Embodiments relate to a nanocellulose support and a method for producing the same.
Cell therapy is a direct and groundbreaking method of treating diseases by injecting living cells into an affected area and has recently gained the most interest in the bio industry. Due to the nature of the treatment method in which cells have to be directly injected into the patient, one of the most important considerations when developing the treatments is the quality of the injected cells, that is, the safety of heteroimmunity and mutation of the cells.
Nanocellulose is a crystalline portion of cellulose, which is the main component of plant cell walls, and is an eco-friendly material with excellent mechanical properties due to hydrogen bonding between molecules. Due to the nature of the material extracted from plants, the nanocellulose is a non-animal and biocompatible material, and thus, there are fewer issues of heteroimmunity or cell mutation compared to the existing animal-derived cell culture materials. In addition, the nanocellulose is known to be advantageous for cell culture because the nanocellulose has a similar structure to the extracellular matrix (ECM). In spite of these advantages, since the nanocellulose is not easy to be attached to cells, it is difficult to mass-produce cells, and it is used only for research purposes as a type of culture in a hydrogel.
An object of embodiments is to solve the above and other problems.
Another object of embodiments is to provide a nanocellulose support suitable for cell culture and a method for producing the same.
Further another object of embodiments is to provide a nanocellulose support capable of mass-producing cells and a method for producing the same.
According to an aspect of an embodiment to achieve the above or other object, a method for producing a nanocellulose support includes: applying a surface-treated nanocellulose solution to a container; drying the applied nanocellulose solution to form a nanocellulose thin film; and modifying surface properties of the nanocellulose thin film by using electron beam irradiation.
According to another aspect of an embodiment, a method for producing a nanocellulose support includes: dropping a plurality of beads into a container; dropping a surface-treated nanocellulose solution into the container; performing a drying process to form a nanocellulose thin film configured to surround an outer circumferential surface of each of the plurality of beads; and modifying surface properties of the nanocellulose thin film by using electron beam irradiation.
The nanocellulose support according to the embodiments and the effects of the method for producing the same are described as follows.
According to at least one of the embodiments, there may be the advantage in that the wettability of the culture solution is improved by the hydrophilizing the nanocellulose thin film formed by the drying process through the electron beam irradiation.
According to at least one of the embodiments, there may be the advantage in that the production of the nanocellulose support using the drying method allows the substrates having various shapes to be coated and has simple processes, thus allowing the mass production and the production of the over-sized support.
According to at least one of the embodiments, there may be the advantage in that the wettability of the culture solution is improved through the hydrophilic treatment to remove the microbubbles in the fiber, thereby improving the cell attachment.
According to at least one of the embodiments, there may be the advantage in that the nanocellulose support is easily decomposed using the plant degradation enzyme to minimize the damage of the cells compared to the method of collecting the cells by cutting the attached site of the cell by using the existing animal degradation enzyme, and the nanocellulose support is decomposed in the short time compared to the existing hydrogel to obtain the cells having the excellent quality in the short time.
According to at least one of the embodiments, there may be the advantage in that it is possible to provide the cell culture method suitable for the mass production in the simple and fast process by overcoming the material limitations of the nanocellulose, thereby significantly contributing to the bio industries in the future.
The additional scope of the applicability of the embodiments will become apparent from the detailed description below. However, the various changes and modifications within the spirit and scope of the embodiments may be clearly understood by those skilled in the art, and thus, specific embodiments such as the detailed description and the preferred embodiments should be understood as given only as examples.
Hereinafter, preferred embodiments of the present invention will be described below in more detail with reference to the accompanying drawings. However, the technical spirit of the present invention is not limited to some embodiments described, but may be implemented in various different forms, and within the technical spirit scope of the present invention, one or more of the components between the embodiments may be selectively coupled and substituted for the use. In addition, terms (including technical and scientific terms) used in the embodiments of the present invention, unless explicitly defined and described, can be generally understood by those skilled in the art to which the present invention pertains, and meanings of the terms, which are commonly used, such as predefined terms may be interpreted by considering the contextual meaning of the related technology. In addition, the terms used in the embodiments of the present invention are used only for explaining a specific exemplary embodiment while not limiting the present invention. In the present specification, a singular form may also include a plural form unless specifically stated in the phrase, and when described as “at least one (or more than one) of B, and (or) C”, a combination of A, B, and C can contain one or more of all possible combinations. In the description of the components of the present invention, the terms first, second, A, B, (a), and (b) may be used. Each of the terms is merely used to distinguish the corresponding component from other components, and does not delimit an essence, an order or a sequence of the corresponding component. In addition, when any component is described as being ‘connected’, ‘coupled’ or ‘linked’ to another component, not only the component is directly connected, coupled, or linked to the other component, but also to the component is ‘connected’, ‘coupled’ or ‘linked’ by another component between the other components. In addition, when described as being formed or disposed in the “upper (top) or below (bottom)” of each component, the upper (top) or below (bottom) is not only when the two components are in direct contact with each other, but also a case in which another component described above is formed or disposed between the two components. In addition, when expressed as “upper (top) or below (bottom)”, it may include the meaning of the downward direction as well as the upward direction based on one component.
Referring to
For example, the nanocellulose solution may be produced by dispersing nanocellulose into a solution.
The nanocellulose is a crystalline portion of cellulose, which is the main component of a plant cell wall, and may be composed of nano-sized particles. For example, the nanocellulose may be produced by crushing raw wood and performing a series of processing procedures. In these processing procedures, a procedure for allowing the nanocellulose to have hydrophilicity may be included.
The nanocellulose may be, for example, nanocellulose surface-treated with anions. The anion may include, for example, a reactive group such as a sulfonic acid group, a phosphonic acid group, a carboxyl group, a sulfuric acid group, or a phosphoric acid group.
The nanocellulose may be, for example, nanocellulose surface-treated with cations. The cation may include, for example, an amine group including an epoxypropyltrimethylammonium group, a diethylaminoethyl group, and a dimethylamionethyl group, an amide group, an amino group, an ammonium group, a phosphonium group, and a sulfonium group.
It is seen that an amount of cell attachment in the nanocellulose thin film formed on the basis of cationic nanocellulose (
Therefore, when the nanocellulose thin film is formed based on the cationic nanocellulose or the anionic nanocellulose according to the first embodiment, mass production of the cells may be possible.
A concentration of the nanocellulose may be 0.01% to 5% by weight. Other concentrations other than these concentrations of nanocellulose may be solutions. For example, the concentration of the nanocellulose may be 0.01% to 2% by weight. For example, the concentration of the nanocellulose may be 0.03% to 1% by weight.
The solution may be, for example, distilled water. The solution may be, for example, sterilized bio-distilled water. The solution may be, for example, a mixed solution in which one of distilled water or sterilized bio-distilled water and ethanol are mixed.
Since the nanocellulose is treated to be hydrophilic, the nanocellulose dispersed in the nanocellulose solution may also be maintained with hydrophilicity. Thus, the nanocellulose solution may be called a surface-treated nanocellulose solution.
For example, the surface-treated nanocellulose solution may be applied to a container.
As illustrated in
The surface-treated nanocellulose solution 112 may be applied to a bottom surface of the container 10. The applied nanocellulose solution may be dried to form the nanocellulose thin film (S214).
For example, the drying process may be performed by a heat source such as a heater or an oven.
A drying temperature may be, for example, 50 degrees to 100 degrees. A drying temperature may be, for example, 60 degrees to 90 degrees.
As illustrated in
Thus, a nanocellulose support in which the nanocellulose thin film 114 is formed directly on the container 10 may be produced.
The nanocellulose support having modified surface properties of the nanocellulose thin film may be produced using electron beam irradiation (S215).
The nanocellulose support may include the container 10 and the nanocellulose thin film 114 disposed in the container 10 to increase in hydrophilicity.
The electron beam irradiation may be, for example, O2 plasma or UV/O3 irradiation.
In the embodiment, the O2 plasma is described for convenience of explanation, but the electron beam irradiation may be performed using other gas plasma.
For example, the O2 plasma irradiation process may be performed with an O2 flow rate of 0.1 sccm to 150 sccm, power of 30 W to 200 W, and a time of 5 seconds to 300 seconds.
It is seen that the cell culture is good at O2 flow rates of 70 sccm (
Thus, the O2 flow rate may be 0.5 sccm to 100 sccm. For example, the O2 flow rate may be 20 sccm to 50 sccm.
Although the cell culture is performed at power of 50 W (
It is seen that the amount of cell attachment significantly increases in the power of 100 W (
Thus, the power may be 50 W to 200 W. For example, the power may be 80 W to 200 W. For example, the power may be 100 W to 200 W.
It is seen that the cell culture is good when an O2 plasma irradiation time is 20 seconds (
Thus, the O2 plasma irradiation time may be 10 seconds to 200 seconds. For example, the O2 plasma irradiation time may be 15 seconds to 100 seconds. For example, the O2 plasma irradiation time may be 20 seconds to 70 seconds.
For example, in an UV/O3 irradiation process, an electron beam is irradiated with UV having a wavelength of 160 nanometers to 300 nanometers for 1 minute to 4 hours, and thus, the hydrophilicity of the surface of the nanocellulose thin film may increase.
For example, in an UV/O3 irradiation process, an electron beam is irradiated with UV having a wavelength of 180 nanometers to 260 nanometers for 3 minute to 2 hours, and thus, the hydrophilicity of the surface of the nanocellulose thin film may increase.
As illustrated in
The surface of the nanocellulose thin film 116 may be modified by the electron beam. For example, the hydrophilicity of the surface of the nanocellulose thin film 116 may increase by the electron beam.
For example, a hydroxyl group (—OH) closed by a hydrogen bond through the drying process may be reactivated by the electron beam irradiation.
As illustrated in
However, as illustrated in
Therefore, as illustrated in
As another example, air bubbles collected inside the nanocellulose thin film 116 by the drying process may be removed due to the increased hydrophilicity by the electron beam irradiation.
As illustrated in
As illustrated in
On the other hand, as in the first embodiment, when the surface treatment process is performed by the electron beam irradiation (S215), the hydrophilicity of the surface of the nanocellulose thin film 116 may increase. Thus, while the nanocellulose solution penetrates into the nanocellulose thin film 116, the bubbles 130 in the nanocellulose thin film 116 are pushed out to remove the air bubbles 130 inside the nanocellulose thin film 116.
As illustrated in
As illustrated in
As illustrated in
As illustrated in
As illustrated in
Therefore, as in the first embodiment, the nanocellulose thin film may have very high hydrophilicity by the electron beam irradiation process (S215 in
The cellulose nano-film according to the first embodiment may be used as one of a water treatment agent, a heavy metal adsorbent, and a fine dust blocking filter.
According to the first embodiment, wettability of the culture solution may be improved by hydrophilizing the nanocellulose thin film formed by the drying process through the electron beam irradiation.
According to the first embodiment, the production of nanocellulose supports using the drying method may allow the container having various shapes to be applied and have simple processes, thus enabling the mass production and production of over-sized supports.
According to the first embodiment, the wettability of the culture solution may be improved through the hydrophilic treatment to remove the microbubbles in the fiber, thereby improving the cell attachment.
According to at least one of the embodiments, the nanocellulose support may be easily decomposed using the plant degradation enzyme to minimize the damage of the cells compared to the method of collecting the cells by cutting the attached site of the cell by using the conventional animal degradation enzyme, and the nanocellulose support may be decomposed in the short time compared to the conventional hydrogel to obtain the cells having the excellent quality in the short time.
According to at least one of the embodiments, it may be possible to provide the cell culture method suitable for the mass production in the simple and fast process by overcoming the material limitations of the nanocellulose, thereby significantly contributing to the bio industries in the future.
Referring to
The nanocellulose solution may be dropped into the container (S223).
For example, the nanocellulose solution may be produced by dispersing nanocellulose into a solution.
The nanocellulose may be, for example, nanocellulose surface-treated with anions. The anion may include, for example, a reactive group such as a sulfonic acid group, a phosphonic acid group, a carboxyl group, a sulfuric acid group, or a phosphoric acid group.
The nanocellulose may be, for example, nanocellulose surface-treated with cations. The cation may include, for example, an amine group including an epoxypropyltrimethylammonium group, a diethylaminoethyl group, and a dimethylamionethyl group, an amide group, an amino group, an ammonium group, a phosphonium group, and a sulfonium group.
Therefore, when the nanocellulose thin film is formed based on the cationic nanocellulose or the anionic nanocellulose according to the second embodiment, mass production of the cells may be possible.
A concentration of the nanocellulose may be 0.01% to 5% by weight. Concentrations other than the concentration of the nanocellulose may be concentrations of the solution. For example, the concentration of the nanocellulose may be 0.01% to 2% by weight. For example, the concentration of the nanocellulose may be 0.03% to 1% by weight.
The solution may be, for example, distilled water. The solution may be, for example, sterilized bio-distilled water. The solution may be, for example, a mixed solution in which one of distilled water or sterilized bio-distilled water and ethanol are mixed.
Since the nanocellulose is treated to be hydrophilic, the nanocellulose dispersed in the nanocellulose solution may also be maintained with hydrophilicity. Thus, the nanocellulose solution may be called a surface-treated nanocellulose solution.
For example, a nanocellulose solution (112 in
Therefore, a minimum amount of nanocellulose solution (112 in
Although not shown, after the nanocellulose solution (112 in
Thereafter, washing may be performed on the plurality of beads (S224).
For example, the nanocellulose solution 112 in the container (10 in
Then, although not shown, sterilized water may be dropped onto the sieve to wash the plurality of beads 150 remaining on the sieve.
Thereafter, a drying process is performed (S225), and a nanocellulose thin film may be formed on the plurality of beads.
Thereafter, an electron beam irradiation process, that is, a surface treatment process (S226) may be performed to form the nanocellulose thin film having the increased hydrophilicity.
As illustrated in
The nanocellulose support may include a plurality of nanocellulose thin films 160 each of which has a bead shape and which have the increased hydrophilicity.
Various thin films each of which has the bead shape will be described with reference to
As illustrated in
The nanocellulose support including the nanocellulose thin film produced according to the second embodiment may be used as microcarriers.
The microcarrier may be a cell culture support for mass-cultivating cells. Thus, the cells may be cultured by filling a culture medium in a cell cultivator (or bioreactor), and suspending microcarriers to which the cells are attached in the culture medium.
Conventionally, a 2D cell cultivator has a small number of culturable cells per unit area, and it is possible to cultivate the cells through many manual processes by the skilled expert. On the other hand, if a 3D microcarrier is used, as in the example, the number of culturable cells per unit area may be large, and automation may be possible, and thus, many cells may be more easily mass-cultivated.
In addition, the nanocellulose support produced according to the second embodiment may be used as a bead for collecting biomaterials. Since modification of surface properties of the nanocellulose thin film acts as a functional group capable of antigen-antibody reaction of a target material, the nanocellulose support according to the second embodiment, that is, a plurality of spherical beads may be suspended in a culture solution containing the cells in the culture solution to collect the target material. Thereafter, the plurality of spherical beads sunk in the culture solution may be collected, and the nanocellulose thin film may be dissolved with a plant degrading enzyme to easily collect the desired target material. For example, cellulase, xylanase, pectinase, hemicellulase, sucrase, amylase, or a combination thereof may be used as the plant degrading enzyme.
Therefore, according to the second embodiment, the nanocellulose thin film may be melted with the plant degrading enzyme to minimize damage of the cells compared to the conventional method of collecting the cells by cutting attachment sites of the cells with an animal degrading enzyme.
According to the second embodiment, it is possible to decompose in a short time compared to the conventional hydrogel, and thus, the cells having excellent quality may be obtained in a short time.
The nanocellulose support produced according to the first and/or second embodiments may be applied as a water treatment bead for removing heavy metals if the functional group capable of removing the heavy metals is attached and may also be used as a fine dust blocking filter.
The nanocellulose is known to have heavy metal and contaminant adsorption properties. Therefore, when developing the nanocellulose filter using the nanocellulose support produced according to the first and/or second embodiment, the wettability to water may be improved, and the entire area of the filter may be activated in a short time to maximize the filter characteristics. Therefore, it is expected that the contribution to the environmental industry will be high.
The detailed description is intended to be illustrative, but not limiting in all aspects. It is intended that the scope according to the embodiment should be determined by the rational interpretation of the claims as set forth, and the modifications and variations according to the embodiment come within the scope of the appended claims and their equivalents.
The embodiment may be applied to various industries such as the bio industry and the environmental pollution industry.
Filing Document | Filing Date | Country | Kind |
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PCT/KR2020/010027 | 7/29/2020 | WO |