CN104856978A - Chitosan spraying film agent and preparation method thereof - Google Patents
Chitosan spraying film agent and preparation method thereof Download PDFInfo
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Abstract
The invention provides a chitosan spraying film agent. The chitosan spraying film agent is prepared from the following components in percentage by weight: 0.05% to 0.5% of chitosan, 0.01% to 0.15% of lactic acid, 0.1% to 0.2% of a surfactant, 0.1% to 0.3% of a cosolvent and the balance of water. The invention further provides a method for preparing the spraying film agent and application of the spraying film agent. The prepared chitosan spraying film agent provided by the invention is little in irritation for skin wounds, and high in film-forming property, biological availability and antibacterial ability, and can effectively promote wound healing.
Description
Technical field
The present invention relates to a kind of chitosan spraying membrane and preparation method thereof, belong to drug world.
Background technology
Along with the development of world industry, wound, has become disease common in daily life, and traumatic infection is then cause dead major reason.According to World Health Organization's statistics, the nineteen ninety whole world various wound lethal number about 5,100,000 people, estimates that the year two thousand twenty can increase to 8,400,000 people.Wound is the very important disease of a class, and treatment and the repairing research of therefore carrying out wound are very necessary.Along with the development that the repair in trauma " moistening healing " of Winter, Hinman and Maibaeh etc. is theoretical, the application of wound repair and protective material adjuvant grows up gradually.
The clinical antibiotic medicine dressing that generally adopts carries out antibacterial therapy to skin wound; but due to the restriction of its structure; though traditional dressing has certain moisture absorption and protective effect; but generally can only be used for the wound emergence therapeutic of small size, non-joint; thus play temporary transient hemostasis; the effect of protection wound surface, and clinical practice is greatly limited.
Chitosan (chitosan) extracts from the film of Carapax Eriocheir sinensis, Crusta Penaeus seu Panulirus or fungus through deacetylated and a kind of polysaccharide that is that formed, is a kind of natural polymer chemical material.Chitosan have hemostasis, antibacterial, promote wound healing, suppress the multiple effect such as cicatrization, there is excellent histocompatibility, inanimate object repellency, do not cause allergic reaction, biodegradability, natural film property is excellent medical dressing and tissue engineering material.The chitosan of high molecular has the advantages such as good film-forming property, antibacterial ability be strong, but because of its degree of crystallinity highly dissoluble poor, cannot preparation, application is greatly limited; Low-molecular-weight chitosan or chitosan derivatives need through degraded, derivatization method preparation, although its solubility problem is improved, its film property, antibacterial ability decline to some extent.
At present, chitosan series products that is commercially available and bibliographical information is all using low-molecular weight chitoglycan (such as CN102846655A, CN104042719A, CN102824308) or chitosan derivatives (such as CN104586753A) as active component, although its dissolubility is better, but antibacterial ability is not good, heavy dose of active component must be used or add multiple antibacterial substance and play synergism, just can reach good antibacterial effect.So just cause the drawbacks such as the supplementary material of preparation expends too much, complicated process of preparation, production cost raising, and harmful effect is produced to the quality of preparation and safety.
Meanwhile, in order to dissolve chitosan well, the reasonable solvent of the multiplex solubility property of current chitosan series products: acetic acid, but there is the strong problem of zest in it, directly affects the safety of medication.
For the drawback of existing chitosan series products, preferred supplementary material, control composition content, provides for clinical treatment skin trauma infects the novel formulation that a kind of zest is little, good film-forming property, antibacterial ability are strong, has very important significance.
Summary of the invention
Technical scheme of the present invention there is provided a kind of chitosan spraying membrane and its production and use.
The invention provides a kind of chitosan spraying membrane, it is prepared from by the component of following weight percentage ratio: chitosan 0.05-0.5%, lactic acid 0.01%-0.15%, surfactant 0.1%-0.2%, cosolvent 0.1%-0.3%, and surplus is water.
Wherein, described chitosan molecule amount is 10w-30w.
Preferably, described chitosan molecule amount is 30w.
Wherein, described surfactant is n-octyl alcohol, water-soluble silicon oil or dimethicone;
Described solubilizing agent is Tween 80, benzoic acid or PEG400.
Preferably, the component of percentage by weight is:
Chitosan 0.1%, lactic acid 0.05%, n-octyl alcohol 0.15%, Tween 80 0.25%, surplus is water.
The invention provides the method for this chitosan of preparation spraying membrane, comprise the steps:
A. each component is taken according to proportioning described in claim 1-5 any one;
B. extracting lactic acid, is prepared into the lactic acid aqueous solution that concentration is 0.1-0.14%;
C. get chitosan, be dispersed in lactic acid solution prepared by b step, swelling, after stirring and dissolving, adjust ph, to 3.5-6.0, is filtered, is obtained chitosan lactic acid solution;
D. cosolvent and surfactant are dissolved in ethanol solution respectively, make the alcoholic solution of cosolvent and the alcoholic solution of surfactant respectively;
E. stir the chitosan lactic acid solution prepared of step c, add the alcoholic solution of the cosolvent that step c obtains, stir to clarify, then add the alcoholic solution of the surfactant that step c obtains, stir to clarify, fill and get final product.
Wherein, in described lactic acid aqueous solution, lactic acid concn is 0.12%.The concentration of the alcoholic solution of described cosolvent and the alcoholic solution of surfactant is 5%.
Chitosan spraying membrane of the present invention reaches more than 99% to colibacillary bacteriostasis rate, reaches 100% to the bacteriostasis rate of staphylococcus aureus.
Present invention also offers the purposes of this chitosan spraying membrane in the medicine preparing the infection for the treatment of skin trauma and/or wound healing.Described medicine is the medicine of Chinese People's Anti-Japanese Military and Political College enterobacteria or staphylococcus aureus.
The present invention is by the particular combination of special component, solvent using solubility property poor: under the condition of lactic acid, dissolve poorly soluble high molecular weight chitosan on the contrary well, obtain with good film-forming property, antibacterial ability is strong, bioavailability is high chitosan spraying membrane, and because do not use acetic acid, its zest is little, safety is good, effect experiment proves, chitosan spraying membrane of the present invention can be effectively antibacterial, treatment skin trauma infects, and promote wound healing, potential applicability in clinical practice is good.
Accompanying drawing explanation
Fig. 1 chitosan spraying of the present invention Membranous preparation processing technology routine figure
Fig. 2 tri-groups of rats the 0th day traumatic wounds figure (being from left to right respectively chitosan spraying membrane group, commercially available prod A group, normal saline group)
Fig. 3 tri-groups of rats the 7th day traumatic wounds figure (being from left to right respectively chitosan spraying membrane group, commercially available prod A group, normal saline group)
Fig. 4 tri-groups of rats the 14th day traumatic wounds figure (being from left to right respectively chitosan spraying membrane group, commercially available prod A group, normal saline group)
Fig. 5 saline rats histopathology slide (being from left to right respectively No. 2 rat skins section (HE200 ×), No. 5 rat skins section (HE200 ×), No. 7 rat skins section (HE200 ×))
Fig. 6 commercially available prod A group rat tissue pathology section (being from left to right respectively No. 3 rat skins section (HE200 ×), No. 6 rat skins section (HE200 ×), No. 8 rat skins section (HE200 ×))
Fig. 7 chitosan spraying membrane group rat tissue pathology section (being from left to right respectively No. 1 rat skin section (HE200 ×) No. 4 rat skins section (HE200 ×) No. 9 rat skins section (HE200 ×))
Fig. 8 negative control group rabbit vagina pathological section (being from left to right respectively rabbit vagina epimere section (HE200 ×), rabbit vagina stage casing section (HE200 ×), rabbit vagina hypomere section (HE200 ×))
Fig. 9 chitosan spraying membrane group matched group rabbit vagina pathological section (being from left to right respectively rabbit vagina epimere section (HE200 ×), rabbit vagina stage casing section (HE200 ×), rabbit vagina hypomere section (HE200 ×))
Below by way of detailed description of the invention, the present invention is described in further detail, but do not limit the present invention, the various change that those skilled in the art make according to the present invention and replacement, only otherwise depart from spirit of the present invention, all should belong to the scope of claims of the present invention.
Detailed description of the invention
The preparation technology of embodiment 1 chitosan spraying membrane
By 0.05g lactic acid dissolution in 100g pure water, make 0.05% lactic acid solution; Weigh 0.2g chitosan (molecular weight is 30w), be dispersed in the 0.05% swelling 30min in lactic acid solution surface, after stirring 1.5h dissolving, regulate pH to 4.5, filter, obtain chitosan lactic acid solution.Tween 80 0.3g and n-octyl alcohol 0.1g is dissolved in alcoholic solution respectively and is made into 5% alcoholic solution, under agitation, slowly add n-octyl alcohol solution with micro-sampling pin, then add tween solution, be stirred to solution clarification, fill and get final product.Encapsulating immediately after bottling, is 100,000 grades of workshop fills at cleanliness factor, and every bottled amount should be no less than sign loading amount 20mL/ bottle.(fabricating technology route map is shown in Fig. 1)
Using method: chitosan spraying membrane evenly sprays in the position of distance wound surface 10-15cm.Each sprinkling pressing 2-3 time, also can increase according to patient's practical situation or reduce consumption.Use 2 every day, using continuously 15 days is a course for the treatment of.
The preparation of embodiment 2 chitosan spraying of the present invention membrane
By 1g lactic acid dissolution in 1000g pure water, make 0.1% lactic acid solution; Weigh 5g chitosan (molecular weight 10W), be dispersed in the 0.1% swelling 50min in lactic acid solution surface, after stirring 2h dissolving, regulate pH to 5, filter, obtain chitosan lactic acid solution.Tween 80 2g and n-octyl alcohol 1.2g is dissolved in alcoholic solution respectively and is made into 5% alcoholic solution, under agitation, slowly add n-octyl alcohol solution with micro-sampling pin, then add tween solution, be stirred to solution clarification, fill and get final product.
The preparation of embodiment 3 chitosan spraying of the present invention membrane
By 5g lactic acid dissolution in 2500g pure water, make 0.2% lactic acid solution; Weigh 3.75g chitosan (molecular weight 30W), be dispersed in the 0.2% swelling 50min in lactic acid solution surface, after stirring 1h dissolving, regulate pH to 4, filter, obtain chitosan lactic acid solution.Tween 80 5g and n-octyl alcohol 4.8g is dissolved in alcoholic solution respectively and is made into 5% alcoholic solution, under agitation, slowly add n-octyl alcohol solution with micro-sampling pin, then add tween solution, be stirred to solution clarification, fill and get final product.
The preparation of embodiment 4 chitosan spraying of the present invention membrane
By 10g lactic acid dissolution in 20kg pure water, make 0.05% lactic acid solution; Weigh 40g chitosan (molecular weight 30W), be dispersed in the 0.05% swelling 1.5h in lactic acid solution surface, after stirring 2h dissolving, regulate pH to 5.3, filter, obtain chitosan lactic acid solution.Tween 80 50g and n-octyl alcohol 36g is dissolved in alcoholic solution respectively and is made into 5% alcoholic solution, under agitation, slowly add n-octyl alcohol solution with micro-sampling pin, then add tween solution, be stirred to solution clarification, fill and get final product.
Beneficial effect of the present invention is proved below by way of specific experiment.
The screening test of surfactant, hydrotropy in experimental example 1 chitosan spraying of the present invention membrane
One, the screening of surfactant
1, experimental technique
Measure chitosan lactic acid solution surface tension value, as the reference index weighing surfactant.Compound concentration is water-soluble silicon oil solution, dimethicone alcoholic solution, each 5mL of n-octyl alcohol alcoholic solution of 0.15%, above solution is added respectively 500mL isocyatic chitosan lactic acid solution and is mixed with three groups of different solution.Under normal temperature and pressure, the solution prepared is injected successively surface tension apparatus measurement ware middle circle connected in star and be about 1mL, record reading, repeats often to organize to average in triplicate.Be evaluation index with surface tension value, select optimum surfactant.
2, experimental result
Table 1 chitosan lactic acid original solution surface tension value
Note: experiment is carried out at normal temperatures and pressures, lower same.
Table 2 three groups of surfactants are on the impact of the surface tension value of chitosan lactic acid solution
Test shows, n-octyl alcohol, water-soluble silicon oil or dimethicone effectively can reduce the surface tension of chitosan lactic acid solution, may be used to preparation the present invention and to spray membrane, wherein, the best results of n-octyl alcohol, therefore, preferred n-octyl alcohol is as surfactant.
Two, the screening of cosolvent
1, experimental technique
Compound concentration is PEG400 alcoholic solution, Tween 80 alcoholic solution, each 5mL of alcohol benzoate solution of 0.25%, above solution is added respectively the isocyatic chitosan lactic acid solution containing n-octyl alcohol alcoholic solution of 500mL, stirs 10min.Whether perusal muddiness disappears, and is measured the change adding cosolvent rear surface tension force at normal temperatures and pressures by surface tension apparatus, chooses optimum cosolvent.
2, experimental result
The pH value situation of change of table 3 three groups of chitosan solutions
Table 4 three groups of cosolvents are on the impact of the surface tension value of chitosan lactic acid solution
Test shows, Tween 80, benzoic acid or PEG400 effectively can reduce the surface tension of chitosan lactic acid solution, and may be used to preparation the present invention and to spray membrane, wherein, the best results of Tween 80, therefore, preferred Tween 80 is as cosolvent.
Three, the present invention sprays the screening of proportioning of membrane
1, experimental technique
Experimental technique: when above-mentioned preferred n-octyl alcohol, Tween 80, fixing chitosan dosage, preparation different ratio Tween 80 and n-octyl alcohol series concentration sample, take surface tension as Testing index, commercial preparation is reference preparation, and screening optimum proportioning, specific experiment the results are shown in following table.
The adjuvant screening test of table 5 different auxiliary ratio
Before not adding cosolvent, the flocculent deposit of group 3-8 sample solution adularescent, liquid is muddy, variable concentrations n-octyl alcohol alcoholic solution is added in group 9-12 sample, liquid clarity is good, especially with group 12, and solution surface tension under group 12 proportioning is minimum, therefore the present invention sprays, the optimum proportioning of membrane is 0.15% n-octyl alcohol alcoholic solution+0.25% tween alcoholic solution+0.1% chitosan lactic acid solution.
The double blind experiment of experimental example 2 chitosan spraying of the present invention membrane film-formation result
(1) experiment material:
Chitosan spraying membrane (being prepared by embodiment 3) of the present invention, commercially available prod A (Jin Fusheng, Guangzhou Run Hong Pharmaceutical Technology Co., Ltd), commercially available prod B (gold wound chitosan wound care film, Hubei Pu Ai pharmaceutcal corporation, Ltd)
(2) experimental technique:
6 experimenters participate in double blind experiment, are sprayed on skin by three kinds of sprays, the evaluation index being film-formation result with mobility, drying property, film property.Wherein mobility standards of grading are: 1 point-be very easy to run off, 2 points-be easier to flowing, 3 points-seldom flowing, 4 points-do not flow completely; Drying property standards of grading are: 1 is divided into >7min drying time, 2 points of drying times are 5-7min, 3 points of drying times are 3-5min, 4 points drying time <3min; Film property standards of grading are: 1 point-imperceptible obvious film, 2 points-can be formed incomplete film, 3 points-can obviously complete film be formed.Cumulative all marks, and calculate the total score (± SD) of all experimenters, score value is higher then represents that film-formation result is better.
(3) experimental result:
Table 6 double blind experiment test result
Table 7 film-formation result appraisal result
As can be seen from film-formation result appraisal result, in double blind experiment, the moulding property of chitosan spraying membrane of the present invention is good, mobility is little, and its film-formation result is better than gold wound chitosan nursing film and Jin Fusheng chitosan anti-bacteria film forming spray.
Experimental result illustrates, the moulding property of chitosan spraying membrane of the present invention is good, mobility is little, is significantly better than existing commercially available prod, effectively can intercept antibacterial, promotes wound healing, and treatment skin trauma infects.
Embodiment 3 chitosan spraying of the present invention membrane film property surface tension, film formation time detect
(1) experiment material:
Chitosan spraying membrane (being prepared by embodiment 3) of the present invention, commercially available prod A (Jin Fusheng, Guangzhou Run Hong Pharmaceutical Technology Co., Ltd)
(2) experimental technique:
A. film formation time detection method: be 20-25 DEG C in temperature, under the condition of relative humidity 70%, about 0.2mL (pressing twice) medicinal liquid be sprayed on curtain coating on the glass plate of dried and clean from distance 5-10cm and pave, natural drying.
B. surface tension test experimental technique: measure the chitosan spraying membrane under the optimum proportioning condition described in embodiment 5 and commercially available prod A surface tension value, as the reference index weighing surfactant.Under normal temperature and pressure, water, chitosan spraying membrane and commercially available prod solution A are injected successively surface tension apparatus measurement ware middle circle connected in star and be about 1mL, record reading, repeats often to organize to average in triplicate.
(3) experimental result:
Table 8 film formation time testing result
Table 9 surface tension comparison and detection result
As can be seen from experimental result, chitosan spraying membrane is little than the surface tension of commercially available prod A, and film formation time is short, and rate of film build is fast.
Experimental result illustrates, the film formation time of chitosan spraying membrane of the present invention is short, and rate of film build is fast, is significantly better than existing commercially available prod, effectively can intercepts antibacterial, promotes wound healing, and treatment skin trauma infects.
Experimental example 4. chitosan spraying of the present invention membrane promotes the test of pesticide effectiveness of wound healing
(1) experiment material
Experiment reagent: chitosan spraying membrane (being prepared by embodiment 3); 10% chloral hydrate solution; Normal saline; Commercially available prod A (Jin Fusheng chitosan anti-bacteria film forming spray, Guangzhou Run Hong Pharmaceutical Technology Co., Ltd, 30mL/ bottle); Medical alcohol.
Laboratory animal: SD rat 30, (purchased from Qinglongshan animal reproduction field, Jiangning county, animal productiong credit number: SCXK (Soviet Union) 2010-0005), age in male and female half and half, 6-8 week.Test and buy for first 1 week, put temperature 25-27 DEG C, in the animal housing of sound insulation, sub-cage rearing, freely ingests, drinks water.
(2) experimental technique:
Model is set up: 30 rats, are divided into 3 groups at random, often organizes 10.Be divided into chitosan spray group, normal saline group, commercially available prod A group.Lumbar injection 10% chloral hydrate 0.7m, electricity consumption is pushed away and shaves off the hair that back is about 3cm × 3cm size, the region of marking pen labelling 2cm × 2cm is used after spraying 75% ethanol disinfection, back skin of unhairing is cut off with eye scissors, wound area is recorded after dry 1h, each group gives corresponding laboratory sample respectively, and medicinal liquid sprays (pressing twice) from distance 5-10cm, daily.
(3) Testing index: the wound situation of observing rat wound site every day, and 3,5,7,9,11,14,17,21 days after surgery dynamically observe Rat Wound Healing situation, measure wound surface area and calculate healing rate, healing rate=(original area-existing wound surface area)/original area, it is generally acknowledged that traumatic wounds dry tack free is not suppurated, blood crusts comes off naturally, and wound is that fair and tender normal skin shape is considered as healing.
(4) experimental result
Wound healing situation is shown in Fig. 2,3,4.
Wound area healing state table:
Table 10 chitosan spraying membrane group wound area
Table 11 normal saline group wound area
Table 12 commercially available prod A group wound area
Table 13 three groups of Rat Wound Healing rates
(note: compare with chitosan spray group, * is P < 0.05, * * is P < 0.01.)
From three groups of Rat Wound Healing rates: compared with normal saline, chitosan spraying membrane of the present invention all significantly improves at the Healing Rate of prometaphase, difference extremely remarkable (P < 0.01); Compared with the A of commercially available prod, chitosan spraying membrane of the present invention significantly improves at the Healing Rate in mid-term, significant difference (P < 0.05).
Experimental result illustrates, it is effective that chitosan spraying membrane wound healing of the present invention, treatment skin trauma infect, and is significantly better than existing commercially available prod.
Experimental example 5 chitosan spraying of the present invention membrane is tested the histopathological study of rat
(1) experiment material
Laboratory animal: get the rat of testing the 21st day in experimental example 4 at random and often organize each 3.
Experiment reagent: chitosan spraying membrane; 10% formalin; Normal saline; Commercially available prod A (Jin Fusheng chitosan anti-bacteria film forming spray, Guangzhou Run Hong Pharmaceutical Technology Co., Ltd, 30mL/ bottle); Medical alcohol;
(2) experimental technique: random execution each group of 3 rats, be separated wound local and surrounding skin tissue thereof, 10% formalin is fixed, and draws materials, dehydration, paraffin embedding.Cut into slices through HE (hematoxylin-eosin staining) dyeing, observation by light microscope is also marked.
(3) standards of grading:
A. epidermis cell is with or without degeneration necrosis, forms rotten to the corn, ulcer, crust: be chosen as 0 point-4 points.0 point, without downright bad, is 1 point-4 points from light to heavy; Downright bad length with " the micro-chi in microscope side " under 40 times of light microscopics (eyepiece 4 ×, object lens 10 ×) measure the length (cm/40 doubly) of ulcer;
B. below slough corium and subcutaneous tissue blood vessel with or without expansion, congested, edema, cell infiltration: be chosen as 0 point-4 points.0 point without obvious pathological changes, 1 point of-4 points of lesion degree are respectively slightly, moderate, severe, pole severe;
C. granulation tissue hyperplasia or fibrosis: be divided into 0-4 to divide, 0 point: ripe granulation tissue, completely fibrosis; 1 point: moderate is ripe, and in granulation tissue, vasculitis cell obviously reduces, the obvious hypertrophy of fibroblast; 2 points: slightly ripe, vasculitis Leukopenia in granulation tissue, fibroblast proliferation; 3 points: granulation tissue is in the early stage inmature stage, and fibrosis is not obvious; 4 points: without granulation tissue, without fibroblast proliferation;
D. around damaging, epithelial tissue is with or without hypertrophy: be divided into 0-3 to divide.0 point: damage local is entirely regeneration, WD epithelial tissue and covers; 1 point: local has epithelial tissue to regenerate covering, the non-differentiation and maturation of epithelium; 2 points: part has epithelial tissue to regenerate; 3 points: without epithelium regeneration.
(4) experimental result
Pathology section examination: see Fig. 5,6,7.
Normal saline group: in 3 skin histologies, there is ulcer on 2 surfaces, and another 1 original damage location is covered by complete epithelium group, and No. 2, No. 7 lesion degrees are similar, rat local epidermis, corium holostrome disappearance, form ulcer, length is respectively 0.7cm or 1.9m/x40, and there is the inflammatory necrosis tissue of volume on surface, the granulation tissue of its lower visible hyperplasia, be made up of the blood capillary of new life and fibroblast, inside have cell infiltration, inflammatory cell type is mainly mononuclear phagocyte and granulocyte.No. 5 injured surfaces are covered by intact skin, keratinization that epidermis is slight, and corium is without adnexa, and by granulation tissue filling, in granulation tissue, blood vessel and inflammatory cell quantity reduce, fibroblast proliferation.
Commercially available prod A group: lesion degree comparatively normal saline group alleviates, No. 6, No. 8 lesion degrees are close, rat local epidermis, corium holostrome lack formation ulcer, length is respectively 1.0cm or 1.8cm/x40, there is inflammatory necrosis tissue on surface, the granulation tissue of its lower visible hyperplasia, is made up of the blood capillary of new life and fibroblast, inside have cell infiltration, inflammatory cell type is the same.No. 3 injured surfaces are covered by intact skin, keratinization that epidermis is slight, and corium is without adnexa, and by granulation tissue filling, in granulation tissue, blood vessel and inflammatory cell quantity reduce, fibroblast proliferation.
Chitosan spraying membrane group: the damage of this group is the lightest, No. 4 rat local epidermises, corium holostrome disappearances, form ulcer, length: 2.9cm/x40, surface is inflammatory necrosis tissue, the granulation tissue of its lower visible hyperplasia, is made up of the blood capillary of new life and fibroblast, inside have cell infiltration, inflammatory cell type is the same.No. 1 injured surface is covered by intact skin, and there is slight keratinization on surface, and corium is without adnexa, and by granulation tissue filling, in granulation tissue, blood vessel and inflammatory cell quantity reduce, and fibroblast is many.No. 9 lesion degrees are lighter than No. 1, substantially without inflammatory cell bottom granulation ulcer, without blood vessel, are filled by the collagen fiber of maturation.
Lesion score: in Table 14-17.
Table 14 normal saline group pathological score table
Table 15 commercially available prod A group pathological score table
Table 16 chitosan spraying membrane group pathological score table
Table 17 rat skin pathological changes light and heavy degree appraisal result
(5) experiment conclusion
Can find out, compared with the A group of commercially available prod, the healing of chitosan spraying membrane of the present invention is better: commercially available prod A group local epidermis, corium holostrome disappearance, form ulcer, skin surface is coated with the inflammatory necrosis tissue of volume, its lower visible granulation tissue hyperplasia.Granulation tissue is made up of the blood capillary of new life and fibroblast, inside has cell infiltration.Damage topical ulcers in chitosan spraying membrane group of the present invention to repair, the epithelium that surface coverage is complete, upper subcutaneous granulation tissue is more ripe, and fibrous tissue increases, and blood vessel number and inflammatory cell quantity reduce.Experimental result illustrates, chitosan of the present invention spraying membrane can wound healing, treatment skin trauma infect, and Be very effective is better than existing commercially available prod.
Experimental example 6 chitosan spraying of the present invention membrane is to rabbit vagina mucous membrane irritation test
(1) experiment material
Experiment reagent: chitosan spraying membrane (being prepared by embodiment 3), normal saline, medical alcohol.
Laboratory animal: first Adult female unpregnancy New Zealand experimental rabbit 6 is (purchased from Qinglongshan animal reproduction field, Jiangning county, animal productiong credit number: SCXK (Soviet Union) 2010-0022), body weight 2.6-3.0kg, animal is all in estrus, vaginal orifice without secretions, without congestion and edema and other damages.Buy the last week in experiment, raise in regular grade Animal House, temperature 20-26 DEG C, relative humidity 40-70%, noise < 60 decibels, drinking-water of freely ingesting.
(2) experimental technique
Model is set up: laboratory animal is divided at random chitosan spraying membrane group and saline control group, often organizes 3.Connect syringe with homemade flexible pipe (flexible pipe head is mellow and full without corner angle), inhaling in syringe has sample liquid, and flexible pipe is gently inserted vagina 4-5cm, injects sample liquid, and slowly extracts flexible pipe out.Successive administration 10 days.In 24h after process in 10th day, put to death laboratory animal, the complete vagina of taking-up of cutting open the belly is placed in 10% formalin internal fixtion, fix fully, draw materials respectively in vagina epimere, stage casing and hypomere, routine paraffin wax embeds, HE (hematoxylin-eosin) dyes, om observation vaginal mucosal epithelium cell is with or without degeneration, necrosis, with or without cell infiltration, upper subcutaneous with or without hyperemia, edema and cell infiltration.
Standards of grading: according to each site morbidity degree from light to heavy, be quantitatively slight or minute quantity " 0.5 point " successively, slight or a small amount of " 1 point ", moderate or more " 2 points ", severe or volume " 3 points ", pole severe or a large amount of " 4 points ", normal " 0 point " cumulative all marks, and calculate and often organize dividing equally of every animal
score value higher expression degree of injury is more serious.
(3) experimental result
Pathology section examination: see Fig. 8, Fig. 9.
Saline control group pathology diagosis: under light microscopic, visible rabbit vagina epimere and stage casing mucosa are coated to simple columnar epithelium, and lamina propria is the connective tissue being rich in blood vessel, and deep is connective tissue and smooth muscle.Hypomere mucous epithelium layer thickens, and part is stratified squamous epithelium.Concrete pathological changes is as follows:
Epimere: 3 epimere mucomembranous epithelial cells are without obvious degeneration, necrosis, and lamina propria mild hyperaemia, and see minute quantity or a small amount of (slight or slight) cell infiltration, inflammatory cell type is mainly neutrophilic granulocyte and mononuclear phagocyte.Other has no obvious pathological changes., there is the cell infiltration of a small amount of, minute quantity or the same type of moderate in stage casing: 3 slight degeneration of mucomembranous epithelial cell; Proper mucous membrane moderate or mild hyperaemia, and see minute quantity (slightly) cell infiltration, inflammatory cell type is the same.Hypomere: lesion degree overweights epimere and 3, stage casing mucomembranous epithelial cell is slight or moderate degeneration, has cell infiltration that is a small amount of or the same type of moderate; Proper mucous membrane moderate or mild hyperaemia, edema, and the cell infiltration of seeing moderate or slight the same type.
Chitosan spraying membrane group pathology diagosis: lesion degree is lighter than saline control group, specific as follows:
Epimere: slightly or slightly congested, other has no obvious pathological changes for 3 epimere proper mucous membranes.Stage casing: 3 slight degeneration of mucomembranous epithelial cell, the wherein cell infiltration of 1 the same type of degree of taking a favourable turn.Proper mucous membrane moderate, slight or slight congested, 1 slight cell infiltration of companion.Hypomere: 3 slight degeneration of mucomembranous epithelial cell, companion is slight or the cell infiltration of slight the same type.3 proper mucous membrane moderates or mild hyperaemia, the slight cell infiltration of companion.
Table 18 chitosan spray group rabbit vagina section pathological score table
Note 1: numeral shows lesion degree: 0.5 point (slight or minute quantity), 1 point (slight or a small amount of), 2 points (moderate or moderate), 3 points (severe or volume), 4 points (pole severe or a large amount of)
Table 19 normal saline group rabbit vagina section pathological score table
Table 20 rabbit vaginosis becomes light and heavy degree appraisal result
From experimental result, degeneration that negative control group vaginal mucosal epithelium cell is slight, there is cell infiltration.Lamina propria is congested, the cell infiltration of edema companion same-type.The most obvious with hypomere pathological changes in 3 sections, upper, middle and lower.All comparatively light with the above-mentioned lesion degree of vaginal mucosa after chitosan spray doses, illustrate that chitosan of the present invention spraying membrane is very little to the stimulation of vaginal mucosa.Compared to similar-type products employing acetic acid is as solvent on the market at present, the present invention uses endogenous material lactic acid instead as solvent, reduces skin irritation, even better than the safety of normal saline.
Experimental result illustrates, the safety of chitosan spraying membrane of the present invention is good, compares normal saline also little to skin irritation.
Experimental example 7 chitosan spraying of the present invention membrane bacteriostatic test
(1) experiment material:
Chitosan spraying membrane (prepared by embodiment 3, lot number is 3,8,14,18,19,24,29,32) of the present invention; Normal saline; Culture medium: nutrient broth medium (Beijing three medicine scientific and technological development company) is by formulated, and regulate pH6.5, add 1.2% agar, after autoclaving, 45 DEG C of constant temperature water baths are for subsequent use.Experimental strain: selection standard strain Escherichia coli ATCC25922, staphylococcus aureus ATCC6538, Candida albicans ATCC10231, is stored in-80 DEG C.Collecting cells: get the fresh slant strains of each test strain, wash lower lawn with sterile saline, with Maxwell opacity tube than turbid, being suitably diluted to concentration, to be about 1-9 × 106cfumL-1 for subsequent use.
(2) experimental technique:
Bacteriostasis rate measures: get each sample 5mL respectively and directly add sterile test tube, prepare 3 parts: in vitro add a kind of experimental bacteria suspension 0.5mL respectively at each, bacterial concentration is made to be about 1-9 × 105cfumL-1, DL instrument mixes, in 25 DEG C place 0,2,10, get 0.5mL after 20min and add normal saline and suitably dilute rear plate method counting.Be measured in the same method in contrast with normal saline simultaneously.Calculate bacteriostasis rate: bacteriostasis rate %=(A-B)/A × 100%A: normal saline negative control count results B: sample count result
(3) experimental result:
Table 21 escherichia coli bacteriostasis rate measurement result
Table 22 staphylococcus aureus bacteriostasis rate measurement result
Table 23 Candida albicans bacteriostasis rate measurement result
(4) experiment conclusion
According to evaluation criterion in " C4: stripping property antimicrobial product bacteriostasis property test method " in " Disposable Sanitary Accessory sanitary standard (GB15979-2002) ", chitosan spraying membrane all shows stronger bacteriostasis to escherichia coli, staphylococcus aureus and Candida albicans, wherein more than 99% is reached to colibacillary suppression ratio, 100% is reached to the suppression ratio of staphylococcus aureus.
Experimental result explanation, chitosan spraying membrane bacteriostasis of the present invention is strong, particularly to clinical common pathogenic bacteria: escherichia coli, staphylococcus aureus and Candida albicans have clear and definite inhibitory action, can treat the infectious disease caused by them, treatment skin trauma infects.
Claims (10)
1. a chitosan spraying membrane, is characterized in that: it is prepared from by the component of following weight percent:
Chitosan 0.05-0.5%, lactic acid 0.01%-0.15%, surfactant 0.1%-0.2%, cosolvent 0.1%-0.3%, surplus is water.
2. spraying membrane according to claim 1, is characterized in that: described chitosan molecule amount is 10w-30w.
3. spraying membrane according to claim 2, is characterized in that: described chitosan molecule amount is 30w.
4. spraying membrane according to claim 1, is characterized in that: described surfactant is n-octyl alcohol, water-soluble silicon oil or dimethicone; Described cosolvent is Tween 80, benzoic acid or PEG400.
5. spraying membrane according to claim 4, is characterized in that: it is prepared from by the component of following weight percent:
Chitosan 0.1%, lactic acid 0.05%, n-octyl alcohol 0.15%, Tween 80 0.25%, surplus are water.
6. prepare the method for the spraying membrane described in claim 1-5 any one, comprise the steps:
A. each component is taken according to proportioning described in claim 1-5 any one;
B. extracting lactic acid, is prepared into the lactic acid aqueous solution that concentration is 0.1-0.14%;
C. get chitosan, be dispersed in lactic acid solution prepared by b step, swelling, after stirring and dissolving, adjust ph, to 3.5-6.0, is filtered, is obtained chitosan lactic acid solution;
D. cosolvent and surfactant are dissolved in ethanol solution respectively, make the alcoholic solution of cosolvent and the alcoholic solution of surfactant respectively;
E. stir the chitosan lactic acid solution prepared of step c, add the alcoholic solution of the cosolvent that step c obtains, stir to clarify, then add the alcoholic solution of the surfactant that step c obtains, stir to clarify, fill and get final product.
7. preparation method according to claim 6, is characterized in that: in described lactic acid aqueous solution, and lactic acid concn is 0.12%.
8. preparation method according to claim 6, is characterized in that: the concentration of the alcoholic solution of described cosolvent and the alcoholic solution of surfactant is 5%.
9. the purposes of the spraying membrane described in claim 1-5 any one in the medicine preparing the infection for the treatment of skin trauma and/or wound healing.
10. purposes according to claim 9, is characterized in that: described medicine is the medicine of Chinese People's Anti-Japanese Military and Political College enterobacteria or staphylococcus aureus.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108079121A (en) * | 2017-12-19 | 2018-05-29 | 高杰 | A kind of chitosan Pterospermi Heterophylli spray promoting blood circulation and removing blood stasis and preparation method thereof |
| CN108498531A (en) * | 2018-06-01 | 2018-09-07 | 广东药科大学 | A kind of chitosan oral solution and preparation method thereof |
| WO2020084416A1 (en) * | 2018-10-27 | 2020-04-30 | Shilpa Medicare Limited | Spray compositions of chitosan for wound healing |
| CN111718687A (en) * | 2019-12-10 | 2020-09-29 | 杭州惠康医疗器械有限公司 | Medical mirror surface anti-fog liquid and preparation method thereof, medical mirror surface anti-fog film and preparation method thereof, anti-fog method and medical instrument |
| CN112438964A (en) * | 2020-11-30 | 2021-03-05 | 威兰德(山东)生物科技有限公司 | Composite antibacterial film spraying agent of lysostaphin and chitosan |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101278948A (en) * | 2008-05-23 | 2008-10-08 | 大连大学 | Biological medical membrane and method of preparing the same |
| CN102670929A (en) * | 2012-06-01 | 2012-09-19 | 山东卫康生物医药科技有限公司 | Composite for injury healing and preparing method thereof |
| CN102824308A (en) * | 2012-08-31 | 2012-12-19 | 广州润虹医药科技有限公司 | Chitosan antibacterial filming sprayer and preparation method |
-
2015
- 2015-05-28 CN CN201510282459.XA patent/CN104856978B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101278948A (en) * | 2008-05-23 | 2008-10-08 | 大连大学 | Biological medical membrane and method of preparing the same |
| CN102670929A (en) * | 2012-06-01 | 2012-09-19 | 山东卫康生物医药科技有限公司 | Composite for injury healing and preparing method thereof |
| CN102824308A (en) * | 2012-08-31 | 2012-12-19 | 广州润虹医药科技有限公司 | Chitosan antibacterial filming sprayer and preparation method |
Non-Patent Citations (2)
| Title |
|---|
| 肖锦等: "《天然高分子絮凝剂》", 30 September 2005, 化学工业出版社 * |
| 黎厚斌: "《包装应用化学》", 31 January 2014, 印刷工业出版社 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108079121A (en) * | 2017-12-19 | 2018-05-29 | 高杰 | A kind of chitosan Pterospermi Heterophylli spray promoting blood circulation and removing blood stasis and preparation method thereof |
| CN108498531A (en) * | 2018-06-01 | 2018-09-07 | 广东药科大学 | A kind of chitosan oral solution and preparation method thereof |
| WO2019227746A1 (en) * | 2018-06-01 | 2019-12-05 | 广东药科大学 | Chitosan oral solution and preparation method therefor |
| WO2020084416A1 (en) * | 2018-10-27 | 2020-04-30 | Shilpa Medicare Limited | Spray compositions of chitosan for wound healing |
| CN112955156A (en) * | 2018-10-27 | 2021-06-11 | 希尔帕医疗保健有限公司 | Chitosan spray composition for wound healing |
| CN111718687A (en) * | 2019-12-10 | 2020-09-29 | 杭州惠康医疗器械有限公司 | Medical mirror surface anti-fog liquid and preparation method thereof, medical mirror surface anti-fog film and preparation method thereof, anti-fog method and medical instrument |
| CN111718687B (en) * | 2019-12-10 | 2023-04-25 | 杭州惠康医疗器械有限公司 | Medical mirror anti-fog liquid and preparation method thereof, medical mirror anti-fog film and preparation method thereof, anti-fog method and medical instrument |
| CN112438964A (en) * | 2020-11-30 | 2021-03-05 | 威兰德(山东)生物科技有限公司 | Composite antibacterial film spraying agent of lysostaphin and chitosan |
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