CN102352051A - Method for preparing collagen-modified bacteria cellulose compound film - Google Patents
Method for preparing collagen-modified bacteria cellulose compound film Download PDFInfo
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Abstract
The invention provides a method for preparing a collagen-modified bacteria cellulose compound film, which relates to relevant fields of biomedical medical materials, medical appliances and cosmetics. The preparation method comprises the following steps of: pretreating and purifying a bacteria cellulose film to obtain purified bacteria cellulose, and functionalizing the hydroxyl of the purified bacteria cellulose; and physically compounding or chemically combining the bacteria cellulose film and a collagen suspension to obtain a collagen-modified bacteria cellulose compound film. Due to the adoption of the collagen-modified bacteria cellulose compound film obtained with the method, the recovery of collagen content in defect tissues can be promoted, the problems of poor water permeability, poor air permeability, poor drainage and poor absorbing effect existing in the conventional collagen medicament are solved simultaneously; a novel method for collagen application is provided, high cell affinity is achieved, and the tissue growth is facilitated. The method has a simple preparation process and a mature process, and is suitable for industrial production of collagen-modified bacteria cellulose compound films with different requirements.
Description
Technical field
The present invention relates to bio-medical material, medicine equipment and makeup association area; Be particularly related to a kind of collagen and modify the preparation method of bacteria cellulose composite membrane, be used to guide fields such as growing film, soft-tissue patch, short skin healing wound dressings, organization engineering skin and health care facial mask.
Background technology
Collagen is to twist into spiral fibrous protein by 3 peptide chains.Exist with the collegen filament form in vivo, glue promptly is dissolved in heating in water.Account for human body protein always more than 1/3.Mainly being distributed in the reticular tissue, is collagen like 70% of skin dry weight.During the human limb activity, collagen protein performance function makes skin have defencive function in the skin, and suitable elasticity and flintiness are arranged again.I, III collagen type are the main types of skin collagen, and both form collagen fibril jointly, and wherein the I Collagen Type VI is a main body, and the III Collagen Type VI holds and periphery.
Find during the detection of skin traumatic wounds that the wound fibroblast contacts prompting I Collagen Type VI with collegen filament and III collagen type product reduces, simultaneously the increased activity of expression of gene and collagenase.In in the reasonable time of after the wound healing, in addition one-year age, tissue tension intensity continues to increase, and collagen content recovers normal level gradually.Therefore, collagen protein replenishes wound repair and improves aging skin function aspect irreplaceable effect is arranged.The application of collagen on skin at present mainly contains transdermal, oral and inject two kinds of methods, and oral mainly the absorption with healthcare products through the food approach is main, and the injection collagen protein is because the cost height is mainly used in beauty industry.What be widely used in wound dressings and facial mask is that transdermal uses, and is main to apply and to stick, and wherein the collagen protein of Tu Fuing very fast effect time length of running off lacks, and the collagen film that sticks use mainly contains pure collagem membrane and composite collagen film.The cost of pure collagem membrane is higher, and the key of composite collagen film is to find suitable matrix membrane, and excellent matrix membrane can provide good wound repairing environment or be used for the high humidity environment of facial mask.
Bacteria cellulose (BC) is claimed micro organism cellulose again.Bacteria cellulose is a kind of natural biopolymer, passes through β-1, a kind of branchiess macromole straight-chain polymer that the 4-glycosidic link is formed by connecting by β-D-glucose.Biologically active, biodegradability, biological fitness have physics, chemistry and the mechanical property of many uniquenesses such as high-crystallinity, high purity, high retentiveness, superfine nano fibre network, high-tensile and Young's modulus.The bacteria cellulose retentiveness is high in addition, and its inside has a lot " duct ", and high ventilation, water permeability are arranged, and can absorb 60-700 doubly to the moisture of its dry weight, and have high wet strength.
Based on the excellent properties of above-mentioned bacteria cellulose, it is with a wide range of applications in biomedical materials field, has had research institution to release the bio-medical goods of some bacteria celluloses abroad at present.Produce artificial skin with bacteria cellulose, the high mechanical strength that under moisture conditions, is had, to the hypertonicity of gas and liquid, these indexs all are better than conventional skin substitute products.This shows that bacteria cellulose film is the excellent matrix membrane that collagen is used for wound and cosmetic use.
Verified to the surface of a wound provides suitable moist environment in a large number over nearly 40 years and helped the healing of wound about the fundamental research of the surface of a wound and clinical treatment.New pattern compress can keep the moistening of the surface of a wound.Can promote the healing of wound again.At present, new pattern compress uses comparatively universally in developed country, and domestic also the beginning is applied in the treatment of the various acute and chronic surface of a wound.The new type functional medical dressing will be the WeiLai Technology main development tendency, some new medical dressing, and for example silver-containing antibacterial medical dressing, biological active dressing, artificial skin etc. have begun to use or be about to come into operation abroad.Present domestic dressing industry kind is single, does not form series product, and functional shortcoming, is main with low-end product, on expensive goods, lacks competitiveness.
Characteristics to collagen protein and bacterial fibers have problems with domestic and international wound dressings separately; The present invention adopts the wound dressings that composite collagen makes in the bacteria cellulose matrix such as solution dipping method and surface coated method; Can replenish the I Collagen Type VI and the III collagen type that lack during the wound healing, thereby promote the healing of wound better.In addition, the collagen modification bacteria cellulose composite membrane of this method preparation also can be used for guiding fields such as growing film, soft-tissue patch, organization engineering skin and health care facial mask.
Summary of the invention
The objective of the invention is to propose the preparation method that a kind of collagen is modified bacteria cellulose composite membrane; The collagen that this method is prepared is modified bacteria cellulose composite membrane and is had excellent mechanical property and good permeable, ventilative or drainage, moisture sorption effect; Simultaneously can promote wound healing, be used to guide fields such as growing film, soft-tissue patch, short skin healing wound dressings, organization engineering skin and health care facial mask.
Technical scheme of the present invention is: prepares a kind of collagen and modifies bacteria cellulose composite membrane, be attached with collagen protein on the Nano grade fibrillar meshwork structure of bacteria cellulose, but even coated bacteria cellulosic fibre.In the surface of bacteria cellulose and network structure, form collagen layer through physics or chemical process; To be collagen protein deposit through physical adsorption in the flourishing surface of bacteria cellulose and network structure the physical method ultimate principle, and the ultimate principle of chemical reaction is aldehyde radical or carboxyl formation chemical bond in collagen protein residue and the functionalization bacteria cellulose film.
The present invention realizes the preparation of collagen modification bacteria cellulose composite membrane by the following method.
Its preparation method may further comprise the steps:
The pre-treatment of step 1, bacteria cellulose film:
Get bacteria cellulose and repeatedly wash, remove striping surface medium and impurity with clear water.Again film is soaked in the NaOH solution of 0.1mol/L, 80 ~ 100 ℃ of following water-bath 60min or more than, wash out thalline and residual substratum in the bacteria cellulose film.Bacteria cellulose film is soaked in the NaOH solution of 0.05mol/L, 80 ~ 100 ℃ of following water-bath 30-60min repeatedly wash with distilled water then again;
The purifying of step 2, bacteria cellulose film:
With distilled water immersion 3-5 time, with the light press mold survey of pH test paper pH value, finally controlling the bacteria cellulose pH value is 7.2 ± 0.2 with the bacteria cellulose film after the above-mentioned pre-treatment, obtains the bacteria cellulose (being designated as BC) behind the purifying, In Shade afterwards preservation;
Step 3, bacteria cellulose radical functino:
Select a series of oxidation systems that are applicable to cellulose materials such as TEMPO, Periodic acid or periodate, nitrogen peroxide etc. for use; Hydroxyl oxygen on the bacteria cellulose is changed into carboxyl or aldehyde radical; With deionized water soaking flushing or remove residual oxidizer repeatedly, obtain the bacteria cellulose film (being designated as OBC) of radical functino then with ion exchange method;
The adjustment of step 4, bacteria cellulose water content and sterilization process:
Pressure with 20-1000N dewaters bacteria cellulose (BC and OBC); Obtain the bacteria cellulose film of different moisture content; Then bacteria cellulose film is passed through high temperature steaming, ultraviolet ray or Co60 radiation sterilization, reach the technology sterilization and require (processing requirement aseptic technique afterwards).
Step 5, collagen are modified the recombining process of bacteria cellulose composite membrane:
Method 1 physics composite algorithm: the recombining process of collagen and BC
The collagen product is made into the solution of 0.01%-20% respectively, above-mentioned collagen solution is coated on BC film surface, the unit coating amount is 10-200 μ L/cm
2Or the collagen product is made into the solution of 0.01%-20% respectively, and above-mentioned BC film is dipped in 1s-24h in the collagen solution, make the physics composite collagen and modify bacteria cellulose composite membrane.
Method 2 chemically composited methods: the recombining process of collagen and OBC
The collagen product is made into the solution of 0.01%-20% respectively, above-mentioned collagen solution is coated on OBC film surface, the unit coating amount is 10-200 μ L/cm
2Or the collagen product is made into the solution of 0.01%-20% respectively, and above-mentioned OBC is dipped in 1s-24h in the collagen solution, make chemically composited collagen and modify bacteria cellulose composite membrane.
Aldehyde radical or carboxyl in the collagen modification bacteria cellulose composite membrane that method 2 makes in collagen protein residue and the functionalization bacteria cellulose film form chemical bond, reach stable Chemical bond.
Step 6, the collagen that will obtain through step 5 are modified bacteria cellulose composite membrane and carried out drying treatment, obtain collagen and modify the compound dry film of bacteria cellulose; Perhaps will modify the bacteria cellulose composite membrane dehydration through the collagen that step 5 obtains, adjustment obtains the compound wet film of collagen modification bacteria cellulose of different water content once more.
Said bacteria cellulose film is to belong in the bacteriums certain by genus acetobacter, Sarcina, Rhodopseudomonas, rhizobium, achromobacter, Alcaligenes, aerobacter, Azotobacter, Agrobacterium etc. nine to produce through fermentation, and the bacteria cellulose that produces has three-dimensional porous network structure.
The method of the drying treatment in the method one said step 5 comprises following 3 kinds of methods:
Normal temperature or 20 ℃ ~ 60 ℃ following freeze-day with constant temperature are handled under a, the ventilation state;
B, with bacteria cellulose drying treatment in-20 ℃ ~-60 ℃ freeze drier, or vacuum drying treatment;
C, the bacterial cellulose wet-coating chilling handled after, drying treatment under Vacuumdrier, thermostat container or normal temperature obtains the lower collagen of fragility and modifies bacteria cellulose composite membrane again.
Collagen by the present invention prepares is modified bacteria cellulose composite membrane, is attached with collagen on the nano-scale fiber network structure of bacteria cellulose, and the collagen content of described composite membrane is the 0.1-20% of gross weight.
Compared with prior art, the present invention has following advantage and beneficial effect:
1, collagen that the present invention obtains is modified bacteria cellulose composite membrane, can promote the recovery of collagen content in the damaged skin;
2, the present invention utilizes unique texture and the character that bacteria cellulose has, and preparation contains the bacteria cellulose film of collagen, make bacteria cellulose obtain excellent biocompatibility and trauma repair ability, and the content of collagen is controlled;
3, the present invention adopts unique recombining process to prepare collagen and modifies bacteria cellulose composite membrane; Permeable, the ventilative or drainage of existing collagen medicament existence, the problem of moisture sorption effect difference have been overcome; The novel method that provides collagen on burn trauma, to use, cellular affinity is good, promotes the wound growth;
4, the chemically composited legal system collagen that is equipped with gained is modified in the bacteria cellulose composite membrane, and the residue in the collagen protein combines through chemical bonds such as imine linkages with functionalization group in the bacteria cellulose, and it is compound more firm.Imine linkage can rupture under certain condition simultaneously, thereby makes collagen reach the purpose of sustained release;
5, preparation process of the present invention is simple; Technical maturity; Be suitable for the industrial production of the collagen modification bacteria cellulose composite membrane of different demands, can be used for guiding fields such as growing film, soft-tissue patch, short skin healing wound dressings, organization engineering skin and health care facial mask.
Description of drawings
Fig. 1 is the micro-structure diagram of bacteria cellulose, has shown the unique nanometer network structure of bacteria cellulose;
Fig. 2 has shown collagen good binding situation in the bacteria cellulose network for the micro-structure diagram of collagen modification bacteria cellulose composite membrane.
Embodiment
Below in conjunction with accompanying drawing, the preparation method who a kind of collagen of the present invention is modified bacteria cellulose composite membrane through specific examples further specifies.
Embodiment one
The commercially available bacteria cellulose film clip that is made by acetobacter xylinum is become the bacteria cellulose sample of 10 * 10cm, and the preparation process that collagen is modified bacteria cellulose composite membrane is following:
The pre-treatment of step 1, bacteria cellulose film:
Get bacteria cellulose and repeatedly wash, remove striping surface medium and impurity with clear water.Again film is soaked in the NaOH solution of 0.1mol/L, more than 90 ℃ of following water-bath 60min, washes out thalline and residual substratum in the bacteria cellulose film.Bacteria cellulose film is soaked in the NaOH solution of 0.05mol/L, 90 ℃ of following water-bath 30min repeatedly wash with distilled water then again;
The purifying of step 2, bacteria cellulose film:
With distilled water immersion 5 times, with the light press mold survey of pH test paper pH value, finally controlling the bacteria cellulose pH value is 7.2 ± 0.2 with the bacteria cellulose film after the above-mentioned pre-treatment, obtains the bacteria cellulose behind the purifying, In Shade afterwards preservation.
The adjustment of step 3, bacteria cellulose water content and sterilization process:
After the bacteria cellulose pre-treatment is adjusted to neutrality; Pressure with 200N dewaters bacteria cellulose; Obtain the bacteria cellulose film of different moisture content, then bacteria cellulose film is sterilized through high temperature steaming, reach the technology sterilization and require (processing requirement aseptic technique afterwards).
The recombining process of step 4, collagen and bacteria cellulose:
The collagen product is made into 1% solution, above-mentioned collagen solution is coated on the bacteria cellulose film surface, the unit coating amount is 20 μ L/cm
2
Step 5, the collagen that will obtain through step 4 are modified bacteria cellulose composite membrane 20 ℃ of freeze-day with constant temperature under ventilation state and are handled, and obtain collagen and modify the compound dry film of bacteria cellulose composite membrane.
Modify bacteria cellulose composite membrane by the collagen that a process for preparing, be attached with collagen on the nano-scale fiber network structure of bacteria cellulose, the collagen content of described composite membrane is 1.2% of a gross weight, and water content is 63.7% of an original bulk.
Embodiment two
The commercially available bacteria cellulose film clip that is made by acetobacter xylinum is become the bacteria cellulose sample of 10 * 10cm, and the preparation process that collagen is modified bacteria cellulose composite membrane is following:
The pre-treatment of step 1, bacteria cellulose film:
Get bacteria cellulose and repeatedly wash, remove striping surface medium and impurity with clear water.Again film is soaked in the NaOH solution of 0.1mol/L, more than 80 ℃ of following water-bath 60min, washes out thalline and residual substratum in the bacteria cellulose film.Bacteria cellulose film is soaked in the NaOH solution of 0.05mol/L, 80 ℃ of following water-bath 60min repeatedly wash with distilled water then again;
The purifying of step 2, bacteria cellulose film:
With distilled water immersion 5 times, with the light press mold survey of pH test paper pH value, finally controlling the bacteria cellulose pH value is 7.2 ± 0.2 with the bacteria cellulose film after the above-mentioned pre-treatment, obtains the bacteria cellulose behind the purifying, In Shade afterwards preservation.
Step 3, bacteria cellulose radical functino:
Select the TEMPO oxidation system for use, the hydroxyl oxygen of C6 position on the bacteria cellulose is changed into carboxyl, with deionized water soaking flushing or remove residual oxidizer repeatedly, obtain the bacteria cellulose film of radical functino then with ion exchange method;
The adjustment of step 4, bacteria cellulose water content and sterilization process:
With the bacteria cellulose in the step 3, with the pressure of 500N bacteria cellulose is dewatered, obtain the bacteria cellulose film of different moisture content, then with bacteria cellulose film through disinfection by ultraviolet light, reach the technology sterilization and require (processing requirement aseptic technique afterwards).
The recombining process of step 5, collagen and bacteria cellulose:
The collagen product is made into 5% solution respectively, above-mentioned collagen solution is coated on the bacteria cellulose film surface, the unit coating amount is 50 μ L/cm
2
Step 6, the collagen that will obtain through step 4 are modified bacteria cellulose composite membrane drying treatment in-50 ℃ freeze drier, obtain collagen and modify the compound dry film of bacteria cellulose composite membrane;
Modify bacteria cellulose composite membrane by the collagen that a process for preparing, be attached with collagen on the nano-scale fiber network structure of bacteria cellulose, the collagen content of described composite membrane is 4.7% of a gross weight, and water content is 35.4% of an original bulk.
Embodiment three
The commercially available bacteria cellulose film clip that is made by acetobacter xylinum is become the bacteria cellulose appearance of 10 * 10cm
Article, the preparation process that collagen is modified bacteria cellulose composite membrane is following:
The pre-treatment of step 1, bacteria cellulose film:
Get bacteria cellulose and repeatedly wash, remove striping surface medium and impurity with clear water.Again film is soaked in the NaOH solution of 0.1mol/L, more than 85 ℃ of following water-bath 60min, washes out thalline and residual substratum in the bacteria cellulose film.Bacteria cellulose film is soaked in the NaOH solution of 0.05mol/L, 85 ℃ of following water-bath 45min repeatedly wash with distilled water then again;
The purifying of step 2, bacteria cellulose film:
With distilled water immersion 5 times, with the light press mold survey of pH test paper pH value, finally controlling the bacteria cellulose pH value is 7.2 ± 0.2 with the bacteria cellulose film after the above-mentioned pre-treatment, obtains the bacteria cellulose behind the purifying, In Shade afterwards preservation.
The adjustment of step 3, bacteria cellulose water content and sterilization process:
After the bacteria cellulose pre-treatment is adjusted to neutrality; Pressure with 50N dewaters bacteria cellulose; Obtain the bacteria cellulose film of different moisture content, then bacteria cellulose film is passed through the Co60 radiation sterilization, reach the technology sterilization and require (processing requirement aseptic technique afterwards).
The recombining process of step 4, collagen and bacteria cellulose:
The collagen product is made into 2% solution respectively, above-mentioned bacteria cellulose is dipped in 60s in the collagen solution.
Step 5, the collagen that will obtain through step 4 are modified the bacteria cellulose composite membrane dehydration, and the adjustment collagen that obtains different water content is modified the bacteria cellulose composite membrane wet film once more.
Modify bacteria cellulose composite membrane by the collagen that a process for preparing, be attached with collagen on the nano-scale fiber network structure of bacteria cellulose, the collagen content of described composite membrane is 2.7% of a gross weight, and water content is 78.9% of an original bulk.
Embodiment four
The commercially available bacteria cellulose film clip that is made by acetobacter xylinum is become the bacteria cellulose sample of 10 * 10cm, and the preparation process that collagen is modified bacteria cellulose composite membrane is following:
The pre-treatment of step 1, bacteria cellulose film:
Get bacteria cellulose and repeatedly wash, remove striping surface medium and impurity with clear water.Again film is soaked in the NaOH solution of 0.1mol/L, more than 90 ℃ of following water-bath 60min, washes out thalline and residual substratum in the bacteria cellulose film.Bacteria cellulose film is soaked in the NaOH solution of 0.05mol/L, 80 ℃ of following water-bath 30min repeatedly wash with distilled water then again;
The purifying of step 2, bacteria cellulose film:
With distilled water immersion 5 times, with the light press mold survey of pH test paper pH value, finally controlling the bacteria cellulose pH value is 7.2 ± 0.2 with the bacteria cellulose film after the above-mentioned pre-treatment, obtains the bacteria cellulose behind the purifying, In Shade afterwards preservation.
Step 3, bacteria cellulose radical functino:
Select the periodate oxidation system for use, the hydroxyl oxygen on bacteria cellulose C2, the C3 position is changed into aldehyde radical, with deionized water soaking flushing or remove residual oxidizer repeatedly, obtain the bacteria cellulose film of radical functino then with ion exchange method;
The adjustment of step 4, bacteria cellulose water content and sterilization process:
With the bacteria cellulose in the step 3, with the pressure of 500N bacteria cellulose is dewatered, obtain the bacteria cellulose film of different moisture content, then with bacteria cellulose film through disinfection by ultraviolet light, reach the technology sterilization and require (processing requirement aseptic technique afterwards).
The recombining process of step 5, collagen and bacteria cellulose:
The collagen product is made into 6% solution respectively, above-mentioned bacteria cellulose is dipped in 1h in the collagen solution.
Step 6, the collagen that will obtain through step 4 are modified the bacteria cellulose composite membrane dehydration, and the adjustment collagen that obtains different water content is modified the bacteria cellulose composite membrane wet film once more.
Modify bacteria cellulose composite membrane by the collagen that a process for preparing, be attached with collagen on the nano-scale fiber network structure of bacteria cellulose, the collagen content of described composite membrane is 6.4% of a gross weight, and water content is 45% of an original bulk.
Embodiment five
The commercially available bacteria cellulose film clip that is made by acetobacter xylinum is become the bacteria cellulose sample of 10 * 10cm, and the preparation process that collagen is modified bacteria cellulose composite membrane is following:
The pre-treatment of step 1, bacteria cellulose film:
Get bacteria cellulose and repeatedly wash, remove striping surface medium and impurity with clear water.Again film is soaked in the NaOH solution of 0.1mol/L, more than 95 ℃ of following water-bath 60min, washes out thalline and residual substratum in the bacteria cellulose film.Bacteria cellulose film is soaked in the NaOH solution of 0.05mol/L, 90 ℃ of following water-bath 30min repeatedly wash with distilled water then again;
The purifying of step 2, bacteria cellulose film:
With distilled water immersion 4 times, with the light press mold survey of pH test paper pH value, finally controlling the bacteria cellulose pH value is 7.2 ± 0.2 with the bacteria cellulose film after the above-mentioned pre-treatment, obtains the bacteria cellulose behind the purifying, In Shade afterwards preservation.
The adjustment of step 3, bacteria cellulose water content and sterilization process:
After the bacteria cellulose pre-treatment is adjusted to neutrality; Pressure with 800N dewaters bacteria cellulose; Obtain the bacteria cellulose film of different moisture content, then bacteria cellulose film is sterilized through high temperature steaming, reach the technology sterilization and require (processing requirement aseptic technique afterwards).
The recombining process of step 4, collagen and bacteria cellulose:
The collagen product is made into 15% solution respectively, above-mentioned collagen solution is coated on the bacteria cellulose film surface, the unit coating amount is 25 μ L/cm
2
After step 5, the collagen that will obtain through step 4 were modified the bacteria cellulose composite membrane chilling and handled, drying treatment in Vacuumdrier obtained collagen and modifies the compound dry film of bacteria cellulose composite membrane again.
Modify bacteria cellulose composite membrane by the collagen that a process for preparing, be attached with collagen on the nano-scale fiber network structure of bacteria cellulose, the collagen content of described composite membrane is 7.2% of a gross weight, and water content is 17.3% of an original bulk.
Embodiment six
The commercially available bacteria cellulose film clip that is made by acetobacter xylinum is become the bacteria cellulose sample of 10 * 10cm, and the preparation process that collagen is modified bacteria cellulose composite membrane is following:
The pre-treatment of step 1, bacteria cellulose film:
Get bacteria cellulose and repeatedly wash, remove striping surface medium and impurity with clear water.Again film is soaked in the NaOH solution of 0.1mol/L, more than 85 ℃ of following water-bath 60min, washes out thalline and residual substratum in the bacteria cellulose film.Bacteria cellulose film is soaked in the NaOH solution of 0.05mol/L, 100 ℃ of following water-bath 45min repeatedly wash with distilled water then again;
The purifying of step 2, bacteria cellulose film:
With distilled water immersion 5 times, with the light press mold survey of pH test paper pH value, finally controlling the bacteria cellulose pH value is 7.2 ± 0.2 with the bacteria cellulose film after the above-mentioned pre-treatment, obtains the bacteria cellulose behind the purifying, In Shade afterwards preservation.
Step 3, bacteria cellulose radical functino:
Select the nitrogen peroxide oxidation system for use, the hydroxyl oxygen of C6 position on the bacteria cellulose is changed into carboxyl, with deionized water soaking flushing or remove residual oxidizer repeatedly, obtain the bacteria cellulose film of radical functino then with ion exchange method;
The adjustment of step 4, bacteria cellulose water content and sterilization process:
With the bacteria cellulose in the step 3; Pressure with 100N dewaters bacteria cellulose; Obtain the bacteria cellulose film of different moisture content, then bacteria cellulose film is passed through the Co60 radiation sterilization, reach the technology sterilization and require (processing requirement aseptic technique afterwards).
The recombining process of step 5, collagen and bacteria cellulose:
The collagen product is made into 1% solution respectively, above-mentioned bacteria cellulose is dipped in 120s in the collagen solution.
Step 6, the collagen that will obtain through step 4 are modified bacteria cellulose composite membrane drying treatment in-45 ℃ freeze drier, obtain collagen and modify the compound dry film of bacteria cellulose composite membrane.
Modify bacteria cellulose composite membrane by the collagen that a process for preparing, be attached with collagen on the nano-scale fiber network structure of bacteria cellulose, the collagen content of described composite membrane is 3.54% of a gross weight, and water content is 70.2% of an original bulk.
Embodiment seven
The commercially available bacteria cellulose film clip that is made by acetobacter xylinum is become the bacteria cellulose sample of 10 * 10cm, and the preparation process that collagen is modified bacteria cellulose composite membrane is following:
The pre-treatment of step 1, bacteria cellulose film:
Get bacteria cellulose and repeatedly wash, remove striping surface medium and impurity with clear water.Again film is soaked in the NaOH solution of 0.1mol/L, more than 100 ℃ of following water-bath 60min, washes out thalline and residual substratum in the bacteria cellulose film.Bacteria cellulose film is soaked in the NaOH solution of 0.05mol/L, 95 ℃ of following water-bath 60min repeatedly wash with distilled water then again;
The purifying of step 2, bacteria cellulose film:
With distilled water immersion 5 times, with the light press mold survey of pH test paper pH value, finally controlling the bacteria cellulose pH value is 7.2 ± 0.2 with the bacteria cellulose film after the above-mentioned pre-treatment, obtains the bacteria cellulose behind the purifying, In Shade afterwards preservation.
The adjustment of step 3, bacteria cellulose water content and sterilization process:
After the bacteria cellulose pre-treatment is adjusted to neutrality; Pressure with 1000N dewaters bacteria cellulose; Obtain the bacteria cellulose film of different moisture content, then bacteria cellulose film is passed through disinfection by ultraviolet light, reach the technology sterilization and require (processing requirement aseptic technique afterwards).
The recombining process of step 4, collagen and bacteria cellulose:
The collagen product is made into 10% solution respectively, above-mentioned collagen solution is coated on the bacteria cellulose film surface, the unit coating amount is 100 μ L/cm
2
Step 5, the collagen that will obtain through step 4 are modified the bacteria cellulose composite membrane dehydration, and the adjustment collagen that obtains different water content is modified the bacteria cellulose composite membrane wet film once more.
Modify bacteria cellulose composite membrane by the collagen that a process for preparing, be attached with collagen on the nano-scale fiber network structure of bacteria cellulose, the collagen content of described composite membrane is 11.2% of a gross weight, and water content is 12.8% of an original bulk.
Embodiment eight
The commercially available bacteria cellulose film clip that is made by acetobacter xylinum is become the bacteria cellulose sample of 10 * 10cm, and the preparation process that collagen is modified bacteria cellulose composite membrane is following:
The pre-treatment of step 1, bacteria cellulose film:
Get bacteria cellulose and repeatedly wash, remove striping surface medium and impurity with clear water.Again film is soaked in the NaOH solution of 0.1mol/L, more than 95 ℃ of following water-bath 60min, washes out thalline and residual substratum in the bacteria cellulose film.Bacteria cellulose film is soaked in the NaOH solution of 0.05mol/L, 95 ℃ of following water-bath 40min repeatedly wash with distilled water then again;
The purifying of step 2, bacteria cellulose film:
With distilled water immersion 4 times, with the light press mold survey of pH test paper pH value, finally controlling the bacteria cellulose pH value is 7.2 ± 0.2 with the bacteria cellulose film after the above-mentioned pre-treatment, obtains the bacteria cellulose behind the purifying, In Shade afterwards preservation.
Step 3, bacteria cellulose radical functino:
Select the periodate oxidation system for use, the hydroxyl oxygen on bacteria cellulose C2, the C3 position is changed into aldehyde radical, with deionized water soaking flushing or remove residual oxidizer repeatedly, obtain the bacteria cellulose film of radical functino then with ion exchange method;
The adjustment of step 4, bacteria cellulose water content and sterilization process:
With the bacteria cellulose in the step 3; Pressure with 150N dewaters bacteria cellulose; Obtain the bacteria cellulose film of different moisture content, then bacteria cellulose film is passed through the Co60 radiation sterilization, reach the technology sterilization and require (processing requirement aseptic technique afterwards).
The recombining process of step 5, collagen and bacteria cellulose:
The collagen product is made into 7.5% solution respectively, above-mentioned bacteria cellulose is dipped in 600s in the collagen solution.
After step 6, the collagen that will obtain through step 4 were modified the bacteria cellulose composite membrane chilling and handled, drying treatment under Vacuumdrier, thermostat container or normal temperature obtained collagen and modifies the compound dry film of bacteria cellulose composite membrane again.
Modify bacteria cellulose composite membrane by the collagen that a process for preparing, be attached with collagen on the nano-scale fiber network structure of bacteria cellulose, the collagen content of described composite membrane is 6.3% of a gross weight, and water content is 70.6% of an original bulk.
More than embodiments of the invention are described in detail, but said content only is preferred embodiment of the present invention, can not be considered to be used to limit practical range of the present invention.All equalizations of doing according to application range of the present invention change and improve etc., all should still belong within the patent covering scope of the present invention.
Claims (4)
1. a collagen is modified the preparation method of bacteria cellulose composite membrane, and said method comprises the steps:
1.1 the pre-treatment of bacteria cellulose film:
Get bacteria cellulose and repeatedly wash, remove striping surface medium and impurity with clear water; Again film is soaked in the NaOH solution of 0.1mol/L, 80 ~ 100 ℃ of following water-bath 60min or more than, wash out thalline and residual substratum in the bacteria cellulose film; Bacteria cellulose film is soaked in the NaOH solution of 0.05mol/L, 80 ~ 100 ℃ of following water-bath 30-60min repeatedly wash with distilled water then again;
1.2 the purifying of bacteria cellulose film:
With distilled water immersion 3-5 time, with the light press mold survey of pH test paper pH value, finally controlling the bacteria cellulose pH value is 7.2 ± 0.2 with the bacteria cellulose film after the above-mentioned pre-treatment, obtains the bacteria cellulose behind the purifying, is designated as BC, In Shade afterwards preservation;
1.3 bacteria cellulose radical functino:
Select a series of oxidation systems that are applicable to cellulose materials such as TEMPO, Periodic acid or periodate, nitrogen peroxide etc. for use; Hydroxyl oxygen on the bacteria cellulose is changed into carboxyl or aldehyde radical; Then with deionized water soaking flushing or remove residual oxidizer repeatedly with ion exchange method; Obtain the bacteria cellulose film of radical functino, be designated as OBC;
1.4 the adjustment of bacteria cellulose water content and sterilization process:
Pressure with 20-1000N dewaters bacteria cellulose BC and OBC, obtains the bacteria cellulose film of different moisture content, then bacteria cellulose film is passed through high temperature steaming, ultraviolet ray or Co60 radiation sterilization, reaches technology sterilization requirement;
1.5 collagen is modified the compound of bacteria cellulose composite membrane;
Carry out drying treatment 1.6 will modify bacteria cellulose composite membrane, obtain collagen and modify the compound dry film of bacteria cellulose through the collagen that step 1.5 obtains; Perhaps will modify the bacteria cellulose composite membrane dehydration through the collagen that step 1.5 obtains, adjustment obtains the compound wet film of collagen modification bacteria cellulose of different water content once more.
2. a kind of collagen according to claim 1 is modified the preparation method of bacteria cellulose composite membrane, and it is characterized in that: said step 1.5 recombining process is specific as follows:
Physics composite algorithm: the recombining process of collagen and BC
The collagen product is made into the solution of 0.01%-20% respectively, above-mentioned collagen solution is coated on BC film surface, the unit coating amount is 10-200 μ L/cm
2Or the collagen product is made into the solution of 0.01%-20% respectively, and above-mentioned BC film is dipped in 1s-24h in the collagen solution, make the physics composite collagen and modify bacteria cellulose composite membrane;
Or chemically composited method: the recombining process of collagen and OBC
The collagen product is made into the solution of 0.01%-20% respectively, above-mentioned collagen solution is coated on OBC film surface, the unit coating amount is 10-200 μ L/cm
2Or the collagen product is made into the solution of 0.01%-20% respectively, and above-mentioned OBC is dipped in 1s-24h in the collagen solution, make chemically composited collagen and modify bacteria cellulose composite membrane.
3. a kind of collagen according to claim 1 is modified the preparation method of bacteria cellulose composite membrane; It is characterized in that: said bacteria cellulose film is to belong in the bacteriums certain by genus acetobacter, Sarcina, Rhodopseudomonas, rhizobium, achromobacter, Alcaligenes, aerobacter, Azotobacter, Agrobacterium etc. nine to produce through fermentation, and the bacteria cellulose that produces has three-dimensional porous network structure.
4. a kind of collagen according to claim 1 is modified the preparation method of bacteria cellulose composite membrane, and it is characterized in that: the method for drying treatment comprises following 3 kinds of methods in the said step 1.6:
Normal temperature or 20 ℃ ~ 60 ℃ following freeze-day with constant temperature are handled under a, the ventilation state;
Or b, with bacteria cellulose drying treatment in-20 ℃ ~-60 ℃ freeze drier, or vacuum drying treatment;
Or c, the bacterial cellulose wet-coating chilling handled after, drying treatment under Vacuumdrier, thermostat container or normal temperature obtains the lower collagen of fragility and modifies bacteria cellulose composite membrane again.
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