CN107412844A - A kind of method that collagen protein sponge is prepared based on air bladder - Google Patents
A kind of method that collagen protein sponge is prepared based on air bladder Download PDFInfo
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- CN107412844A CN107412844A CN201710266808.8A CN201710266808A CN107412844A CN 107412844 A CN107412844 A CN 107412844A CN 201710266808 A CN201710266808 A CN 201710266808A CN 107412844 A CN107412844 A CN 107412844A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0036—Porous materials, e.g. foams or sponges
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/0005—Ingredients of undetermined constitution or reaction products thereof
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Abstract
The present invention relates to a kind of method that collagen protein sponge is prepared based on air bladder, belong to medical material preparation field.The present invention is used as base-material by the use of air bladder, black fungus is as auxiliary material, after drying and crushing, boiling degreasing is carried out to it using sodium carbonate liquor, go the removal of impurity, recycle the microorganism enriched in the stone of a dog's gallbladder, kidney or bladder and enzyme, microorganism extraction is carried out to degreasing thing, extract is modified using the sour and beneficial thing in the stone of a dog's gallbladder, kidney or bladder simultaneously, increase bioactivity, change collagen structure, form space net structure, increase water imbibition, in extraction process, oxidation processes are carried out to wherein cellulose, increase hemostatic factor, improve hemostatic, it is combined again by increasing titanium with albumen thing, increase mechanical property, finally by saltouing, dialysis purification, molding is molded, so as to which collagen protein sponge be made.Collagen protein sponge rate of water absorption prepared by the present invention is fast, and water absorbent rate is high, and mechanical strength is high and bioactivity is strong, and safety coefficient is high.
Description
Technical field
The present invention relates to a kind of method that collagen protein sponge is prepared based on air bladder, belong to medical material preparation field.
Background technology
Collagen is the main component in animal connective tissue, its accounted in mammal body total protein 25~
30%.Collagen is also one of most critical raw material of biotech industry, in medical material, cosmetics, food industry
Etc. equal extensive application.Collagen（Also referred to as collagen）It is a kind of structural protein of extracellular matrix, is prevalent in dynamic
In the bone of object, cartilage, skin, tendon and ligament, body and internal organs are played support, protection, combination and formed boundary every etc.
Effect.In addition in terms of biomethanics, also there is the work(such as transport of signal transduction, growth factor and cell factor
Energy.
Collagen has obtained the consistent accreditation of people in field of food with the application of cosmetic field.And collagen
Research and application in terms of biomaterial for medical purpose also have relevant report, and the research of hemostatic material is used as collagen
Make some progress.Because collagen is substantially exactly a kind of natural tissues timbering material, therefore collagen turns into
Build tissue engineering bracket ideal material, such as skin, bone tissue, the support of blood vessel organ, afterwards in-vivo tissue cell climb
Enter to grow with it in support and merge, guide tissue regeneration.Pure collagen tissue engineering rack have good biocompatibility,
Plasticity, easy processing and can effectively facilitate cell adherence and propagation the advantages that.But collagen is as pure natural macromolecule,
The shortcomings that maximum is exactly that mechanical strength is poor, and degradation speed is too fast, in vivo using when be difficult to there is enough intensity to carry out supporting tissue
Rebuild, prevent it from reaching the requirement as cytoskeleton, and, water suction slow using the collagen protein sponge rate of water absorption of its preparation
Multiplying power is low, and mechanical strength is low, and bioactivity is poor.Therefore, it is fast to research and develop a kind of absorption speed, Water absorption and mechanical strength
Height, the good collagen protein sponge of bioactivity have important practical significance.
The content of the invention
The technical problems to be solved by the invention：Slow for current collagen protein sponge rate of water absorption, water absorbent rate is low, power
Low intensity is learned, the problem of bioactivity difference.Provide a kind of method that collagen protein sponge is prepared based on air bladder.
In order to solve the above technical problems, the present invention uses technical scheme as described below：
A kind of method that collagen protein sponge is prepared based on air bladder, specific preparation process are：
（1）By air bladder and black fungus in mass ratio 5:1~7:1 is well mixed, is dried, and crushes, and crosses 100 mesh sieves, collected
Sieve particle；
（2）By the sodium carbonate liquor in mass ratio 1 that particle and mass fraction are 10% that sieves:4, it is well mixed, 105~110
20~30min is heated at DEG C, is freeze-dried after naturally cooling to room temperature, obtains freeze-drying thing；
（3）Meter takes fermentation material in parts by weight, takes 50~55 parts of distilled water, 20~25 parts of above-mentioned dried objects, 12~16 portions of grapes
Sugar and 7~9 part of 150 mesh stone of a dog's gallbladder, kidney or bladder powder；
（4）Fermentation material is put into fermentation tank, then into fermentation tank add lactic acid until fermentation tank in fermentation material pH to 1.5~
2.5, oxygen is filled with into fermentation tank, ferment 16~19h, collects fermentation cylinder for fermentation mixture；
（5）Fermenting mixture in fermentation tank is put into filter press press filtration is carried out with 20MPa, pressing filtering liquid is collected, to pressing filtering liquid
It is centrifuged, collects supernatant；
（6）Supernatant, sodium chloride, the titanium valve that purity is 99.4% and 1, 8-Cineole are well mixed, are placed in 4~7 DEG C of standings
Overnight, then filtered, collect filter residue；
（7）Filter residue and 0.1M and pH are pressed into quality 1 for 7 phosphate buffer:2 is well mixed, is put into bag filter, dialysis temperature
Spend for 6~8 DEG C, using 0.4mol/L acetic acid dialysis 24h, reuse distilled water dialysis 48h, the residue in bag filter is put into
In mould, and mould is placed in cooler, design temperature is -30~-25 DEG C, after freeze overnight, mould is taken out and is placed in purple
Sterilizing in outer sterilizer, is stripped, and collects demoulding thing, you can obtain collagen protein sponge.
The step（3）Middle fermentation material can also contain 9~12 parts of yeast extracts.
The step（4）Middle fermentation jar temperature is 35~40 DEG C.
The step（4）In to be filled with the speed of oxygen be 20~25mL/min.
The step（6）The titanium valve and the mass ratio of 1, 8-Cineole that middle supernatant, sodium chloride, purity are 99.4% are 95
~100:14~16:2~4:1.3~2.3.
The step（7）The molecular cut off of middle bag filter is 7000~9000.
Compared with other method, advantageous effects are the present invention：
（1）The present invention after drying and crushing, utilizes sodium carbonate liquor by the use of air bladder as base-material, black fungus as auxiliary material
Boiling degreasing is carried out to it, goes the removal of impurity, recycles the microorganism enriched in the stone of a dog's gallbladder, kidney or bladder and enzyme, carrying out microorganism to degreasing thing carries
Take, while extract be modified using the sour and beneficial thing in the stone of a dog's gallbladder, kidney or bladder, increase bioactivity, change collagen structure,
Space net structure is formed, increases water imbibition, and then improve collagen protein sponge rate of water absorption and water absorbent rate；
（2）The present invention carries out oxidation processes in extraction process to cellulose, increases hemostatic factor, improves hemostatic while black
The extract of agaric, increase the restorative of collagen, then be combined by increasing titanium with albumen thing, increase mechanical property,
And then improve the mechanical strength of collagen protein sponge；
（3）Collagen protein sponge rate of water absorption prepared by the present invention is fast, and water absorbent rate is high, and mechanical strength is high and bioactivity is strong,
Low manufacture cost；
（4）Collagen protein sponge safety coefficient prepared by the present invention is high, low to human immunity rejection, can be widely applied to cure
Field.
Embodiment
By air bladder and black fungus in mass ratio 5:1~7:1, mixing, which is put into 60 DEG C of drying boxes, dries 10~12h, then will be dry
Mixture after dry is put into pulverizer, 30~40min is crushed with 1100r/min, then by the crushed mixture in pulverizer
100 mesh sieves are crossed, collect sieving particle；By the sodium carbonate liquor in mass ratio 1 that particle and mass fraction are 10% that sieves:4 mixing
Uniformly, it is placed in heater, design temperature is 105~110 DEG C, heats 20~30min, naturally cools to room temperature, use liquid
Chilled nitrogen machine is freeze-dried to it, obtains freeze-drying thing；In parts by weight meter take fermentation material, take 50~55 parts of distilled water,
20~25 parts of above-mentioned dried objects, 12~16 parts of glucose and 7~9 part of 150 mesh stone of a dog's gallbladder, kidney or bladder powder；Fermentation material is put into fermentation tank,
Lactic acid is added into fermentation tank again up to the mixture pH to 1.5~2.5 in fermentation tank, with 20~25mL/min into fermentation tank
Oxygen is filled with, sets fermentation jar temperature as 35~40 DEG C, 16~19h of fermentation is stirred with 180r/min, collects fermentation cylinder for fermentation
Mixture；Fermenting mixture in fermentation tank is put into filter press, press filtration is carried out with 20MPa, collects pressing filtering liquid, then by press filtration
Liquid is put into centrifuge, is centrifuged 5~10min with 900r/min, is collected supernatant；In mass ratio 95~100:14~16:2~4:
1.3~2.3, by supernatant, sodium chloride, purity be 99.4% titanium valve and 1, 8-Cineole be well mixed, be placed in 4~7 DEG C it is quiet
Put overnight, then filtered, collect filter residue；Filter residue and 0.1M and pH are pressed into quality 1 for 7 phosphate buffer:2 mixing are equal
It is even, it is put into the bag filter that molecular cut off is 7000~9000, dialysis temperature is 6~8 DEG C, is dialysed using 0.4mol/L acetic acid
24h, distilled water dialysis 48h is reused, the residue in bag filter is put into mould, and mould is placed in cooler, if
Constant temperature degree is -30~-25 DEG C, after freeze overnight, mould is taken out and is placed in sterilizing in ultraviolet sterilization device, is stripped, received
Collection demoulding thing, you can obtain collagen protein sponge.
Example 1
By air bladder and black fungus in mass ratio 5:1, mixing, which is put into 60 DEG C of drying boxes, dries 10h, then by dried mixture
It is put into pulverizer, 30min is crushed with 1100r/min, the crushed mixture in pulverizer is then crossed into 100 mesh sieves, collected
Sieve particle；By the sodium carbonate liquor in mass ratio 1 that particle and mass fraction are 10% that sieves:4 is well mixed, is placed in heater
In, design temperature is 105 DEG C, heats 20min, naturally cools to room temperature, it is freeze-dried using liquid nitrogen freezers, obtained
It is freeze-dried thing；Meter takes fermentation material in parts by weight, takes 50 parts of distilled water, 20 parts of above-mentioned dried objects, 12 parts of glucose and 7 parts
150 mesh stone of a dog's gallbladder, kidney or bladder powder；Fermentation material is put into fermentation tank, then lactic acid is added up to the mixture pH in fermentation tank into fermentation tank
To 1.5, oxygen is filled with into fermentation tank with 20mL/min, sets fermentation jar temperature as 35 DEG C, is stirred and fermented with 180r/min
16h, collect fermentation cylinder for fermentation mixture；Fermenting mixture in fermentation tank is put into filter press, pressed with 20MPa
Filter, pressing filtering liquid is collected, then pressing filtering liquid is put into centrifuge, 5min is centrifuged with 900r/min, collects supernatant；In mass ratio
95:14:2:1.3, by supernatant, sodium chloride, purity be 99.4% titanium valve and 1, 8-Cineole be well mixed, be placed in 4 DEG C it is quiet
Put overnight, then filtered, collect filter residue；Filter residue and 0.1M and pH are pressed into quality 1 for 7 phosphate buffer:2 mixing are equal
It is even, it is put into the bag filter that molecular cut off is 7000, dialysis temperature is 6 DEG C, using 0.4mol/L acetic acid dialysis 24h, then is made
Dialysed 48h, the residue in bag filter be put into mould, and mould is placed in cooler with distilled water, design temperature for-
30 DEG C, after freeze overnight, mould is taken out and is placed in sterilizing in ultraviolet sterilization device, is stripped, collect demoulding thing, you can
Collagen protein sponge.
Example 2
By air bladder and black fungus in mass ratio 7:1, mixing, which is put into 60 DEG C of drying boxes, dries 12h, then by dried mixture
It is put into pulverizer, 40min is crushed with 1100r/min, the crushed mixture in pulverizer is then crossed into 100 mesh sieves, collected
Sieve particle；By the sodium carbonate liquor in mass ratio 1 that particle and mass fraction are 10% that sieves:4 is well mixed, is placed in heater
In, design temperature is 110 DEG C, heats 30min, naturally cools to room temperature, it is freeze-dried using liquid nitrogen freezers, obtained
It is freeze-dried thing；In parts by weight meter take fermentation material, take 55 parts of distilled water, 25 parts of above-mentioned dried objects, 12 parts of yeast extracts, 16 parts
Glucose and 9 part of 150 mesh stone of a dog's gallbladder, kidney or bladder powder；Fermentation material is put into fermentation tank, then lactic acid is added until fermentation tank into fermentation tank
In mixture pH to 2.5, oxygen is filled with into fermentation tank with 25mL/min, fermentation jar temperature is set as 40 DEG C, with 180r/
Min stirring fermentation 19h, collect fermentation cylinder for fermentation mixture；Fermenting mixture in fermentation tank is put into filter press, with
20MPa carries out press filtration, collects pressing filtering liquid, then pressing filtering liquid is put into centrifuge, centrifuges 10min with 900r/min, collects supernatant
Liquid；In mass ratio 100:16:4:2.3, supernatant, sodium chloride, the titanium valve that purity is 99.4% and 1, 8-Cineole are mixed equal
It is even, it is placed in 7 DEG C and stands overnight, then filtered, collect filter residue；Filter residue and 0.1M and pH are pressed into matter for 7 phosphate buffer
Amount 1:2 is well mixed, is put into the bag filter that molecular cut off is 9000, and dialysis temperature is 8 DEG C, saturating using 0.4mol/L acetic acid
24h is analysed, distilled water dialysis 48h is reused, the residue in bag filter is put into mould, and mould is placed in cooler,
Design temperature is -25 DEG C, after freeze overnight, mould is taken out and is placed in sterilizing in ultraviolet sterilization device, is stripped, and is collected de-
Mould thing, you can obtain collagen protein sponge.
Example 3
By air bladder and black fungus in mass ratio 6:1, mixing, which is put into 60 DEG C of drying boxes, dries 11h, then by dried mixture
It is put into pulverizer, 35min is crushed with 1100r/min, the crushed mixture in pulverizer is then crossed into 100 mesh sieves, collected
Sieve particle；By the sodium carbonate liquor in mass ratio 1 that particle and mass fraction are 10% that sieves:4 is well mixed, is placed in heater
In, design temperature is 108 DEG C, heats 25min, naturally cools to room temperature, it is freeze-dried using liquid nitrogen freezers, obtained
It is freeze-dried thing；Meter takes fermentation material in parts by weight, takes 53 parts of distilled water, 23 parts of above-mentioned dried objects, 4 parts of glucose and 8 part 150
Mesh stone of a dog's gallbladder, kidney or bladder powder；Fermentation material is put into fermentation tank, then into fermentation tank add lactic acid until fermentation tank in mixture pH extremely
2.0, oxygen is filled with into fermentation tank with 23mL/min, sets fermentation jar temperature as 38 DEG C, is stirred and fermented with 180r/min
17.5h, collect fermentation cylinder for fermentation mixture；Fermenting mixture in fermentation tank is put into filter press, pressed with 20MPa
Filter, pressing filtering liquid is collected, then pressing filtering liquid is put into centrifuge, 8min is centrifuged with 900r/min, collects supernatant；In mass ratio
98:15:3:1.8, by supernatant, sodium chloride, purity be 99.4% titanium valve and 1, 8-Cineole be well mixed, be placed in 5 DEG C it is quiet
Put overnight, then filtered, collect filter residue；Filter residue and 0.1M and pH are pressed into quality 1 for 7 phosphate buffer:2 mixing are equal
It is even, it is put into the bag filter that molecular cut off is 8000, dialysis temperature is 7 DEG C, using 0.4mol/L acetic acid dialysis 24h, then is made
Dialysed 48h, the residue in bag filter be put into mould, and mould is placed in cooler with distilled water, design temperature for-
28 DEG C, after freeze overnight, mould is taken out and is placed in sterilizing in ultraviolet sterilization device, is stripped, collect demoulding thing, you can
Collagen protein sponge.
Collagen protein sponge currently in use is done into performance test, the results showed that：The water absorption rate of collagen is
42.3%, porosity 87.6%, bleeding stopping period 92s, expansion multiple 1.19, and by the collagen protein sponge in example 1~3
Performance test is done, as a result as shown in Table 1.
Table one
| Example | Water absorption rate/% | Porosity/% | Expansion multiple | Bleeding stopping period/s |
| 1 | 44.32 | 90.56 | 1.25 | 89 |
| 2 | 49.96 | 92.84 | 1.36 | 83 |
| 3 | 47.03 | 91.50 | 1.29 | 85 |
Claims (6)
- A kind of 1. method that collagen protein sponge is prepared based on air bladder, it is characterised in that specifically preparation process is：（1）By air bladder and black fungus in mass ratio 5:1~7:1 is well mixed, is dried, and crushes, and crosses 100 mesh sieves, collected Sieve particle；（2）By the sodium carbonate liquor in mass ratio 1 that particle and mass fraction are 10% that sieves:4, it is well mixed, 105~110 20~30min is heated at DEG C, is freeze-dried after naturally cooling to room temperature, obtains freeze-drying thing；（3）Meter takes fermentation material in parts by weight, takes 50~55 parts of distilled water, 20~25 parts of above-mentioned dried objects, 12~16 portions of grapes Sugar and 7~9 part of 150 mesh stone of a dog's gallbladder, kidney or bladder powder；（4）Fermentation material is put into fermentation tank, then into fermentation tank add lactic acid until fermentation tank in fermentation material pH to 1.5~ 2.5, oxygen is filled with into fermentation tank, ferment 16~19h, collects fermentation cylinder for fermentation mixture；（5）Fermenting mixture in fermentation tank is put into filter press press filtration is carried out with 20MPa, pressing filtering liquid is collected, to pressing filtering liquid It is centrifuged, collects supernatant；（6）Supernatant, sodium chloride, the titanium valve that purity is 99.4% and 1, 8-Cineole are well mixed, are placed in 4~7 DEG C of standings Overnight, then filtered, collect filter residue；（7）Filter residue and 0.1M and pH are pressed into quality 1 for 7 phosphate buffer:2 is well mixed, is put into bag filter, dialysis temperature Spend for 6~8 DEG C, using 0.4mol/L acetic acid dialysis 24h, reuse distilled water dialysis 48h, the residue in bag filter is put into In mould, and mould is placed in cooler, design temperature is -30~-25 DEG C, after freeze overnight, mould is taken out and is placed in purple Sterilizing in outer sterilizer, is stripped, and collects demoulding thing, you can obtain collagen protein sponge.
- A kind of 2. method that collagen protein sponge is prepared based on air bladder according to claim 1, it is characterised in that：The step Suddenly（3）Middle fermentation material also contains 9~12 parts of yeast extracts.
- A kind of 3. method that collagen protein sponge is prepared based on air bladder according to claim 1, it is characterised in that：The step Suddenly（4）Middle fermentation jar temperature is 35~40 DEG C.
- A kind of 4. method that collagen protein sponge is prepared based on air bladder according to claim 1, it is characterised in that：The step Suddenly（4）In to be filled with the speed of oxygen be 20~25mL/min.
- A kind of 5. method that collagen protein sponge is prepared based on air bladder according to claim 1, it is characterised in that：The step Suddenly（6）The titanium valve and the mass ratio of 1, 8-Cineole that middle supernatant, sodium chloride, purity are 99.4% are 95~100:14~16:2 ~4:1.3~2.3.
- A kind of 6. method that collagen protein sponge is prepared based on air bladder according to claim 1, it is characterised in that：The step Suddenly（7）The molecular cut off of middle bag filter is 7000~9000.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103007336A (en) * | 2013-01-14 | 2013-04-03 | 福建省博特生物科技有限公司 | Fish skin collagen based composite sponge and preparation method thereof |
| CN103205480A (en) * | 2013-04-15 | 2013-07-17 | 武汉工业学院 | Method for producing high-quality collagen oligopeptide by using fish skin or fishbone |
| CN103451258A (en) * | 2013-06-19 | 2013-12-18 | 浙江海洋学院 | Preparation technology of croaker fish bladder collagen with high anti-fatigue activity |
| CN104726527A (en) * | 2015-03-24 | 2015-06-24 | 宁德市夏威食品有限公司 | Preparation technology for producing collagen through enzymolysis of fish scales |
| CN105985996A (en) * | 2015-02-14 | 2016-10-05 | 上海拜瑞曼克生物科技有限公司 | Preparation method of collagens |
-
2017
- 2017-04-21 CN CN201710266808.8A patent/CN107412844A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103007336A (en) * | 2013-01-14 | 2013-04-03 | 福建省博特生物科技有限公司 | Fish skin collagen based composite sponge and preparation method thereof |
| CN103205480A (en) * | 2013-04-15 | 2013-07-17 | 武汉工业学院 | Method for producing high-quality collagen oligopeptide by using fish skin or fishbone |
| CN103451258A (en) * | 2013-06-19 | 2013-12-18 | 浙江海洋学院 | Preparation technology of croaker fish bladder collagen with high anti-fatigue activity |
| CN105985996A (en) * | 2015-02-14 | 2016-10-05 | 上海拜瑞曼克生物科技有限公司 | Preparation method of collagens |
| CN104726527A (en) * | 2015-03-24 | 2015-06-24 | 宁德市夏威食品有限公司 | Preparation technology for producing collagen through enzymolysis of fish scales |
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Application publication date: 20171201 |