CN1943796A - The preparation method of the radiant crosslink porous reticular cells amnionic hydro gel dressing and its preparation method - Google Patents
The preparation method of the radiant crosslink porous reticular cells amnionic hydro gel dressing and its preparation method Download PDFInfo
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- CN1943796A CN1943796A CN 200610122993 CN200610122993A CN1943796A CN 1943796 A CN1943796 A CN 1943796A CN 200610122993 CN200610122993 CN 200610122993 CN 200610122993 A CN200610122993 A CN 200610122993A CN 1943796 A CN1943796 A CN 1943796A
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Abstract
The invention discloses a preparation method of an irradiative crosslink network cellular porous hydrogel dressing of amniotic membrane, the specific methods are: put the hydrogel solutions comprising of PVA, chitosan and acrylic acid-acrylamide copolymer on the porous stainless steel plate used to put the netty cellular amniotic membrane, add a small amount of agar, agitate uniformly, evacuate, and the hydrogel prototype was obtained; then press the hydrogel to film; then expose the membrane to 60Coy- ray irradiation in room temperature to crosslink, then dehydrate, freeze-dried, radiative sterilized, packed under aseptic conditions. The invention possesses good protection treatments, such as alleviate pain, resist to phlogosis and bacterium, and promote healing. It also has good hygroscopic properties, water retention, permeability and flexibility, further more we can have a selection of drugs according to sensitive bacteria of wound, pain, the need of arristing or activzte bleeding to and to help achieve sustained-release drug treatment purposes.
Description
Technical field:
The present invention relates to medical biological dressing field, refer in particular to the manufacture method of the cross-linking radiation holey human acellular amniotic membrane aerogel dressing of a kind of energy absorption and the new compounding pharmaceutical of slow release.
Background technology:
Modern burn treating not only pays attention to improve patient's survival rate, emphasizes the recovery of function simultaneously, seeks ideal Graftskin, and the early stage covering and the later stage reduction of large-area burns wound surface all seemed extremely important.Ideal Graftskin should meet following condition: (1) cost is low; (2) long shelf-life; (3) can use after the resurrection; (4) no antigen; (5) tool persistency; (6) tool pliability; (7) can prevent loss of moist; (8) antibacterial had barrier action; (9) compliance to irregular wound surface is good; (10) be easy to fix; (11) with the childhood development expansion of correspondingly to grow of growing up; (12) once migration process is promptly finished in operation; (13) can hyperplasia.Still the incompetent at present desirable Graftskin that satisfies above-mentioned condition fully.But, to the research of this substitute, existing both at home and abroad many reports.From practical standpoint, Graftskin can be divided into temporary and nonvolatil two big classes from designing.In the research of numerous temporary Graftskins, amniotic membrane has nontoxic, nonirritant, no antigen, is easy to sterilization and preserves because of it, also has good pliability, necessarily permeable, breathability and wound surface bonding force, cover the back microorganism and can not penetrate intrusion, waterproof and draw materials easily, characteristics such as cost is low and being used widely.Amniotic membrane is the innermost layer of people's Placenta Hominis.It with amniotic cavity in amniotic fluid contact with fetus.Normal amniotic membrane is thin and transparent, and certain toughness is arranged, and thickness is 0.02～0.5mm, no blood vessel, nerve and lymphatic vessel tissue.Amniotic membrane can be divided into five layers successively, i.e. epithelial layer, basement membrane layer, compacted zone, fibroblast layer and spongy layer.Epithelial layer derives from ectoderm, and connective tissue derives from mesoderm.The collagen composition of amniotic membrane also has a spot of laminin and elastin laminin based on I, II and III Collagen Type VI.It is generally acknowledged that amniotic membrane does not almost have antigenicity.Amniotic membrane is acknowledged as the improved biomaterial of a kind of revolutionary character as a kind of biomaterial, and practice proves that also amniotic membrane is that a kind of good wound surface covers dressing, and it can relieve the pain, again can antiinflammatory, infection, wound surface there are extraordinary therapeutical effect and protective effect.And intact, the no cicatrix of skin repair, promptly attractive in appearance economical again, it preserves convenient, and low price is worthy to be popularized.
What research of the prior art was maximum is amniotic membrane preparation and preservation aspect, as patent publication No. be CN1522568A and patent publication No. though but to be the described bioamnion of CN1201697A preserve for room temperature, do not make to take off cell because of it and handle and in clinical use, limited to; Patent publication No. is the deficiency that has overcome above-mentioned preparation in the technology of CN1757717A, adopted take off cell technology make monolayer take off the dry amniotic membrane of cell more preceding two kinds more practical and preserve, but still be monolayer amniotic membrane, a common deficiency of these amniotic membranes is: thin, mechanicalness is poor, drag is easily torn, and easily curls, be difficult to plant and be cut into shape, easily dissolve behind the flap coverage etc.For overcoming these shortcomings, have in the prior art adopt the multilamellar fresh amnion simple overlapping be applied to ophthalmologic operation many reports are also arranged, patent publication No. is to adopt two-layer in the described technology of CN1799558A or multilamellar fresh amnion or preserve amniotic membrane by obtaining thicker glue adhesion amnion with the adherent method of Fibrin Glue, though having overcome monolayer amniotic membrane approaches and easily tears, the deficiency that difficulty is cut out, but also reduced the inherent transparency of amniotic membrane own simultaneously, just mechanically increase the thickness of amniotic membrane, but the good water absorption that does not have hydrogel, water-retaining property, breathability and pliability more do not have the function of slow releasing pharmaceutical; Other has an invention is to carry out the crosslinked compound bio amniotic membrane of deriving that obtains with the amniotic membrane of special preparation and collagen solution for mentioning the amniotic membrane preparation method in " biologically-derived amniotic membrane, compound bio derive amniotic membrane and preparation method thereof " (patent publication No. is CN1618954A).This has mentioned a kind of newer method that is cross-linked to form the biological dressing film about amniotic membrane and high-molecular biologic first, novelty is arranged very much, but wherein amniotic membrane prepares normal configuration and the biological characteristics that the method that adopts organic chemistry material confrontation amniotic membrane to carry out defat, de-sludging, digestion has been destroyed amniotic membrane fully, only stay supporting structure, the large number of biological active factorses such as EGF, BFGF that are present in the amnion cell layer structure are lost, influence its biological activity, also destroyed its flexible physics characteristic simultaneously.And collagen do not have several big advantage of hydrogel, as good water absorption, and water-retaining property, the function of breathability and pliability and slow releasing pharmaceutical etc.So also limit its use.
Simple property aerogel dressing is a kind of hydrophilic net high-polymer swelling body that contains large quantity of moisture because of it, has good water absorption (from 10% to several thousand times water absorption of self dry weight), smooth surface, good biocompatibility is so obtained extensive studies and application at biomedical engineering field.Aerogel dressing can fit tightly but adhesion with irregular wound surface, reduces the chance of bacteria breed.Therefore the hydrogel burn dressing, all carries out primary study and has obtained encouraging progress the hydrogel burn dressing both at home and abroad near the requirement of desirable burn dressing.The commodity of some hydrogel burn dressing also engender and have obtained extensive use clinically.The hydrogel burn dressing that various functions are also arranged, as patent publication No. is that CN1679972A is by name: " a kind of burn dressing of chitin gel with gradient structure and preparation method thereof " and patent of invention " a kind of hollow acetate fiber antibiotic aerogel dressing and preparation thereof " (patent publication No. is CN1706503A) by name and " containing medicine; polyvinyl alcohol hydrogel dressing of chitosan and preparation method thereof " (patent publication No. is CN1579559A) etc., all mention a kind of water content height that has, the intensity height, breathability and pliability good in addition can slow releasing pharmaceutical the preparation method of aerogel dressing of function.But also only only limit to use hydrogel separately, do not see that hydrogel and allosome or heterogeneity biological material are arranged is crosslinked, as mentioned in this article the amniotic membrane material.So it is good that its clinical effectiveness does not combine the dressing of amniotic membrane of biologically active.And the way of its slow releasing pharmaceutical is immersed in the drug solution of having chosen in advance and having fixed repress finished product after the swelling earlier, and have can not be at the deficiency of wound surface concrete condition symptomatic treatment, and simultaneously, as untimely use, pharmaceutical efficacy can weaken.
Summary of the invention:
The present invention is intended to invent a kind of excellent protection therapeutic effect that both possessed; as pain relieving; anti-inflammatory, antibacterial; promote healing etc.; has good water absorption again; water-retaining property, breathability and flexible dressing, but also can be according to sensitive bacterial, the pain situation of wound surface, stop blooding or the needs of invigorating blood circulation etc. are selected corresponding medicine selectively and can slow releasing pharmaceutical be reached the manufacture method of the purpose cross-linking radiation holey human acellular amniotic membrane aerogel dressing of assisting treatment.
Technical scheme provided by the invention for achieving the above object is: the concrete manufacture method of holey human acellular amniotic membrane aerogel dressing is:
One, the making of mesh structural porous human acellular amniotic membrane: Placenta Hominis that will be after Medical Treatment, under aseptic condition, operate, passivity is separated in the potential gap of amniotic membrane and chorion, obtain smooth, transparent amniotic membrane, clean with balance liquid, be soaked in above-mentioned another part solution sterilization again 25～35 minutes, then with amniotic membrane, be tiled on the celluloid filter paper that has micropore, epithelial surface is put into trypsin and the EDTA mixed liquor digests up again, gently scrapes epithelial cell with cotton rod, normal saline flushing repeatedly, the filter paper that will be covered with amniotic membrane as required is cut into the size of different size, holes on the filter paper of amniotic membrane with puncher, removes filter paper, dry under the room temperature, place the interior cold preservation of pure glycerin aseptic bottle more than 95% standby;
Two, hydrogel material is prepared: by the degree of polymerization is that 1500-3000, alcoholysis degree are the polyvinyl alcohol of 98-100%, molecular quantum 100-1000gmol
-1, the chitosan of deacetylating degree of chitosan 80-98% and acrylic acid-acrylamide copolymer form, each constituent mass percentage composition is as follows: polyvinyl alcohol 5～20%, chitosan 0.5～10%, acrylic acid-acrylamide copolymer 2～15%;
Polyvinyl alcohol is main solid polymer, has stronger mechanical strength; Acrylic acid-acrylamide copolymer has good water absorption, and suction back hold facility is stronger; Chitosan is a wetting agent, can biodegradation, and the small molecule bioactive height, water-retaining property is strong.
Three, manufacturing process:
1), polyvinyl alcohol be dissolved in make solution A in the normal saline, acrylic acid-acrylamide copolymer is dissolved in makes solution B in the normal saline, merge solution A and B, it is even to add wetting agent chitosan heated and stirred, makes it to dissolve fully shape.Form solution C, and room temperature leaves standstill;
2), be placed in size and the amniotic membrane stainless steel disc of the same size after will previous prepared mesh structural porous human acellular amniotic membrane rehydration cleaning, slough epithelial one to face down;
3), prepared solution C is filled in the stainless steel disc that is placed with amniotic membrane, adds a small amount of agar and fully shake all, leave standstill 8～12 hours, and be evacuated, obtain hydrogel original shape D;
4), with the diaphragm that D is pressed into thick about 3～8mm, obtain hydrogel diaphragm E;
5), E is utilized
60Coy-X-ray room X relaxing the bowels with purgatives of warm nature cross-linking radiation, irradiation dose are 10～80kGy;
6), with the E of irradiation dehydration, lyophilization, radiosterilization, in aseptic condition encapsulation down, 0～4 ℃ of cryopreservation is finished the making of dressing.
During clinical use, prepare the solution that contains antibiotics according to the wound surface needs, or contain hemorrhage solution, or blood circulation promoting medicine solution, or the solution of analgesic drug product, the holey human acellular amniotic membrane aerogel dressing for preparing is taken out, tear packing off, put into one or more immersions of above-mentioned solution, directly overlay on the wound surface after allowing its swelling suck medicine.Can add that binder is fixed or fixing, also can change once or allow its natural solution absorption a week.
Applicable scope of the present invention is: be applicable to that 1, a variety of causes burns wound repair and temporary skin substitutes that (scalding) hinders, especially be fit to II degree or dark II degree wound surface, the temporary transient covering of wound surface after also can making III degree wound surface or cutting crust.2, be applicable to skin tissue defects and decubital ulcer, chronic ulcer of skin wound surface.
Advantage of the present invention and beneficial effect are:
1), under the prerequisite of doing substrate with amniotic membrane crosslinked hydrogel, it is not high to have overcome single amniotic membrane biomaterial mechanical strength, the deficiency of poor compliance.Make the more convenient clinical use of this dressing.
2), make crosslinked hydrogel under the prerequisite of substrate at amniotic membrane, make this dressing possess the strong water absorption of hydrogel, moisture retention, breathability again, the amniotic membrane that makes wound surface be in the no rejection of an existing similar human body skin covers, and similar normal human's moist environment is arranged again.Be the support that human acellular amniotic membrane provides a kind of epithelial cell to creep, hydrogel has guaranteed that epithelial cell is creeped and the moist environment of wound repair.
3), human acellular amniotic membrane is made mesh structural porous as substrate, help the drain of wound secretion and adsorbed by hydrogel, help keeping the gnotobasis of wound surface, accelerate epithelium regeneration.Simultaneously hydrogel is outside amniotic membrane, only to water and oxygen can by and do not allow extraneous antibacterial, dust to pass through, can prevent effectively that external environment from infecting.
4), be substrate in the hydrogel with the human acellular amniotic membrane, given full play to can relieving the pain of human acellular amniotic membrane, again can antiinflammatory, infection, wound surface is had extraordinary therapeutical effect and protective effect.
5), the spacial framework in the hydrogel, drug slow can be discharged into wound surface constantly.Play continued treatment, reduce dressing change frequency, alleviate the effect of medical worker's workload and patient's financial burden.
6), make not swollen dry finished product, before clinical need to use again according to the sensitive bacterial situation of wound surface, the pain situation, the needs that stop blooding or invigorate blood circulation etc. select corresponding drug solution to soak swelling selectively, suck medicine, it is with strong points, be difficult for causing drug resistance, can also reach pain relieving according to actual needs, invigorate blood circulation or the hemostatic purpose, overcome traditional method and be immersed in the drug solution of having chosen in advance and fixing repress finished product after the swelling earlier, have and simultaneously, to use immediately at the deficiency of wound surface concrete condition symptomatic treatment, instant preparation has overcome as untimely use the deficiency that pharmaceutical efficacy can weaken.
The specific embodiment:
Specific embodiments of the invention are provided below:
One, the making of mesh structural porous human acellular amniotic membrane:
Get cesarean anemia of pregnant woman's Placenta Hominis, the capable serology procuratorial work of antenatal parent is got rid of and is contained HIV, HBV, HCV and syphilis.Get its Placenta Hominis after the cesarean immediately, under aseptic condition, operate, passivity is separated in the potential gap of amniotic membrane and chorion, obtains smooth, transparent amniotic membrane, cleans with the balance liquid that contains 50mg/L penicillin, 50mg/L streptomycin, 100mg/L neomycin, 2.5mg/L amphotericin B.Be soaked in above-mentioned another part solution sterilization again 30 minutes.With amniotic membrane, be tiled on the celluloid filter paper that has 0.45 μ m micropore then.Epithelial surface is put into the 0.25wt% trypsin and the 0.02wt%EDTA mixed liquor digests up again, gently scrapes epithelial cell with cotton rod, normal saline flushing repeatedly, and the filter paper that will be covered with amniotic membrane as required is cut into the size of different size.On the filter paper of amniotic membrane, hole (the about 3～5mm in aperture, the about 10mm of every span) with puncher, remove filter paper, dry under the room temperature, place the interior 4 ℃ of cold preservations of pure glycerin aseptic bottle more than 95% standby.
Two, hydrogel material is prepared:
(degree of polymerization is 1500-3000, and alcoholysis degree is 98-100% by polyvinyl alcohol.) and chitosan (molecular quantum 100-1000gmol
-1, deacetylating degree of chitosan 80-98%), acrylic acid-acrylamide copolymer is formed.Each constituent mass percentage composition is as follows: polyvinyl alcohol 5～20%, chitosan 0.5～10%, acrylic acid-acrylamide copolymer 2～15%.
Three, manufacturing process:
1), polyvinyl alcohol be dissolved in make solution A in the normal saline, acrylic acid-acrylamide copolymer is dissolved in makes solution B in the normal saline, merge solution A and B, it is even to add wetting agent chitosan heated and stirred, makes it to dissolve fully shape.Form solution C, and room temperature leaves standstill.
2), to be placed on special size after will previous prepared mesh structural porous human acellular amniotic membrane rehydration cleaning consistent with the amniotic membrane size, in the square rustless steel square plate of the about 1cm of height, slough epithelial one and face down.
3), prepared solution C is filled in the rustless steel square plate that is placed with amniotic membrane, adds a small amount of agar and fully shake all, leave standstill 10 hours, and be evacuated.Obtain hydrogel original shape D.
4), with the diaphragm that D is pressed into thick about 3～8mm, obtain hydrogel diaphragm E.
5), E is utilized
60Coy-X-ray room X relaxing the bowels with purgatives of warm nature cross-linking radiation, irradiation dose are 10～80kGy
6), with the E of irradiation dehydration, lyophilization, radiosterilization, in aseptic condition encapsulation down, 0～4 ℃ of cryopreservation is finished the making of dressing.
The embodiment of the above is preferred embodiment of the present invention only, is not to limit practical range of the present invention with this, so the equivalence that all shapes according to the present invention, structure and principle are done changes, all should be covered by in protection scope of the present invention.
Claims (1)
1, the manufacture method of cross-linking radiation holey human acellular amniotic membrane aerogel dressing, it is characterized in that: concrete manufacture method is:
One, the making of mesh structural porous human acellular amniotic membrane: Placenta Hominis that will be after Medical Treatment, under aseptic condition, operate, passivity is separated in the potential gap of amniotic membrane and chorion, obtain smooth, transparent amniotic membrane, clean with balance liquid, be soaked in above-mentioned another part solution sterilization again 25～35 minutes, then with amniotic membrane, be tiled on the celluloid filter paper that has micropore, epithelial surface is put into trypsin and the EDTA mixed liquor digests up again, gently scrapes epithelial cell with cotton rod, normal saline flushing repeatedly, the filter paper that will be covered with amniotic membrane as required is cut into the size of different size, holes on the filter paper of amniotic membrane with puncher, removes filter paper, dry under the room temperature, place the interior cold preservation of pure glycerin aseptic bottle more than 95% standby;
Two, hydrogel material is prepared: by the degree of polymerization is that 1500-3000, alcoholysis degree are the polyvinyl alcohol of 98-100%, molecular quantum 100-1000g mol
-1, the chitosan of deacetylating degree of chitosan 80-98% and acrylic acid-acrylamide copolymer form, each constituent mass percentage composition is as follows: polyvinyl alcohol 5～20%, chitosan 0.5～10%, acrylic acid-acrylamide copolymer 2～15%;
Three, manufacturing process:
1), polyvinyl alcohol be dissolved in make solution A in the normal saline, acrylic acid-acrylamide copolymer is dissolved in makes solution B in the normal saline, merge solution A and B, it is even to add wetting agent chitosan heated and stirred, makes it to dissolve fully shape.Form solution C, and room temperature leaves standstill;
2), be placed in size and the amniotic membrane stainless steel disc of the same size after will previous prepared mesh structural porous human acellular amniotic membrane rehydration cleaning, slough epithelial one to face down;
3), prepared solution C is filled in the stainless steel disc that is placed with amniotic membrane, adds a small amount of agar and fully shake all, leave standstill 8～12 hours, and be evacuated, obtain hydrogel original shape D;
4), with the diaphragm that D is pressed into thick about 3～8mm, obtain hydrogel diaphragm E;
5), E is utilized
60Coy-X-ray room X relaxing the bowels with purgatives of warm nature cross-linking radiation, irradiation dose are 10～80kGy;
6), with the E of irradiation dehydration, lyophilization, radiosterilization, in aseptic condition encapsulation down, 0～4 ℃ of cryopreservation is finished the making of dressing.
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| CN102382238A (en) * | 2011-08-19 | 2012-03-21 | 宁波超能科技股份有限公司 | Preparation method for radiopolymerized slow-release high-polymer material |
| CN103480034A (en) * | 2013-09-29 | 2014-01-01 | 金陵科技学院 | Irradiation crosslinked chitosan/gelatin/polyvinyl alcohol hydrogel dressing as well as preparation method and application thereof |
| CN104224842A (en) * | 2014-09-03 | 2014-12-24 | 苗九昌 | Preparation method of compound amniotic membrane powder and compound amniotic membrane powder prepared thereby |
| CN104971380A (en) * | 2014-04-11 | 2015-10-14 | 烟台隽秀生物科技有限公司 | Acellular matrix repairing gel and new method for preparing the same |
| CN106693033A (en) * | 2015-08-03 | 2017-05-24 | 罗静峰 | Preparation method of sterilizing and blooding-stopping medical dressing with low cost |
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| CN110585486A (en) * | 2019-08-28 | 2019-12-20 | 湖南光琇医院有限公司 | Amnion composite material and preparation method and application thereof |
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| CN102382238A (en) * | 2011-08-19 | 2012-03-21 | 宁波超能科技股份有限公司 | Preparation method for radiopolymerized slow-release high-polymer material |
| CN102382238B (en) * | 2011-08-19 | 2013-09-25 | 宁波超能科技股份有限公司 | Preparation method for radiopolymerized slow-release high-polymer material |
| CN103480034A (en) * | 2013-09-29 | 2014-01-01 | 金陵科技学院 | Irradiation crosslinked chitosan/gelatin/polyvinyl alcohol hydrogel dressing as well as preparation method and application thereof |
| CN104971380A (en) * | 2014-04-11 | 2015-10-14 | 烟台隽秀生物科技有限公司 | Acellular matrix repairing gel and new method for preparing the same |
| CN104224842A (en) * | 2014-09-03 | 2014-12-24 | 苗九昌 | Preparation method of compound amniotic membrane powder and compound amniotic membrane powder prepared thereby |
| CN106693034A (en) * | 2015-08-03 | 2017-05-24 | 罗静峰 | Preparation method of pollution-free surgical dressing for sterilizing and stopping bleeding |
| CN106693033A (en) * | 2015-08-03 | 2017-05-24 | 罗静峰 | Preparation method of sterilizing and blooding-stopping medical dressing with low cost |
| CN106729935A (en) * | 2015-08-03 | 2017-05-31 | 罗静峰 | A kind of preparation method of the bactericidal haemostatic medical dressing using safety |
| CN106822981A (en) * | 2015-08-03 | 2017-06-13 | 罗静峰 | A kind of preparation method of the bactericidal haemostatic medical dressing having no side effect |
| CN107441547A (en) * | 2016-05-30 | 2017-12-08 | 四川大学华西医院 | A kind of wound repair material and its production and use |
| CN107441547B (en) * | 2016-05-30 | 2020-11-20 | 四川大学华西医院 | Wound repair material and preparation method and application thereof |
| CN110585486A (en) * | 2019-08-28 | 2019-12-20 | 湖南光琇医院有限公司 | Amnion composite material and preparation method and application thereof |
| CN110585486B (en) * | 2019-08-28 | 2021-11-30 | 湖南光琇医院有限公司 | Amnion composite material and preparation method and application thereof |
| WO2021114005A1 (en) * | 2019-12-02 | 2021-06-17 | 江南大学 | Method of preparing amnion-spongy chitosan composite double-layer wound dressing |
| CN115845141A (en) * | 2023-03-02 | 2023-03-28 | 健诺维(成都)生物科技有限公司 | Preparation method and application of dry amnion |
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