CN106563157A - Preparation method of compound collagen sponge - Google Patents
Preparation method of compound collagen sponge Download PDFInfo
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- CN106563157A CN106563157A CN201610990442.4A CN201610990442A CN106563157A CN 106563157 A CN106563157 A CN 106563157A CN 201610990442 A CN201610990442 A CN 201610990442A CN 106563157 A CN106563157 A CN 106563157A
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Abstract
The invention discloses a preparation method of compound collagen sponge, belonging to the technical field of preparation of medicinal auxiliary materials. The preparation method of the compound collagen sponge comprises the following steps: removing scales on eel fish skin, carrying out degreasing aftertreatment, carrying out enzymolysis, carrying out salting out, and carrying out concentrating, thus obtaining eel fish skin collagen; fermenting acetobacter xylinum, thus preparing a bacterial cellulose membrane; and finally, pouring the eel fish skin collagen and the bacterial cellulose membrane into a mould together with citric acid solution for freeze-drying forming, thus obtaining the compound collagen sponge, wherein the content of collagen in the eel fish skin is very high, basically, no impure protein exists, collagen is firmly embedded in the net structure by adopting bacterial cellulose, and after freeze-drying, the density is increased, the mechanical property is enhanced, and deformation seldom occurs. The prepared compound collagen sponge has high density, the collagen molecules are combined tightly, the pore size and distribution are uniform, so that the compound collagen sponge has very high mechanical strength and good in-vivo and degradation stability, and in addition, the sponge in good in adhesion, has high shape integrity rate in the use process, and has the wide application prospect.
Description
Technical field
The invention discloses a kind of preparation method of composite collagen sponge, belongs to medical preparing technical field.
Background technology
Collagen protein is the main component of extracellular matrix, is that mammal in-vivo content is most, is distributed most wide function
Property albumen, collagen itself is excellent tissue engineering bracket material as a kind of fibrous protein, is the life being utilized earliest
One of thing material, was widely applied in recent years in wound repair field.
Collagen protein sponge main component is collagen protein, and it is similar to human collagen protein structure to be well suited for human organ
Reparation and regeneration.Collagen protein can promote the growth of wound healing and granulation tissue, with good hemostasis and filling effect,
Can be used for wound healing, hemostasis, operation residual cavity filling etc., collagen protein sponge has been acknowledged as a kind of the medical of high-quality
Material.
Be presently used for quick-acting haemostatic powder, repair in trauma and cavity filling commercialization medical collagen sponge material essentially from
The collagen protein extracted in mammal.But, the collagen resource of mammal source is limited and collagen protein is extracted,
Purifying procedure is loaded down with trivial details, medical collagen valuable product, significantly limit its range of application.Further, since there are people and animals being total to
Suffer from the disease（Such as bovine spongiform encephalopathy, foot and mouth disease）Potential risk, the collagen product in this kind of source is in terms of biological safety
Queried by user.In recent years, Fish collagen resource just progressively receives publicity.With traditional mammal collagen phase
Than Fish collagen is respectively provided with bigger advantage at aspects such as the popularity, biological safety and product cost originated.Therefore, enclose
Get most of the attention around the performance study of fish source collagen albumen and its application in field of biomedical materials.
Compared with marine fishes, freshwater fish collagen protein has bigger actual application value, one side fresh-water fishes money
Source is enriched, and collagen protein has more preferable thermal stability in another aspect fresh-water fishes, is particularly suited for needing to have complete three
The manufacture of the collagen biomaterial for medical purpose of helical molecular structure.Even so, as the filling of quick-acting haemostatic powder, repair in trauma and cavity
The freshwater fish Collagen sponges of effect, compared with mammal collagen, in heat stability, vivo degradation stability and body
The aspects such as interior material shape stability still suffer from obvious deficiency：As the Collagen sponges density after lyophilizing is less, easily formed compared with
Big, heterogeneity sponge aperture, therefore product mechanical performance is poor, and when under the conditions of human body temperature, thermal denaturation easily occur, inhale
The phenomenon such as the water-swellable intrinsic form forfeiture of rear material and internal fast degradation, causes that its biological function can not be given full play to, and
The tightness degree for improving and combining between the density size of collagen-based materials and tropocollagen molecule of these performances is closely related.Therefore,
The tightness degree combined between density and tropocollagen molecule that freshwater fish Collagen sponges are improved using technological means and then raising
Material property becomes the task of top priority for expanding freshwater fish collagen application to meet using needs.
The content of the invention
Present invention mainly solves technical problem：For existing collagen protein, density is less, machinery after sponge lyophilizing
Poor-performing, and when under the conditions of human body temperature, the intrinsic form of material after thermal denaturation, water absorption and swelling easily occur and lose and internal
Fast degradation phenomenon, causes that the defect of its biological function can not be given full play to, there is provided one kind is using collagen protein in anguilla japonica fish skin
It is combined the method for preparing collagen protein sponge with Bacterial cellulose, the present invention first scales on anguilla japonica fish skin, after defat post processing, then
Digest, saltout, concentrate, obtain anguilla japonica collagen protein of fish skin, then acetobacter xylinum is carried out into fermentation preparing bacteria cellulose film, finally
Pour mould lyophilizing shaping into citric acid solution etc. again, you can obtain composite collagen sponge, the collagen egg in anguilla japonica fish skin
Bai Hanliang is very high, almost without foreign protein, is firmly embedded in collagen protein in its network structure using Bacterial cellulose, is freezing
Density increase after dry, and mechanical performance enhancing, are unlikely to deform, and the obtained composite collagen sponge of the present invention has higher
Density, is tightly combined between tropocollagen molecule, even pore distribution, stable with higher mechanical strength and preferable vivo degradation
Property, and sponge adhesiveness is good, form percentage of head rice is high during use, has broad application prospects.
In order to solve above-mentioned technical problem, the technical solution adopted in the present invention is：
（1）200～300g anguilla japonica fish skin is taken, the fish skin after scaling is washed with deionized 3 by the fish scale on its surface of artificial removal
After～5 times, in being placed in beaker, 400～500mL mass fractions are added for 10% sodium hydroxide solution, stirring immersion 30～
40min post-heating boils 1～2h, makes fish skin defat, by after defat fish skin take out, be placed in meat grinder rub 5～
10min, obtains broken fish skin, and broken fish skin is added in enzymatic vessel, adds 1～2g pepsin, stirs at 35～40 DEG C
3～4h of enzymolysis, enzymolysis terminates, and collects the material in enzymatic vessel, in being placed in centrifuge, is centrifuged under 5000～6000r/min
20～30min, collects supernatant；
（2）Above-mentioned supernatant is added in beaker, 3～5g Sodium Chloride is added, after 5～10min of stirring, 1～2h is stood, is made
Collagen protein of fish skin is salted out and, and end of saltouing, liquid of saltouing is filtered, and collects filtering residue, and filtering residue is added to into interception for 12000
In the bag filter of～15000 dalton, with distilled water as dialysis solution, dialyse 24～26h at 0～4 DEG C, changes every 3～4h
Bag filter once, after the completion of dialysis, is placed in 6～8h of concentration in Polyethylene Glycol by dialysis solution, obtains anguilla japonica fish skin collagen concentrated solution,
And be spray-dried, dried object, i.e. anguilla japonica collagen protein of fish skin are obtained, it is standby；
（3）Weigh 10～20g sucrose, 6～8g peptones, 8～12g yeast powders, 3～5g dipotassium hydrogen phosphates, 0.3～0.5g Fructus Citri Limoniaes
Acid and 600～700mL deionized waters, in being added to culture dish, after stirring 3～5min of mixing, in moving into sterilization tank, 100～
At 110 DEG C after 5～10min of high temperature sterilize, in being transferred to fermentation tank, then acetobacter xylinum is inoculated into into fermentation by 8～10% inoculum concentrations
In tank, anaerobic fermentation 3～4 days, fermentation ends, bacteria cellulose film is taken out at 25～35 DEG C, and as in beaker, is added
After deionized water is by its submergence, then it is carried out to be heated to boiling, after 1～2h of insulation, bacteria cellulose film is taken out and spent
Ion water washing 3～5 times, obtains Bacterial cellulose；
（4）Weigh the above-mentioned Bacterial cellulose of 100～200g, 50～60g steps（2）Standby anguilla japonica collagen protein of fish skin and 3～
5g xanthan gum is added in beaker, adds 300～400g mass fractions for 1% citric acid solution, 5～10min of magnetic agitation
Afterwards, pour in mould, 24～26h of lyophilization at -45～-35 DEG C, you can obtain composite collagen sponge.
The present invention application process be：Patient's wound is cleaned and routine disinfection first, will be multiple obtained in the present invention
Close collagen protein sponge and blood oozing from the wound surface region is completely covered, and gently press sponge it is closely attached with wound surface, while sponge edge
0.5～0.8cm of edge of wound need to slightly be exceeded, subsequently daily replacing sponge 1 time, with 7 days as a course for the treatment of, while visual wound healing feelings
Condition, proper extension sponge uses the time instead 2～3 days, until wound heals completely.After testing, the obtained composite collagen egg of the present invention
Bai Haimian density is 24～26mg/cm3, aperture size is 105～120 μm, and aperture is consistent, is evenly distributed, in vivo using after 24h
It is 50～60% that sponge form percentage of head rice is 92～99%, 24h vitro enzymes degradation rate, and stability to degradation is preferable, sponge thermal deformation temperature
Spend for 45～50 DEG C, with preferable heat stability, and collagen protein sponge of the present invention has stronger water absorbing capacity, water suction times
Number is 50～60 times of sponge itself weight.
The invention has the beneficial effects as follows：
（1）Obtained composite collagen sponge of the invention has preferably internal stability, resistance to enzymatic degradation performance and material
Morphological stability, while water absorbing capacity and swelliong power are preferably, form percentage of head rice is high during use, and efficiently solves product
In chemical residual problem, Product Safety is higher；
（2）Obtained composite collagen sponge of the invention has good porous form structure and higher density, and quality is soft
Soft, high resilience is bent, effectively increases its drug permeability, while its adhesiveness is preferably, collagen content, water suction
Multiplying power is higher, with preferable anastalsis, improves clinical application effect.
Specific embodiment
200～300g anguilla japonica fish skin, the fish scale on its surface of artificial removal, the fish skin deionized water after scaling are taken first
After washing 3～5 times, in being placed in beaker, 400～500mL mass fractions are added for 10% sodium hydroxide solution, stirring immersion 30
～40min post-heating boils 1～2h, makes fish skin defat, by after defat fish skin take out, be placed in meat grinder rub 5～
10min, obtains broken fish skin, and broken fish skin is added in enzymatic vessel, adds 1～2g pepsin, stirs at 35～40 DEG C
3～4h of enzymolysis, enzymolysis terminates, and collects the material in enzymatic vessel, in being placed in centrifuge, is centrifuged under 5000～6000r/min
20～30min, collects supernatant；Subsequently above-mentioned supernatant is added in beaker, 3～5g Sodium Chloride is added, stirring 5～
After 10min, 1～2h is stood, salt out collagen protein of fish skin and come, end of saltouing, liquid of saltouing is filtered, and collects filtering residue, will be filtered
Slag is added in the bag filter that interception is 12000～15000 dalton, with distilled water as dialysis solution, is dialysed at 0～4 DEG C
24～26h, dialysis solution is changed once every 3～4h, after the completion of dialysis, bag filter is placed in into 6～8h of concentration in Polyethylene Glycol, is obtained
To anguilla japonica fish skin collagen concentrated solution, and it is spray-dried, is obtained dried object, i.e. anguilla japonica collagen protein of fish skin, it is standby；Again
Weigh 10～20g sucrose, 6～8g peptones, 8～12g yeast powders, 3～5g dipotassium hydrogen phosphates, 0.3～0.5g citric acids and 600
～700mL deionized waters, in being added to culture dish, after stirring 3～5min of mixing, in moving into sterilization tank, at 100～110 DEG C
After 5～10min of high temperature sterilize, in being transferred to fermentation tank, then acetobacter xylinum is inoculated in fermentation tank by 8～10% inoculum concentrations,
Anaerobic fermentation 3～4 days, fermentation ends, bacteria cellulose film is taken out at 25～35 DEG C, and as in beaker, adds deionization
After water is by its submergence, then it is carried out to be heated to boiling, after 1～2h of insulation, bacteria cellulose film is taken out and deionized water
Washing 3～5 times, obtains Bacterial cellulose；Finally weigh the above-mentioned Bacterial cellulose of 100～200g, 50～60g above-mentioned steps standby
Anguilla japonica collagen protein of fish skin and 3～5g xanthan gum be added in beaker, add 300～400g mass fractions for 1% citric acid
Solution, after 5～10min of magnetic agitation, in pouring mould into, 24～26h of lyophilization at -45～-35 DEG C, you can be combined
Collagen protein sponge.
Example 1
200g anguilla japonica fish skin is taken first, and the fish skin after scaling is washed with deionized 3 times by the fish scale on its surface of artificial removal
Afterwards, it is placed in beaker, it is 10% sodium hydroxide solution to add 400mL mass fractions, and stirring immersion 30min post-heating boils 1h,
Fish skin defat is made, the fish skin after defat is taken out, be placed in rubbing 5min in meat grinder, obtain broken fish skin, broken fish skin is added
To in enzymatic vessel, 1g pepsin is added, the stirring enzymolysis 3h at 35 DEG C, enzymolysis terminates, and collects the material in enzymatic vessel, and
It is placed in a centrifuge, 20min is centrifuged under 5000r/min, collects supernatant；Subsequently above-mentioned supernatant is added in beaker,
3g Sodium Chloride is added, after stirring 5min, 1h is stood, collagen protein of fish skin is salted out and is come, end of saltouing, liquid mistake of saltouing
Filter, collects filtering residue, filtering residue is added in the bag filter that interception is 12000 dalton, with distilled water as dialysis solution, in 0 DEG C
Lower dialysis 24h, dialysis solution is changed once every 3h, after the completion of dialysis, bag filter is placed in Polyethylene Glycol and concentrates 6h, obtains eel
Eel fish skin collagen concentrated solution, and be spray-dried, dried object, i.e. anguilla japonica collagen protein of fish skin are obtained, it is standby；Weigh again
10g sucrose, 6g peptones, 8g yeast powders, 3g dipotassium hydrogen phosphates, 0.3g citric acids and 600mL deionized waters, are added to culture dish
In, after stirring mixing 3min, in moving into sterilization tank, after high temperature sterilize 5min at 100 DEG C, in being transferred to fermentation tank, then by 8%
Inoculum concentration is inoculated into acetobacter xylinum in fermentation tank, and anaerobic fermentation 3 days, fermentation ends, bacteria cellulose film is taken at 25 DEG C
Go out, and as in beaker, after addition deionized water is by its submergence, then it is carried out to be heated to boiling, it is after insulation 1h, antibacterial is fine
The plain film of dimension takes out and is washed with deionized 3 times, obtains Bacterial cellulose；Finally weigh on the above-mentioned Bacterial cellulose of 100g, 50g
State the standby anguilla japonica collagen protein of fish skin of step and 3g xanthan gum is added in beaker, add 300g mass fractions for 1% Fructus Citri Limoniae
Acid solution, after magnetic agitation 5min, in pouring mould into, the lyophilization 24h at -45 DEG C, you can obtain composite collagen sea
It is continuous.
This example operation is easy, when using, patient's wound is cleaned and routine disinfection first, by obtained in the present invention
Composite collagen sponge is completely covered blood oozing from the wound surface region, and gently presses sponge it is closely attached with wound surface, while sponge side
Edge need to slightly exceed edge of wound 0.5cm, subsequently daily replacing sponge 1 time, with 7 days as a course for the treatment of, while visual wound healing situation,
Proper extension sponge uses the time instead 2 days, until wound heals completely.After testing, the obtained composite collagen sponge of the present invention
Density is 24mg/cm3, aperture size is 105 μm, and aperture is consistent, is evenly distributed, in vivo using sponge form percentage of head rice after 24h
It is 50% for 92%, 24h vitro enzymes degradation rate, preferably, sponge heat distortion temperature is 45 DEG C to stability to degradation, steady with preferable heat
It is qualitative, and collagen protein sponge of the present invention has stronger water absorbing capacity, Water absorption is 50 times of sponge itself weight.
Example 2
250g anguilla japonica fish skin is taken first, and the fish skin after scaling is washed with deionized 4 times by the fish scale on its surface of artificial removal
Afterwards, it is placed in beaker, it is 10% sodium hydroxide solution to add 450mL mass fractions, and stirring immersion 35min post-heating boils 1h,
Fish skin defat is made, the fish skin after defat is taken out, be placed in rubbing 8min in meat grinder, obtain broken fish skin, broken fish skin is added
To in enzymatic vessel, 1g pepsin is added, the stirring enzymolysis 3h at 38 DEG C, enzymolysis terminates, and collects the material in enzymatic vessel, and
It is placed in a centrifuge, 25min is centrifuged under 5500r/min, collects supernatant；Subsequently above-mentioned supernatant is added in beaker,
4g Sodium Chloride is added, after stirring 8min, 1h is stood, collagen protein of fish skin is salted out and is come, end of saltouing, liquid mistake of saltouing
Filter, collects filtering residue, filtering residue is added in the bag filter that interception is 13500 dalton, with distilled water as dialysis solution, in 2 DEG C
Lower dialysis 25h, dialysis solution is changed once every 3h, after the completion of dialysis, bag filter is placed in Polyethylene Glycol and concentrates 7h, obtains eel
Eel fish skin collagen concentrated solution, and be spray-dried, dried object, i.e. anguilla japonica collagen protein of fish skin are obtained, it is standby；Weigh again
15g sucrose, 7g peptones, 10g yeast powders, 4g dipotassium hydrogen phosphates, 0.4g citric acids and 650mL deionized waters, are added to culture
In ware, after stirring mixing 4min, in moving into sterilization tank, after high temperature sterilize 8min at 105 DEG C, in being transferred to fermentation tank, then press
9% inoculum concentration is inoculated into acetobacter xylinum in fermentation tank, anaerobic fermentation 3 days at 30 DEG C, fermentation ends, by bacteria cellulose film
Take out, and as in beaker, after addition deionized water is by its submergence, then it is carried out to be heated to boiling, after insulation 1h, by antibacterial
Cellulose membrane takes out and is washed with deionized 4 times, obtains Bacterial cellulose；Finally weigh the above-mentioned Bacterial cellulose of 150g, 55g
The standby anguilla japonica collagen protein of fish skin of above-mentioned steps and 4g xanthan gum are added in beaker, add 350g mass fractions for 1% lemon
Lemon acid solution, after magnetic agitation 8min, in pouring mould into, the lyophilization 25h at -40 DEG C, you can obtain composite collagen
Sponge.
This example operation is easy, when using, patient's wound is cleaned and routine disinfection first, by obtained in the present invention
Composite collagen sponge is completely covered blood oozing from the wound surface region, and gently presses sponge it is closely attached with wound surface, while sponge side
Edge need to slightly exceed edge of wound 0.7cm, subsequently daily replacing sponge 1 time, with 7 days as a course for the treatment of, while visual wound healing situation,
Proper extension sponge uses the time instead 2 days, until wound heals completely.After testing, the obtained composite collagen sponge of the present invention
Density is 25mg/cm3, aperture size is 113 μm, and aperture is consistent, is evenly distributed, in vivo using sponge form percentage of head rice after 24h
It is 55% for 95%, 24h vitro enzymes degradation rate, preferably, sponge heat distortion temperature is 48 DEG C to stability to degradation, steady with preferable heat
It is qualitative, and collagen protein sponge of the present invention has stronger water absorbing capacity, Water absorption is 55 times of sponge itself weight.
Example 3
300g anguilla japonica fish skin is taken first, and the fish skin after scaling is washed with deionized 5 times by the fish scale on its surface of artificial removal
Afterwards, it is placed in beaker, it is 10% sodium hydroxide solution to add 500mL mass fractions, and stirring immersion 40min post-heating boils 2h,
Fish skin defat is made, the fish skin after defat is taken out, be placed in rubbing 10min in meat grinder, obtain broken fish skin, broken fish skin is added
To in enzymatic vessel, 2g pepsin is added, the stirring enzymolysis 4h at 40 DEG C, enzymolysis terminates, and collects the material in enzymatic vessel, and
It is placed in a centrifuge, 30min is centrifuged under 6000r/min, collects supernatant；Subsequently above-mentioned supernatant is added in beaker,
5g Sodium Chloride is added, after stirring 10min, 2h is stood, collagen protein of fish skin is salted out and is come, end of saltouing, liquid mistake of saltouing
Filter, collects filtering residue, filtering residue is added in the bag filter that interception is 15000 dalton, with distilled water as dialysis solution, in 4 DEG C
Lower dialysis 26h, dialysis solution is changed once every 4h, after the completion of dialysis, bag filter is placed in Polyethylene Glycol and concentrates 8h, obtains eel
Eel fish skin collagen concentrated solution, and be spray-dried, dried object, i.e. anguilla japonica collagen protein of fish skin are obtained, it is standby；Weigh again
20g sucrose, 8g peptones, 12g yeast powders, 5g dipotassium hydrogen phosphates, 0.5g citric acids and 700mL deionized waters, are added to culture
In ware, after stirring mixing 5min, in moving into sterilization tank, after high temperature sterilize 10min at 110 DEG C, in being transferred to fermentation tank, then press
10% inoculum concentration is inoculated into acetobacter xylinum in fermentation tank, anaerobic fermentation 4 days at 35 DEG C, fermentation ends, by bacteria cellulose film
Take out, and as in beaker, after addition deionized water is by its submergence, then it is carried out to be heated to boiling, after insulation 2h, by antibacterial
Cellulose membrane takes out and is washed with deionized 5 times, obtains Bacterial cellulose；Finally weigh the above-mentioned Bacterial cellulose of 200g, 60g
The standby anguilla japonica collagen protein of fish skin of above-mentioned steps and 5g xanthan gum are added in beaker, add 400g mass fractions for 1% lemon
Lemon acid solution, after magnetic agitation 10min, in pouring mould into, the lyophilization 26h at -35 DEG C, you can obtain composite collagen
Sponge.
This example operation is easy, when using, patient's wound is cleaned and routine disinfection first, by obtained in the present invention
Composite collagen sponge is completely covered blood oozing from the wound surface region, and gently presses sponge it is closely attached with wound surface, while sponge side
Edge need to slightly exceed edge of wound 0.8cm, subsequently daily replacing sponge 1 time, with 7 days as a course for the treatment of, while visual wound healing situation,
Proper extension sponge uses the time instead 3 days, until wound heals completely.After testing, the obtained composite collagen sponge of the present invention
Density is 26mg/cm3, aperture size is 120 μm, and aperture is consistent, is evenly distributed, in vivo using sponge form percentage of head rice after 24h
It is 60% for 99%, 24h vitro enzymes degradation rate, preferably, sponge heat distortion temperature is 50 DEG C to stability to degradation, steady with preferable heat
It is qualitative, and collagen protein sponge of the present invention has stronger water absorbing capacity, Water absorption is 60 times of sponge itself weight.
Claims (1)
1. a kind of preparation method of composite collagen sponge, it is characterised in that concrete preparation process is：
（1）200～300g anguilla japonica fish skin is taken, the fish skin after scaling is washed with deionized 3 by the fish scale on its surface of artificial removal
After～5 times, in being placed in beaker, 400～500mL mass fractions are added for 10% sodium hydroxide solution, stirring immersion 30～
40min post-heating boils 1～2h, makes fish skin defat, by after defat fish skin take out, be placed in meat grinder rub 5～
10min, obtains broken fish skin, and broken fish skin is added in enzymatic vessel, adds 1～2g pepsin, stirs at 35～40 DEG C
3～4h of enzymolysis, enzymolysis terminates, and collects the material in enzymatic vessel, in being placed in centrifuge, is centrifuged under 5000～6000r/min
20～30min, collects supernatant；
（2）Above-mentioned supernatant is added in beaker, 3～5g Sodium Chloride is added, after 5～10min of stirring, 1～2h is stood, is made
Collagen protein of fish skin is salted out and, and end of saltouing, liquid of saltouing is filtered, and collects filtering residue, and filtering residue is added to into interception for 12000
In the bag filter of～15000 dalton, with distilled water as dialysis solution, dialyse 24～26h at 0～4 DEG C, changes every 3～4h
Bag filter once, after the completion of dialysis, is placed in 6～8h of concentration in Polyethylene Glycol by dialysis solution, obtains anguilla japonica fish skin collagen concentrated solution,
And be spray-dried, dried object, i.e. anguilla japonica collagen protein of fish skin are obtained, it is standby；
（3）Weigh 10～20g sucrose, 6～8g peptones, 8～12g yeast powders, 3～5g dipotassium hydrogen phosphates, 0.3～0.5g Fructus Citri Limoniaes
Acid and 600～700mL deionized waters, in being added to culture dish, after stirring 3～5min of mixing, in moving into sterilization tank, 100～
At 110 DEG C after 5～10min of high temperature sterilize, in being transferred to fermentation tank, then acetobacter xylinum is inoculated into into fermentation by 8～10% inoculum concentrations
In tank, anaerobic fermentation 3～4 days, fermentation ends, bacteria cellulose film is taken out at 25～35 DEG C, and as in beaker, is added
After deionized water is by its submergence, then it is carried out to be heated to boiling, after 1～2h of insulation, bacteria cellulose film is taken out and spent
Ion water washing 3～5 times, obtains Bacterial cellulose；
（4）Weigh the above-mentioned Bacterial cellulose of 100～200g, 50～60g steps（2）Standby anguilla japonica collagen protein of fish skin and 3～
5g xanthan gum is added in beaker, adds 300～400g mass fractions for 1% citric acid solution, 5～10min of magnetic agitation
Afterwards, pour in mould, 24～26h of lyophilization at -45～-35 DEG C, you can obtain composite collagen sponge.
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| CN201610990442.4A CN106563157A (en) | 2016-11-10 | 2016-11-10 | Preparation method of compound collagen sponge |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108752619A (en) * | 2018-05-30 | 2018-11-06 | 广东医科大学 | A kind of simple and easy method improving collagen protein sponge mechanical performance |
| CN108904861A (en) * | 2018-07-23 | 2018-11-30 | 天津市长江医疗器械有限公司 | Collagen protein sponge promotes application and its physicochemical property detection method in wound healing drug in preparation |
| CN110665053A (en) * | 2019-10-31 | 2020-01-10 | 河南亚都实业有限公司 | Medical composite collagen sponge dressing |
-
2016
- 2016-11-10 CN CN201610990442.4A patent/CN106563157A/en not_active Withdrawn
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108752619A (en) * | 2018-05-30 | 2018-11-06 | 广东医科大学 | A kind of simple and easy method improving collagen protein sponge mechanical performance |
| CN108904861A (en) * | 2018-07-23 | 2018-11-30 | 天津市长江医疗器械有限公司 | Collagen protein sponge promotes application and its physicochemical property detection method in wound healing drug in preparation |
| CN110665053A (en) * | 2019-10-31 | 2020-01-10 | 河南亚都实业有限公司 | Medical composite collagen sponge dressing |
| CN110665053B (en) * | 2019-10-31 | 2022-04-15 | 瑞研创展(清远)科技有限公司 | Medical composite collagen sponge dressing |
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Application publication date: 20170419 |