TW201420764A - Manufacturing method of fabricated 3-D composites bio-film - Google Patents

Abstract

The manufacturing method of fabricated 3-D composites bio-film, which utilizes bacterial cellulose to form the main component of the fabricated bio-film, and makes the bio-film possesses mechanical strength, permeability and plasticity by adding natural polymers and cross-linking agent, enabling the bio-film to prepare 3-D film with control-release ability. Moreover, the 3-D bio-film exhibits more desirable overlying skin attached effect to offer the isolation and prevention from secondary infection.

Description

Method for preparing stereoscopic composite biofilm

The invention relates to a method for preparing a biofilm, in particular to a method for preparing a wet or dry stereorecombination composite biofilm.

At present, the method for producing a bacterial fiber biofilm is a batch fermentation culture of glucose xyloacetic acid bacteria, and a vinegar film is produced in one plate and then cut into a desired thickness and shape by cutting, but in biology. In the film process, it needs to be statically set and expanded for several days, so there is a problem of insufficient uniformity of finished products, and it is impossible to automate continuous production, which leads to a low yield and production speed, and thus the cost is high; and after the film is cut, It is impossible to prepare a three-dimensional film, and the film cannot be used for each curved portion of the human body; in addition, in order to cut a film having uniformity, a large amount of scrap is generated.

In addition, the crystallinity of bacterial cellulose increases during the drying process, which leads to the inability of the dehydrated film to rehydrate rapidly. Therefore, the bacterial cellulose biofilm is not yet stored and transported in a dehydrated state, so its convenience is lowered, resulting in difficulty in commercialization. .

It can be seen that the above-mentioned conventional methods for preparing biofilms still have many defects, which is not a good design, and needs to be improved.

In view of this, the inventors of the present invention have developed a technique for overcoming the disadvantages of the above-mentioned conventional methods for preparing biofilms, and the technique utilizes bacteria secreted by Gluconobacter oxysporum. The cellulose, or the waste vinegar film of the vinegar industry, or the waste material of the bio-mask industry as the main raw materials, after pre-treatment such as washing, fine crushing and derivatization, and then mixing the hydrophilic colloid and polymer compound, and finally The cross-linking technology was used to carry out the cross-linking reaction, and finally the method for preparing the stereo-recombinant composite biofilm was successfully developed.

The object of the present invention is to provide a method for preparing a stereorecombination composite biofilm, which overcomes the problem that the traditional fermentation culture is difficult to produce a three-dimensional biofilm with high rehydration ability, high coating property and low price.

The invention provides a preparation method of a wet stereo-recombination composite biofilm, which comprises the steps of obtaining bacterial cellulose, homogenizing after removing impurities, and then derivatizing it to prepare derivatized cellulose; then adding derivatized cellulose to water , colloid, heating and stirring uniformly to prepare a wet film glue; then, the wet film glue is injected into the three-dimensional mold to form a film, and the three-dimensional mold containing the wet film glue is statically placed at a predetermined temperature and Then, the three-dimensional mold containing the wet film glue is immersed in the cross-linking agent solution for cross-linking reaction; after the cross-linking reaction, the wet film glue liquid forms a wet stereo-recombination composite biofilm in the three-dimensional mold.

Wherein, the step of preparing the derivatized cellulose is to oxidize the carboxyl group to decompose the carboxyl group; the step of preparing the wet film glue liquid is to heat and stir the water, the alginate and the gelatin to form a wet film glue liquid, wherein the wet The moisture content of the film glue is 70~99.9% (w/v), the concentration of alginate is 0.1~5.0% (w/v), the concentration of gelatin is 0.1~5.0% (w/v); film forming step The wet film glue is separately injected into the three-dimensional mold; the step of standing is to hold the three-dimensional mold containing the wet film glue at 0-20 ° C for 0.5-24 hours; the crosslinking reaction step will contain the wet state. The three-dimensional mold of the film glue is immersed in a crosslinking agent solution having a concentration of 0.5-20% (w/v) for crosslinking reaction, wherein the crosslinking agent is calcium chloride; finally, after the crosslinking reaction, the wet film The glue liquid forms a wet stereoscopic composite composite biofilm in a three-dimensional mold.

The invention further provides a preparation method of a dry stereoscopic composite biofilm, the steps of which are first to obtain bacterial cellulose, homogenize after removing impurities, and then derivatized to prepare derivatized cellulose; then, the derivatized cellulose is added. Water, colloid, plasticizer and chitosan, heated and stirred evenly, adjust the pH value to prepare dry film glue; then dry film glue is injected into the three-dimensional mold to form a film, and will contain dry state The three-dimensional mold of the film glue is statically placed at a predetermined temperature and time; then the three-dimensional mold containing the dry film glue is immersed in the crosslinking agent solution for crosslinking reaction; finally, the film is removed, and the three-dimensional film is removed. The recombinant composite biofilm is freeze-dried to form a dry stereoscopic composite biofilm.

Wherein, the step of preparing the derivatized cellulose is to oxidize the carboxyl group to derivatize the carboxyl group; the step of preparing the dry film glue liquid is to add the derivatized cellulose to water, alginate, chitosan and a plasticizer, and heat and stir. Evenly, finally adjust the pH value to 4~10 to make it a dry film glue. The moisture content of the dry film glue is 70~99.9% (w/v) and the alginate concentration is 0.1~5.0% (w /v), chitosan concentration is 0.1~5.0% (w/v), plasticizer concentration is 0.1~5.0% (w/v), and the chitosan has molecular weight greater than 10kDa and deacetylation degree More than 75% of chitosan powder; the film forming step is to inject the dry film glue into the three-dimensional mold; the step of standing is to place the three-dimensional mold containing the dry film glue at 0~20 °C for 0.5. ~24 hours; the crosslinking reaction step is to immerse a three-dimensional mold containing a dry film glue in a crosslinking agent solution having a concentration of 0.5 to 20% (w/v) for crosslinking reaction, wherein the crosslinking agent is chlorinated Calcium; Finally, after cross-linking reaction, the dry film glue liquid forms a dry stereo-recombination composite biofilm in a three-dimensional mold.

Although biofilm production technology is currently available in China, most of the bacterial cellulose membranes are directly fermented using glucose xyloacetate. The invention can also utilize the vinegar film of the vinegar industry or the bottom material of the nata or the biological mask as the basic material of the biofilm, and the substrates are all bacterial cellulose having a nano-structure, which not only solves the processing. The problem of waste and the creation of high added value, its characteristics are: (1) The three-dimensional film can be prepared to overcome the shortcomings of the current biofilm or dressing flat products, for example, it can be used as a dressing for a special part such as a "finger film" to increase the conformability; (2) the raw materials are all bio-friendly. Degradable substances, and the source is wide, the price is low (or even no cost); (3) the rehydration ability is superior to the general commercially available bacterial fiber film; (4) the network structure can effectively coat and release functional substances (such as Essence, antibacterial substances, anti-inflammatory substances, etc.).

In addition, chitosan is a natural biopolymer present in the shells of crustaceans and arthropods, usually extracted from the shell of shrimps and crabs. Chitosan has the characteristics of tissue compatibility and biological activity, which can reduce environmental pollution and increase economic value. Chitosan and its derivatives have antibacterial properties, can effectively inhibit the growth of Gram-negative bacteria and some Gram-positive bacteria, and can be used to prevent the growth of food pathogens and extend the shelf life. The potential for natural fungicides. By analyzing the molecular weight of chitosan and the effect of different deacetylation on the antibacterial ability of chitosan, it is shown that the minimum inhibitory concentration of chitosan on S. aureus increases with the degree of deacetylation. It is reduced, so the chitosan with high degree of deacetylation has a lower antibacterial effect than the lower one. Therefore, when preparing a stereoscopic biofilm, the addition of chitosan having a high degree of deacetylation can effectively increase the antibacterial property of the stereoscopic biofilm.

The present invention is exemplified by the following examples, but the present invention is not limited by the following examples.

Embodiments of the present invention provide a method for preparing a wet stereoscopic composite biofilm, which is shown in FIG. 1 and includes the following steps:

Step S11: preparing a derivatized cellulose; the bacterial cellulose is a vinegar film produced by acetic acid bacteria or under it The waste material is rinsed to remove impurities and residual bacterial liquid, and then homogenized by a stirrer. After dehydration, the oxidation reaction is carried out for 6-24 hours, and the oxidized liquid is repeatedly washed and removed to obtain a derivatized cellulose having a functional group such as a carboxyl group. When the bacterial cellulose is oxidized to form a carboxyl group, the fiber bundle becomes wider and the network voids are larger, which provides more space for adsorbing other gel materials, forms a mixed gel, and improves the mechanical properties of the film.

Step S12: preparing a wet film glue; adding alginate and gelatin powder to water and heating and dissolving in a hot water bath, and then adding the derivatized cellulose to the mixer to uniformly mix to prepare a moisture content of 70.0 to 99.9% (w/v). State film glue; wherein the concentration of the alginate is 0.1~5.0% (w/v), and the concentration of gelatin is 0.1~5.0% (w/v); considering the film forming step, if the moisture is too high, the wet state The film glue is not easy to gel set during the standing process; too little mixing is uneven during the mixing process, and the viscosity is too high, which may cause easy adhesion to the mold, and it is difficult to form a flat film, so it is necessary to select both good flow properties. And the range of moisture content of mechanical strength.

Step S13: forming a film; injecting the wet film glue into the three-dimensional mold, so that the wet film glue adheres to the three-dimensional mold, and finally forms a three-dimensional film.

Step S14: standing; the three-dimensional mold containing the wet film glue is allowed to stand in a 0 to 20 ° C and low humidity environment for 0.5 to 24 hours.

Step S15: crosslinking reaction; after standing, the three-dimensional mold containing the wet film glue is immersed in a crosslinking agent solution having a concentration of 0.5-20% (w/v), and the crosslinking reaction is carried out for 0.5 to 24 hours. The cross-linking agent is calcium oxide; after the cross-linking reaction, the wet film glue liquid forms a wet stereo-recombination composite biofilm in a three-dimensional mold.

Step S16: removing the film; after removing the mold, the wet stereoscopic composite biofilm is obtained; When the proportion of derivatized cellulose decreases and the proportion of colloid increases, the mechanical strength of the resulting film will increase and the hardness will increase (tensile force up to 100 kPa and elongation rate of more than 200%), so in order to obtain proper film comfort and adhesion Sexuality and texture, the ratio between the derived cellulose and the colloid can be adjusted moderately.

Another embodiment of the present invention provides a method for preparing a dry stereoscopic composite biofilm, which is shown in FIG. 2 and includes the following steps:

Step S21: preparing derivatized cellulose; the bacterial cellulose is a vinegar film produced by acetic acid bacteria or a waste thereof, and after rinsing to remove impurities and residual bacterial liquid, homogenizing by a mixer, dehydrating and then performing an oxidation reaction for 6 to 24 hours, and repeatedly washing After removing the oxidizing solution, it is a derivatized cellulose having a functional group such as a carboxyl group. When the bacterial cellulose is oxidized to form a carboxyl group, the fiber bundle becomes wider and the network voids are larger, which provides more space for adsorbing other gel materials, forms a mixed gel, and improves the mechanical properties of the film.

Step S22: preparing a dry film glue; adding the alginate, the chitosan powder and the plasticizer to the water and heating and dissolving in a hot water bath, and then adding the derivatized cellulose to the mixer and stirring uniformly to prepare a moisture content of 70.0 to 99.9. %(w/v) dry film glue; the concentration of alginate in the dry film glue is 0.1~5.0% (w/v), and the concentration of chitosan is 0.1~5.0% (w/v) The concentration of the plasticizer is 0.1 to 5.0% (w/v); wherein the chitosan is a chitosan powder having a molecular weight of more than 10 kDa and a deacetylation degree of more than 75%; If the moisture is too high, the dry film glue is not easy to gel set during the standing process; too little is unevenly mixed during the stirring process, and the viscosity is too high, which may cause easy adhesion to the mold, and it is difficult to form a flat film. Choose a moisture content that combines good flow properties with mechanical strength range.

Step S23: adjusting the pH value; adjusting the pH of the dry film glue to a range of 4 to 10.

Step S24: forming a film; injecting the dry film glue into the three-dimensional mold, causing the dry film glue to adhere to the three-dimensional mold, and finally forming a three-dimensional film.

Step S25: standing; the three-dimensional mold containing the dry film glue liquid is allowed to stand in the environment of 0-20 ° C and low humidity for 0.5 to 24 hours.

Step S26: crosslinking reaction; after standing, the three-dimensional mold containing the dry film glue is immersed in a crosslinking agent solution having a concentration of 0.5-20% (w/v), and the crosslinking reaction is carried out for 0.5 to 24 hours. Wherein the crosslinking agent is calcium oxide.

Step S27: stripping; after the cross-linking reaction, the stereo-recombination composite biofilm can be obtained by removing the mold.

Step S28: freeze-drying; the stereo-recombination composite biofilm after the film is removed, and then freeze-drying, and finally form a dry stereo-recombination composite biofilm; when the colloid ratio is increased, the cross-linking effect of the film can be fully exerted, and after rehydration The tensile strength can reach more than 500 kPa.

It can be seen from the above that the wet stereo-recombination composite biofilm prepared by the invention takes the cellulose derived from bacterial cellulose as a main component and forms a new carboxyl group on the surface of the fiber, so that the fiber is more easily compounded with other polymers, and the application is improved. Therefore, it can be used to prepare a wet stereocomplex composite biofilm to overcome the shortcomings of the conventional biofilm produced by direct fermentation of the bacteria, which is difficult to prepare stereotypes and continuous production. The dry stereo-recombination composite biofilm prepared by the invention has the advantages of good gas permeability, and is different from the biofilm made of commercially available bacterial cellulose, which cannot be rehydrated after drying, and the dry stereoscopic recombination compound Biofilms have the ability to rehydrate quickly and after rehydration The film still has a certain mechanical strength and texture; in addition, the dry stereoscopic composite biofilm has the advantages of easy storage, transportation and convenient use.

The above-mentioned embodiments and the drawings are only the preferred embodiments of the present invention, and the scope of the present invention is not limited thereto, that is, the equivalent variations and modifications of the scope of the present invention should be covered by the present invention. Within the scope.

In summary, the method for preparing a stereorecombination composite biofilm provided by the present invention is innovative, and can improve the above-mentioned plurality of functions by the conventional method for preparing a biofilm, and should fully meet the statutory invention patent requirements of novelty and progress.提出 Submit an application in accordance with the law, and ask your office to approve the application for this invention patent, in order to invent invention, to the sense of virtue.

S11‧‧‧Preparation of derived cellulose

S12‧‧‧Preparation of wet film glue

S13‧‧‧ film

S14‧‧‧Stay

S15‧‧‧ cross-linking reaction

S16‧‧‧ release film

S21‧‧‧Preparation of derived cellulose

S22‧‧‧ Preparation of dry film glue

S23‧‧‧ Adjust pH

S24‧‧‧ film

S25‧‧‧Stay

S26‧‧‧ cross-linking reaction

S27‧‧‧ release film

S28‧‧‧ freeze drying

FIG. 1 is a flow chart of the preparation of the wet stereoscopic composite composite biofilm of the present invention.

2 is a flow chart of the dry stereo composite composite biofilm of the present invention.

S11‧‧‧Preparation of derived cellulose

S12‧‧‧Preparation of wet film glue

S13‧‧‧ film

S14‧‧‧Stay

S15‧‧‧ cross-linking reaction

S16‧‧‧ release film

Claims (26)

The invention relates to a method for preparing a wet stereo-recombination composite biofilm, which comprises the following steps: (1) preparing a derivatized cellulose: taking bacterial cellulose, homogenizing and derivatizing after removing impurities, thereby becoming a derivatized cellulose; Preparing a wet film glue liquid: adding the derivatized cellulose to water, colloid, heating and stirring uniformly to become a wet film glue liquid; (3) film forming: injecting the wet film glue into a three-dimensional mold; (4) Resting: the stereo mold containing the wet film glue is statically placed at a predetermined temperature and time; (5) cross-linking reaction: after standing, the three-dimensional mold containing the wet film glue is impregnated in the cross-linking The agent solution is subjected to a crosslinking reaction; after the crosslinking reaction, the wet film glue liquid forms a wet stereoscopic composite biofilm in a three-dimensional mold. The method for preparing a wet stereo-recombination composite biofilm according to claim 1, wherein the derivatization treatment deoxidizes the bacterial cellulose to a carboxyl group. The method for preparing a wet stereocomplex composite biofilm according to claim 1, wherein the moisture film has a moisture content of 70-99.9% (w/v). The preparation method of the wet stereoscopic composite composite biofilm according to claim 1, wherein the colloid comprises alginate and gelatin. The preparation method of the wet stereoscopic composite composite biofilm according to claim 4, wherein the concentration of the alginate is 0.1 to 5.0% (w/v). The preparation method of the wet stereoscopic composite composite biofilm according to claim 4, wherein the gelatin concentration is 0.1 to 5.0% (w/v). The method for preparing a wet stereoscopic composite composite biofilm according to claim 1, wherein the predetermined temperature is 0 to 20 °C. The preparation method of the wet stereoscopic composite composite biofilm according to claim 1, wherein the predetermined time is 0.5 to 24 hours. The method for preparing a wet stereocomplex composite biofilm according to claim 1, wherein the crosslinking reaction time is 0.5 to 24 hours. The method for preparing a wet stereocomplex composite biofilm according to claim 1, wherein the crosslinking agent is calcium oxide. The method for preparing a wet stereocomplex composite biofilm according to claim 10, wherein the calcium oxide concentration is 0.5-20% (w/v). The invention relates to a method for preparing a dry stereoscopic composite biofilm, which comprises the following steps: (1) preparing a derivatized cellulose: taking bacterial cellulose, homogenizing and derivatizing after removing impurities, thereby becoming a derivatized cellulose; Preparing a dry film glue: adding the derivatized cellulose to water, colloid, chitosan and plasticizer, heating and stirring uniformly, and finally adjusting the pH value to become a dry film glue; (3) film formation: The dry film glue is injected into the three-dimensional mold; (4) standing: the three-dimensional mold containing the dry film glue is statically placed at a predetermined temperature and time; (5) crosslinking reaction: after standing, The three-dimensional mold containing the dry film glue is immersed in the cross-linking agent solution for crosslinking reaction; (6) freeze-drying: after the cross-linking reaction, the stereo-recombination composite biofilm can be obtained by removing the mold, and the three-dimensional reconstituted biofilm is obtained. The recombinant composite biofilm is freeze-dried, and finally a dry stereoscopic composite biofilm is formed. The method for preparing a dry stereo-recombination composite biofilm according to claim 12, wherein the derivatization treatment deoxidizes the bacterial cellulose to a carboxyl group. A method for preparing a dry stereoscopic composite biofilm according to claim 12 of the patent application, The moisture content of the dry film glue is 70 to 99.9% (w/v). The method for preparing a dry stereo-recombination composite biofilm according to claim 12, wherein the colloid is alginate. The method for preparing a dry stereoscopic composite biofilm according to claim 15 wherein the concentration of the alginate is 0.1 to 5% (w/v). The method for preparing a dry stereo-recombination composite biofilm according to claim 12, wherein the chitosan is a chitosan powder having a molecular weight of more than 10 kDa and a deacetylation degree of more than 75%. The method for preparing a dry stereo-recombination composite biofilm according to claim 17 wherein the concentration of the chitosan is 0.1 to 5.0% (w/v). The method for preparing a dry stereo-recombination composite biofilm according to claim 12, wherein the concentration of the plasticizer is 0.1 to 5% (w/v). The method for preparing a dry stereo-recombination composite biofilm according to claim 12, wherein the pH is 4-10. The method for preparing a dry stereoscopic composite biofilm according to claim 12, wherein the predetermined temperature is 0 to 20 °C. The method for preparing a dry stereoscopic composite biofilm according to claim 12, wherein the predetermined time is 0.5 to 24 hours. The method for preparing a dry stereo-recombination composite biofilm according to claim 12, wherein the crosslinking reaction time is 0.5 to 24 hours. The method for preparing a dry stereo-recombination composite biofilm according to claim 12, wherein the crosslinking agent is calcium oxide. The method for preparing a dry stereo-recombination composite biofilm according to claim 24, wherein the calcium oxide concentration is 0.5-20% (w/v). A method for preparing a dry stereoscopic composite biofilm according to claim 12 of the patent application, The moisture content of the dry stereoscopic composite biofilm is 1% to 20%.