CN102108138B - Method for preparing chitosan collagen gel - Google Patents
Method for preparing chitosan collagen gel Download PDFInfo
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- CN102108138B CN102108138B CN2009102488220A CN200910248822A CN102108138B CN 102108138 B CN102108138 B CN 102108138B CN 2009102488220 A CN2009102488220 A CN 2009102488220A CN 200910248822 A CN200910248822 A CN 200910248822A CN 102108138 B CN102108138 B CN 102108138B
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Abstract
The invention discloses a method for preparing chitosan collagen gel, which comprises 5 to 15 percent of chitosan, 1 to 10 percent of collagen, 0.1 to 1 percent of gel substrate and the balance of water, wherein the chitosan is water-soluble and has a molecular weight of 3,000 to 400,000 Daltons; the molecular weight of the collagen is 5,000 to 3 million Daltons; and the gel substrate is one or two of Carbomer, polyethylenes, and cellulose derivatives. The chitosan collagen gel is used for stopping bleeding on acute or chronic wound faces, absorbing seepage, cleaning wounds in an autolytic manner, protecting exposed nerve endings, relieving pains, protecting wet environments of the wounds, accelerating the healing of the wounds and inhibiting the generation of scar.
Description
Technical field
The invention belongs to the biomaterial for medical purpose field, be specifically related to a kind of preparation method of chitosan collagen protein gelatin.
Background technology
Chitosan is a kind of natural polymers.Be the unique positively charged tunicin that occurring in nature is found up to now, undersaturated cation group is arranged in the molecule, electronegative all kinds of objectionable impuritiess such as bacterium, fungi and virus etc. are had powerful adsorption; Like streptococcus aureus; Intestinal bacteria, Candida albicans etc., its mechanism is amino positively charged; Can with bacterial adhesion together, the metabolism of interfere with bacterial outer wall; The positive charge on chitosan surface reduces negative charge with the acceptor interaction of erythrocyte surface amino-acid residue, through the hemostasis, the analgesic activity that red corpuscle obviously and are directly sticked and the congregation realization is good; The migration of leukin and scavenger cell in can promoting promotes wound healing; Through promoting the generation of granulation tissue and epithelium, reduce the contraction of wound in the wound, play the effect that suppresses scar; Because its Degradation can be absorbed by human body, thereby eliminate bleeding and pain when removing, and also can not delay the healing of wound in addition because staying chip, and can not produce the strong impulse effect to tissue on every side.
Collagen protein is a kind of biological polymer substance; Collagen has important biological property-mechanical property height, promotes cell growth, hemostasis, biocompatibility and biological degradability, can combine closely with wound, infiltrates in the middle of the cambium; Support when growing as cell; Can stop blooding rapidly, the promotion granulation grows, prevents hemorrhage, promotes wound healing.
The present invention adopts the gel formulation, smears with its formulation characteristic and protects wound, can keep wound moistening, creates the environment of hypoxemia, little acid, accelerating wound healing, and can absorb sepage, from the dissolubility debridement, protection NE tip eases the pain.
Summary of the invention
The preparation method who the purpose of this invention is to provide a kind of chitosan collagen protein gelatin.
The invention provides a kind of chitosan collagen protein gelatin, this gel comprises chitosan 1-15%, collagen protein 1-20%, and gel matrix 0.1-1%, all the other are water.
Chitosan collagen protein gelatin provided by the invention, said chitosan is a water-soluble chitosan, its molecular weight is 3000 dalton～400,000 dalton; The molecular weight of said collagen protein is 5000 dalton～3,000,000 dalton; Said gel matrix is one or both in carbomer, polyethylene kind, the derivatived cellulose.
The present invention also provides the preparation method of chitosan collagen protein gelatin, and step is following: using dissolved in distilled water to become mass percent water-soluble chitosan is 1～15% solution; Using dissolved in distilled water to become mass percent collagen protein is 01～20% solution; Gel matrix become 0.2～3% solution with dissolved in distilled water; Above-mentioned three kinds of solution are mixed by a certain percentage, and the mass percent that makes chitosan is 0.5～15%, and the mass percent of collagen protein is 1～10%, and the mass percent of gel matrix is 0.1～1%; Transfer pH value to 5～8, stirring makes it all promptly get gelifying agent; The gelifying agent deaeration is got final product.
The preparation method of chitosan collagen protein gelatin provided by the invention, the dissolving of said gel matrix is under agitation carried out, and stirring velocity is 100-1000r/m, and churning time is 1-10 hour.
The preparation method of chitosan collagen protein gelatin provided by the invention, said deaeration mode adopts a kind of in supercentrifuge, the vacuum emulsification; Wherein during supercentrifuge, centrifugal rotational speed is 1000-5000r/m, and centrifugation time is 5-30 minute; During vacuum emulsification, stirring velocity is 20-100r/m, and churning time is 5-300 minute, and vacuum tightness is 0.08-0.6Mpa.
Chitosan collagen protein gelatin provided by the invention is applied to the hemostasis of the acute and chronic surface of a wound, absorbs sepage, and from the dissolubility debridement, protection NE tip eases the pain, and keeps the moist environment of wound simultaneously, and accelerating wound healing suppresses scar and generates.
Characteristics of the present invention: it is residual that chitosan collagen protein gelatin can improve the sponge foam chip that collagen sponge, diaphragm etc. cause; Absorb shortcomings such as not thorough, demoulding difficulty, increase scar; Chitosan and collagen protein are used jointly; Bring into play the effect of two kinds of biomaterials simultaneously, avoid the drawback of single therapy, to the surface of a wound airtight, moistening healing environment is provided simultaneously.Water-soluble chitosan has also further improved the restriction of chitosan on production technique, has avoided the use of pungency solvent, prevents that pungent is residual.
Embodiment
Following embodiment will further explain the present invention, but therefore not limit the present invention.
Embodiment 1
The preparation molecular weight is the 3000 daltonian water-soluble chitosan aqueous solution, and making its mass percent is 10%.The preparation mass percent is 5% Collagen Hydrolysate solution.Two solution were mixed by 1: 1.Add mass percent and be 0.3% carbomer, transfer PH to 6.5, stir gel.The centrifugal 10-20 of whizzer 3000-4000r/m minute.
Embodiment 2
The preparation molecular weight is the 10000 daltonian water-soluble chitosan aqueous solution, and making its mass percent is 10%.The preparation mass percent is 5% Collagen Hydrolysate solution.Two solution were mixed by 1: 2.Add mass percent and be 0.3% carbomer, transfer PH to 6.5, stir gel.The vacuum emulsification deaeration gets final product.Rotating speed 50r/min stirred 60 minutes.
The experiment of embodiment 3 gel dressing trauma repair effects
1. material:
The dressing of chitosan collagen protein gelatin, sterile gauze
2. animal:
Healthy guinea pig, body weight 200 grams
3. trauma repair effect test:
Get four of cavys, be divided into A group and organize two groups with B, 2 every group, with the guinea pig back cropping, alcohol disinfecting, the holostrome skin of excision 2 * 2cm at the back, the A group with gel dressing be applied on the surface of a wound, sterile gauze wraps up; The B group is directly wrapped up the surface of a wound with sterile gauze.Observed the reparation situation of gel dressing in 1 day, 3 days, 5 days, 7 days, 14 days in postoperative, record wound healing situation (seeing table 1) to the surface of a wound.
Table 1 wound healing situation
Postoperative was formed mouthful all healings, no swelling phenomenon in 14 days two.A group skin color is normal, and slight scar is arranged.B group skin color is light rubescent, and big scar is arranged.
4. result and discussion:
A forms mouthful healing state and obviously is better than the B group, and A group sample (gel dressing) has the promotion wound healing, reduces the effect of scar.
The test of embodiment 4 gel dressing haemostatic effects
1. material:
The dressing of chitosan collagen protein gelatin, sterile gauze (control group)
2. animal:
Healthy rabbits, body weight 2.0kg～2.5kg.
3. rabbit ear wound hemostasis test:
Get 8 of rabbit, be divided into two groups (A group and B groups) at random, 4 every group, left and right sides ear is labeled as the A group respectively: A1, A2, B group: B1, B2.With the alignment of rabbit left and right sides ear, in ear edge 1/3rd places, the long wound of the about 1cm of crosscut, wound is from ear edge crosscut rabbit ear, and all ears are done same treatment of wounds.A group: smear the dressing of chitosan collagen protein gelatin in 5 seconds, cover sterile gauze and pushed one minute; The B group was pushed one minute with gauze.Begin to observe the hemorrhage situation of wound in one minute later on, and the oozing of blood that picks up counting no longer, hemorrhagely be bleeding stopping period, record result's (seeing table 2).
The bleeding stopping period of table 2 wound
| Group | The animal number of elements | Bleeding stopping period (branch) |
| The A group | 4 | 2.10±0.10 |
| The B group | 4 | 3.35±0.10 |
4. result and discussion:
The result shows that A group, B group all have anastalsis in various degree, and wherein A group effect obviously is better than the B group.Show that the dressing of chitosan collagen protein gelatin has anthemorrhagic performance preferably.
Embodiment 5 water-soluble chitosan extracorporeal bacteria inhibitor tests
This experiment has been carried out bacteriostatic test to water-soluble chitosan.Chitosan is made into 5% the aqueous solution, is diluted to 2.5%, 1%, 0.5%, 0.25% with sterile distilled water then, do blank with sterile distilled water simultaneously, intestinal bacteria, streptococcus aureus are done bacteriostatic test with the agar diffusion paper disk method.The result shows that chitosan all has restraining effect to intestinal bacteria, streptococcus aureus, and the chitosan aqueous solution of different concns is different to the inhibition strength of bacterial classification.
Experiment material and method
1. water-soluble chitosan.
2. test strain
Intestinal bacteria, streptococcus aureus.
3. substratum
Beef-protein medium.
4. the preparation of water-soluble chitosan solution
The 5g molecular weight of getting sterilization is that 10,000 daltonian water-soluble chitosans are dissolved in the 100ml water, processes 5% chitosan aqueous solution, again with sterile distilled water be diluted to 2.5%, 1%, more than 0.5% kind of concentration.Prepare sterile distilled water simultaneously and do controlled trial.
5. preparation strain liquid
Have a try in following various confessions of aseptic technique and to test bacterial classification and carry out the strain inclined plane activation respectively with suitable solid slant culture base.Every kind of bacterial classification difference picking one ring respectively is mixed with the bacteria suspension that contains the about 106/ml of viable count with SPSS then, and is for use.
6. the preparation of filter paper
Select bibulous high-quality filter paper, use punch tool to break into the circular filter paper sheet of some diameters as 6mm, subsequent use behind dry sterilization.
7. antibacterial testing method (antibacterial around-France)
Respectively various bacteria suspensions are evenly spread upon substratum plate surface; The scraps of paper with aseptic nipper difference gripping sterilization; Fully dip in the chitosan solution of getting various concentration and be attached to various containing on the bacterium plate, do blank to dip in the sterile distilled water scraps of paper of getting respective concentration.Then plate is put into 37 ℃ of incubators and cultivated 24 hours, take out the size that each inhibition zone diameter is measured in the back.
8. result and analysis
The chitosan of table 3 different concns is to the bacteriostatic diameter (mm) of various confession examination bacterium
Experimental result shows that chitosan all has certain restraining effect to intestinal bacteria, streptococcus aureus.Chitosan concentration is can play certain bacteriostatic action at 0.25% o'clock.The restraining effect of the chitosan of different concns is different, considers cost and formulation needs, and the present invention takes 1%～15% concentration chitosan; In antibacterial useful range, reduce working concentration as far as possible, and guarantee the formation of gel formulation; At present; A large amount of research shows that the fungistatic effect of water-soluble chitosan is better, and molecular weight is also influential to the chitosan bacteriostasis property, descends gradually with molecular weight rising fungistatic effect.Particularly to intestinal bacteria, the chitosan molecule amount is more little, and bacteriostatic action more obviously.With regard to the α-chitosan of several kinds of different molecular weights the inhibition research of several frequently seen bacterium (intestinal bacteria, streptococcus aureus, Bacillus subtilus, Clostridium perfringens) is shown that the fungistatic effect of low-molecular-weight α-chitosan is superior to high-molecular weight α-chitosan among the Song Xianzhou etc. " bacteriostatic action of the α-chitosan of different molecular-weight average ".Summer Wenshuis etc. adopt E.coli as test strain, and it is the strongest to record molecular weight and be 1500 chitooligosaccharide-fungistatic effect.The corresponding adjusting of chitosan on concentration and molecular weight in the present invention prescription, the fungistatic effect of assurance product reaches the purpose of clinical diagnosis and treatment.
Comparative example 1 clinical relatively chitosan collagen protein gelatin dressing and aquagel trauma repair effect
1. material
The dressing of chitosan collagen protein gelatin: molecular weight 3000 daltonian water-soluble chitosans 1%, molecular weight 5000 daltonian collagen proteins 12%, Acritamer 940 0.6%, all the other are water.
Aquagel dressing: chitosan 5%, 0.7% glacial acetic acid solution dissolving.
2. postoperative patient 8 people are divided into two groups of A, B, every group 4 people.
3. the trauma repair effect relatively
(1) to after the wound disinfection, by aseptic technique the dressing of chitosan collagen protein gelatin is applied on the A group patient surface of a wound, the sterile gauze wrapping is changed the next day of according to circumstances.Aquagel dressing is applied on the B group patient surface of a wound after the processing equally by aseptic technique, the sterile gauze wrapping, the next day, changed.Observed the reparation situation of two kinds of dressing in 1 day, 3 days, 7 days, 14 days in postoperative, record wound healing situation to the surface of a wound.
(2) observations:
A group: postoperative obviously rubescent, the obvious swelling of skin color in 1 day, obviously pain, slight oozing of blood, hinder not bonding, the obvious infection of edge; Postoperative slightly rubescent, the slight swelling of skin color in 3 days, mild pain, no oozing of blood, hinder bonding, the slight infection of edge part; Postoperative slightly rubescent, the no swelling of skin color in 5 days, mild pain, no oozing of blood, hinder the edge major part bonding, do not have to infect; 7 days skin colors of postoperative are normal, do not have swelling, do not have pain, do not have oozing of blood, hinder edge bonding, do not have and infect.
B group: postoperative obviously rubescent, the obvious swelling of skin color in 1 day, obviously pain, slight oozing of blood, hinder not bonding, the obvious infection of edge; Postoperative obviously rubescent, the slight swelling of skin color in 3 days, mild pain, no oozing of blood, hinder not bonding, the slight infection of edge; Postoperative slightly rubescent, the slight swelling of skin color in 5 days, mild pain, no oozing of blood, hinder not bonding, the slight infection of edge; 7 days skin colors of postoperative are slightly rubescent, do not have swelling, do not have pain, do not have oozing of blood, hinder the edge part bonding, do not have and infect.
In sum, the dressing of chitosan collagen protein gelatin is compared with aquagel dressing, on prescription, selects for use water-soluble chitosan to replace the solubility in acid chitosan, and has increased collagen protein.Water-soluble chitosan forms through special process is refining, has improved the solubility property of chitosan, has kept the primary characteristic of chitosan, has prevented to stimulate the use of solvent, avoids surface of a wound acidity residual.Collagen protein has characteristics such as nonantigenic, biological degradability, bio-compatibility, bioresorbable, in the stage of wound healing, is in skin tissue recovering and regrouping process, matter between the important cells of performance assist in functions.Collagen protein can slow down the infectation of bacteria phenomenon, reduce wound suppurates, increases the wound of the formation of granulation tissue, healing and can not produce the common use of contraction and chitosan and better quickened wound healing.
Claims (7)
1. chitosan collagen protein gelatin, it is characterized in that: this gel comprises chitosan 0.5-15%, collagen protein 1-10%, gel matrix 0.1-1%, all the other are water;
Said chitosan is a water-soluble chitosan;
Said gel matrix is a carbomer.
2. according to the said chitosan collagen of claim 1 protein gelatin, it is characterized in that: the molecular weight of said collagen protein is 5000 dalton～3,000,000 dalton.
3. according to the said chitosan collagen of claim 1 protein gelatin, it is characterized in that: the molecular weight of said chitosan is 3000 dalton～400,000 dalton.
4. the preparation method of the said chitosan collagen of claim 1 protein gelatin, it is characterized in that: step is following:
(1) using dissolved in distilled water to become mass percent water-soluble chitosan is 1～15% solution;
(2) using dissolved in distilled water to become mass percent collagen protein is 1～20% solution;
(3) gel matrix is become 0.2～3% solution with dissolved in distilled water;
(4) above-mentioned three kinds of solution are mixed by a certain percentage, the mass percent that makes chitosan is 0.5～15%, and the mass percent of collagen protein is 1～10%, and the mass percent of gel matrix is 0.1～1%;
(5) adjust pH to 5～8, stirring makes it all promptly get gelifying agent;
(6) the gelifying agent deaeration is got final product.
5. according to the preparation method of the said chitosan collagen of claim 4 protein gelatin, it is characterized in that: the dissolving of the gel matrix in the said step (3), under agitation to carry out, stirring velocity is 100-1000r/m, churning time is 1-10 hour.
6. according to the preparation method of the said chitosan collagen of claim 4 protein gelatin, it is characterized in that: the deaeration mode in the said step (6), adopt a kind of in supercentrifuge, the vacuum emulsification.
7. according to the preparation method of the said chitosan collagen of claim 6 protein gelatin, it is characterized in that: said supercentrifuge, centrifugal rotational speed are 1000-5000r/m, and centrifugation time is 5-30 minute; Said vacuum emulsification, stirring velocity are 20-100r/m, and churning time is 5-300 minute, and vacuum tightness is 0.08-0.6Mpa.
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| CN2009102488220A CN102108138B (en) | 2009-12-28 | 2009-12-28 | Method for preparing chitosan collagen gel |
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| CN2009102488220A CN102108138B (en) | 2009-12-28 | 2009-12-28 | Method for preparing chitosan collagen gel |
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| CN102108138B true CN102108138B (en) | 2012-07-25 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102526799A (en) * | 2011-12-31 | 2012-07-04 | 杭州市第一人民医院 | Chitosan collagen membrane for guiding regeneration of peripheral bones of dental implant |
| CN106334209A (en) * | 2015-07-08 | 2017-01-18 | 北京师范大学 | Polydopamine modified chitosan hemostatic dressing |
| CN107441479B (en) * | 2017-08-08 | 2018-12-21 | 广东海洋大学 | Marine active peptide/chitin burn ointment for treating scald and preparation method thereof |
| CN114796160A (en) * | 2021-01-27 | 2022-07-29 | 深圳市百吉因生物科技有限公司 | Antibacterial material and application thereof |
| US11806430B1 (en) | 2023-02-24 | 2023-11-07 | King Faisal University | Rectal gel with date palm phenolics and vanillic acid |
| CN116173291A (en) * | 2023-03-08 | 2023-05-30 | 重庆医科大学附属第二医院 | Chitosan recombinant humanized collagen gel and preparation method and application thereof |
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| CN1380109A (en) * | 2002-04-17 | 2002-11-20 | 苗九昌 | Chitosan collagen and calcium alginate compounded spongy biological dressing and its preparation process |
| CN1526764A (en) * | 2003-09-24 | 2004-09-08 | 山东大学 | Composite collagen-base rack material and its pepn and use |
| CN1554448A (en) * | 2003-12-23 | 2004-12-15 | 张淑荣 | Chitosan gelatine polyvinyl alcohol biological hemostatic dressing |
| CN101260191A (en) * | 2008-04-01 | 2008-09-10 | 武汉大学 | Temperature sensitive type chitosan/glutin hydrogel and its preparation method and use |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1380109A (en) * | 2002-04-17 | 2002-11-20 | 苗九昌 | Chitosan collagen and calcium alginate compounded spongy biological dressing and its preparation process |
| CN1526764A (en) * | 2003-09-24 | 2004-09-08 | 山东大学 | Composite collagen-base rack material and its pepn and use |
| CN1554448A (en) * | 2003-12-23 | 2004-12-15 | 张淑荣 | Chitosan gelatine polyvinyl alcohol biological hemostatic dressing |
| CN101260191A (en) * | 2008-04-01 | 2008-09-10 | 武汉大学 | Temperature sensitive type chitosan/glutin hydrogel and its preparation method and use |
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Effective date of registration: 20160715 Address after: 110000 No. 2, Chaoyang Industrial Park, Shenyang Economic Development Zone, Liaoning 4, China Patentee after: Shenyang Xing Xing Biological Technology Co., Ltd. Address before: 110179 Liaoning province Shenyang Hunnan New Long Street No. 203 room 10-1 Patentee before: Shenyang Xinsheng Biology Tachnology Co.,Ltd. |
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