CN117686661A - Application of bionic skin in cosmetic test - Google Patents
Application of bionic skin in cosmetic test Download PDFInfo
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Abstract
The invention belongs to the technical field of tissue engineering skin and cosmetic testing, and particularly relates to application of bionic skin in cosmetic testing. The method comprises the steps of 3D printing a plurality of three-dimensional bionic skin which comprises a dermis cell layer, a epidermis cell layer and a collagen skeleton layer and utilizes different 3D printing planning schemes on a printing substrate by using epidermis stem cells and dermis stem cells which are separated and cultured by human skin tissues and matching with collagen skeleton materials, and dividing a plurality of samples to be tested with the same size on each three-dimensional bionic skin through tissue culture and fusion to obtain a plurality of groups of 3D printing bionic skin samples which can be used for efficacy tests of moisturizing, sun screening, wrinkle removing and the like of cosmetics. And then, replacing the human skin to be tested or the cell sample to be tested by adopting the 3D printed bionic skin sample for testing, improving the testing efficiency and accuracy, and optimizing the evaluation result by utilizing the planability of the 3D printing process and reversely optimizing the 3D printing planning scheme of the bionic skin according to the evaluation result.
Description
Technical Field
The invention relates to application of bionic skin in cosmetic testing, in particular to application of 3D printing bionic skin in cosmetic testing, and belongs to the technical field of tissue engineering skin and cosmetic testing.
Background
Cosmetics generally have the effects of moisturizing, sun protection, wrinkle removal and the like, and according to the supervision and management regulations and guidelines of the cosmetic industry, the cosmetic effects claim to be supported by sufficient evidence, for example, the moisturizing effects claim to be evaluated by a certain moisturizing effect measuring method. The current method for measuring the moisturizing effect of the cosmetics comprises a traditional in-vitro weighing method, a skin test method, a horny layer water content test method, a cell biological method and the like. In addition, the method and the device for measuring the moisturizing efficacy of the cosmetics, disclosed in CN202110941436 by the team Wu Jianxin of Chinese medical science, simulate the water loss of biological skin cells by adopting HaCaT cells, and measure the moisturizing efficacy of the cosmetics based on an MTT colorimetric method, so that the accuracy of a measurement result is improved to a certain extent.
With the gradual maturation of the 3D printing bionic skin technology, the physiological and functional characteristics of the technology are very similar to those of real skin, and the models can simulate the hierarchical structure, cell type and physiological response of the real skin. Taking a biological 3D printing full-custom skin with a school team earlier research result application number of CN201810463765 and a preparation method thereof as an example, a three-dimensional scanning modeling technology is adopted, a method of layered printing and layer-by-layer stacking is adopted, and skin tissues with specific positions, specific thickness and specific shape of a human body are printed, so that the 3D printing bionic skin according to actual needs is realized, and the biological 3D printing full-custom skin can be applied to skin repair of patients with skin diseases or skin injuries.
The technology not only points out that the 3D printing bionic skin can be used for repairing human skin, but also refers to the application of the 3D printing bionic skin in the research aspect of an in-vitro skin model, but also does not provide specific feasibility measures, and at present, no mature scheme is provided for applying the 3D printing bionic skin model and the planning advantages of the 3D printing scheme in the preparation of the 3D printing bionic skin model to the test of the effects of moisturizing, sun screening, wrinkle removing and the like of cosmetics.
Disclosure of Invention
The invention provides an application method of bionic skin in cosmetic testing, in particular to an application method of 3D printing bionic skin in cosmetic testing, which aims to overcome the defects in the prior art and realizes new breakthrough in the aspects of testing efficiency and testing accuracy.
The specific technical scheme provided by the invention is as follows:
the application method of the bionic skin in the cosmetic test adopts a plurality of bionic skin samples printed in 3D to replace the human skin or cell sample to be tested for the cosmetic efficacy test.
Preferably, in the above application method, the 3D printed multiple bionic skin samples are used for efficacy tests of cosmetic moisturizing, sun protection, wrinkle removal and the like.
Preferably, in the above application method, the 3D printed multiple bionic skin samples are used for cosmetic moisturizing efficacy test, which includes the steps of:
s1, 3D printing bionic skin preparation
Using epidermis stem cells and dermis stem cells which are separated and cultured by human skin tissues and are matched with collagen skeleton materials, printing the epidermis stem cells and the dermis stem cells on a printing substrate in a 3D layer-by-layer printing mode to obtain 1 st-M (M is more than or equal to 2) three-dimensional bionic skin containing dermis cell layers, epidermis cell layers and collagen skeleton layers, and dividing a plurality of samples to be tested with the same size on each three-dimensional bionic skin through tissue culture and fusion to obtain a plurality of groups of 3D printing bionic skin samples which can be used for testing the moisturizing efficacy of cosmetics, wherein each group contains the plurality of samples to be tested with the same size;
wherein, the 1-M three-dimensional bionic skins are printed by adopting different 3D printing planning schemes;
s2, testing moisturizing efficacy of cosmetics
S2-1, selecting three identical tested samples from each group of tested samples in the 1 st to M groups of 3D printing bionic skin tested samples obtained in the step S1: sample group a, sample group B and sample group C; the sample group A is an experimental sample group, the sample group B is a comparison sample group I, and the sample group C is a comparison sample group II;
s2-2, detecting and recording skin water content data of each tested sample in the sample group A, the sample group B and the sample group C;
s2-3, smearing a solution containing a basic water-retaining agent and a cosmetic efficacy raw material to be tested on each tested sample of the sample group A, smearing a solution containing only the basic water-retaining agent on each tested sample of the sample group B and the sample group C, wherein the concentration of the basic water-retaining agent smeared on the tested sample of the sample group B is equal to the concentration of the basic water-retaining agent smeared on the tested sample of the sample group A, and the concentration of the basic water-retaining agent smeared on the tested sample of the sample group C is equal to the total concentration of the basic water-retaining agent smeared on the tested sample of the sample group A and the cosmetic efficacy raw material to be tested;
s2-4, after the set time is reached, detecting and recording new skin water content data of each tested sample in the sample group A, the sample group B and the sample group C;
s2-5, calculating skin moisture content change data of the tested sample according to the skin moisture content data measured in the step S2-2 and the skin moisture content data measured in the step S2-4, and evaluating the moisturizing effect of the cosmetic to be tested according to the skin moisture content change data of the tested sample;
s2-6, performing inter-group comparison on the cosmetic moisturizing effect evaluation results obtained by the 1 st to M th groups of 3D printing bionic skin tested samples to obtain the optimal cosmetic moisturizing effect evaluation results and the optimal 3D printing scheme of the three-dimensional bionic skin.
Preferably, in the above application method, step S1 includes the steps of:
s1-1, cell extraction
Crushing, carrying out tissue enzymolysis, filtering and centrifuging on human skin tissues to obtain aggregated cells, and inoculating and culturing the cells to obtain primary skin cells;
s1-2, cell culture
Adding ROCK inhibitor into a part of primary skin cells to perform subculture to obtain the required amount of epidermal stem cells, and performing subculture without adding ROCK inhibitor to a part of primary skin cells to obtain the required amount of dermal stem cells;
s1-3, 3D printing bionic skin
Printing the epidermis stem cells and the dermis stem cells obtained in the step S1-2 on the same printing substrate in a 3D layer-by-layer printing mode by matching with collagen skeleton materials to obtain 1 st-M three-dimensional bionic skin which comprises dermis cell layers, epidermis cell layers and collagen skeleton layers and has different 3D printing planning schemes, and carrying out tissue culture on the obtained three-dimensional bionic skin;
s1-4, preparation of sample to be tested
After tissue culture and fusion of the three-dimensional bionic skin obtained in the step S1-3, a plurality of samples to be measured with the same size are segmented on each three-dimensional bionic skin, so that a plurality of groups of 3D printing bionic skin samples which can be used for testing the moisturizing efficacy of cosmetics are obtained, and each group comprises the plurality of samples to be measured with the same size.
Preferably, in the above application method, in step S1, the different 3D printing schemes are different layer thickness schemes and/or different initial cell concentration schemes.
Preferably, in the above application method, in step S1, the multilayered collagen scaffold materials distributed in the three-dimensional bionic skin are all fixed by a crosslinking agent.
Preferably, in the above application method, in step S1, the step of dividing the plurality of samples to be measured with the same size on each three-dimensional bionic skin is to select a region with uniform thickness and quality on each three-dimensional bionic skin to divide the plurality of samples to be measured with the same size.
Preferably, in the above application method, in step S2-3, the base water-retaining agent is a component other than the component contained in the cosmetic to be measured itself, or the base water-retaining agent has the same component as the cosmetic to be measured itself.
Preferably, in the above application method, in step S2-6, performing the group-to-group comparison of the cosmetic moisturizing efficacy evaluation results obtained from the 3D printed simulated skin test samples of groups 1 to M includes: i, eliminating the evaluation result with obvious distortion from each group of evaluation results; and/or II, selecting a result which is closer to an empirical value or a theoretical value from the evaluation results of each group.
Preferably, in the above application method, in step S2-6, at least one set of evaluation results is checked and verified by a second cosmetic moisturizing efficacy test method based on 3D printing of the biomimetic skin.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, a plurality of 3D printed bionic skin samples are adopted to replace tested human skin or tested cell samples, and are used for moisturizing, sun-screening, wrinkle-removing and other effects of cosmetics, especially moisturizing effect tests, so that more reliable test results can be provided. Moreover, due to the planability of the 3D printing process, the tested sample design of various printing planning schemes can be used, the test evaluation result is continuously optimized in the test process, and the 3D printing planning scheme of the bionic skin can be reversely optimized according to the test evaluation result.
Drawings
FIG. 1 is a flow chart of an application method of the present invention.
Detailed Description
The present invention will be described in further detail with reference to specific embodiments, but the scope of the invention is not limited to these embodiments, and all changes and equivalents that do not depart from the spirit of the invention are intended to be included in the scope of the invention.
The 3D printing bionic skin prepared by the invention can be applied to the tests of moisturizing, sun-screening, wrinkle-removing and other effects of cosmetics according to the needs, and the embodiment takes the application in the tests of the moisturizing effects of cosmetics as an example, and describes an application method of the bionic skin in the tests of cosmetics in detail. Specifically, the method adopts a plurality of bionic skin samples printed in 3D to replace the skin of a tested human body or the tested cell sample for the moisture-preserving efficacy test of cosmetics. The procedure for obtaining a plurality of biomimetic skin samples for 3D printing according to the present invention is first described in example 1 below.
Example 1
3D printing bionic skin preparation
According to the embodiment, the epidermis stem cells and the dermis stem cells which are separated and cultured by human skin tissues are used, the collagen skeleton materials are matched, the 1 st to M (M is more than or equal to 2) three-dimensional bionic skin containing the dermis cell layers, the epidermis cell layers and the collagen skeleton layers is printed on a printing substrate in a 3D layer-by-layer printing mode, a plurality of samples to be measured with the same size are segmented on each three-dimensional bionic skin through tissue culture and fusion, and a plurality of groups of 3D printing bionic skin samples which can be used for testing the moisturizing efficacy of cosmetics are obtained, wherein each group contains the plurality of samples to be measured with the same size.
In the above scheme, the method for 3D printing three-dimensional bionic skin on a printing substrate by using human skin tissue to separate and culture epidermal stem cells and dermal stem cells and matching with collagen skeleton materials can be implemented by referring to the school team advanced research result CN 201810463765.
In this embodiment, multiple three-dimensional bionic skins need to be printed according to a 3D printing planning scheme designed in advance, so that a high verification success rate and an optimal test result can be obtained as much as possible in a one-time batch test, and more research rules are expected to be found in the comprehensive comparison of the transverse direction and the longitudinal direction. According to the 3D printing factor analysis which can influence the test evaluation result in advance, different 3D printing planning schemes can be different layer thickness planning schemes or different initial cell concentration planning schemes.
The different layer thickness planning schemes not only refer to the printing layer number or the total layer thickness of the epidermis cell layer or the dermis cell layer, but also can refer to the relative thickness among the layers, and the initial cell concentration refers to the concentration during printing, and besides the control of the jet quantity or the walking speed of a printer, the control can also be realized by planning the printing layer number of the cell layer. The specific process comprises the following steps:
s1-1, cell extraction
Human skin tissue is crushed, subjected to tissue enzymolysis, filtered and centrifuged to obtain aggregated cells, and the cells are inoculated and cultured to obtain primary skin cells.
S1-2, cell culture
And adding a ROCK inhibitor into a part of the primary skin cells to perform subculture to obtain the required amount of epidermal stem cells, and performing subculture without adding the ROCK inhibitor into a part of the primary skin cells to obtain the required amount of dermal stem cells.
S1-3, 3D printing bionic skin
And (3) printing the epidermis stem cells and the dermis stem cells obtained in the step (S1-2) on the same printing substrate in a 3D layer-by-layer printing mode by matching with collagen skeleton materials to obtain 1 st-M three-dimensional bionic skin which comprises dermis cell layers, epidermis cell layers and collagen skeleton layers and has different 3D printing planning schemes, and carrying out tissue culture on the obtained three-dimensional bionic skin. Referring to CN201810463765 in the printing process, the multi-layer collagen skeleton materials distributed in the three-dimensional bionic skin are all fixed through a cross-linking agent.
S1-4, preparation of sample to be tested
After tissue culture and fusion of the three-dimensional bionic skin obtained in the step S1-3, selecting a region with uniform thickness and quality on each three-dimensional bionic skin, and dividing the region into a plurality of samples to be measured with the same size, thereby obtaining a plurality of groups of 3D printing bionic skin samples which can be used for testing the moisturizing efficacy of cosmetics, wherein each group comprises the plurality of samples to be measured with the same size.
Example 2
The method of the invention can replace the tested human skin or the tested cell sample with a plurality of bionic skin samples printed in 3D for the moisture preserving efficacy test of cosmetics. The scheme of replacing human skin with 3D printing bionic skin can be that the 3D printing bionic skin is directly applied to the moisture preservation efficacy test evaluation based on skin moisture content and transdermal water dispersion detection implemented by human body before. The scheme of replacing the detected cell sample with the 3D printing bionic skin can be to apply the 3D printing bionic skin to the detected cell simulating the biological skin cell to the cosmetic moisturizing efficacy measurement based on the MTT colorimetric method.
In the former example, the present embodiment provides a method for testing the moisturizing efficacy of cosmetics.
S2-1, selecting three test samples with the same quantity from each group of test samples in the 1 st to M groups of 3D printing bionic skin test samples obtained in the embodiment 1: sample group a, sample group B and sample group C; the sample group A is an experimental sample group, the sample group B is a comparison sample group I, and the sample group C is a comparison sample group II.
S2-2, detecting and recording skin water content data of each tested sample in the sample group A, the sample group B and the sample group C.
S2-3, smearing a solution containing a basic water-retaining agent and a cosmetic efficacy raw material to be tested on each tested sample of the sample group A, and smearing a solution containing only the basic water-retaining agent on each tested sample of the sample group B and the sample group C, wherein the concentration of the basic water-retaining agent smeared on the tested sample of the sample group B is equal to the concentration of the basic water-retaining agent smeared on the tested sample of the sample group A, and the concentration of the basic water-retaining agent smeared on the tested sample of the sample group C is equal to the total concentration of the basic water-retaining agent smeared on the tested sample of the sample group A and the cosmetic efficacy raw material to be tested.
The three sample groups are the lower limit of the number of the sample groups, which is set by the invention, and often cannot be obtained, the comparison results of the sample group A and the sample group B can reflect the performance of the cosmetics to be tested in the aspect of absolute water retention performance, the comparison results of the sample group A and the sample group C can reflect the performance of the cosmetics to be tested in the aspect of relative water retention performance, and more sample groups can be set according to the requirements so as to facilitate comparison, for example, three identical sample groups are additionally set after the basic water retention agent or the basic water retention agent formula is replaced.
It should be noted that the basic water-retaining agent herein may select not only components other than the components contained in the cosmetic itself to be measured but also the same components as the cosmetic itself to be measured.
S2-4, after the set time is reached, detecting and recording new skin water content data of each tested sample in the sample group A, the sample group B and the sample group C.
S2-5, calculating skin moisture content change data of the tested sample according to the skin moisture content data measured in the step S2-2 and the skin moisture content data measured in the step S2-4, and evaluating the moisturizing effect of the cosmetic to be tested according to the skin moisture content change data of the tested sample. The water content change data detected by the invention not only refer to the change data of the water content value in the detection field, but also contain related data of percutaneous water loss, and all the data can be acquired by corresponding detection instruments.
S2-6, performing inter-group comparison on the cosmetic moisturizing effect evaluation results obtained by the 3D printing bionic skin tested samples of the 1 st to M th groups to obtain the optimal cosmetic moisturizing effect evaluation results. The evaluation result may be a quantized skin moisture content increase and/or quantized percutaneous moisture loss decrease, etc., and is preferably comprehensively evaluated by various kinds of data.
The group comparison of the cosmetic moisturizing efficacy evaluation results obtained from the 3D printing bionic skin test samples of groups 1 to M can obtain high verification success rate and optimized test results as much as possible in disposable batch test data, and more rules are found, and the process is generally carried out by the following two means without limiting the sequence:
and I, eliminating the evaluation result with obvious distortion from each group of evaluation results.
The evaluation result of obvious distortion can be an evaluation result which is obtained by the bias of the threshold setting of the 3D printing planning scheme of the tested sample and has larger difference with other groups of data, or an evaluation result which is obtained by the reasons of temporary uncertainty and does not accord with the expected rule, and the like, and the part of data possibly has no moisture preservation efficacy evaluation significance but has research contrast significance.
And II, selecting a result which is closer to an empirical value or a theoretical value from the evaluation results of each group. Empirical or theoretical values are basic insights or basic data of various verification works and are derived from different approaches such as rule of thumb, software simulation, early experiments and the like.
According to the method provided by the embodiment, the above-mentioned evaluation result is not necessarily a single numerical parameter, but may be a data set, for example, when the tested sample obtained by printing is enough, the functional raw materials of the cosmetics to be tested in the sample group a may be set to different raw material ratios, so that the obtained data set can reflect the advantages and disadvantages of the functional raw material ratio relationship of the cosmetics to be tested, and thus, a better functional raw material ratio scheme may be conveyed to the designer.
The method has the important significance that by performing the group-to-group comparison on the cosmetic moisturizing effect evaluation results obtained by the 1 st to M th groups of 3D printing bionic skin tested samples, the optimal 3D printing scheme of the three-dimensional bionic skin can be obtained simultaneously while the optimal cosmetic moisturizing effect evaluation results are obtained, and the obtained optimal 3D printing scheme is the scheme directly adopted in the next batch verification work to a large extent.
Example 3
The scheme of replacing the tested cell sample with 3D printed biomimetic skin as described in the beginning of example 2 above is briefly described in this example.
Specifically, in this example, the 3D printed bionic skin obtained in example 1 was applied to the measurement of the moisturizing efficacy of cosmetics based on the MTT colorimetric method instead of the measured cells simulating biological skin cells, and the method may be performed as follows:
s3-1, taking partial 3D printing bionic skin test samples obtained in the embodiment 1, and dividing the partial 3D printing bionic skin test samples into 3 groups: sample group D, sample group E and sample group F, and three samples to be tested were placed in MEM medium of 10% FBS at 37℃with 5% CO 2 Culturing.
S3-2, taking the tested sample of the sample group D as a control group, taking the tested sample of the sample group E, removing the culture medium, and replacing the culture medium with MEM culture solution prepared by 10% sterile NaCl, wherein the concentration of the NaCl system is controlled at 3%,37 ℃ and 5% CO 2 Culturing for 0.5h to obtain the dehydrated bionic skin.
S3-3, removing the culture medium from the tested sample of the sampling group F, and changing to 9:1 bodyMEM culture solution and cosmetic to be tested in volume ratio at 37deg.C, 5% CO 2 After 16h of culture, the culture medium is removed, MEM culture solution prepared by 10% sterile NaCl which is consistent with the step S3-2 is added, and the culture is carried out for 0.5h under the same condition, so as to obtain the bionic skin repaired by the cosmetics to be detected.
S3-4, respectively measuring absorbance values of bionic skin of the sample group D, the sample group E and the sample group F at 570nm by an MTT method and using an ELISA detector, calculating cell activity values, and performing comparative analysis.
Compared with the method of the embodiment 2, the MTT colorimetric method has slightly complicated process and the period is generally more than 20 hours, but the verification result with more pertinence can be obtained by verifying the method aiming at part of tested products, so that the MTT colorimetric method can be used as a moisturizing efficacy measurement scheme aiming at the part of tested products and can also be used as a comparison verification scheme to be matched with the method of the embodiment 2. For example, in step S2-6 of the application method described in example 2, if there is an uncertainty in the evaluation results obtained based on the current batch of the sample under test, at least one set of evaluation results in example 2 may be compared or verified by using the MTT colorimetric method of example 3 as a comparison test method. In this case, the sample to be taken in step S3-1 may be a sample to be measured prepared based on only one or a few of the 3D printing schemes.
Example 4
The present embodiment provides an industrial application verification for the application method of the 3D printing bionic skin in cosmetic testing described in the previous embodiments.
Some commercially available Z brand skin cosmetics have two products G1 and G2, containing different plant extract formulas, each of which is declared to have moisturizing efficacy and give declared human body test indexes. According to the application protocol for the above products G1 and G2, 5 different ratios of epidermal cell layer to dermal cell layer thickness 1 were designed in example 1: 1. 3: 2. 2: 1. 5: 2. 3:1 as 5 printing planning schemes, the three-dimensional bionic skin obtained by each printing planning scheme is tested according to the sample preparation and smearing schemes of the sample group A, the sample group B and the sample group C, which are described above, and the test conditions are as follows:
independent sample groups a, B, C were set for each of cosmetic product G1 and cosmetic product G2 and the moisturizing efficacy test and moisturizing efficacy evaluation were performed according to the procedure of example 2, but the verification process for products G1 and G2 was performed simultaneously.
According to the skin moisture content amplification and the skin moisture loss amplitude reduction data, the plant extract formulas of the products G1 and G2 have moisturizing effects, the moisturizing effects of the product G1 are superior to those of the product G2 in absolute moisturizing performance and relative moisturizing performance, verification results are different from the declared human body test indexes, and the fact that the absorption and moisturizing performances of the 3D printing bionic skin and the real human body skin are different is shown, but the verification results are correct is shown. Wherein the ratio of the thickness of the epidermis cell layer to the dermis cell layer is 2:1 and 5:2 has smaller difference between the verification data obtained by the three-dimensional bionic skin sample and the declared human body test index, and can be used as a reference basis for the subsequent sample preparation. In addition, it was found in the test that the plant extract formulation of the product G2 was not as much as the added amount of the base water-retaining agent (sample group a exhibited a lower moisturizing efficacy than sample group C) in terms of the relative moisturizing properties, indicating that its main efficacy is not limited to water retention and moisturizing.
The above description is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above examples, and all technical solutions belonging to the concept of the present invention belong to the protection scope of the present invention. It should be noted that variations or modifications can be made by those skilled in the art without departing from the principles of the present invention, which is also considered as being within the scope of the present invention.
Claims (10)
1. The application method of the bionic skin in the cosmetic test is characterized in that a plurality of 3D printed bionic skin samples are adopted to replace the tested human skin or the tested cell samples for the cosmetic efficacy test.
2. The method of claim 1, wherein the 3D printed plurality of biomimetic skin samples are used for cosmetic moisturizing, sun protection, wrinkle removal efficacy testing.
3. The method of application according to claim 2, wherein the 3D printed plurality of biomimetic skin samples are used for cosmetic moisturizing efficacy testing, comprising the steps of:
s1, 3D printing bionic skin preparation
Using epidermis stem cells and dermis stem cells which are separated and cultured by human skin tissues and are matched with collagen skeleton materials, printing the epidermis stem cells and the dermis stem cells on a printing substrate in a 3D layer-by-layer printing mode to obtain 1 st-M (M is more than or equal to 2) three-dimensional bionic skin containing dermis cell layers, epidermis cell layers and collagen skeleton layers, and dividing a plurality of samples to be tested with the same size on each three-dimensional bionic skin through tissue culture and fusion to obtain a plurality of groups of 3D printing bionic skin samples which can be used for testing the moisturizing efficacy of cosmetics, wherein each group contains the plurality of samples to be tested with the same size;
wherein, the 1-M three-dimensional bionic skins are printed by adopting different 3D printing planning schemes;
s2, testing moisturizing efficacy of cosmetics
S2-1, selecting three identical tested samples from each group of tested samples in the 1 st to M groups of 3D printing bionic skin tested samples obtained in the step S1: sample group a, sample group B and sample group C; the sample group A is an experimental sample group, the sample group B is a comparison sample group I, and the sample group C is a comparison sample group II;
s2-2, detecting and recording skin water content data of each tested sample in the sample group A, the sample group B and the sample group C;
s2-3, smearing a solution containing a basic water-retaining agent and a cosmetic efficacy raw material to be tested on each tested sample of the sample group A, smearing a solution containing only the basic water-retaining agent on each tested sample of the sample group B and the sample group C, wherein the concentration of the basic water-retaining agent smeared on the tested sample of the sample group B is equal to the concentration of the basic water-retaining agent smeared on the tested sample of the sample group A, and the concentration of the basic water-retaining agent smeared on the tested sample of the sample group C is equal to the total concentration of the basic water-retaining agent smeared on the tested sample of the sample group A and the cosmetic efficacy raw material to be tested;
s2-4, after the set time is reached, detecting and recording new skin water content data of each tested sample in the sample group A, the sample group B and the sample group C;
s2-5, calculating skin moisture content change data of the tested sample according to the skin moisture content data measured in the step S2-2 and the skin moisture content data measured in the step S2-4, and evaluating the moisturizing effect of the cosmetic to be tested according to the skin moisture content change data of the tested sample;
s2-6, performing inter-group comparison on the cosmetic moisturizing effect evaluation results obtained by the 1 st to M th groups of 3D printing bionic skin tested samples to obtain the optimal cosmetic moisturizing effect evaluation results and the optimal 3D printing scheme of the three-dimensional bionic skin.
4. A method of application according to claim 3, wherein step S1 comprises the steps of:
s1-1, cell extraction
Crushing, carrying out tissue enzymolysis, filtering and centrifuging on human skin tissues to obtain aggregated cells, and inoculating and culturing the cells to obtain primary skin cells;
s1-2, cell culture
Adding ROCK inhibitor into a part of primary skin cells to perform subculture to obtain the required amount of epidermal stem cells, and performing subculture without adding ROCK inhibitor to a part of primary skin cells to obtain the required amount of dermal stem cells;
s1-3, 3D printing bionic skin
Printing the epidermis stem cells and the dermis stem cells obtained in the step S1-2 on the same printing substrate in a 3D layer-by-layer printing mode by matching with collagen skeleton materials to obtain 1 st-M three-dimensional bionic skin which comprises dermis cell layers, epidermis cell layers and collagen skeleton layers and has different 3D printing planning schemes, and carrying out tissue culture on the obtained three-dimensional bionic skin;
s1-4, preparation of sample to be tested
After tissue culture and fusion of the three-dimensional bionic skin obtained in the step S1-3, a plurality of samples to be measured with the same size are segmented on each three-dimensional bionic skin, so that a plurality of groups of 3D printing bionic skin samples which can be used for testing the moisturizing efficacy of cosmetics are obtained, and each group comprises the plurality of samples to be measured with the same size.
5. The method of claim 3 or 4, wherein in step S1, the different 3D printing schemes are different layer thickness schemes and/or different initial cell concentration schemes.
6. The method of claim 3 or 4, wherein in step S1, the multilayered collagen scaffold material distributed in the three-dimensional simulated skin is immobilized by a cross-linking agent.
7. The method according to claim 3 or 4, wherein in step S1, the plurality of samples to be measured with the same size are segmented on each three-dimensional bionic skin by selecting a region with uniform thickness and quality on each three-dimensional bionic skin.
8. The method according to claim 3 or 4, wherein in step S2-3, the basic water-retaining agent is a component other than the component contained in the cosmetic to be measured itself, or the basic water-retaining agent has the same component as the cosmetic to be measured itself.
9. The method according to claim 3 or 4, wherein in step S2-6, performing group-to-group comparison on the cosmetic moisturizing efficacy evaluation results obtained from the 3D printed simulated skin test samples of groups 1 to M comprises: i, eliminating the evaluation result with obvious distortion from each group of evaluation results; and/or II, selecting a result which is closer to an empirical value or a theoretical value from the evaluation results of each group.
10. The method of claim 9, wherein in step S2-6, at least one set of evaluation results is verified by a second cosmetic moisturizing efficacy test method based on 3D printed biomimetic skin.
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| CN202311698447.6A CN117686661A (en) | 2023-12-12 | 2023-12-12 | Application of bionic skin in cosmetic test |
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