negative charge, make the negative charge skewness on bacteria cell wall and cell membrane, the synthesis of interference cell wall, break Cell wall synthesis in its natural state and dissolution equilibrium, cell wall is made to trend towards dissolving, cell membrane deformation fracture because osmotic pressure can not be born, content such as water, the protein etc. of cell ooze out, and cause bacterolysis and dead.(3) dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride has antibacterial, the bactericidal action of wide spectrum.
Inventive gel agent has the mechanism promoting wound healing: aminoacid and chitosan can produce synergism, activation aged cells, make it to be converted into healthy cell, the effect of weak cell or immunologic cellular activity can also be promoted simultaneously, thus play the healing promoting wound.
The amino acid whose kind concertedness that can affect between itself and chitosan promotes effect and the antibacterial action of 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 the present invention's aminoacid used is not suitable for, pH value due to most of antibacterial isoelectric point, IP is 3 ~ 4, and the character of bacterial cell surface electric charge controls by pH value in water, namely, when the pH value of water is lower than antibacterial isoelectric point, IP, bacterial cell surface is positively charged, otherwise then electronegative.Antimicrobial fluid during test prepared by the present invention is close to neutral water, therefore bacterial cell surface is electronegative.Therefore the aminoacid of polarity neutral aminoacid and polarity zone negative charge all can not react with electronegative antibacterial fully, therefore only have the antibacterial of the effective absorption of aminoacid ability with negative charge of polarity zone positive charge, thus play the effect of bacteriostasis and sterilization.
Simultaneously, whether amino acid whose kind also can affect between itself and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride can produce synergitic antibacterial effect, carboxyl in polarity neutral and electronegative aminoacid easily and between dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, charging neutrality occurs and react, therefore the antibacterial effect of dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride can be affected, and polarity zone positive charge aminoacid itself is positively charged, respective antibacterial effect can not be affected after being combined with dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride.
Therefore, in preferred version of the present invention, amino acid classes is polarity zone positive charge aminoacid, and most preferably, aminoacid is one or more in lysine, arginine and histidine.
Compared with prior art, the present invention has following beneficial effect:
(1) not only high-efficiency antimicrobial, bacteriostasis property can have been stopped blooding, promoted wound healing but also have, bio-safety is nontoxic, stable performance, to gynaecopathias such as vaginitis (bacterial vaginosis, monilial vaginitis, Combination vaginitis, vulvitis) and cervical erosion, there is therapeutical effect, and can the antibacterial and wound healing effect of cleaning-nursing female private part, hemostasis and pain-relieving, antiinflammatory;
(2) simple, with low cost, the environmental protection of technique, with short production cycle, be applicable to commercial production in enormous quantities, there is great application prospect.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention will be further described, but therefore do not limit the invention in described example ranges.The test method used in following embodiment if no special instructions, is conventional method; The raw material used, reagent etc., if no special instructions, being can from raw material that commercial sources obtains and the reagent such as routine is commercial.
embodiment 1
Antibacterial effect contrast during different antibacterial raw material independent role
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 100 mL sterilized water respectively, do blank with sterile distilled water simultaneously, with agar diffusion paper disk method, bacteriostatic experiment is done to escherichia coli and Candida albicans.
As follows to the antibiotic rate experimental result of escherichia coli and Candida albicans: 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%.Meanwhile, as can be seen from independent amino acid whose data, the antibacterial effect of surface band positive charge amino acid lysine, arginine, histidine is obviously better than electronegative amino acid aspartic acid, the antibacterial effect of glutamic acid.In addition, nonpolar amino acid, owing to being comparatively insoluble in water, is not here considered.
0.9g dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is dissolved in 100 mL sterilized water with 0.1g lysine, 0.1g arginine, 0.1g histidine, 0.1g aspartic acid, 0.1g glutamic acid respectively after mixing, do blank with sterile distilled water simultaneously, with agar diffusion paper disk method, bacteriostatic experiment is done to escherichia coli and Candida albicans.Wherein, the antibacterial effect of each component and consumption proportional.
Dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride and the antibiotic rate experimental result of all kinds of amino acid whose mixture to escherichia coli and Candida albicans as follows:
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.
As can be seen from above data, after dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride mixes with positively charged amino acid lysine, arginine, histidine, its antibacterial effect had collaborative increasing, and after dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride mixes with electronegative amino acid aspartic acid and glutamic acid, its antibacterial effect reduces on the contrary.
embodiment 2:
0.2g chitosan being added 100mL mass concentration is in the acetic acid solution of 3%, and stirred at ambient temperature evenly dissolves completely to chitosan; Under agitation, in chitosan acetic acid solution, add the lysine of 0.2g, stir and make it dissolve completely; Equally under agitation, in above-mentioned mixed solution, add 0.1g dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, stir, make it be uniformly dissolved.Be dissolved in by 15g gelatin in 100mL pure water, under 30 DEG C of conditions, heated and stirred is to being uniformly dissolved.The mixed solution of chitosan, lysine and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is added in the gelatin solution after dissolving, stir, then leave standstill and be cooled to room temperature, obtain product, regulate pH to be 6.5 with lactic acid solution, and product sterilizing is encapsulated.
Carry out anti-microbial property detection to the pluralgel agent prepared under above-mentioned condition, assay proves: the antibiotic rate of this pluralgel agent to escherichia coli and Candida albicans reaches 90% and 95% respectively.
Carry out repair in trauma test to the gel that meets prepared under above-mentioned condition, 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 experimental
Get Cavia porcellus four, be divided into A group and B group two groups, often organize 2, by guinea pig back cropping, alcohol disinfecting, at the full thickness skin of back excision 2 × 2cm, plural gel dressing is applied on wound surface by A group, sterile gauze wrapping; The direct sterile gauze of B group wraps up wound surface.Gel dressing is observed to the reparation situation of wound surface, record wound healing situation in postoperative 1 day, 2 days, 4 days, 6 days, 12 days, as shown in table 1.
Table 1 wound healing experimental result
| Group | Animal number of elements | Postoperative 1 day | Postoperative 2 days | Postoperative 4 days | Postoperative 6 days |
| A group | 2 | Obviously rubescent | Slightly rubescent | Slightly rubescent | Normally |
| Obvious tumefaction | Slight swelling | Without swelling | Without swelling | ||
| Do not bond | Do not bond | Slight bonding | Bonding | ||
| Without oozing of blood | Without oozing of blood | Without oozing of blood | Without oozing of blood | ||
| B group | 2 | Obviously rubescent | Obviously rubescent | Slightly rubescent | Slightly rubescent |
| Obvious tumefaction | Slight swelling | Slight swelling | Slight swelling | ||
| Do not bond | Do not bond | Do not bond | Slight bonding | ||
| Oozing of blood | Slight oozing of blood | Without oozing of blood | Without oozing of blood |
Within postoperative 12 days two, form a mouthful all healed, without swelling phenomenon.A group skin color is normal, has slight cicatrix.B group skin color is light rubescent, has larger cicatrix.
Experiment conclusion: A forms mouthful healing state and is obviously better than B group, and A group sample (pluralgel) has promotion wound healing, reduces effect of cicatrix.And compared with application number 200910248822.0 chitosan collagen gel used, the repairing effect of pluralgel prepared by the present invention to wound is more excellent.
embodiment 3:
0.9g chitosan being added 100mL mass concentration is in the acetic acid solution of 5%, and stirred at ambient temperature evenly dissolves completely to chitosan; Under agitation, in chitosan acetic acid solution, add 2g arginine, stir and make it dissolve completely; Equally under agitation, in above-mentioned mixed solution, add 1.5g dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, stir, make it be uniformly dissolved.Be dissolved in by 15g gelatin in 100mL pure water, under 40 DEG C of conditions, heated and stirred is to being uniformly dissolved.The mixed solution of chitosan, arginine and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is added in the gelatin solution after dissolving, stir, then leave standstill and be cooled to room temperature, pH is regulated to be 6 with lactic acid solution, obtain product, and product sterilizing is encapsulated.
Carry out anti-microbial property detection to the pluralgel agent prepared under above-mentioned condition, assay proves: the antibiotic rate of this pluralgel agent to escherichia coli and Candida albicans reaches 96% and 98% respectively.
Carry out haemostatic effect test to the pluralgel agent prepared under above-mentioned condition, testing procedure referenced patent application number is 200910248822.0 " a kind of preparation methoies of chitosan collagen gel ":
1. material
Composite antibacterial gel, sterile gauze (matched group)
2. animal
Healthy rabbits, body weight 2.0kg ~ 2.5 kg
3. rabbit ear wound hemostasis experiment
Get rabbit 8, be divided at random two groups (A group and B groups), often organize 4, left and right ear is labeled as A group respectively: A1, A2, B group: B1, B2.By rabbit left and right ear to it, in ear edge 1/3rd place, crosscut is about the long wound of 1cm, and wound is from ear edge crosscut rabbit ear, and all ears do same treatment of wounds.A group: smear chitosan collagen gel dressing in 5 seconds, covers sterile gauze and presses 1 minute; B group gauze presses one minute.Within one minute, start the bleeding observing wound later, and start timing to no longer oozing of blood, hemorrhage be bleeding stopping period, record result is as shown in table 2.
The haemostatic effect test of table 2 wound
| Group | Animal number of elements | Bleeding stopping period |
| A group | 4 | 1.90±0.10 |
| B group | 4 | 3.10±0.10 |
Experiment conclusion: result display A group, B group all have anastalsis in various degree, and wherein A group successful is better than B group.This illustrates that composite antibacterial gel has good anthemorrhagic performance, and bleeding stopping period A group 2.10 ± 0.10, the B group 3.35 ± 0.10 being simultaneously the chitosan collagen gel of 200910248822.0 with application number is compared shorter, and anthemorrhagic performance is more excellent.
embodiment 4:
0.5g chitosan being added 100mL mass concentration is in the acetic acid solution of 5%, and stirred at ambient temperature evenly dissolves completely to chitosan; Under agitation, in chitosan acetic acid solution, add 1g histidine, stir and make it dissolve completely; Equally under agitation, in above-mentioned mixed solution, add 1g dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, stir, make it be uniformly dissolved.Be dissolved in by 30g gelatin in 100mL pure water, under 40 DEG C of conditions, heated and stirred is to being uniformly dissolved.The mixed solution of chitosan, histidine and dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is added in the gelatin solution after dissolving, stir, then leave standstill and be cooled to room temperature, pH is regulated to be 6.5 with lactic acid solution, obtain product, and product sterilizing is encapsulated.
Carry out anti-microbial property detection to the pluralgel agent prepared under above-mentioned condition, assay proves: the antibiotic rate of this pluralgel agent to escherichia coli and Candida albicans reaches 92% and 95% respectively.
Claims (2)
1. a composite antibacterial gel, is characterized in that, containing chitosan and aminoacid, described aminoacid is one or more in lysine, arginine and histidine,
described chitosan and amino acid whose mass fraction are respectively 0.2 ~ 6% and 0.2 ~ 13%; Or described chitosan and amino acid whose mass ratio are 1:1 ~ 1:2.5,
described composite antibacterial gel contains dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride;
the mass fraction of described dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride is 0.1 ~ 10%; Or described dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride and described amino acid whose mass ratio are 1:1 ~ 1:2;
described composite antibacterial gel contains gelatin, and the mass fraction of described gelatin is 75 ~ 98%,
the pH value of described composite antibacterial gel is 6.
2. a preparation method for composite antibacterial gel, is characterized in that, comprises following processing step:
S1. taking chitosan, to add mass concentration be in the acetic acid solution of 3 ~ 8%, and stirred at ambient temperature evenly dissolves completely to chitosan;
S2. under agitation, in chitosan acetic acid solution, add aminoacid, stir, make it dissolve completely;
S3. under agitation, in above-mentioned mixed solution, add dimethyl stearyl [3-(trimethoxy is silica-based) propyl group] ammonium chloride, be stirred to and be uniformly dissolved;
S4. taking gelatin adds in pure water, and under 30 ~ 40 DEG C of conditions, heated and stirred is dissolved;
S5. the mixed solution of S3 step is added in the gelatin solution after dissolving, stir, regulate 6 ~ 7 with lactic acid solution, leave standstill sterilizing after being cooled to room temperature and encapsulate;
Described aminoacid is one or more in lysine, arginine and histidine;
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.
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| CN110583755A (en) * | 2019-08-30 | 2019-12-20 | 江苏大学 | Double-histidine/dendrobium officinale polysaccharide composite antibacterial agent and preparation method and application thereof |
| CN110812471A (en) * | 2019-10-31 | 2020-02-21 | 南阳师范学院 | Antibacterial composite gel for wound surface |
| CN112315899B (en) * | 2020-11-04 | 2022-11-25 | 湖南天根乐微君科技有限公司 | Biological gel dressing for preventing/treating HPV and gynecological inflammation and preparation method and application thereof |
| CN112369427A (en) * | 2020-11-13 | 2021-02-19 | 南京威尔凯勒生物技术有限公司 | Broad-spectrum antiviral antibacterial disinfectant and preparation method thereof |
| CN112940285B (en) * | 2021-02-02 | 2024-01-16 | 浙江工商大学 | Hydrogel convenient to clean and application |
| CN114456403A (en) * | 2021-12-31 | 2022-05-10 | 南京神奇科技开发有限公司 | Temperature-sensitive gel containing organosilicon quaternary ammonium salt and preparation method thereof |
| CN115252640B (en) * | 2022-06-23 | 2023-08-29 | 中国科学院长春应用化学研究所 | Chitosan-N-arginine nanoparticle, and preparation method and application thereof |
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