CN113528290A - Device and method for preparing bacterial cellulose composite material with core-shell structure through dynamic fermentation - Google Patents
Device and method for preparing bacterial cellulose composite material with core-shell structure through dynamic fermentation Download PDFInfo
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- C08J2333/04—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides, or nitriles thereof; Derivatives of such polymers esters
- C08J2333/06—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides, or nitriles thereof; Derivatives of such polymers esters of esters containing only carbon, hydrogen, and oxygen, the oxygen atom being present only as part of the carboxyl radical
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- C08J2383/00—Characterised by the use of macromolecular compounds obtained by reactions forming in the main chain of the macromolecule a linkage containing silicon with or without sulfur, nitrogen, oxygen, or carbon only; Derivatives of such polymers
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- C08J2423/00—Characterised by the use of homopolymers or copolymers of unsaturated aliphatic hydrocarbons having only one carbon-to-carbon double bond; Derivatives of such polymers
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- C08J2477/00—Characterised by the use of polyamides obtained by reactions forming a carboxylic amide link in the main chain; Derivatives of such polymers
Abstract
The invention provides a device and a method for preparing a bacterial cellulose composite material with a core-shell structure through dynamic fermentation. The device comprises a fermentation culture container, two rollers and two heating guide plates, wherein the two rollers and the two heating guide plates are arranged in the fermentation culture container; two ends of the rotating shafts of the two rollers are respectively movably connected with the inner wall of the fermentation culture container and are arranged in parallel in the horizontal direction, a gap is arranged between the rollers, and the distance between the rotating shafts of the rollers is adjustable; the two heating guide plates are parallel to the rotating shafts of the two rollers, one end of one heating guide plate is movably connected with the fermentation culture container, and the other end of the other heating guide plate extends obliquely downwards to the upper part of the gap of the two rollers and is abutted against one roller; one end of the other heating guide plate is movably connected with the fermentation culture container, and the other end of the other heating guide plate extends to the upper part of the gap of the two rollers in a downward inclined way and is abutted against the other roller. The device can realize dynamic fermentation and coating, and the bacterial cellulose composite material with controllable shape and size, good biocompatibility and good safety is obtained.
Description
Technical Field
The invention belongs to the technical field of dynamic fermentation, and relates to a device and a method for preparing a bacterial cellulose composite material with a core-shell structure through dynamic fermentation.
Background
Cellulose is the most abundant biopolymer with huge development potential on the earth, not only is the traditional raw material of textile industry and paper industry, but also can be used for manufacturing high molecular composite materials and high performance materials, and plays an important role in a plurality of high and new technological fields. The bacterial cellulose is natural cellulose obtained by microbial fermentation, and is a high molecular compound formed by connecting glucose by beta-1, 4-glucoside chains. Bacterial cellulose and natural cellulose produced by plants or seaweeds have the same molecular building blocks and have many unique properties: high crystallinity and degree of polymerization. Compared with plant cellulose, the bacterial cellulose has no associated products such as lignin, pectin, hemicellulose and the like, and has high crystallinity (up to 95 percent and 65 percent of plant cellulose) and high degree of polymerization (DP value is 2000-8000); ② hyperfine nanometer net structure. The bacterial cellulose fiber is a fiber bundle with the thickness of 40-60 nanometers and is formed by combining microfibers with the diameter of 3-4 nanometers, and the fibers are mutually interwoven to form a developed hyperfine nano network structure; high elastic modulus and tensile strength. The elastic modulus of the bacterial cellulose is several times to more than ten times of that of common plant fiber, and the tensile strength is high; water Retention Values (WRV). The WRV value of the bacterial cellulose which is not dried is as high as over 1000%, and the water holding capacity after freeze drying is still over 600%. The re-swelling capacity of the bacterial cellulose dried at 100 ℃ in water is equivalent to that of cotton linters; excellent biocompatibility, adaptability and biodegradability; sixthly, the synthesis controllability is realized. Due to the excellent properties of the bacterial cellulose, the bacterial cellulose can be widely applied to the fields of food, biomedicine, medical apparatus and instruments, tissue engineering materials and the like.
Because the bacterial cellulose has strong hydrophilicity, viscosity and stability, the bacterial cellulose can be used as a food forming agent, a thickening agent, a dispersing agent and an anti-dissolving agent, can improve the mouthfeel and can be used as a sausage casing and a framework of certain foods, and becomes a novel important food base material and dietary fiber. For example, the product is widely applied to food such as jelly, milk tea, jam, desserts and the like, and is one of popular raw materials in commercial food at present. Due to good biocompatibility, high mechanical strength in a wet state and good liquid and gas permeability of the bacterial cellulose, the bacterial cellulose can be used as an excellent biological material to be applied to various fields of biomedicine. Biofill and Gengiflex, for example, are two typical bacterial cellulose products that have been widely used as surgical and dental materials. Biofil has been successfully used as a temporary substitute for artificial skin for secondary and tertiary burns, ulcers, etc. Gengiflex has been used for repair of periodontal tissue. A novel biomaterial BASYC designed based on in-situ plasticity of bacterial cellulose is expected to be used as an artificial blood vessel in microsurgery. Meanwhile, reported applications also include the repair of tissues such as bone, cornea, cartilage, tendon, etc.
At present, the fermentation preparation technology of bacterial cellulose mainly comprises static fermentation and dynamic fermentation, wherein the preparation of the bacterial cellulose composite material mainly comprises the static fermentation. The static fermentation process is to add various water-soluble polymers such as carboxymethyl cellulose, hemicellulose, chitosan, gelatin and the like into a culture solution to obtain different types of composite materials. Meanwhile, by adopting the air-permeable material as a mold, bacterial cellulose materials with different shapes can be obtained in the static fermentation process. For example, in the UK Patent 12, 169, 543, an oxygen-permeable hand-shaped mold is used to prepare an artificial skin in the shape of a glove; patent EP Patent 0,396,344; patent JP Patent 3,272,772, utilizes oxygen permeable hollow circular tubes to prepare artificial blood vessels by injecting culture solution containing bacteria under static fermentation conditions. In the aspect of dynamic fermentation, the conventional paddle type stirring can only prepare granular bacterial cellulose. The rotary disk fermentation apparatus (Rotating disk fermentation device) designed by Krystinowicz A et al solves the problem that the bacterial cellulose is difficult to form a film during dynamic culture, and obtains a disk-shaped bacterial cellulose film (Journal of Industrial Microbiology & Biotechnology 2002(29): 189-195). On the basis of the above, Chinese patent CN2937138Y discloses a rotating disc type fermentation reaction device, but the device of the type can only obtain a disc-shaped bacterial cellulose membrane. In order to further obtain the bacterial cellulose material with controllable shape, Chinese patent CN101914434A designs a method for dynamically preparing heterotype cavity bacterial cellulose material, which can obtain cavity bacterial cellulose material with certain cross-sectional shape. However, in this method, the mold must pass through the rotating shaft, and a completely closed bacterial cellulose-coated material cannot be obtained.
In conclusion, the bacterial cellulose fermentation culture device which is reasonable, simple and feasible in design can enable the method for preparing the composite material which has a certain shape and is completely coated with the required composite material to have great practical significance and commercial prospect.
Disclosure of Invention
One purpose of the invention is to provide a device for preparing a bacterial cellulose composite material with a core-shell structure by dynamic fermentation; the invention also aims to provide a method for preparing the bacterial cellulose composite material with the core-shell structure by dynamic fermentation. The invention further aims to provide the bacterial cellulose composite material with the core-shell structure prepared by the method.
The purpose of the invention is realized by the following technical scheme:
on one hand, the invention provides a device for preparing a bacterial cellulose composite material with a core-shell structure by dynamic fermentation, which comprises:
the fermentation culture container and two rollers and two heating guide plates arranged in the fermentation culture container;
two ends of the rotating shaft of the two rollers are respectively movably connected with the inner wall of the fermentation culture container, the two rollers are arranged in parallel in the horizontal direction, a gap is arranged between the rollers, and the distance between the rotating shafts of the rollers is adjustable;
the two heating guide plates are parallel to the rotating shafts of the two rollers, one end of one heating guide plate is movably connected with the fermentation culture container (preferably connected through a fastener), and the other end of the heating guide plate extends obliquely downwards to the upper part of the gap of the two rollers and is abutted against one roller; one end of the other heating guide plate is movably connected with the fermentation culture container, and the other end of the other heating guide plate extends to the upper part of the gap of the two rollers in a downward inclined mode and is abutted against the other roller.
The device provided by the invention can realize dynamic fermentation and coating, can coat and obtain the bacterial cellulose composite material with controllable shape and size, is simple, convenient to operate and high in yield, and is suitable for industrial production; the obtained coated bacterial cellulose composite material can be widely applied to the fields of biological medicines and medical apparatuses such as surface bacterial cellulose coating of implant medical apparatuses and apparatuses, coating of slow-release medicines or micro apparatuses and the like, and can also be used for coating of foods or food materials and the like.
In the above apparatus, preferably, the apparatus further comprises a driving part for driving the two rollers to rotate; more preferably, the driving member is configured to drive the two rollers to rotate in the same direction (which may be clockwise or counterclockwise in the same direction).
In the above apparatus, preferably, the apparatus further includes a heating member for heating the two heating baffles. The heating component can be an external heating device which makes the heating guide plate generate heat or a heating component which is carried by the heating guide plate.
In the above apparatus, the two rollers are preferably cylindrical rollers having the same shape and size.
In the above device, preferably, the two heating guide plates respectively form an angle of 15-60 degrees with the inner side wall of the fermentation culture container. The two heating guide plates are movable and detachable movable connecting fittings, can be fixed at the top of the fermentation culture container by using fasteners, and adjust the inclination angles of the two heating guide plates by using the fasteners so as to adjust the flow rate of the heating liquid of the thermoplastic macromolecules.
On the other hand, the invention also provides a method for preparing the bacterial cellulose composite material with the core-shell structure by dynamic fermentation, which adopts the device for dynamic fermentation and comprises the following steps:
adjusting the distance between the rollers to ensure that the minimum width of the gap is smaller than the diameter or the size of the core material (the size is satisfied that the core material can be clamped above two rollers), sterilizing the core material to be coated, and placing the core material above the gap of the two rollers to ensure that the core material can be respectively abutted against the two rollers above the gap; the rotation speed and the rotation direction of the roller are set, so that the core material can realize vibration (including rotation) without horizontal displacement along with the rotation of the roller;
the length of the heating plates and the angle of the heating plates are adjusted, so that one ends of the two heating plates are positioned at a gap where the core material is abutted to the roller (thereby ensuring that the roller drives the core material to rotate, and the thermoplastic polymer flows down along the heating guide plate and is coated on the surface of the core material through the rotation of the roller);
preparing a bacterial cellulose fermentation culture solution, carrying out high-pressure sterilization, and then mixing the bacterial cellulose fermentation culture solution with strain seed mash to obtain a fermentation mixed solution;
pouring the fermentation mixed liquid into a fermentation culture container, immersing the core material, starting a roller to rotate in the same direction for dynamic fermentation, discharging fermentation liquid after the fermentation is finished, and coating the outer layer of the core material with bacterial cellulose obtained by bacterial strain fermentation to form a core material-bacterial cellulose compound; starting the heating guide plates, enabling the heating liquid of the thermoplastic macromolecules to respectively flow down along the heating guide plates at two ends, and uniformly coating the thermoplastic macromolecules on the surface of the core material-bacterial cellulose compound along with the rotation of the roller, so as to prepare the bacterial cellulose composite material with the core-shell structure; alternatively, the first and second electrodes may be,
starting the heating guide plates, enabling the heating liquid of the thermoplastic macromolecules to respectively flow down along the heating guide plates at two ends, and uniformly coating the thermoplastic macromolecules on the surface of the core material along with the rotation of the roller to obtain a core material-thermoplastic macromolecule compound; pouring the fermentation mixed liquid into a fermentation culture container, immersing a roller, starting the roller to rotate in the same direction for dynamic fermentation, discharging the fermentation liquid after the fermentation is finished, and covering the outer layer of the core material-thermoplastic polymer compound with bacterial cellulose obtained by bacterial strain fermentation so as to prepare the bacterial cellulose composite material with the core-shell structure.
In the invention, the coating amount of the bacterial cellulose and the coating amount of the thermoplastic high polymer material are reasonably set according to actual needs.
In the dynamic fermentation method of the invention, the obtained core-shell structure bacterial cellulose composite material can be as follows from inside to outside: a core material-bacterial cellulose-thermoplastic polymer structure, or a core material-thermoplastic polymer-bacterial cellulose structure.
In the above method, preferably, the method further includes repeating the step of dynamically fermenting the coated bacterial cellulose and/or the step of coating the thermoplastic polymer to obtain a bacterial cellulose composite material with a more hierarchical core-shell structure. The bacterial cellulose composite materials with different hierarchical structures can be obtained by adopting the repeated steps; for example, the structure may be a "core material-bacterial cellulose-thermoplastic polymer" structure, a "core material-thermoplastic polymer-bacterial cellulose" structure, or the like from the inside to the outside.
In the above method, preferably, the core material includes one or a combination of plural kinds of inorganic materials, organic polymer materials, and metal materials.
In the above method, preferably, the shape of the core material is a spherical shape, a spheroidal shape, a cylindrical shape, a rod-like body, or any irregular body.
In the above method, preferably, the thermoplastic polymer material includes one or more of polyethylene, polypropylene, polystyrene, polymethyl methacrylate, nylon, polyurethane, polyester and polylactic acid.
In the above method, preferably, the heating temperature of the heating baffle is 50 to 300 ℃.
In the above method, preferably, the strain comprises a combination of one or more of acetobacter xylinum, rhizobium, sarcina, pseudomonas, achromobacter, alcaligenes, aerobacter and azotobacter.
In the method, preferably, the addition amount of the strain seed mash is 1-5 wt% of the fermentation culture solution.
In the above method, preferably, the time for performing the dynamic fermentation is 3 to 30 days; the fermentation temperature is 20-30 ℃.
In the above method, preferably, the two drums have the same rotation speed, both of which are 0.1 to 60rpm, when performing dynamic fermentation; preferably 4 to 20 rpm.
The flow rate of the culture solution of the bacterial cellulose in the dynamic culture process must be controlled within a certain range, strains with over-high flow rate are easy to change, the bacterial cellulose is difficult to attach to the core material, and only granular bacterial cellulose can be obtained in the culture solution. When the flow rate is low, a large amount of bacterial cellulose is generated on the gas-liquid interface of the culture container, and the coating of the bacterial cellulose on the core material is influenced. In addition, the flow velocity of the culture solution also influences the yield of the bacterial cellulose, the flatness of the coating on the surface of the core material and the three-dimensional network structure of the nano-fibers forming the bacterial cellulose; preferably, the rotation speed of the roller is controlled within a range of 4 to 20 rpm.
In the method, preferably, during the dynamic fermentation, 0.1-5 wt% of soluble additive is added into the fermentation mixed liquor; the soluble additive comprises one or more of gelatin, sodium hyaluronate, starch, pectin, chitosan, sodium alginate and soluble cellulose derivatives.
The inventor finds that the physicochemical properties of the bacterial cellulose can be improved by adding the soluble polymer in situ in the bacterial cellulose culture process, and the composite material meeting various applications is obtained. The dynamic fermentation method adopted by the invention can be compounded with soluble polymers in the fermentation culture process while the bacterial cellulose coats the core material, thereby expanding the application range of the bacterial cellulose composite material.
In the above method, preferably, the method further comprises a step of purifying the compound coated with bacterial cellulose after the dynamic fermentation is finished, specifically: washing the mixture for 4-6 hours in a NaOH aqueous solution with the mass percentage of 4-8% at the temperature of 70-100 ℃, and repeatedly washing the mixture to be neutral by using distilled water. Thallus protein in the fermentation product and residual culture medium adhered to the bacterial cellulose membrane are removed through purification, so that the coated bacterial cellulose composite material reaches medical or edible conditions.
On the other hand, the invention also provides the bacterial cellulose composite material with the core-shell structure, which is prepared by the method, wherein the inner core layer is a core material, and the core material is coated with a single layer or multiple layers of bacterial cellulose and/or thermoplastic polymer materials.
The invention has the beneficial effects that:
(1) the device provided by the invention can realize dynamic fermentation and coating, can coat and obtain the bacterial cellulose composite material with controllable shape and size, is simple, convenient to operate and high in yield, and is suitable for industrial production; the obtained coated bacterial cellulose composite material can be widely applied to the fields of biological medicines and medical apparatuses such as surface bacterial cellulose coating of implant medical apparatuses and apparatuses, coating of slow-release medicines or micro apparatuses and the like, and can also be used for coating of foods or food materials and the like.
(2) The dynamic fermentation method of the invention does not need any toxic solvent, does not bring about the problems of environmental pollution, ecological hazard and the like, and meets the use requirements of medical use or eating; various water-soluble polymers are added in the fermentation process, so that various bacterial cellulose composite materials with different properties can be obtained, and the bacterial cellulose composite material has excellent biocompatibility and biological safety.
(3) The dynamic fermentation method can be used for quickly obtaining the bacterial cellulose composite material with core-shell structures of different levels, and can be widely applied to various fields such as biological medicines, medical instruments, foods and the like.
Drawings：
FIG. 1 is a schematic structural diagram of an apparatus for preparing a bacterial cellulose composite material with a core-shell structure by dynamic fermentation in an embodiment of the present invention.
Description of the symbols of the drawings:
1. fermentation culture container, 2, roller, 3, fermentation culture solution, 4, core material to be coated, 5, heating guide plate, 6 and fastener.
Detailed Description
The technical solutions of the present invention will be described in detail below in order to clearly understand the technical features, objects, and advantages of the present invention, but the present invention is not limited to the practical scope of the present invention.
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1
The embodiment provides a device for preparing a bacterial cellulose composite material with a core-shell structure through dynamic fermentation, as shown in fig. 1, the device comprises:
a fermentation culture vessel 1, two rollers 2 and two heating guide plates 5 which are arranged in the fermentation culture vessel. Two ends of a rotating shaft of the two rollers 2 are respectively movably connected with the inner wall of the fermentation culture container, the two rollers are arranged in parallel in the horizontal direction, a gap is arranged between the rollers, and the distance between the rotating shafts of the rollers is adjustable; the two heating guide plates 5 are parallel to the rotating shafts of the two rollers 2, one end of one heating guide plate 5 is movably connected with the fermentation culture container 1 through a fastener 6, and the other end extends obliquely downwards to the upper part of the gap of the two rollers and is abutted against one of the two rollers; one end of the other heating guide plate is movably connected with the fermentation culture container through a fastener 6, and the other end of the other heating guide plate extends to the upper part of the gap of the two rollers in a downward inclined mode and is abutted against the other roller 2. The two rollers are preferably cylindrical rollers of the same shape and size.
During dynamic fermentation, the core material 4 to be coated is positioned above the gap between the two rollers 2 and is abutted against the rollers; the fermentation culture 3 is loaded in the fermentation container 1. The device also comprises a driving part for driving the two rollers to rotate, and the driving part is used for driving the two rollers to rotate in the same direction. The device also comprises a heating component for heating the two heating guide plates.
On the other hand, the embodiment also provides a method for preparing a bacterial cellulose composite material with a core-shell structure by dynamic fermentation, which adopts the apparatus for implementing the method to perform dynamic fermentation, and includes the following steps:
(1) two cylindrical glass rollers which are same in shape and size and can rotate clockwise at a constant speed are arranged in parallel in the fermentation culture container with an upward opening. A spherical polyurethane material (core) having a diameter of 30mm was placed between two rolls, and the distance between the rolls was adjusted to 26mm and the rotation speed was adjusted to 4rpm as shown in FIG. 1. The spherical polyurethane can rotate along with the roller to realize vibration without horizontal displacement.
(2) Two heating guide plates are fixed at the top of the fermentation culture container, the angles of the two heating guide plates and the side surfaces of the fermentation culture container are adjusted to be 30 degrees, and one ends of the two heating plates are located in the gap where the spherical polyurethane is abutted to the roller.
(3) Selecting acetobacter xylinum capable of secreting bacterial cellulose to activate and prepare seed mash, and then mixing the seed mash with the strain concentration of 1wt% with a fermentation culture medium to obtain a fermentation mixed solution; wherein the fermentation medium is a high-temperature sterilized medium, and the components of the medium are the common components for bacterial cellulose fermentation in the field.
(4) Pouring the fermentation mixed liquid into a fermentation culture container, immersing a core material, starting a roller to rotate in the same direction for dynamic fermentation, fermenting for 3 days at 35 ℃, discharging fermentation liquid after the fermentation is finished, coating the outer layer of spherical polyurethane with bacterial cellulose obtained by bacterial strain fermentation, soaking the product in NaOH aqueous solution with the mass percentage of 4%, heating for 6 hours at the temperature of 100 ℃, and repeatedly washing the product to be neutral by using distilled water to form the spherical polyurethane-bacterial cellulose compound.
(5) And starting the heating guide plates, adjusting the heating temperature to 190 ℃, enabling the heating liquid of the thermoplastic high-molecular polyethylene to flow down along the heating guide plates at two ends respectively, and uniformly coating the thermoplastic high-molecular polyethylene on the surface of the core material-bacterial cellulose compound along with the rotation of the roller, thereby preparing the core-shell structure composite material with the spherical polyurethane material surface uniformly coated with the polyethylene/bacterial cellulose.
Example 2
The embodiment provides a method for preparing a core-shell structure bacterial cellulose composite material by dynamic fermentation, which adopts the device of the embodiment 1 to perform dynamic fermentation, and comprises the following steps:
(1) two cylindrical glass rollers which are same in shape and size and can rotate clockwise at a constant speed are arranged in parallel in the fermentation culture container with an upward opening. A bioceramic (core) 50mm in diameter and 70mm in length was placed between two rollers, the spacing between which was adjusted to 40mm and the rotation rate was adjusted to 8rpm, as shown in FIG. 1. The biological ceramics can realize vibration without horizontal displacement along with the rotation of the roller.
(2) Two heating guide plates are fixed at the top of the fermentation culture container, the angles of the two heating guide plates and the side surfaces of the fermentation culture container are adjusted to be 45 degrees, and one ends of the two heating plates are located in the gap where the biological ceramics and the roller are abutted.
(3) Selecting rhizobium capable of secreting bacterial cellulose, activating to prepare seed mash, and mixing the seed mash with the strain concentration of 2wt% with a fermentation culture medium to obtain a fermentation mixed solution; wherein the fermentation medium is sterilized at high temperature, the components of the medium are common components for bacterial cellulose fermentation in the field, and the fermentation mixed solution also comprises 0.1wt% of gelatin.
(4) Pouring the fermentation mixed solution into a fermentation culture container, immersing a core material, starting a roller to rotate in the same direction for dynamic fermentation, fermenting for 30 days at 20 ℃, discharging fermentation liquor after the fermentation is finished, coating bacterial cellulose obtained by bacterial strain fermentation on the outer layer of the biological ceramic, soaking the product in NaOH aqueous solution with the mass percentage of 5%, heating for 5 hours at the temperature of 90 ℃, and repeatedly washing with distilled water until the product is neutral to form the biological ceramic-bacterial cellulose compound.
(5) And starting the heating guide plates, adjusting the heating temperature to 230 ℃, enabling the heating liquid of the thermoplastic high-molecular polypropylene to flow down along the heating guide plates at two ends respectively, and uniformly coating the thermoplastic high-molecular polypropylene on the surface of the biological ceramic-bacterial cellulose compound along with the rotation of the roller, thereby preparing the core-shell structure composite material with the biological ceramic surface uniformly coated with the polypropylene/bacterial cellulose.
Example 3
The embodiment provides a method for preparing a core-shell structure bacterial cellulose composite material by dynamic fermentation, which adopts the device of the embodiment 1 to perform dynamic fermentation, and comprises the following steps:
(1) two cylindrical stainless steel rollers which have the same shape and size and can rotate clockwise at a constant speed are arranged in parallel in the fermentation culture container with an upward opening. A titanium alloy orthopedic implant screw (core) of 6.5mm was placed between two rollers, as shown in fig. 1, with the spacing between the rollers adjusted to 5mm and the rotation rate adjusted to 12 rpm. The screw can rotate along with the roller to realize vibration without horizontal displacement.
(2) Two heating guide plates are fixed at the top of the fermentation culture container, the angles of the two heating guide plates and the side surfaces of the fermentation culture container are adjusted to be 45 degrees, and one ends of the two heating plates are located in the gap where the screw abuts against the roller.
(3) Selecting sarcina capable of secreting bacterial cellulose, activating to prepare seed mash, and mixing the seed mash with the strain concentration of 3wt% with a fermentation culture medium to obtain a fermentation mixed solution; the fermentation medium is sterilized at high temperature, the components of the medium are common components for bacterial cellulose fermentation in the field, the fermentation mixed solution also comprises 1wt% of sodium hyaluronate and sodium alginate, and the mass ratio of the sodium hyaluronate to the sodium alginate is 1: 1.
(4) pouring the fermentation mixed liquid into a fermentation culture container, immersing a core material, starting a roller to rotate in the same direction for dynamic fermentation, fermenting for 5 days at 25 ℃, discharging fermentation liquid after the fermentation is finished, coating the outer layer of the screw with bacterial cellulose obtained by bacterial strain fermentation, soaking the product in NaOH aqueous solution with the mass percentage of 6%, heating for 4 hours at the temperature of 80 ℃, and repeatedly washing with distilled water until the product is neutral to form the screw-bacterial cellulose compound.
(5) Starting the heating guide plate, adjusting the heating temperature to 200 ℃, enabling the heating liquid of thermoplastic high polymer polypropylene to flow down along the heating guide plates at two ends respectively, rotating along with the roller, and uniformly coating the thermoplastic high polymer on the surface of the screw-bacterial cellulose compound, thereby preparing the core-shell structure composite material with the screw surface uniformly coated with the polypropylene/bacterial cellulose.
Example 4
The embodiment provides a method for preparing a core-shell structure bacterial cellulose composite material by dynamic fermentation, which adopts the device of the embodiment 1 to perform dynamic fermentation, and comprises the following steps:
(1) two cylindrical polytetrafluoroethylene rollers which are same in shape and size and can rotate clockwise at a constant speed are arranged in parallel in the fermentation culture container with an upward opening. A rod-like material (core material) of polymethyl methacrylate filled with nano-hydroxyapatite having a diameter of 2mm was placed between two rollers, and as shown in FIG. 1, the distance between the two rollers was adjusted to 1.5mm and the rotation speed was adjusted to 2 rpm. The bar can rotate along with the roller to realize vibration without horizontal displacement.
(2) Two heating guide plates are fixed at the top of the fermentation culture container, the angles of the two heating guide plates and the side surfaces of the fermentation culture container are adjusted to be 60 degrees, and one ends of the two heating plates are located in the gap where the bar is abutted to the roller.
(3) Selecting pseudomonas capable of secreting bacterial cellulose, activating to prepare seed mash, and mixing the seed mash with a strain concentration of 4wt% with a fermentation culture medium to obtain a fermentation mixed solution; wherein the fermentation medium is sterilized at high temperature, the components of the medium are common components for bacterial cellulose fermentation in the field, and the fermentation mixed solution also comprises 2wt% of soluble starch.
(4) Pouring the fermentation mixed liquid into a fermentation culture container, immersing a core material, starting a roller to rotate in the same direction for dynamic fermentation, fermenting for 7 days at 30 ℃, discharging fermentation liquid after the fermentation is finished, coating bacterial cellulose obtained by bacterial strain fermentation on the outer layer of the bar, soaking the product in NaOH aqueous solution with the mass percentage of 7%, heating for 6 hours at the temperature of 70 ℃, and repeatedly washing with distilled water until the bar-bacterial cellulose compound is neutral to form the bar-bacterial cellulose compound.
(5) And starting the heating guide plates, adjusting the heating temperature to 270 ℃, enabling the heating liquid of the thermoplastic polymer nylon to flow down along the heating guide plates at two ends respectively, and uniformly coating the thermoplastic polymer on the surface of the bar-bacterial cellulose compound along with the rotation of the roller, thereby preparing the core-shell structure composite material with the bar surface uniformly coated with the nylon/bacterial cellulose.
Example 5
The embodiment provides a method for preparing a core-shell structure bacterial cellulose composite material by dynamic fermentation, which adopts the device of the embodiment 1 to perform dynamic fermentation, and comprises the following steps:
(1) two cylindrical plastic rollers which are same in shape and size and can rotate clockwise at a constant speed are arranged in parallel in the fermentation culture container with an upward opening. The irregular-body breast-filling silicone rubber (core material) was placed between the two rolls, and the distance between the two rolls was adjusted to 70mm and the rotation rate was adjusted to 30rpm, as shown in FIG. 1. The silicon rubber can rotate along with the roller to realize vibration without horizontal displacement.
(2) Two heating guide plates are fixed at the top of the fermentation culture container, the angles of the two heating guide plates and the side surfaces of the fermentation culture container are adjusted to be 45 degrees, and one ends of the two heating plates are located in the gap where the silicon rubber is abutted to the roller.
(3) Mixing and activating achromobacter and alcaligenes capable of secreting bacterial cellulose to prepare seed mash, and then mixing the seed mash with the strain concentration of 5wt% with a fermentation culture medium to obtain a fermentation mixed solution; wherein the fermentation medium is sterilized at high temperature, the components of the medium are common components for bacterial cellulose fermentation in the field, and the fermentation mixed solution also comprises 3wt% of pectin.
(4) Pouring the fermentation mixed liquid into a fermentation culture container, immersing a core material, starting a roller to rotate in the same direction for dynamic fermentation, fermenting for 15 days at 31 ℃, discharging the fermentation liquid after the fermentation is finished, coating the outer layer of the silicon rubber with bacterial cellulose obtained by bacterial strain fermentation, soaking the product in NaOH aqueous solution with the mass percentage of 8%, heating for 5 hours at the temperature of 70 ℃, and repeatedly washing with distilled water until the product is neutral to form the silicon rubber-bacterial cellulose compound.
(5) And starting the heating guide plate, adjusting the heating temperature to 190 ℃, enabling the heating liquid of the thermoplastic high-molecular polyethylene to flow down along the heating guide plates at two ends respectively, and uniformly coating the thermoplastic high-molecular polyethylene on the surface of the silicon rubber-bacterial cellulose compound along with the rotation of the roller, thereby preparing the core-shell structure composite material with the silicon rubber surface uniformly coated with the polyethylene/bacterial cellulose.
Example 6
The embodiment provides a method for preparing a core-shell structure bacterial cellulose composite material by dynamic fermentation, which adopts the device of the embodiment 1 to perform dynamic fermentation, and comprises the following steps:
(1) two cylindrical plastic rollers which are same in shape and size and can rotate clockwise at a constant speed are arranged in parallel in the fermentation culture container with an upward opening. An irregularly shaped cobalt chromium molybdenum alloy artificial hip head (core) was placed between two rollers, as shown in fig. 1, with the spacing between the rollers adjusted to 50mm and the rotation rate adjusted to 15 rpm. The alloy can rotate along with the roller to realize vibration without horizontal displacement.
(2) Two heating guide plates are fixed at the top of the fermentation culture container, the angles of the two heating guide plates and the side surfaces of the fermentation culture container are adjusted to be 60 degrees, and one ends of the two heating plates are located in the gap where the alloy is abutted to the roller.
(3) Selecting aerobacter and azotobacter capable of secreting bacterial cellulose, mixing and activating to prepare seed mash, and then mixing the seed mash with a strain concentration of 3wt% with a fermentation culture medium to obtain a fermentation mixed solution; wherein the fermentation medium is sterilized at high temperature, the components of the medium are common components for bacterial cellulose fermentation in the field, the fermentation mixed solution also comprises 5wt% of chitosan and sodium carboxymethyl cellulose, and the mass ratio of the chitosan to the sodium carboxymethyl cellulose is 1: 3.
(4) pouring the fermentation mixed solution into a fermentation culture container, immersing a core material, starting a roller to rotate in the same direction for dynamic fermentation, fermenting for 10 days at 32 ℃, discharging fermentation liquor after the fermentation is finished, coating the outer layer of the alloy with bacterial cellulose obtained by bacterial strain fermentation, soaking the product in NaOH aqueous solution with the mass percentage of 6%, heating for 4 hours at the temperature of 90 ℃, and repeatedly washing with distilled water until the product is neutral to form the alloy-bacterial cellulose composite.
(5) And starting the heating guide plates, adjusting the heating temperature to 190 ℃, enabling the heating liquid of the thermoplastic high-molecular polyethylene to flow down along the heating guide plates at two ends respectively, and uniformly coating the thermoplastic high-molecular polyethylene on the surface of the alloy-bacterial cellulose compound along with the rotation of the roller, thereby preparing the core-shell structure composite material with the alloy surface uniformly coated with the polyethylene/bacterial cellulose.
Performance test experiments:
the core-shell structure composite material prepared in example 1 and having the surface uniformly coated with the polyethylene/bacterial cellulose was subjected to the following performance tests:
biocompatibility experiment: the composite material (example 1) and the polyurethane material (core material of example 1) were evaluated for cytotoxicity, delayed contact sensitization of guinea pigs, skin irritation, and the like, respectively, with reference to biological evaluation of GB/T16886 medical devices.
The results show that: the polyurethane material (core material of example 1) has cytotoxicity of grade 2 and has skin sensitization reaction; the composite material (example 1) has cytotoxicity less than 2 grade, no skin sensitization reaction and no skin irritation reaction, and has good biological safety. It is demonstrated that the use of this patent improves the biocompatibility properties of the material.
Claims (13)
1. A device for preparing a bacterial cellulose composite material with a core-shell structure through dynamic fermentation comprises:
the fermentation culture container and two rollers and two heating guide plates arranged in the fermentation culture container;
two ends of the rotating shaft of the two rollers are respectively movably connected with the inner wall of the fermentation culture container, the two rollers are arranged in parallel in the horizontal direction, a gap is arranged between the rollers, and the distance between the rotating shafts of the rollers is adjustable;
the two heating guide plates are parallel to the rotating shafts of the two rollers, one end of one heating guide plate is movably connected with the fermentation culture container, and the other end of the other heating guide plate extends obliquely downwards to the upper part of the gap of the two rollers and is abutted against one of the two rollers; one end of the other heating guide plate is movably connected with the fermentation culture container, and the other end of the other heating guide plate extends to the upper part of the gap of the two rollers in a downward inclined mode and is abutted against the other roller.
2. The device of claim 1, further comprising a driving member for driving the two rollers to rotate, preferably, the driving member is used for driving the two rollers to rotate in the same direction.
3. The device of claim 1, further comprising a heating element that heats both heating baffles.
4. The apparatus of claim 1, wherein the two rollers are cylindrical rollers of the same shape and size.
5. The device of claim 1, wherein the two heating guide plates are respectively arranged at an angle of 15-60 degrees with the inner side wall of the fermentation culture container.
6. A method for preparing a bacterial cellulose composite material with a core-shell structure by dynamic fermentation, which is to perform dynamic fermentation by adopting the device of any one of claims 1 to 5, and comprises the following steps:
adjusting the distance between the rollers to enable the minimum width of the gap to be smaller than the diameter or the length of the core material, placing the core material to be coated above the gap between the two rollers after sterilizing, and ensuring that the core material can be respectively abutted against the two rollers above the gap; the rotation speed and the rotation direction of the roller are set, so that the core material can realize vibration without horizontal displacement along with the rotation of the roller;
setting the length of the heating plates and adjusting the angles of the heating plates to ensure that one ends of the two heating plates are positioned at a gap where the core material is abutted to the roller;
preparing a bacterial cellulose fermentation culture solution, carrying out high-pressure sterilization, and then mixing the bacterial cellulose fermentation culture solution with strain seed mash to obtain a fermentation mixed solution;
pouring the fermentation mixed liquid into a fermentation culture container, immersing the core material, starting a roller to rotate in the same direction for dynamic fermentation, discharging fermentation liquid after the fermentation is finished, and coating the outer layer of the core material with bacterial cellulose obtained by bacterial strain fermentation to form a core material-bacterial cellulose compound; starting the heating guide plates, enabling the heating liquid of the thermoplastic macromolecules to respectively flow down along the heating guide plates at two ends, and uniformly coating the thermoplastic macromolecules on the surface of the core material-bacterial cellulose compound along with the rotation of the roller, so as to prepare the bacterial cellulose composite material with the core-shell structure; alternatively, the first and second electrodes may be,
starting the heating guide plates, enabling the heating liquid of the thermoplastic macromolecules to respectively flow down along the heating guide plates at two ends, and uniformly coating the thermoplastic macromolecules on the surface of the core material along with the rotation of the roller to obtain a core material-thermoplastic macromolecule compound; pouring the fermentation mixed liquid into a fermentation culture container, immersing a roller, starting the roller to rotate in the same direction for dynamic fermentation, discharging the fermentation liquid after the fermentation is finished, and covering the outer layer of the core material-thermoplastic polymer compound with bacterial cellulose obtained by bacterial strain fermentation so as to prepare the bacterial cellulose composite material with the core-shell structure.
7. The method according to claim 6, further comprising repeating the step of dynamically fermenting the coated bacterial cellulose and/or the step of coating the thermoplastic polymer to obtain a further hierarchical core-shell structure bacterial cellulose composite material.
8. The method of claim 6, wherein the core material comprises a combination of one or more of an inorganic material, an organic polymer material, and a metal material;
preferably, the shape of the core material is a sphere, a sphere-like shape, a cylinder, a rod or any irregularity.
9. The method of claim 6, wherein the thermoplastic polymer material comprises a combination of one or more of polyethylene, polypropylene, polystyrene, polymethylmethacrylate, nylon, polyurethane, polyester, and polylactic acid;
preferably, the heating temperature of the heating guide plate is 50-300 ℃.
10. The method of claim 6, wherein the strain comprises a combination of one or more of Acetobacter xylinum, Rhizobium, Sphaerococcus, Pseudomonas, Achromobacter, Alcaligenes, Aerobacter, and Azotobacter;
preferably, the addition amount of the strain seed mash is 1-5 wt% of the fermentation culture solution;
preferably, the dynamic fermentation is carried out for 3-30 days; the fermentation temperature is 20-30 ℃;
preferably, when the dynamic fermentation is carried out, the rotating speeds of the two rollers are the same and are both 0.1-60 rpm; preferably 4 to 20 rpm.
11. The method according to claim 6, wherein in the dynamic fermentation process, 0.1-5 wt% of soluble additives are added into the fermentation mixed liquor; the soluble additive comprises one or more of gelatin, sodium hyaluronate, starch, pectin, chitosan, sodium alginate and soluble cellulose derivatives.
12. The method according to claim 6, wherein the step of purifying the bacterial cellulose-coated complex after the dynamic fermentation is completed comprises: washing the mixture for 4-6 hours in a NaOH aqueous solution with the mass percentage of 4-8% at the temperature of 70-100 ℃, and repeatedly washing the mixture to be neutral by using distilled water.
13. The bacterial cellulose composite material with the core-shell structure, which is prepared by the method of any one of claims 6 to 12, has an inner core layer which is a core material, and the core material is coated with a single layer or multiple layers of bacterial cellulose and/or thermoplastic polymer materials.
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