CN109966566B - Integrated bacterial cellulose patch and preparation method thereof - Google Patents
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Abstract
The invention discloses an integrated bacterial cellulose patch which is of a double-layer structure, wherein one layer of the patch is bacterial cellulose with a nano-pore structure, the other layer of the patch is bacterial cellulose containing a micro-pore structure, and the proportion of the thicknesses of the two layers in the double-layer structure is controllable. The preparation of the patch is to spray a culture medium containing a pore-forming agent on the surface of a bacterial cellulose membrane containing a nanopore structure obtained by static culture for 6-50 times, so as to realize membrane-liquid contact culture, and the pore-forming agent and bacteria are removed to obtain the integrated bacterial cellulose patch. In the preparation process, the aperture and the thickness of the nanopore structure layer are controlled by controlling the particle size and the adding amount of the pore-foaming agent added into the non-inoculated culture medium; the thickness and the pore size of the membrane can be regulated and controlled in a micron scale by controlling the spraying amount of the culture medium sprayed on the nanopore structure layer in the process of forming the bacterial cellulose membrane with the micron pore structure. The invention is expected to provide a research foundation for biomaterials such as hernia repair patches, cartilage scaffolds and the like.
Description
Technical Field
The invention relates to preparation of a hernia repair patch material, in particular to an integrated bacterial cellulose patch and a preparation method thereof.
Background
Extraabdominal hernias are one of the most common hernia diseases. According to statistics, the number of newly-increased extraabdominal hernia patients is 2000 ten thousand every year in the world, and the number of newly-increased patients is 200-400 thousand every year in China. The groin part is adjacent to the urogenital system, so that the light patient causes local distending pain and the life quality is influenced; in severe cases, when the small intestine protrudes from the hernia ring and is stuck and unable to return to the abdominal cavity, severe abdominal pain, nausea, vomiting, constipation, abdominal distension and the like may be associated with the severe cases, and in addition, hernia may induce other diseases, which may lead to death of the patient in severe cases.
Patches are commonly used clinically to repair hernias. This requires that the patch material have certain performance requirements: the composite material has certain tensile strength, proper micron pore size on the abdominal wall side, favorable cell growth to reach excellent biocompatibility, excellent antibacterial property, adhesion resistance on the side near the visceral organs, etc. However, current hernia repair patch materials are prone to varying degrees of complications. For example, polypropylene patches contain only micro-macro-pores, which are easy for cells to grow in, but can cause severe adhesion to the abdominal cavity, obstruction of the digestive tract and even intestinal fistula. The expanded polytetrafluoroethylene patch has the anti-adhesion performance due to the small pore size, but macrophages and fibroblasts are difficult to enter, the firmness and the anti-infection capability after the patch is repaired are poor, and the patch material must be removed to control infection once the infection occurs. Structurally uniform patch materials lose advantage in the repair of specific extraabdominal hernias. The polypropylene and expanded polytetrafluoroethylene composite material has a double-layer structure, the double layers are bonded by physical adsorption or an adhesive, and the possibility of layering is caused when external force is applied, so that the polypropylene and expanded polytetrafluoroethylene composite material is not integrated in the true sense. Therefore, the research and development of the integrated patch material are urgently needed, so that the integrated patch material not only has micron macropores and can enable tissues to grow in smoothly, but also has the advantages of infection resistance, adhesion resistance, good biocompatibility and tensile strength, and is an important direction in the field of hernia repair materials.
In recent years, bacterial cellulose is widely applied to the field of biological materials due to the characteristics of hyperfine three-dimensional network structure, high water absorption performance, higher biocompatibility, adaptability and degradability, simple preparation, greenness, no pollution and the like. Particularly, the bacterial cellulose only contains nanometer-scale pores but does not have micron-scale macropores, and can inhibit the tissue from growing in, so that when the bacterial cellulose is used as a patch, the adhesion between the material and abdominal organs can be prevented. However, this structure does not satisfy the micron-sized fibroblast and macrophage ingrowth, and thus, when used as a patch, the patch is poor in repair firmness and infection resistance. Therefore, the natural nano-pore diameter of the bacterial cellulose is required to be used for preventing adhesion at one side close to the abdominal viscera, and the micro-pore diameter is constructed at the other side for the cells to grow in. There is the report in the literature that the laser drilling can be used for conveniently constructing the micro-macropore on the bacterial cellulose membrane, but because the laser drilling pore-forming depth is difficult to control, the whole bacterial cellulose membrane is easy to punch through, so that a double-layer structure with one layer being the nanometer pore diameter and the other layer being the micrometer pore diameter cannot be constructed, and the laser drilling cannot regulate and control the parameters such as the pore size, the pore density and the like of each layer of bacterial cellulose membrane, and meanwhile, the prepared pores cannot be communicated in the longitudinal direction and are not beneficial to the tissue repair.
In consideration of the defects of the existing hernia repair patch material, the hernia repair patch has limitation on the development of biological materials and tissue engineering, and the prior art cannot realize a bacterial cellulose membrane with one layer of nano-pores and the other layer of micro-macropores and can regulate and control the parameters of the micro-pore size, the pore density and the like.
Disclosure of Invention
Aiming at the prior art, the invention provides an integrated bacterial cellulose patch which is expected to provide a research basis for biomaterials such as hernia repair patches, cartilage scaffolds and the like.
In order to solve the technical problems, the integrated bacterial cellulose patch provided by the invention is of a double-layer structure, wherein one layer of the patch is bacterial cellulose with a nano-pore structure, the other layer of the patch is bacterial cellulose containing a micro-pore structure, and the proportion of the thicknesses of the two layers in the double-layer structure is controllable.
The preparation method of the integrated bacterial cellulose patch provided by the invention sprays culture medium containing pore-forming agent on the surface of the bacterial cellulose membrane containing the nanopore structure obtained by static culture for 6-50 times to realize membrane-liquid contact culture, and removes the pore-forming agent and bacteria to obtain the integrated bacterial cellulose patch; the method comprises the following specific steps:
step one, preparing a culture medium: preparing a culture medium, placing the culture medium in a sterilization pot, and sterilizing for 30-60 min at high temperature and high pressure;
step two, preparing a nanopore bacterial cellulose membrane: inoculating the bacterial liquid into a culture medium in an aseptic environment; statically culturing the inoculated culture medium at the temperature of 30 ℃ for 1-2 d to obtain a bacterial cellulose membrane containing a nanopore structure and with the thickness of 1.0-3.0 mm; wherein the ratio of the amount of the inoculated culture medium to the area of the bacterial cellulose membrane is 0.10-0.30 mL/cm2；
Step three, forming a bacterial cellulose membrane with a micro-pore structure on the nano-pore bacterial cellulose membrane, thereby forming an integrated bacterial cellulose membrane with a double-layer structure, wherein one layer of the integrated bacterial cellulose membrane comprises nano-pores and the other layer of the integrated bacterial cellulose membrane comprises micro-macropores in the thickness direction; the method comprises the following steps:
step 3-1), adding the pore-foaming agent which is subjected to ultraviolet sterilization and has the particle size range of 50-300 mu m into a culture medium of the sterile strain, and uniformly mixing to obtain the culture medium containing the pore-foaming agent; wherein the mass volume concentration of the pore-foaming agent and the culture medium is 0.05-0.20 mg/mL;
step 3-2), in a mist form, according to the volume ratio of 0.010-0.031 mL/cm2Spraying the culture medium containing the pore-forming agent obtained in the step 3-1) on the bacterial cellulose membrane obtained in the step two;
step 3-3), after the culture medium containing the pore-forming agent on the bacterial cellulose membrane is completely consumed, performing treatment according to the concentration of 0.010-0.031 mL/cm2Spraying the culture medium containing the pore-forming agent obtained in the step 3-1) on the bacterial cellulose membrane;
and 3-4) repeating the steps 3-3) for 6-50 times to finally form a bacterial cellulose membrane with the thickness of 1.0-10.0 mm and containing the pore-foaming agent, removing the pore-foaming agent and bacteria, and cleaning to be neutral to obtain the integrated bacterial cellulose patch.
In the second step of the preparation method of the integrated bacterial cellulose patch, the strains of the bacterial liquid include but are not limited to acetobacter xylinum, acetobacter aceti, acetobacter gluconicum, acetobacter acetogenins and acetobacter pasteurianus.
In the third step, the pore-forming agent includes but is not limited to one of gelatin microspheres, silicon dioxide microspheres, sodium alginate microspheres and paraffin microspheres.
Compared with the prior art, the integrated bacterial fiber patch is prepared by adopting a combined membrane liquid interface culture method and a particle filtration method, the patch is of a double-layer structure, one layer is bacterial cellulose with a nano-pore structure, the other layer is bacterial cellulose containing micro-macropores, the thicknesses of the two layers are controllable, an integrated and inseparable integral structure is formed, and the thickness and the pore size of the membrane can be regulated and controlled in a micron scale. The preparation method is simple and convenient, the process is safe, green and environment-friendly, the obtained material not only keeps the natural nano-aperture of the bacterial cellulose, but also can generate structures with different aperture sizes and pore densities so as to be suitable for different tissue engineering. It can provide materials for the preparation of materials for hernia repair patch, osteochondral tissue repair and the like.
Drawings
Fig. 1 is a microscopic photograph of a micro-macro-pore based on a bacterial cellulose nano-pore in the integrated bacterial cellulose patch prepared in example 1 of the present invention.
Fig. 2 is a magnified photograph of a portion of a micron macropore in the integrated bacterial cellulose patch prepared in example 1 of the present invention.
Fig. 3 is a photomicrograph of a cross-section of the integrated bacterial cellulose patch prepared in example 1 of the present invention.
Fig. 4 is a microscopic photograph of bacterial cellulose natural fibers and nanopores in the integrated bacterial cellulose patch prepared in embodiment 1 of the present invention.
Detailed Description
The integrated bacterial cellulose patch provided by the invention is of a double-layer structure, wherein one layer is bacterial cellulose with a nano-pore structure, the other layer is bacterial cellulose containing a micro-pore structure, and the proportion of the thicknesses of the two layers in the double-layer structure is controllable in the preparation process. The preparation method of the patch mainly comprises the following steps: preparing a culture medium; preparing a nano-pore bacterial cellulose membrane by static culture; adding a pore-forming agent into a culture medium, spraying the culture medium containing the pore-forming agent on a bacterial cellulose membrane containing nano pores in a mist form to realize membrane liquid contact culture, spraying again when the culture medium is completely consumed, repeating for many times, and finally removing the pore-forming agent and bacteria, thereby forming an integrated bacterial cellulose membrane with one layer containing the nano pores and the other layer containing the micro-macro pores in the thickness direction. The invention is expected to provide a research foundation for biomaterials such as hernia repair patches, cartilage scaffolds and the like.
The invention will be further described with reference to the following figures and specific examples, which are not intended to limit the invention in any way.
Example 1, preparation of an integrated bacterial cellulose patch, the steps are as follows:
s1, preparing a culture medium from deionized water, anhydrous glucose, peptone, disodium hydrogen phosphate and yeast powder, wherein the pH of the culture medium is 4-5, and placing the culture medium in a sterilization pot for sterilization for 30min at 115 ℃ and 0.1MPa for later use;
s2, inoculating a bacterial liquid into a culture medium under an aseptic environment, wherein the strain of the bacterial liquid is acetobacter xylinum, and statically culturing a proper amount of the inoculated culture medium at the temperature of 30 ℃ for 1d to obtain a bacterial cellulose membrane containing a nanopore structure and with the thickness of 1.0 mm; wherein the ratio of the amount of inoculated culture medium to the area of the bacterial cellulose membrane is 0.10mL/cm2；
S3, adding the gelatin microspheres with the average particle size of 125 μm into the non-inoculated culture medium prepared in the step S1 after ultraviolet sterilization, wherein the mass volume concentration of the gelatin microspheres and the culture medium is about 0.10mg/mL, and uniformly mixing for later use;
s4, spraying the culture medium containing the gelatin microspheres prepared in the step S3 on the bacterial cellulose membrane prepared in the step S2 in a mist form, wherein the amount of the sprayed culture medium is about 0.010mL/cm in the spraying process2Spraying again after the culture medium is completely consumed, repeating for 20 times to prepare a bacterial cellulose membrane containing gelatin microspheres with the thickness of 5 mm;
s5, sequentially placing the bacterial cellulose membrane containing the gelatin microspheres obtained in the step S4 in deionized water and a sodium hydroxide solution (0.5mol/L) at 80 ℃ for 24 hours respectively, removing the gelatin microspheres and bacteria, and cleaning to be neutral to obtain the integrated bacterial cellulose patch.
FIG. 1 is a photomicrograph of a micro-macropore based on the nano-pores of the bacterial cellulose in the integrated bacterial cellulose patch obtained in example 1; FIG. 2 is a magnified photograph of a portion of a micron macropore in the integrated bacterial cellulose patch obtained in example 1; FIG. 3 is a photomicrograph of a cross-section of the integrated bacterial cellulose patch obtained in example 1; fig. 4 is a microscopic photograph of the bacterial cellulose natural fibers and the nanopores in the integrated bacterial cellulose patch obtained in example 1.
Example 2, preparation of an integrated bacterial cellulose patch, comprising the following steps:
s1, preparing a culture medium from deionized water, anhydrous glucose, peptone, disodium hydrogen phosphate and yeast powder, wherein the pH of the culture medium is 4-5, and placing the culture medium in a sterilization pot for sterilization for 30min at 115 ℃ and 0.1MPa for later use;
s2, inoculating a bacterial liquid into a culture medium under an aseptic environment, wherein the strain of the bacterial liquid is acetobacter aceti, and statically culturing the inoculated culture medium at 30 ℃ for 1.5d to obtain a bacterial cellulose membrane containing a nanopore structure and with the thickness of 1.5 mm; wherein the ratio of the amount of inoculated culture medium to the area of the bacterial cellulose membrane is 0.16mL/cm2；
S3, adding paraffin wax microspheres with average grain diameter of 250 mu m into the non-inoculated culture medium after ultraviolet sterilization, wherein the content of the paraffin wax microspheres is about 0.05mg/cm2Mixing well for later use
S4, spraying the culture medium containing the paraffin microspheres obtained in the step S3 on the bacterial cellulose membrane obtained in the step S2 in a mist form, wherein the dosage of the sprayed culture medium is about 0.020mL/cm in the spraying process2Spraying again after the culture medium is completely consumed, repeating for 15 times to prepare a bacterial cellulose membrane with the thickness of 3.0mm and containing paraffin microspheres;
s5, sequentially placing the obtained bacterial cellulose membrane containing the paraffin microspheres in deionized water and a sodium hydroxide solution (0.5mol/L) at 80 ℃ for heating for 24 hours respectively, removing the paraffin microspheres and bacteria, and cleaning to be neutral to obtain the integrated bacterial cellulose patch.
Example 3 preparation of an integrated bacterial cellulose patch, the steps are as follows:
s1, preparing a culture medium from deionized water, anhydrous glucose, peptone, disodium hydrogen phosphate and yeast powder, wherein the pH of the culture medium is 4-5, and placing the culture medium in a sterilization pot for sterilization for 30min at 115 ℃ and 0.1 MPa;
and S2, inoculating the bacterial liquid into a culture medium under an aseptic environment, wherein the bacterial liquid is acetobacter pasteurianus. Statically culturing the inoculated culture medium at 30 ℃ for 2d to obtain a bacterial cellulose membrane with a thickness of 2.0mm and a nanopore structure; wherein the ratio of the amount of inoculated culture medium to the area of the bacterial cellulose membrane is 0.21mL/cm2；
S3, after ultraviolet sterilization, adding the silicon dioxide microspheres with the average particle size of 200 mu m into the non-inoculated culture medium, wherein the content of the silicon dioxide microspheres is about 0.20mg/mL, and uniformly mixing.
S4, spraying the culture medium containing the silica microspheres obtained in the step S3 on the bacterial cellulose membrane obtained in the step S2 in a mist form, wherein the amount of the sprayed culture medium is about 0.025mL/cm in the spraying process2Spraying again after the culture medium is completely consumed, repeating for 35 times to prepare a bacterial cellulose membrane containing silicon dioxide microspheres with the thickness of 7 mm;
s5, sequentially placing the obtained bacterial cellulose membrane containing the silicon dioxide microspheres in deionized water and a sodium hydroxide solution (0.5mol/L) at 80 ℃ for heating for 24 hours respectively, removing the silicon dioxide microspheres and bacteria, and cleaning to be neutral to obtain the integrated bacterial cellulose patch.
Example 4 preparation of an integrated bacterial cellulose patch, the steps are as follows:
s1, preparing a culture medium from deionized water, anhydrous glucose, peptone, disodium hydrogen phosphate and yeast powder, wherein the pH of the culture medium is 4-5, and placing the culture medium in a sterilization pot for sterilization for 30min at 115 ℃ and 0.1MPa for later use;
s2, inoculating the bacterial liquid into a culture medium under an aseptic environment, wherein the bacterial liquid is acetobacter xylinum, and taking the inoculated culture medium with the concentration of 0.30mL/cm2Statically culturing at 30 deg.C for 2d to obtain the product with thickness of 3.0mm and containing nanopore structureA bacterial cellulose membrane; wherein the ratio of the amount of inoculated culture medium to the area of the bacterial cellulose membrane is 0.30mL/cm2；
S3, adding the gelatin microspheres with the average particle size of 150 μm into the non-inoculated culture medium prepared in the step S1 after ultraviolet sterilization, wherein the mass volume concentration of the gelatin microspheres and the culture medium is about 0.10mg/mL, and uniformly mixing for later use;
s4, spraying the culture medium containing the gelatin microspheres prepared in the step S3 on the bacterial cellulose membrane prepared in the step S2 in a mist form, wherein the amount of the sprayed culture medium is about 0.010mL/cm in the spraying process2Spraying again after the culture medium is completely consumed, repeating for 20 times to prepare a bacterial cellulose membrane containing gelatin microspheres with the thickness of 5 mm;
s5, sequentially placing the bacterial cellulose membrane containing the gelatin microspheres obtained in the step S4 in deionized water and a sodium hydroxide solution (0.5mol/L) at 80 ℃ for 24 hours respectively, removing the gelatin microspheres and bacteria, and cleaning to be neutral to obtain the integrated bacterial cellulose patch.
Example 5 preparation of an integrated bacterial cellulose patch, the steps are as follows:
s1, preparing a culture medium from deionized water, anhydrous glucose, peptone, disodium hydrogen phosphate and yeast powder, wherein the pH of the culture medium is 4-5, and placing the culture medium in a sterilization pot for sterilization for 30min at 115 ℃ and 0.1MPa for later use;
s2, inoculating a bacterial liquid into a culture medium under an aseptic environment, wherein the strain of the bacterial liquid is acetobacter xylinum, and statically culturing the inoculated culture medium at 30 ℃ for 1d to obtain a bacterial cellulose membrane containing a nanopore structure and with the thickness of 1.0 mm; wherein the ratio of the amount of inoculated culture medium to the area of the bacterial cellulose membrane is 0.30mL/cm2；
S3, adding the gelatin microspheres with the average particle size of 150 μm into the non-inoculated culture medium prepared in the step S1 after ultraviolet sterilization, wherein the mass volume concentration of the gelatin microspheres and the culture medium is about 0.10mg/mL, and uniformly mixing for later use;
s4, spraying the culture medium containing the gelatin microspheres prepared in the step S3 in a mist form on the bacterial cellulose membrane prepared in the step S2In the spraying process, the amount of the spraying medium is about 0.010mL/cm2Spraying again after the culture medium is completely consumed, repeating for 20 times to prepare a bacterial cellulose membrane containing gelatin microspheres with the thickness of 5 mm;
s5, sequentially placing the bacterial cellulose membrane containing the gelatin microspheres obtained in the step S4 in deionized water and a sodium hydroxide solution (0.5mol/L) at 80 ℃ for 24 hours respectively, removing the gelatin microspheres and bacteria, and cleaning to be neutral to obtain the integrated bacterial cellulose patch.
In conclusion, the integrated bacterial fiber patch prepared by the combined membrane-liquid interface culture method and the particle filtration method is of a double-layer structure, wherein one layer is bacterial cellulose with a nano-pore structure, the other layer is bacterial cellulose containing micro-macropores, and the thicknesses of the two layers are controllable, so that an integrated and inseparable integral structure is formed. In the step of preparing the nanopore bacterial cellulose membrane by static culture in the preparation process, the aperture and the thickness of a nanopore structure layer are controlled by controlling the particle size and the adding amount of a pore-forming agent added into a non-inoculated culture medium; the thickness and the pore size of the membrane can be regulated and controlled in a micron scale by controlling the spraying amount of the culture medium sprayed on the nanopore structure layer in the process of forming the bacterial cellulose membrane with the micron pore structure. The preparation method of the integrated bacterial fiber patch with the nano/micron double-layer structure is simple and convenient, the process is safe, green and environment-friendly, the obtained material not only keeps the natural nano-aperture of the bacterial cellulose, but also can generate structures with different aperture sizes and pore densities so as to be suitable for different tissue engineering. It can provide materials for the preparation of materials for hernia repair patch, osteochondral tissue repair and the like.
Although the present invention has been described in detail with reference to the drawings, the present invention is not limited to the above embodiments, which are only illustrative and not restrictive, and those skilled in the art can make many modifications without departing from the spirit of the present invention, which falls within the protection of the present invention.
Claims (3)
1. The preparation method of the integrated bacterial cellulose patch is characterized in that the patch is of a double-layer structure, one layer is bacterial cellulose with a nano-pore structure, the other layer is bacterial cellulose containing a micro-pore structure, and the proportion of the thicknesses of the two layers in the double-layer structure is controllable; the preparation process comprises the following steps: spraying a culture medium containing a pore-forming agent on the surface of the bacterial cellulose membrane containing the nanopore structure obtained by static culture for 6-50 times to realize membrane liquid contact culture, and removing the pore-forming agent and bacteria to obtain the integrated bacterial cellulose patch; the method comprises the following specific steps:
step one, preparing a culture medium: preparing a culture medium, placing the culture medium in a sterilization pot, and sterilizing for 30-60 min at high temperature and high pressure;
step two, preparing a nanopore bacterial cellulose membrane: inoculating the bacterial liquid into a culture medium in an aseptic environment; statically culturing the inoculated culture medium at the temperature of 30 ℃ for 1-2 d to obtain a bacterial cellulose membrane containing a nanopore structure and with the thickness of 1.0-3.0 mm; wherein the ratio of the amount of the inoculated culture medium to the area of the bacterial cellulose membrane is 0.10-0.30 mL/cm2；
Step three, forming a bacterial cellulose membrane with a micro-pore structure on the nano-pore bacterial cellulose membrane, thereby forming an integrated bacterial cellulose membrane with a double-layer structure, wherein one layer of the integrated bacterial cellulose membrane comprises nano-pores and the other layer of the integrated bacterial cellulose membrane comprises micro-macropores in the thickness direction; the method comprises the following steps:
step 3-1), adding the pore-foaming agent which is subjected to ultraviolet sterilization and has the particle size range of 50-300 mu m into a culture medium of the sterile strain, and uniformly mixing to obtain the culture medium containing the pore-foaming agent; wherein the mass volume concentration of the pore-foaming agent and the culture medium is 0.05-0.20 mg/mL;
step 3-2), in a mist form, according to the volume ratio of 0.010-0.031 mL/cm2Spraying the culture medium containing the pore-forming agent obtained in the step 3-1) on the bacterial cellulose membrane obtained in the step two;
step 3-3), after the culture medium containing the pore-forming agent on the bacterial cellulose membrane is completely consumed, performing treatment according to the concentration of 0.010-0.031 mL/cm2Spraying the culture medium containing the pore-forming agent obtained in the step 3-1) on the bacterial cellulose membrane;
and 3-4) repeating the steps 3-3) for 6-50 times to finally form a bacterial cellulose membrane with the thickness of 1.0-10.0 mm and containing the pore-foaming agent, removing the pore-foaming agent and bacteria, and cleaning to be neutral to obtain the integrated bacterial cellulose patch.
2. The method for preparing the integrated bacterial cellulose patch as claimed in claim 1, wherein the bacterial strain of the bacterial liquid in the second step includes but is not limited to acetobacter xylinum, acetobacter aceti, acetobacter xylinum, acetobacter acetogenium and acetobacter pasteurianus.
3. The preparation method of the integrated bacterial cellulose patch as claimed in claim 2, wherein in the third step, the pore-forming agent includes but is not limited to one of gelatin microspheres, silica microspheres, sodium alginate microspheres and paraffin microspheres.
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| CN102274549A (en) * | 2011-07-10 | 2011-12-14 | 东华大学 | Method for preparing bacteria cellulose bracket material and product thereof |
| CN102973985B (en) * | 2012-12-26 | 2014-08-13 | 东华大学 | Porous bacterial cellulose skin repair material with density structure and preparation method thereof |
| CN109097420A (en) * | 2018-08-06 | 2018-12-28 | 华东交通大学 | Graphene/bacterial cellulose composite material and preparation method with gradient-structure |
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| CN102274549A (en) * | 2011-07-10 | 2011-12-14 | 东华大学 | Method for preparing bacteria cellulose bracket material and product thereof |
| CN102973985B (en) * | 2012-12-26 | 2014-08-13 | 东华大学 | Porous bacterial cellulose skin repair material with density structure and preparation method thereof |
| CN109097420A (en) * | 2018-08-06 | 2018-12-28 | 华东交通大学 | Graphene/bacterial cellulose composite material and preparation method with gradient-structure |
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