CN101869727B - Reinforced anti-adhesion composite gel, preparation method and application - Google Patents
Reinforced anti-adhesion composite gel, preparation method and application Download PDFInfo
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Abstract
The invention relates to the field of biological medicinal materials, and discloses a reinforced anti-adhesion composite gel. The reinforced anti-adhesion composite gel comprises the following components in mass/volume ratio: 0.01 to 8 percent of polysaccharide with anionic groups, 0.0000001 to 1 percent of functional polypeptide with positive charge groups, and 0.05 to 5 percent of polysaccharide with cationic groups or cationic polymerized polypeptide or mixture of the two in any ratio. The invention also provides a preparation method for the reinforced anti-adhesion composite gel. The reinforced anti-adhesion composite gel can effectively reduce the occurrence of adhesion by adding the anti-adhesion functional polypeptide with polyelectrolyte interaction and the polysaccharide with cationic groups and/or cationic polymerized polypeptide into materials.
Description
Technical field
The invention belongs to biomedical materials field, relate to a kind of pluralgel for having the reinforced anti-adhesion function behind the surgical operation and its preparation method and application.
Background technology
Adhesion behind the surgical operation between tissue and the organ is the disease that often occurs clinically, and adhesion is that connective fiber band and adjacent tissue or organ combine and the anomalous structure that forms.Adhesion formation has universality, it is reported that 50%～100% abdominal cavity and operation on pelvis can cause adhesion in various degree.The clinical severe complication that adhesion causes comprises intestinal obstruction, infertility, chronic pelvic pain etc., has increased again the difficulty of operation and the potentiality of further complications.Therefore, the reason of further investigation Adhesion formation and prevention and the adhesion of minimizing surgical postoperative have become primary study and the application direction of current surgical field.
The method of minimizing adhesion commonly used has two kinds at present: 1) directly pour into various materials, such as dextran 70, heparin sodium, cortisone, antibiotic and enzyme etc., reduce or suppress fibrous connective tissue forming; 2) use interleaving agent, between wound tissue, provide mechanical barrier, such as expanded polytetrafluoroethylsealing (Gore-Tex) and oxidized regenerated cellulose (Interceed TC7) surgical membrane etc.
Pay close attention in recent years the more macromolecular material polyvinyl alcohol that has, polylactic acid etc., the biomaterial cellulose family, animal derived material such as hyaluronate sodium, the chitosan class, collagen protein, fibroin albumen, mucous membrane of animal and valve etc., they have preferably biocompatibility, some albumen also remains with some active groups, help organization healing, but obvious defective is arranged also, as produce acid degradation product (poly-lactic acid material), gel and the film used are held time shorter in vivo, poor mechanical property etc., and the use of some foreign proteins brings the immunogenicity risk inevitably, in order to improve the mechanical property of these products, vivo degradation time and reduction immunogenicity, have to use cross-linking agent, cross-linking agent commonly used is glutaraldehyde (GA), GA is cross-linked material fast and effectively, reduce antigenicity, improve stability and the mechanical property of material, but the calcification phenomenon often occurs in the material after GA processes, this is extremely harmful, and more serious cell-cytotoxic reaction usually occurs in the material part that GA processes.
As the maximum natural polysaccharide derivant of occurring in nature quantity, be widely used in a plurality of fields with anionic group polysaccharide carboxymethyl cellulose, carboxymethyl chitosan, hyaluronic acid.Carboxymethyl cellulose, carboxymethyl chitosan, hyaluronic acid all are a kind of anionic linear polymeric polysaccharose substances, outward appearance is white in color or micro-yellow powder, tasteless, nontoxic, the toughness solution that becomes soluble in water, have unique physicochemical property, and the functions such as tool thickening, suspension, stable emulsifying, rheological behavior.Have good film property, high viscosity have high strength and high flexibility after carboxymethyl cellulose, carboxymethyl chitosan, the hyaluronic acid of high molecule mass are made thin film relatively, and the solution of high concentration still has good transparency.
Sodium carboxymethyl cellulose, carboxymethyl chitosan, hyaluronic acid confirm through serial toxicological experiment result: have no side effect, to skin and mucosa nonirritant, in vivo non-immunogenicity and pyrogen reaction, haemolysis not, without mutagenicity and lethal mutation effect, be that the good body of histocompatibility is implanted into biomaterial.Sodium carboxymethyl cellulose, carboxymethyl chitosan, the most frequently used form of hyaluronic acid are solution, gel and film, solution or gel are applied in wound surface, and solution can be uniformly distributed in the art district, plays " aquation is floating " effect, intercept art district and surrounding tissue, prevent Adhesion formation.As a mechanical separator, form with glue or film covers tissue or organ wound surface, have absorbability and well reach the characteristics such as biocompatibility is good, but be absorbed degraded in about about 4 days in vivo, yet damage the finishing the time more than 7 days of taking of posterior synechiae, so carboxymethyl cellulose, carboxymethyl chitosan, hyaluronic acid use separately the Film with Preventing Adhesion effect unsatisfactory.
The existing product that respectively carboxymethyl cellulose, carboxymethyl chitosan, hyaluronic acid sugar is used for Film with Preventing Adhesion clinically is solution or gel in recent years, understands with position Flowing Hard behind the body to guarantee that it is in concentration and the time of wound site but put into.
Summary of the invention
The object of the present invention is to provide a kind of reinforced anti-adhesion composite gel, to overcome existing band anionic group polysaccharide such as carboxymethyl cellulose, carboxymethyl chitosan, hyaluronic acid and derivant thereof, pectin and derivant thereof, the deficiency of the one pack system preventing adhesiving effects such as chondroitin sulfate.
The present invention also provides the preparation method of above-mentioned reinforced anti-adhesion composite gel.
Another object of the present invention is to provide the application of above-mentioned reinforced anti-adhesion composite gel.
A kind of reinforced anti-adhesion composite gel, for comprising the hydrogel of functional polypeptide, band cation group polysaccharide and/or cationic polymerization polypeptide (polylysine) with anionic group polysaccharide, positively charged group, wherein the mass/volume of each component ratio is:
Band anionic group polysaccharide: 0.01%～8%;
The functional polypeptide of positively charged group: 0.0000001%～1%;
Band cation group polysaccharide or cationic polymerization polypeptide, or the mixture of both arbitrary proportions: 0.05%～5%;
Described band anionic group polysaccharide is selected from sodium carboxymethyl cellulose, the derivant of carboxymethyl chitosan, hyaluronic acid, hyaluronic derivant, pectin, pectin or one or more the mixture in the chondroitin sulfate;
The functional polypeptide of described positively charged group is the functional polypeptide that one or both ends connect positively charged group, and positively charged group can be preferably one to five lysine that side chain is positively charged and/or arginine; Functional polypeptide be preferably contain-arginine-glycine-aspartic acid-polypeptide, contain-valine-Gly-Val-Pro-glycine-polypeptide or contain-isoleucine-lysine-valine-alanine-valine-polypeptide in the mixed polypeptide of one or both formation; Connect positively charged group by the one or both ends at functional polypeptide, make its with the reaction of the carboxyl generation polyelectrolyte of anionic group polysaccharide; Functional polypeptide can obtain by chemical synthesis process or gene engineering expression purification process;
Described band cation group polysaccharide is water-soluble chitosan, and its molecular weight is 500～25000;
Described cationic polymerization polypeptide is polylysine, and its molecular weight is 500～300000.
The functional polypeptide and the band cation group polysaccharide that in anionic group polysaccharide such as carboxymethyl cellulose and/or carboxymethyl chitosan, hyaluronic acid material, add positively charged group among the present invention, the positive charge group can be ingenious expediently with the materials such as band anionic group polysaccharide such as carboxymethyl cellulose, carboxymethyl chitosan, hyaluronic acid in the reaction of negative charge group carboxyl generation polyelectrolyte, namely polyelectrolyte interacts.Form by the polyelectrolyte reaction with anionic group polysaccharide, functional polypeptide, with cation group polysaccharide and/or cationic polymerization polypeptide and to have the complex of strengthening the anti function.
The preparation method of reinforced anti-adhesion composite gel of the present invention is: will be mixed in proportion with functional polypeptide, band cation group polysaccharide and/or cationic polymerization polypeptide and the water of anionic group polysaccharide, positively charged group, stir, to forming without cotton-shaped even gel, reinforced anti-adhesion composite gel is processed to get in degerming.
Strengthening the compound anti-adhesion gel can be the aqueous gel preparation, also can be dried to gel film or sponge membrane, and film can be further crosslinked, is conducive to increase degradation time in film strength and the extension body.
The present invention is by adding an amount of interactional anti functional polypeptide of polyelectrolyte being arranged and be with cation group polysaccharide and/or cationic polymerization polypeptide in material, on the one hand, crosslinked action by the polyelectrolyte reaction, strengthen the molecular weight of material, prolong degradation time in vivo, strengthen its mechanical isolation effect; On the other hand, functional polypeptide can promote the quickening reparation recovery from illness of tissue or organ wound surface, avoids adhesion to occur, and can stop fibrin deposition, stops platelet aggregation and activation, stops albumin exudation, finally prevents Adhesion formation.Reinforced anti-adhesion composite gel of the present invention can effectively reduce the generation of adhesion phenomenon.
The specific embodiment
Embodiment 1
Take by weighing pharmaceutical grade sodium carboxymethyl cellulose 2.5 grams, add first 80 milliliters of water for injection heated water bath (30～60 ℃) stirring and dissolving; Add 50 milligrams of functional polypeptides, its amino acid sequence is: lysine-lysine-threonine-Vitro By Serine/arginine-Gly-Asp-serine stirs; Add 1.5 gram chitosans (molecular weight 10000～20000), the limit edged stirs, after inject water to 100 milliliters, stir to forming without cotton-shaped even gel, reinforced anti-adhesion composite gel is processed to get in degerming.
Embodiment 2
Take by weighing pharmaceutical grade carboxymethyl chitosan 2 grams, add first 80 milliliters of water for injection heated water bath (30～60 ℃) stirring and dissolving; Add 60 microgram functional polypeptides, its amino acid sequence is: lysine-lysine-cysteine-Vitro By Serine/arginine-Gly-Asp-serine-cysteine-lysine-lysine (can be condensed into ring-type) stirs; Add 2.0 gram chitosans (molecular weight 2000～3000), the limit edged stirs, after inject water to 100 milliliters and stir to forming without cotton-shaped even gel, reinforced anti-adhesion composite gel is processed to get in degerming.
Embodiment 3
Take by weighing pharmaceutical grade hyaluronic acid 1.5 grams, add first 80 milliliters of water for injection heated water bath (30～60 ℃) stirring and dissolving; Add 100 microgram functional polypeptides, its amino acid sequence is: lysine-lysine-threonine-Vitro By Serine/arginine-Gly-Asp-serine stirs; Add 0.05 gram polylysine (molecular weight is 3000-5000), the limit edged stirs, after inject water to 100 milliliters and stir to forming without cotton-shaped even gel, reinforced anti-adhesion composite gel is processed to get in degerming.
Embodiment 4
Take by weighing pharmaceutical grade sodium carboxymethyl cellulose 2.5 grams, add first 80 milliliters of water for injection heated water bath (30～60 ℃) stirring and dissolving; Add 50 microgram functional polypeptides, its amino acid sequence is: lysine-arginine-threonine-Vitro By Serine/arginine-Gly-Asp-serine-lysine-lysine stirs; Add 1.0 gram chitosans (molecular weight 6000～8000) and 0.5 gram polylysine (molecular weight is 1000-5000), the limit edged stirs, after inject water to 100 milliliters and stir to forming without cotton-shaped even gel, reinforced anti-adhesion composite gel is processed to get in degerming.
Embodiment 5
Take by weighing pharmaceutical grade sodium hydroxyethlcellulose 6 grams, add first 80 milliliters of water for injection heated water bath (30～60 ℃) stirring and dissolving; Add 5 milligrams of functional polypeptides, its amino acid sequence is: lysine-arginine-threonine-Vitro By Serine/arginine-Gly-Asp-serine-lysine-lysine stirs; Add 1.0 gram chitosans (molecular weight 6000～8000) and 0.3 gram polylysine (molecular weight is 10000-30000), the limit edged stirs, after inject water to 100 milliliters and stir to forming without cotton-shaped even gel, reinforced anti-adhesion composite gel is processed to get in degerming.
Embodiment 6
Take by weighing aching and limp ossein 0.2 gram of pharmaceutical grade bright sulfur, add first 80 milliliters of water for injection heated water bath stirring and dissolving; Add 1 milligram of functional polypeptide, its amino acid sequence is: lysine-lysine-threonine-Vitro By Serine/arginine-Gly-Asp-serine stirs; Add 0.5 gram polylysine (molecular weight is 150000-300000), the limit edged stirs, after inject water to 100 milliliters and stir to forming without cotton-shaped even gel, reinforced anti-adhesion composite gel is processed to get in degerming.
Embodiment 7
Take by weighing pharmaceutical grade sodium carboxymethyl cellulose 1.5 grams and pectin 2 grams, add first 80 milliliters of water for injection heated water bath (30～60 ℃) stirring and dissolving; Add 100 microgram functional polypeptides, its amino acid sequence is: lysine-arginine-threonine-Vitro By Serine/arginine-Gly-Asp-serine-lysine-lysine stirs; Add 0.05 gram polylysine (molecular weight is 1000-5000), the limit edged stirs, after inject water to 100 milliliters and stir to forming without cotton-shaped even gel, reinforced anti-adhesion composite gel is processed to get in degerming.
The comparison of embodiment Final 8 compound anti-adhesion gel and existing single component anti-adhesion gel preventing adhesiving effect:
36 of Wister rats, body weight (220 ± 40) gram, male and female are regardless of, and are divided at random 3 groups: the A group is matched group, and B group and C group respectively are 12 for experimental group, and all animals adopts etherization, anaesthetizes safe rear dorsal position, is fixed on the operating-table.Take off median abdominal incision, be about 4 centimetres, successively enter abdomen, propose caecum, in 20 centimetres at ileum end, be interrupted with knife blade and peel off ileum serous coat band, wide 0.3 centimetre, be about 4 centimetres, totally 3 places.Then form face at A and do not do other processing, close abdomen.B form face inject the sodium carboxymethyl cellulose gel (concentration 2.5%, 3.5ml); C forms the reinforced anti-adhesion composite gel 3.5ml that face injects embodiment 1.
The postoperative sub-cage rearing, the equal fasting of postoperative 10 hours, feedstuff is rat pellet of the same race.10 days all animals of postoperative are put to death.With reference to adhesion grade staging: 0 grade without adhesion, and the ileum serosal surface is repaired fully; The I level is loose a small amount of adhesion only, and is easily separated, without oozing of blood, and large reparation of ileum serous coat; The adhesion of II level is slightly close than the I level, oozing of blood during separation, and the ileum serous coat is repaired about half; The adhesion of III level become bulk or with the extensive adhesion of other internal organs, but without blocking, ileum serous coat small part is repaired; The adhesion of IV level is extensive, and densification has intestinal obstruction, and the near-end dilatation of intestine is obvious.The intra-abdominal adhesions situation is evaluated classification.
The result shows: each organizes the adhesion significant difference, the A group is the III---IV level, the B group is take the I level as main, the C group is take the 0---I level as main, B group adhesion grade obviously alleviates than the A group, two groups of differences have significant (P＜0.01), and C group adhesion grade obviously alleviates than the B group, and two groups of differences have significant (P＜0.01).Embodiment 9 strengthens the comparison of compound anti-adhesion gel film and existing single component anti-adhesion gel preventing adhesiving effect
Get the reinforced anti-adhesion composite gel 4ml of embodiment 1, place 3 centimetres culture dish sialorrhea film forming, the lyophilization anti-adhesion gel sponge membrane that strengthened in 24 hours carries out the adhesion grade relatively with said method.
A group is matched group, and B group and C group are experimental group, B form face inject the sodium carboxymethyl cellulose gel (concentration 2.5%, 3.5ml); C forms face and uses above-mentioned reinforced anti-adhesion gel sponge film.The result shows, each organizes the adhesion significant difference, the A group is the III---IV level, the B group is take the I level as main, the C group is take the 0---I level as main, and B group adhesion grade obviously alleviates than the A group, and two groups of differences have significant (P＜0.01), C group adhesion grade obviously alleviates than the B group, and two groups of differences have significant (P＜0.01).
Claims (8)
1. a reinforced anti-adhesion composite gel is characterized in that, for comprise functional polypeptide with anionic group polysaccharide, positively charged group, with the hydrogel of cation group polysaccharide and/or cationic polymerization polypeptide, wherein the mass/volume of each component ratio is:
Band anionic group polysaccharide: 0.01%～8%;
The functional polypeptide of positively charged group: 0.0000001%～1%;
Band cation group polysaccharide or cationic polymerization polypeptide, or the mixture of both arbitrary proportions: 0.05%～5%;
Describedly be selected from one or more mixture in sodium carboxymethyl cellulose and derivant, carboxymethyl chitosan and derivant, hyaluronic acid and derivant, pectin and derivant or the chondroitin sulfate with the anionic group polysaccharide;
The functional polypeptide of described positively charged group is that one or both ends connect one to five lysine that side chain is positively charged and/or arginic functional polypeptide.
2. a kind of reinforced anti-adhesion composite gel claimed in claim 1, it is characterized in that, described functional polypeptide be contain-arginine-glycine-aspartic acid-polypeptide, contain-valine-Gly-Val-Pro-glycine-polypeptide or contain-isoleucine-lysine-valine-alanine-valine-polypeptide in the mixed polypeptide of one or both formation.
3. a kind of reinforced anti-adhesion composite gel claimed in claim 1 is characterized in that, described band cation group polysaccharide is water-soluble chitosan.
4. a kind of reinforced anti-adhesion composite gel claimed in claim 3 is characterized in that, described water-soluble chitosan molecular weight is 500～250000.
5. a kind of reinforced anti-adhesion composite gel claimed in claim 1 is characterized in that, described cationic polymerization polypeptide is polylysine.
6. a kind of reinforced anti-adhesion composite gel claimed in claim 5 is characterized in that, described polylysine molecular weight is 500～300000.
7. the preparation method of the described reinforced anti-adhesion composite gel of claim 1, it is characterized in that, functional polypeptide, band cation group polysaccharide and/or cationic polymerization polypeptide and water with anionic group polysaccharide, positively charged group are mixed in proportion, stir, to forming without cotton-shaped even gel, reinforced anti-adhesion composite gel is processed to get in degerming.
8. the application of reinforced anti-adhesion composite gel claimed in claim 1 aspect preparation reinforced anti-adhesion gel film or reinforced anti-adhesion sponge membrane.
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| WO2012057381A1 (en) * | 2010-10-29 | 2012-05-03 | 주식회사 휴메딕스 | Adhesion barrier containing hyaluronic acids and l-arginine |
| CN102614551B (en) * | 2012-03-31 | 2013-12-18 | 山西伯朗生物技术有限公司 | Preparation method for medical absorbable anti-adhesion film |
| CN102772821B (en) * | 2012-08-01 | 2015-07-01 | 苏州博创同康生物工程有限公司 | Absorbable and hemostatic multifunctional particle with tissue induction and preparation and application of multifunctional particle |
| WO2014186937A1 (en) * | 2013-05-20 | 2014-11-27 | Gao Min | Preparation method of mussel adhesive protein gel, mussel adhesive protein gel and use thereof |
| CN106038516A (en) * | 2016-03-14 | 2016-10-26 | 苏州博创康源生物技术有限公司 | Compound drug loading tissue pasting membrane and preparation method thereof |
| CN106038517A (en) * | 2016-03-14 | 2016-10-26 | 苏州博创康源生物技术有限公司 | Polysaccharide medicine loading tissue adhesion film and preparing method thereof |
| CN105879127A (en) * | 2016-04-25 | 2016-08-24 | 东莞市联洲知识产权运营管理有限公司 | Method for preparing postoperative adhesion prevention materials |
| CN106220873A (en) * | 2016-08-16 | 2016-12-14 | 潘忠宁 | A kind of synthetic method of microgel base composite aquogel |
| CN107823701A (en) * | 2017-10-27 | 2018-03-23 | 温州生物材料与工程研究所 | A kind of poly glycosyl styptic sponge, preparation technology and application with active hemostatic function |
| CN113243548A (en) * | 2021-05-18 | 2021-08-13 | 河南卷烟工业烟草薄片有限公司 | Preparation method of low-viscosity low-irritation reconstituted tobacco of heated cigarette |
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| CN101036780A (en) * | 2007-04-27 | 2007-09-19 | 四川大学 | Application of self-assembled short peptide in the medicine for treating burn and face wound |
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| US7279177B2 (en) * | 2002-06-28 | 2007-10-09 | Ethicon, Inc. | Hemostatic wound dressings and methods of making same |
| EP2026846A4 (en) * | 2006-06-05 | 2010-09-29 | Bactiguard Ab | A polymer matrix, uses thereof and a method of manufacturing the same |
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| CN1685829A (en) * | 2005-04-30 | 2005-10-26 | 武汉大学 | Preparation method of chitin/zinc compound bactericide |
| CN101036780A (en) * | 2007-04-27 | 2007-09-19 | 四川大学 | Application of self-assembled short peptide in the medicine for treating burn and face wound |
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