KR20230077388A - Wrinkle forming apparatus for artificial skin - Google Patents

Abstract

Provided is an apparatus for forming artificial skin wrinkles, which includes an upper surface and a lower surface facing the upper surface, and the lower surface includes one or more uneven portions, and the uneven portion includes a stamp of which the entire surface is coated with a parylene coating layer. Using the apparatus for forming the artificial skin wrinkles including the stamp according to one embodiment, it is possible to manufacture wrinkled artificial skin that is most similar to real skin with wrinkles at the interface between a dermal layer and an epidermal layer.

Description

Artificial skin wrinkle forming device {WRINKLE FORMING APPARATUS FOR ARTIFICIAL SKIN}

The present disclosure relates to an artificial skin wrinkle forming device including a stamp capable of forming wrinkles on artificial skin.

Skin is an organ that covers the outside of the body and is composed of three layers: epidermis, dermis, and subcutaneous fat layer from the outside. The epidermis is dominated by keratinocytes of stratified squamous epithelium. The dermis composed of matrix proteins such as collagen fibers and elastic fibers is located under the epidermis, and the dermis contains blood vessels, nerves, sweat glands, and the like. The subcutaneous fat layer is composed of fat cells. The skin maintains its shape by interacting with various cells and components as described above, and exhibits various functions such as regulating body temperature and functioning as a barrier against the external environment.

Artificial skin is a three-dimensional reconstruction of the skin using skin cells and skin constituents such as collagen and elastin. Because it shows functional characteristics, it is also called skin equivalent or reconstructed skin. Artificial skin is not only used for replacement (permanent engraftment type) or regeneration (temporary covering type) of damaged skin such as burns and trauma, but also in various fields such as skin physiology research, skin irritation evaluation, and skin efficacy evaluation. .

However, since the artificial skin has a flat structure at the interface between the dermal layer and the epidermal layer (DEJ), unlike real skin, there is a limit to conducting anti-aging experiments with only the artificial skin. This is because the interface between the dermal and epidermal layers of real skin is not flat but curved.

Unfortunately, not much research has been actively conducted on how to manufacture artificial skin that mimics the curves or wrinkles of the skin. is one of the reasons for the large drop in

Accordingly, the inventors of the present invention, after long research, have developed a stamp capable of producing a new wrinkled artificial skin that has not been reported so far, and an artificial skin wrinkle forming device including the same.

Modifications of artificial skin models to date have mainly relied on cell biological methods that change cells or add or remove specific substances, and these methods wrinkle the structure of the interface between the dermal layer and the epidermal layer constituting the artificial skin. Since it is not required, there are limitations in many aspects in applying the experimental results of artificial skin models so far to actual skin. One embodiment is to provide an artificial skin wrinkle-forming device including a stamp for the artificial skin wrinkle-forming device so that the artificial skin has wrinkles like real skin.

According to one embodiment, artificial skin wrinkle formation comprising a stamp comprising an upper surface and a lower surface facing the upper surface, wherein the lower surface includes one or more concave-convex portions, and the entire surface of the concave-convex portion is coated with a parylene coating layer. provide the device.

The parylene coating layer may have a thickness of 2 μm or less.

The parylene coating layer may have a thickness of 1 μm to 2 μm.

The stamp may have a width of 20 mm or less.

The uneven portion may have a triangular shape.

The lower surface may include one or more triangular concave-convex portions, and widths and lengths of the triangular concave-convex portions may be the same.

The lower surface includes one or more triangular concave-convex portions, and widths and lengths of the triangular concave-convex portions may be different from each other.

The stamp may satisfy Equation 1 below.

[Equation 1]

(w + h) x 2 ≤ S ≤ (w + h) x 5

In Equation 1 above,

S is the width of the stamp,

w is the average width of the concavo-convex portion included in the lower surface of the stamp,

h is the average length of the concavo-convex part included in the lower surface of the stamp.

While the lower surface of the stamp includes one or more uneven portions, the stamp may satisfy Equation 2 below.

[Equation 2]

G x 2 < S

In Equation 2 above,

S is the width of the stamp,

G is an average distance between one or more uneven portions included in the lower surface of the stamp.

The uneven portion may have a semicircular shape.

The lower surface of the stamp may include one or more semicircular concave-convex portions, and the semicircular concave-convex portion may have the same width and length.

The lower surface of the stamp includes one or more semicircular concave-convex portions, and the width and length of the semicircular concave-convex portion may be different from each other.

While the lower surface of the stamp includes one or more concave-convex portions, the stamp may satisfy Equation 3 below.

[Equation 3]

w x 24 < S < w x 45

In Equation 3 above,

S is the width of the stamp,

w is the average width of the concavo-convex portion included in the lower surface of the stamp.

The lower surface includes three or more convex-convex portions, and the concave-convex portion includes a first concave-convex portion, a second concave-convex portion, and a third concave-convex portion, and the first concave-convex portion and the third concave-convex portion have the same width and length, A width of the second concave-convex portion is greater than a width of the first concave-convex portion, a length of the second concave-convex portion is longer than a length of the first concave-convex portion, and the second concave-convex portion is positioned between the first concave-convex portion and the third concave-convex portion. can do.

The stamp may be a stamp for wrinkle formation on artificial skin.

The artificial skin wrinkle forming device may include a mold fixing frame and a mold mounted on the mold fixing frame, and the mold may include a mounting portion mounted on the mold fixing frame and the stamp connected to the mounting portion.

The artificial skin wrinkle forming device may further include a holding device to which the mold fixing frame is attached.

The mold fixing frame and the mold may be independently made of acrylonitrile-butadiene-styrene (ABS) resin or polylactic acid (PLA) resin.

Using the artificial skin wrinkle forming device including a stamp according to an embodiment, wrinkled artificial skin having wrinkles at the interface between the dermal layer and the epidermal layer can be manufactured in a form most similar to real skin.

1 and 2 are each independently a photograph of an artificial skin wrinkle forming device (artificial skin wrinkle forming device attached to a holding device) according to an embodiment.
3 is a photograph of a mold fixing frame constituting an artificial skin wrinkle forming device according to an embodiment.
4 is a photograph of an artificial skin wrinkle forming device (a mold fixing frame and a mold mounted thereto) according to an embodiment.
5 is a photograph showing a state in which wrinkles are formed in the artificial dermal layer by stamping with an artificial skin wrinkle forming device after the non-wrinkled artificial dermis layer is supported on the shoulder strap.
6 to 13 are cross-sectional views of a stamp (of a mold) independently included in an artificial skin wrinkle forming device according to an embodiment.
14 is a digital camera photograph of general artificial skin without forming wrinkles with a stamp.
FIG. 15 is a digital camera photograph of an artificial skin having the shape of FIG. 10, after stamping the dermis layer with a stamp (Comparative Example 1) without any coating on the concave-convex portion, and then culturing to the epidermal layer.
16 is a 3D optical coherence tomography (OCT) photograph of an upper surface of general artificial skin on which wrinkles are not formed with a stamp.
17 is a 3D optical coherence tomography (OCT) image of the upper surface of artificial skin after stamping the dermal layer with a stamp (Comparative Example 1) having the shape of FIG. ; Optical Coherence Tomography).
18 is an H&E staining photograph of a cross-section of general artificial skin without wrinkles formed by a stamp.
FIG. 19 is an H&E staining photograph of a cross-section of artificial skin having the shape of FIG. 10, but after stamping the dermis layer with a stamp (Comparative Example 1) without any coating on the concave-convex portion, and then culturing to the epidermal layer.
20 is a digital camera photograph of artificial skin after stamping the dermal layer with the stamp according to Example 1 and then culturing the epidermal layer.
21 is a digital camera photograph of artificial skin after stamping the dermal layer with a stamp according to Comparative Example 2 and then culturing to the epidermal layer.
22 is a digital camera photograph of artificial skin after stamping the dermal layer with the stamp according to Example 2 and then culturing the epidermal layer.
23 is a digital camera photograph of artificial skin after stamping the dermal layer with a stamp according to Comparative Example 3 and then culturing to the epidermal layer.
24 is a digital camera photograph of artificial skin after stamping the dermal layer with the stamp according to Example 3 and then culturing to the epidermal layer.
25 is a digital camera photograph of artificial skin after stamping the dermal layer with a stamp according to Comparative Example 4 and then culturing to the epidermal layer.
26 is a 3D optical coherence tomography (OCT) photograph of the upper surface of artificial skin after stamping the dermal layer with the stamp according to Example 1 and then culturing to the epidermal layer.
27 is a 3D optical coherence tomography (OCT) photograph of the upper surface of artificial skin after stamping the dermis layer with a stamp according to Comparative Example 2 and then culturing to the epidermal layer.
28 is a 3D optical coherence tomography (OCT) photograph of the upper surface of artificial skin after stamping the dermal layer with the stamp according to Example 2 and then culturing to the epidermal layer.
29 is a 3D optical coherence tomography (OCT) photograph of the upper surface of artificial skin after stamping the dermal layer with a stamp according to Comparative Example 3 and then culturing to the epidermal layer.
30 is a 3D optical coherence tomography (OCT) photograph of the upper surface of artificial skin after stamping the dermis layer with the stamp according to Example 3 and then culturing to the epidermal layer.
31 is a 3D optical coherence tomography (OCT) photograph of the upper surface of artificial skin after stamping the dermis layer with a stamp according to Comparative Example 4 and then culturing to the epidermal layer.
Fig. 32 is a diagram explaining the width of the stamp, the length of the concave-convex portion, and the width of the concave-convex portion.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement the present invention. However, the present invention may be embodied in many different forms and is not limited to the embodiments set forth herein.

In the present specification, "artificial skin" means a three-dimensional reconstruction of skin using skin cells and collagen, a skin component, and a polymer composite that exhibits structural and functional characteristics similar to real skin. It is the widest meaning that includes all ramen.

In the present specification, "stamping" may refer to an operation of applying force so that the concavo-convex part of the lower surface of the stamp constituting the artificial skin wrinkle forming device touches a simulated layer such as an artificial dermis layer, and after applying the force, the artificial skin An operation of removing the pleating device (or stamp) may also be included.

In this specification, when a part such as a layer, film, region, plate, etc. is said to be “on” another part, this includes not only the case where it is “directly on” the other part, but also the case where there is another part in the middle. Conversely, when a part is said to be "directly on" another part, it means that there is no other part in between.

Hereinafter, an artificial skin wrinkle forming device including a stamp according to an embodiment will be described.

Real skin has curved wrinkles rather than a flat interface between the epidermal layer and the dermis layer. Conventional artificial skin has a flat interface between the epidermal layer and the dermis layer (see FIG. 18), so the actual skin and conventional artificial skin have different shapes, It was difficult to apply the results of experiments with artificial skin to real skin. In fact, most of the substances judged to be effective in improving skin wrinkles through artificial skin test results do not actually have a significant skin wrinkle improvement effect, which is because artificial skin with a flat interface between the epidermal layer and the dermal layer has almost no wrinkles, This is because it is not an appropriate model to judge the improvement effect.

Even if it is not possible to easily confirm the wrinkle improvement effect of actual skin with conventional artificial skin models, it is not possible to conduct animal experiments according to the Cosmetic Animal Experiment Prohibition Act. There is a problem in that it is difficult to easily predict the wrinkle improvement effect on the actual skin.

In addition, in order to solve the above problems, a technique of forming wrinkles on conventional artificial skin by stretching the conventional artificial skin has been introduced, but this is because the shape of the wrinkles is different each time depending on the stretching conditions, so that the wrinkles of the desired shape are formed. It was impossible to manufacture artificial skin, and above all, there is a problem in that it is not easy to easily form wrinkles on conventional artificial skin by stretching.

However, according to one embodiment, since the stamp can form wrinkles on the artificial skin by a stamping method rather than a stretching method, wrinkles are not formed on the artificial skin already manufactured as a subsequent operation, but from the initial stage of artificial skin production. Since the step of forming wrinkles can be performed and the shape or shape to be stamped can be easily changed, artificial skin having wrinkles of a desired shape can be easily produced in large quantities.

According to one embodiment, the stamp includes an upper surface and a lower surface facing the upper surface, the lower surface includes one or more concave-convex portions, and the entire surface of the concave-convex portion is coated with a Parylene coating layer. .

Specifically, since the entire surface of the concave-convex portion of the lower surface of the stamp is coated with a parylene coating layer, including the end portion, the wrinkled artificial skin is very similar to the actual skin, compared to the case where the concave-convex portion is not coated with anything or is coated with another material. can be easily manufactured.

For example, if nothing is coated on the concave-convex part of the lower surface of the stamp or if another material is coated, or if the parylene coating layer is not coated on the end part even though it has a parylene coating layer, after the concave-convex part is inserted into the artificial skin When it comes off again, the keratinocytes present in the epidermal layer of the artificial skin adhere to the concave-convex portion, which inevitably causes tissue deformation or damage to the epidermal layer.

The present inventors have continuously studied and researched, and even if it is a stamp, the degree to which keratinocytes present in the epidermal layer of artificial skin stick to the concave-convex part is different depending on what material is coated on the concave-convex part and in what form It was confirmed through continued research that when parylene was coated on the entire surface including the end of the concave-convex portion, it was confirmed that the above-described problem does not occur, and the present invention was completed.

The parylene coating may mean that parylene is thermally decomposed in a vacuum state at room temperature to form a polymer and then coated.

For example, the parylene coating layer may have a thickness of 2 μm or less, for example, 1 μm to 2 μm. When the parylene coating layer has a thickness of more than 2 μm, the thickness of the coating layer becomes too thick, making it difficult to form wrinkles in artificial skin similar to those of actual skin. The effect of the rylene coating layer, that is, the effect of preventing the keratinocytes present in the epidermal layer of the spherical skin from adhering to the concave-convex portion may be reduced, which may be undesirable.

For example, the stamp may have a width of 20 mm or less, for example, 3 mm to 20 mm. When the width of the stamp is less than 3 mm, it is difficult to form concavo-convex portions in the stamp, and when stamping, collagen constituting the artificial dermal mixture may protrude around the stamp, which is undesirable, and the width of the stamp is greater than 20 mm In this case, since the stamp is not easily inserted into the shoulder strap constituting the artificial skin wrinkle forming device described later, stamping is not performed at a desired height, which may be undesirable because it is difficult to form fine wrinkles in the artificial dermis layer.

For example, the concave-convex portion may have various shapes, for example, a polygonal shape such as a triangle, a quadrangle, a pentagon, a hexagon, or a semicircular shape. It may have a width and a length, or may have different widths and lengths.

Meanwhile, wrinkles may be easily formed on the artificial dermal layer only when the concave-convex portion included in the lower surface of the stamp satisfies a specific parameter according to the shape of the concave-convex portion.

Specifically, when the uneven portion has a triangular shape, the stamp may satisfy Equation 1 below.

[Equation 1]

(w + h) x 2 ≤ S ≤ (w + h) x 5

In Equation 1 above,

S is the width of the stamp,

w is the average width of the concavo-convex portion included in the lower surface of the stamp,

h is the average length of the concavo-convex part included in the lower surface of the stamp.

More specifically, when the uneven portion has a triangular shape and the width S of the stamp is 10 mm or more, for example, 10 mm to 12 mm, the stamp may satisfy Equation 1-1 below.

[Equation 1-1]

(w + h) x 2.2 ≤ S ≤ (w + h) x 4

In Equation 1-1 above,

S is the width of the stamp,

w is the average width of the concavo-convex portion included in the lower surface of the stamp,

h is the average length of the concavo-convex part included in the lower surface of the stamp.

When a stamp including a triangular concave-convex portion satisfies Equation 1, or when a stamp including a triangular concave-convex portion and having a width of 10 μm or more satisfies Equation 1-1, the stamp is easily artificially formed. Not only can wrinkles be formed on the dermal layer, but also the shape of fine wrinkles can be reproduced.

In addition, when the concave-convex portion has a triangular shape, the stamp may include one or more, for example, two or more concave-convex portions on the lower surface and simultaneously satisfy Equation 2 below.

[Equation 2]

G x 2 < S

In Equation 2 above,

S is the width of the stamp,

G is an average distance between one or more, for example, two or more concavo-convex portions included in the lower surface of the stamp.

When the stamp including the triangular concave-convex portion satisfies Equation 2 above, the stamp may include several concave-convex portions, so that more various types of wrinkles may be formed on the artificial dermal layer.

Meanwhile, when the concave-convex portion has a semicircular shape rather than a triangular shape, the stamp may include one or more, for example, two or more semicircular concave and convex portions on the lower surface, and simultaneously satisfy Equation 3 below.

[Equation 3]

w x 24 < S < w x 45

In Equation 3 above,

S is the width of the stamp,

w is the average width of the concavo-convex portion included in the lower surface of the stamp.

When the concavo-convex portion has a round semicircular shape rather than a polygonal shape such as a triangle, it may be easier to form wrinkles in the artificial dermis layer by including several concave-convex portions on the lower surface of the stamp. For example, when the relationship between the width of the stamp and the average width of the semicircular concave-convex portion satisfies Equation 3, fine and various wrinkles equivalent to those in the case where the concave-convex portion has a polygonal shape can be reproduced in the artificial dermal layer.

For example, the lower surface constituting the stamp includes three or more convex-convex portions, the concave-convex portion includes a first concave-convex portion, a second concave-convex portion, and a third concave-convex portion, and the first concave-convex portion and the third concave-convex portion have a width and lengths are equal to each other, the width of the second concave-convex portion is two ? part and the third concavo-convex part (see FIG. 10). In this case, it is possible to form wrinkles in a form very similar to real human skin on the artificial skin, and the artificial skin wrinkle forming device having a stamp having a lower surface including the first concave-convex portion, the second concave-convex portion, and the third concave-convex portion When the skin wrinkle improvement compound is treated on the wrinkled artificial skin prepared using the above, the wrinkles formed by the first and third concave-convex portions disappear, and only the wrinkles formed by the second concave-convex portion remain. It can be very useful for screening anti-wrinkle compounds. Specifically, 'wrinkled artificial skin manufactured using an artificial skin wrinkle forming device having a stamp having a lower surface including the first concave-convex part, the second concave-convex part, and the third concave-convex part as a compound having a skin wrinkle improvement effect. In the case of treatment, the wrinkles formed by the first concave-convex portion and the third concave-convex portion are not deep and are easily restored (wrinkle smoothing), but the wrinkles formed by the second concave-convex portion are deep and not easily restored In bar, by comparing the degree of restoration of wrinkles formed by the first and third concave and convex portions and the degree of restoration of wrinkles formed by the second concave and convex portion, it is easy to determine whether the treated specific compound has an effect on improving skin wrinkles. can be screened.

For example, the stamp according to one embodiment may be a stamp for wrinkle formation on artificial skin.

For example, the artificial skin wrinkle forming device may include a mold holding frame and a mold mounted on the mold holding frame, and the mold may include a mounting portion mounted on the mold holding frame and the stamp connected to the mounting portion.

FIG. 3 shows a photograph of the mold fixing frame, and the artificial skin wrinkle forming device composed of the mold and the mold fixing frame can be manufactured by inserting the mounting part of the mold into the central hole of the mold fixing frame (see FIG. 4 ). In the artificial skin wrinkle forming device having such a shape, the height can be adjusted by pulling or pushing the mounting portion of the mold, and wrinkles can be formed on the artificial skin by pushing the mounting portion in the stamp direction.

More specifically, the artificial skin wrinkle forming device according to one embodiment may further include a holding device to which the mold fixing frame is attached (see FIGS. 1 and 2). At this time, the artificial skin in the holding device, specifically An artificial dermis layer may be supported. That is, after the artificial dermis layer is supported in the holding device, the artificial skin wrinkle forming device is fixed by holding the mold fixing frame of the artificial skin wrinkle forming device composed of the mold and the mold fixing frame to the holding device, and then the mounting part of the mold After adjusting the height by pulling or pushing, by pushing the mounting part of the mold in the direction in which the artificial dermal layer is supported, it is possible to easily form wrinkles of a desired shape in the artificial dermal layer. (See FIG. 5) For example, Transwell ^® sold by Corning may be used as the holding device, but the type of the holding device is not necessarily limited thereto.

For example, the mold fixing frame and the mold may be independently made of acrylonitrile-butadiene-styrene (ABS) resin or polylactic acid (PLA) resin. When the mold fixing frame and the mold are made of the above type of resin, it may be easy to reproduce even fine wrinkles without contaminating the artificial dermal layer even when directly touching the artificial dermal layer. Furthermore, the above kind of resin is a material that can be manufactured with a 3D printer, and it may be possible to mass-produce the artificial skin wrinkle forming device according to one embodiment at a low price.

The embodiments of the present invention described above will be described in more detail through the following examples. However, the following examples are for illustrative purposes only and do not limit the scope of the present invention.

Manufacture of artificial skin wrinkle forming device

Example 1

An ABS resin was molded with a 3D printer (Printed by Fortus 250mc, Stratasys) to manufacture an artificial skin wrinkle forming device including a stamp having the same structure as that shown in FIG. 11 . At this time, the entire surface of the uneven portion of the stamp was coated with parylene.

Example 2

An ABS resin was molded with a 3D printer (Printed by Fortus 250mc, Stratasys) to manufacture an artificial skin wrinkle forming device including a stamp having the same structure as that shown in FIG. 12 . At this time, the entire surface of the uneven portion of the stamp was coated with parylene.

Example 3

An ABS resin was molded with a 3D printer (Printed by Fortus 250mc, Stratasys) to manufacture an artificial skin wrinkle forming device including a stamp having the same structure as that shown in FIG. 13 . At this time, the entire surface of the uneven portion of the stamp was coated with parylene.

Comparative Example 1

An ABS resin was molded with a 3D printer (Printed by Fortus 250mc, Stratasys) to manufacture an artificial skin wrinkle forming device including a stamp having the same structure as that shown in FIG. 10 . At this time, the entire surface of the concavo-convex portion of the stamp was not coated with anything.

Comparative Example 2

It was the same as in Example 1, except that the entire surface of the concavo-convex part of the stamp was not coated with parylene.

Comparative Example 3

It was the same as in Example 2, except that the entire surface of the concave-convex part of the stamp was coated with Teflon instead of parylene coating.

Comparative Example 4

It was the same as in Example 3, except that the entire surface of the concave-convex part of the stamp was coated with ceramic instead of parylene coating.

Manufacture of wrinkled artificial skin

control group

After loading a mixture of collagen (Nutragen, Advanced Biomatrix) and fibroblasts (NDFn, ThermoFisher) in a volume of 400ul on Transwell ^® (Corning), an artificial dermal layer was formed. Thereafter, a fibroblast culture medium (M106, Gibco) was put on the artificial dermal layer and cultured for 7 days to stabilize the artificial dermal layer. Next, keratinocytes (HEKn, ThermoFisher) were seeded on the artificial dermal layer, cultured for 1 day in a keratinocyte culture medium (EpiLife, Gibco), and epidermal layer production medium (CnT-3D- Non-wrinkled artificial skin was prepared by culturing in PR, (CellnTEC) for 1 day, and then culturing in the same epidermal layer production medium while being exposed to air for 14 days to form an artificial epidermal layer, which was shown in FIGS. 14 and 16 and shown in FIG. 18 .

Comparative Example 1-1

After supporting a mixture of collagen (Nutragen, Advanced Biomatrix) and fibroblasts (NDFn, ThermoFisher) in a volume of 400ul on Transwell ^® (Corning), the artificial skin wrinkle-forming device of Comparative Example 1 was applied to the Transwell ^® (Corning ), and stamped so that the stamp constituting the artificial skin wrinkle forming device touches the artificial dermal layer. In a 37°C, 5% CO ₂ incubator for cell culture, the stamp was left in contact with the mixture for 2 hours to harden the mixture (gelation), and then the artificial skin wrinkle forming device was removed to prepare a wrinkled artificial dermal layer. . Thereafter, a fibroblast culture medium (M106, Gibco) was put on the artificial dermal layer and cultured for 7 days to stabilize the artificial dermal layer. Next, keratinocytes (HEKn, ThermoFisher) were seeded on the artificial dermal layer, cultured for 1 day in a keratinocyte culture medium (EpiLife, Gibco), and epidermal layer production medium (CnT-3D- Wrinkled artificial skin was prepared by culturing in PR, (CellnTEC) for 1 day, and then culturing in the same epidermal layer production medium while being exposed to air for 14 days to form an artificial epidermal layer, which was shown in FIGS. 15, 17 and 17. 19.

(Rating 1)

From the control group and Comparative Example 1-1, when stamping artificial skin using a stamp including concave-convex portions uncoated with anything, the concave-convex portion adheres to the collagen and epidermal layer of the dermal layer, resulting in tissue deformation or destruction, resulting in wrinkled artificial skin. It can be confirmed that there may be problems with skin production.

Example 1-1

After supporting a mixture of collagen (Nutragen, Advanced Biomatrix) and fibroblasts (NDFn, ThermoFisher) in a volume of 400ul on Transwell ^® (Corning), the artificial skin wrinkle forming device of Example 1 was applied to the Transwell ^® (Corning ), and stamped so that the stamp constituting the artificial skin wrinkle forming device touches the artificial dermal layer. In a 37°C, 5% CO ₂ incubator for cell culture, the stamp was left in contact with the mixture for 2 hours to harden the mixture (gelation), and then the artificial skin wrinkle forming device was removed to prepare a wrinkled artificial dermal layer. . Thereafter, a fibroblast culture medium (M106, Gibco) was put on the artificial dermal layer and cultured for 7 days to stabilize the artificial dermal layer. Next, keratinocytes (HEKn, ThermoFisher) were seeded on the artificial dermal layer, cultured for 1 day in a keratinocyte culture medium (EpiLife, Gibco), and epidermal layer production medium (CnT-3D- Wrinkled artificial skin was prepared by culturing in PR, (CellnTEC) for 1 day, and then culturing in the same epidermal layer production medium while exposed to air for 14 days to form an artificial epidermal layer, which is shown in FIGS. 20 and 26 was

Example 2-1

Wrinkled artificial skin was prepared in the same manner as in Example 1-1, except that the artificial skin wrinkle-forming apparatus of Example 2 was used instead of the artificial skin wrinkle-forming apparatus of Example 1, which is shown in FIGS. 22 and 28 showed up

Example 3-1

Wrinkled artificial skin was prepared in the same manner as in Example 1-1, except that the artificial skin wrinkle-forming apparatus of Example 3 was used instead of the artificial skin wrinkle-forming apparatus of Example 1, which is shown in FIGS. 24 and 30 showed up

Comparative Example 2-1

Wrinkled artificial skin was prepared in the same manner as in Example 1-1, except that the artificial skin wrinkle-forming apparatus of Comparative Example 2 was used instead of the artificial skin wrinkle-forming apparatus of Example 1, which is shown in FIGS. 21 and 27 showed up

Comparative Example 3-1

Wrinkled artificial skin was prepared in the same manner as in Example 1-1, except that the artificial skin wrinkle-forming apparatus of Comparative Example 3 was used instead of the artificial skin wrinkle-forming apparatus of Example 1, which is shown in FIGS. 23 and 29 showed up

Comparative Example 4-1

Wrinkled artificial skin was prepared in the same manner as in Example 1-1, except that the artificial skin wrinkle-forming apparatus of Comparative Example 3 was used instead of the artificial skin wrinkle-forming apparatus of Example 1, which is shown in FIGS. 25 and 31 showed up

(Rating 2)

From Example 1-1 to Example 3-1 and Comparative Example 1-1 to Comparative Example 4-1, a stamp including a concave-convex portion that is not coated with anything, a stamp including a concave-convex portion coated with Teflon on the entire surface, and a ceramic Compared to the case of stamping the artificial skin using a stamp including concave-convex portions coated on the entire surface, when stamping the artificial skin using a stamp including concave-convex portions coated on the entire surface of parylene, wrinkles are more formed. It was clear and clear, and at the same time, it was natural like wrinkles in the actual skin, and it could be confirmed that the phenomenon of the epidermal layer sticking to the concavo-convex part and falling off entirely was not observed.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements of those skilled in the art using the basic concept of the present invention defined in the following claims are also present. It falls within the scope of the right of invention.

Claims (18)

Including an upper surface and a lower surface facing the upper surface,
The lower surface includes one or more concavo-convex portions,
Artificial skin wrinkle forming device comprising a stamp coated with a parylene coating layer on the entire surface of the uneven portion. In paragraph 1,
The parylene coating layer has a thickness of 2 μm or less. In paragraph 2,
The parylene coating layer has a thickness of 1 μm to 2 μm artificial skin wrinkle forming device. In paragraph 1,
The stamp is an artificial skin wrinkle forming device having a width of 20 mm or less. In paragraph 1,
The artificial skin wrinkle forming device having a triangular shape. In paragraph 5,
The lower surface includes one or more triangular concave-convex portions, and the width and length of the triangular concave-convex portion are the same as each other. In paragraph 5,
The lower surface includes one or more triangular concave-convex portions, and the width and length of the triangular concave-convex portions are different from each other. In paragraph 5,
The stamp is an artificial skin wrinkle forming device that satisfies Equation 1 below:
[Equation 1]
(w + h) ≤ S ≤ (w + h) x 2.5
In Equation 1 above,
S is the width of the stamp,
w is the width of the concavo-convex part included in the lower surface of the stamp,
h is the length of the uneven portion included in the lower surface of the stamp. In paragraph 5,
The lower surface of the stamp includes one or more concave-convex portions, and the artificial skin wrinkle-forming device that satisfies Equation 2 below:
[Equation 2]
G x 2 < S
In Equation 2 above,
S is the width of the stamp,
G is an average distance between one or more uneven portions included in the lower surface of the stamp. In paragraph 1,
The artificial skin wrinkle forming device having a semicircular shape. In paragraph 10,
The lower surface of the stamp includes at least one semicircular concave-convex portion, and the width and length of the semicircular concave-convex portion are the same as each other. In paragraph 10,
The lower surface of the stamp includes at least one semicircular concave-convex portion, and the width and length of the semicircular concave-convex portion are different from each other. In paragraph 10,
The lower surface of the stamp includes one or more concave-convex portions, and the artificial skin wrinkle forming device satisfying Equation 3 below:
[Equation 3]
w x 12 < S < w x 22.5
In Equation 3 above,
S is the width of the stamp,
w is the width of the concavo-convex portion included in the lower surface of the stamp. In paragraph 1,
The lower surface includes three or more concave-convex portions,
The uneven portion is composed of a first uneven portion, a second uneven portion, and a third uneven portion,
The first concave-convex portion and the third concave-convex portion have the same width and length,
The width of the second concave-convex portion is greater than the width of the first concave-convex portion,
The length of the second concave-convex portion is longer than the length of the first concave-convex portion,
The second convex-convex portion is positioned between the first concave-convex portion and the third concave-convex portion. In paragraph 1,
The stamp is an artificial skin wrinkle forming device that is a stamp for artificial skin wrinkle formation. In paragraph 1,
The artificial skin wrinkle forming device
It includes a mold holding frame and a mold mounted on the mold holding frame,
The mold includes a mounting portion mounted on the mold fixing frame and the stamp connected to the mounting portion.
Artificial skin wrinkle forming device. In clause 16,
The artificial skin wrinkle forming device further comprises a holding device to which the mold fixing frame is attached. In clause 16,
The mold fixing frame and the mold are each independently made of acrylonitrile-butadiene-styrene (ABS) resin or polylactic acid (PLA) resin.

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