CN104345146A - Cosmetic selection device - Google Patents
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- CN104345146A CN104345146A CN201410256890.2A CN201410256890A CN104345146A CN 104345146 A CN104345146 A CN 104345146A CN 201410256890 A CN201410256890 A CN 201410256890A CN 104345146 A CN104345146 A CN 104345146A
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6881—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from skin
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Abstract
The disclosure relates to a droplet manipulating device and a method for manipulating a droplet. The droplets manipulating device includes a first magnetic field generator, a second magnetic field generator, and a flow channel. The first magnetic field generator produces a first magnetic field on the droplet, so that the droplet has the direction of magnetic field corresponding to the first magnetic field. Further, the second magnetic field generator produces a second magnetic field on the droplet so as to drive the droplet to be in motion in the flow channel.
Description
Technical field
The present invention relates to the system of selection of cosmetics.
Background technology
Human skin often with extraneous contact, and be subject to various stimulation.Its representative example stimulated is ultraviolet.Even if ultraviolet, when not causing too large change to skin in appearance, still can bring out the DNA break of Skin Cell or the sex change etc. of collagen, and become the reason of skin aging.Ultraviolet often routinely causes skin stress.In addition, for women, wash one's face, the daily behavior such as cosmetic also can cause stimulation to skin, these stimulations can as potential stress and accumulate in skin.And, for these stimulate (stress) if resistibility reduce because of certain reason, namely think potential coarse skin or susceptibility skin.Then become and be considered to skin coarse skin state during the environmental stimuli accumulation of such skin.Skin is coarse presents temporary transient state, but its state continued just is called " coarse skin ".
" coarse skin " is the general name of dry and crude skin condition, and its most significant clinical manifestation is the scales of skin that peel off.Known exist close relationship between so coarse skin state and the barrier function of skin, the more serious objective indicator as barrier function of coarse skin (following through skin water loss, be called TEWL) higher, the index as the skin roughened state produced because of physical chemical factor measures TEWL (non-patent literature 1: day skin meeting will, the 92nd volume, the 1001st page of-1003 pages, nineteen eighty-two).
Think that foregoing ultraviolet, chemical substance or other various stimulation can bring impact to skin internal environment, hinder the result of the barrier function of epidermis can produce this coarse skin.When skin presents above-mentioned situation, the prostaglandin Proliferation, Differentiation of cell being brought to impact can be produced in skin inside, rising (the non-patent literature 2:JOURNAL OF INVESTIGATIVE DERMATOLOGY of cytokine concentration, 81st volume, 519-523 page, 1983), hyperfunction (the non-patent literature 3:Journal of Clinical Investigation of nucleic acid (DNA) complex functionality, 87th volume, 1668-1673 page, 1991), rising (the non-patent literature 4:Jounal of Lipid Research of the TEWL of close relationship is there is with barrier function, 26th volume, 418th page-427 pages, 1985 and the 30th volume, 323rd page-333 pages, 1989) etc. change.But the report about the correlativity of above-mentioned phenomenon is little, and its detailed mechanism is also indefinite.Therefore, the index of coarse skin relies on my statement, finding of naked eye and do not have clear and definite index up to now.
In addition, also there is the people of potential coarse skin, but naked eyes detect that coarse skin potential like this has difficulties.In addition, also there is the situation myoplasm comprising potential coarse skin being called susceptibility skin.
Present people's attention that needs are careful when selecting cosmetics of coarse skin or potential coarse skin.Reason is that the skin of coarse skin its state because of cosmetics often occurs worsens further.Therefore, should be noted that following item, namely select the cosmetics of low irritant as far as possible, by using the cosmetics of low stimulation, the development preventing skin coarse, potential coarse skin are to the deterioration of coarse skin.
But reality is up to now, the people of coarse skin or recognize that the people of potential coarse skin or susceptibility skin selects the benchmark of cosmetics only to have the countermeasure of the cosmetics that stimulation during choice for use is few as mentioned above.Therefore, be following method when selecting cosmetics usually, namely the buyer of cosmetics and sellers pour out skin state, the experience according to oneself and the cosmetics information of accumulation to these sellers or consultant face-to-face, select it to think to be suitable for the cosmetics of skin.
In recent years to skin bring stress research obtain remarkable progress, identified become stress protein, the gene of index.As this kind of protein, galactose agglutinin receives publicity.
Galactose agglutinin refer to can identify, linked galactose (β ?galactoside structure) or the general name of protein of each sugar chain crosslinked.Known its with grow, break up, the biological phenomena such as form is formed, metastases, Apoptosis (cell is dead) is relevant.At present, as mammiferous galactose agglutinin, there will be a known Galectin-1 to-15 15 kinds.In the case of human, be not equivalent to-5 ,-6 ,-14, the galactose agglutinin of-15, have 10 kinds at present but have the possibility increased further from now on.Particularly Galectin-7 as skin stress index and receive publicity.
Galectin-7 detects in mammiferous stratified epithelium, in keratinocyte from basalis to cuticula in detect.In patent documentation 1 (Japanese Unexamined Patent Publication 2006-182744 publication), disclose Galectin-7 and express the screening technique of promotion material and coordinated the composition of Galectin-7 composition by coating and played moisture-keeping function, skin symptom improvement result.And then, in patent documentation 2 (Japanese Unexamined Patent Publication 9-216833 publication), disclose the one kind or two or more external preparation comprising galactose agglutinin there is prevention, improve wrinkle of skin, the low inferior the symptoms of aging of elastic force and wound healing effect.
Present inventor finds that Galectin-7 can become the objective indicator of coarse skin, thus proposes patented claim.
Prior art document
Patent documentation
Patent documentation 1 Japanese Unexamined Patent Publication 2006-182744 publication
Patent documentation 2 Japanese Unexamined Patent Publication 9-216833 publication
Non-patent literature
Non-patent literature skin on the 1st understands will, the 92nd volume, the 1001st page of-1003 pages, nineteen eighty-two
Non-patent literature 2J.Invest.Dermatol, Vol.81,519-523,1983
Non-patent literature 3Journal of Clinical Investigation, Vol.87,1668-1673,1991
Non-patent literature 4J.Lipid.Res., Vol.26,418-427,1985
Summary of the invention
The people that problem of the present invention is to be supplied to coarse skin or potential coarse skin or susceptibility skin selects the method for suitable cosmetics.
The present invention is by following Composition of contents.
(1) a kind of method selecting cosmetics, it is characterized in that, when selecting cosmetics to coarse skin, Galectin-7 expression is high when measuring the expression of Galectin-7 of keratoderma, select the cosmetics having coordinated the composition with the effect reducing Galectin-7.
(2) method of the selection cosmetics Gen Ju (1), is characterized in that, the method measuring the expression of the Galectin-7 of keratoderma is undertaken by enzyme-linked immunosorbent assay.
(3) system of selection of basis (1) or the cosmetics described in (2), it is characterized in that, the composition with the effect reducing Galectin-7 is the composition selected by following method, even if tested substance contact people dimensional culture epidermis model, the amount of the Galectin-7 in described model cell is fewer than the amount of the Galectin-7 do not contacted in the described model cell of tested substance, be judged as that tested substance has the effect reducing Galectin-7.
(4) system of selection of basis (1) or the cosmetics described in (2), it is characterized in that, having the composition of effect reducing Galectin-7 is any one in the potpourri of Salix alba Extract, dog rose extract, galactose, Cer NS and ceramide 5.
(5) a kind of method, it is characterized in that, what Galectin-7 quantitative determination information and the Galectin-7 in cosmetics of contrast client skin suppressed composition contains information, selects the cosmetics of the galactose agglutinin suppression composition containing the Galectin-7 concentration being suitable for client.
According to the present invention, the people that can make pours out one's woes has a coarse skin with the Galectin-7 amount of skin for index is to select suitable cosmetics.Because the mensuration of Galectin-7 gathers evaluation sample by easy methods of operating such as tape stripping methods, and due to burden less, no matter who measures all can obtain identical result, so do not need to be seeked advice from by the mode of interviewing with expert as in the past.
And then, as long as in advance the skin Galectin-7 concentration of client is made into database and possesses, suppress the kind of composition, content, inhibition data to contrast by the Galectin-7 in the cosmetics that make with other approach, then cosmetics sales person just can select the coarse skin being suitable for client apply some make up moment.
Accompanying drawing explanation
Fig. 1 is the figure of the expression of the Galectin-7 measuring people's different parts.
Fig. 2 is the figure of the expression of the Galectin-7 measuring the coarse skin model of pressure made by SDS.
Fig. 3 is the figure of the expression of the Galectin-7 measuring the coarse skin model of pressure made by acetone.
Fig. 4 is the Galectin-7 expression of 608 experimenters and the figure of the frequency of occurrences.
Fig. 5 is for confirming Galectin-7 and the figure through the correlativity of skin water loss (TEWL).
Fig. 6 represents that the extract of white willow and dog rose reduces the figure of TEWL in the test of end user's dimensional culture epidermis model.
Fig. 7 represents that the extract of white willow and dog rose reduces the figure of the generation of Galectin-7 in the test of end user's dimensional culture epidermis model.
Fig. 8 represents that galactose reduces the figure of TEWL in the test of end user's dimensional culture epidermis model.
Fig. 9 represents that galactose suppresses the figure of the generation of Galectin-7 in the test of end user's dimensional culture epidermis model.
Figure 10 represents that lactoferrin does not suppress the figure of the generation of Galectin-7 in the test of end user's dimensional culture epidermis model.
Figure 11 represents that Cer NS and ceramide 5 reduce the figure of TEWL in the test of end user's dimensional culture epidermis model.
Figure 12 represents that Cer NS and ceramide 5 suppress the figure of the generation of Galectin-7 in the test of end user's dimensional culture epidermis model.
Embodiment
Below, be described in detail about the present invention.
The system of selection of cosmetics of the present invention with the expression of the Galectin-7 in keratoderma for index.
As long as can obtain cuticular part, the galactose agglutinin measurement site in the present invention can be any part.And, from the object that cosmetics are selected, preferably face, neck, upper arm parts etc. are routinely exposed to the many positions of extraneous chance as determination object as main portions.In addition, the preferred tape stripping of cuticular collection, scraping etc. can obtain cuticular method easily.
The expression of the Galectin-7 of each sample gathered measures by the past known method.Such as, can use according to the method with the enzyme-linked immunosorbent assay (enzyme immunoassay), radio immunoassay (radioimmunoassay), immunoblotting (western blotting method) etc. of the reaction of anti-Galectin-7 antibody.These mensuration means have commercially available, the representatively enzyme-linked immunosorbent assay kit of Galectin-7, can illustrate the ELISA Kit of Galeectin7Human ELISA Kit that Ai Bokang company (Abcam) makes, R & D systems Inc..
From each sample, Galectin-7 is passed through himself known biochemical method, such as freeze-thaw method, ultrasonic fragmentation, homogeneous method etc. prepare soluble component.After extraction, promptly measure.
As have accumulated from the external world stress the coarse skin of skin or susceptibility skin cuticular Galectin-7 expression compared with healthy skin significantly high, therefore, using the number of the Galectin-7 expression in cuticula as index, the degree of coarse skin can be judged easily or whether be potential coarse skin or susceptibility skin.The straightforward procedure that cuticula used in the present invention can use tape stripping method etc. only to be gathered cuticular surface part by cutin adhesive tape gathers.In addition, the present invention can measure the expression of Galectin-7 under do not cause ultraviolet or chemical substance etc. to stimulate to skin, therefore, burden can not be caused to evaluate the degree of the coarse skin of user to user, suitable cosmetics can be selected according to this result.
Method of the present invention is made up of following operation: gather operation, gathers cuticula; Measure operation, measure the expression of the Galectin-7 in the cuticula of described collection operation collection; With selection operation, the expression according to the Galectin-7 of described mensuration operation mensuration selects cosmetics.
First, the Galectin-7 expression in the Galectin-7 expression of mensuration and the cuticula of healthy skin is contrasted.In addition, also can measure the Galectin-7 expression in the cuticula at a certain evaluation object position of experimenter, and by measured Galectin-7 expression with have healthy skin people same evaluation object position cuticula in Galectin-7 expression contrast.The cuticular Galectin-7 expression with the people of healthy skin, by using from the several mean value with the data of the people of healthy skin, can carry out more objective appraisal.Specifically, the distribution of healthy skin can be set from the expression distribution of the Galectin-7 of the experimenter randomly drawed.
Result after contrast like this, when measured Galectin-7 expression is significantly greater than the Galectin-7 expression of healthy skin, can be evaluated as stress accumulation large, and the possibility that can be evaluated as coarse skin or potential coarse skin or susceptibility skin is high.
In addition, when Galectin-7 expression in the cuticula at a certain evaluation object position of experimenter is greater than the Galectin-7 expression in the cuticula at the same evaluation object position of the people with healthy skin significantly, be also evaluated as stress accumulation large.In addition, when this numerical value remarkable high time, be judged as coarse skin or potential coarse skin or susceptibility skin.
When being evaluated as coarse skin, as cosmetics, select the Galectin-7 amount corresponding to skin containing be improved skin coarse, reduce the cosmetics of composition that Galectin-7 produces.But, known to reducing the cosmetics of composition that Galectin-7 produces and indefinite.Therefore, the generation that clearly whether can reduce the Galectin-7 of skin for the composition of cosmetics is needed in advance.This kind of composition is selected by the screening of end user's dimensional culture epidermis model.
In screening technique when selecting to reduce the composition that Galectin-7 produces, people's dimensional culture epidermis model of using refers to " cultivating the lower epidermal tissue maintained ", for by the skin histology from application on human skin collection or embryonic stem cell or adult stem cell are broken up obtained skin histology in vitro and be maintained the tissue cultivated on multilayer structure one side in vitro.Particularly about during from people, LabCyte EPI-MODEL (Japan Tissue Engineering Co., Ltd.) or the such commercially available people's dimensional culture epidermis mold product of EPI model 24 can be used.
When making tested substance contact people's dimensional culture epidermis model, hatch add the tested substance of dissolving with any concentration with the damping fluids such as PBS etc. in the cultivation cup that people's dimensional culture epidermis model is equipped with after, then measure Galectin-7 and evaluate its effect.
When implementing the test of end user dimensional culture epidermis model, when adding the composition that can confirm to make the Galectin-7 of people's cell to increase in advance, can be easy to that detecting sends as an envoy to becomes the effect of the generation reduction of the Galectin-7 of target tested substance.As this kind of material, surfactant can be illustrated.Particularly preferably lauryl sodium sulfate (following, to be recited as SDS).
When the composition that SDS etc. makes Galectin-7 increase being added in people's dimensional culture epidermis model, composition Galectin-7 being increased tested substance and SDS etc. in people's dimensional culture epidermis model adds with Time to be cultivated, and measures the amount of the Galectin-7 in cell.
The amount of the Galectin-7 in dimensional culture epidermis model cell measures by the past known method.Such as, can use according to the method with the enzyme-linked immunosorbent assay, radio immunoassay, immunoblotting etc. of the reaction of anti-Galectin-7 antibody.These mensuration means have commercially available.The representatively enzyme-linked immunoassay kit of Galectin-7, can illustrate the Galeectin7Human ELISA Kit of Human Galectin-7DuoSet Economy Pack (R & D Systems Inc.), the anti-Inc. of Ai Bo.When extracting Galectin-7 from dimensional culture epidermis model cell, can be passed through himself known biochemical method, such as freeze-thaw method, ultrasonic fragmentation, homogeneous method etc. prepare soluble component.After extraction, rapid test Galectin-7.
The cosmetics of the people as the coarse skin that is suitable for pouring out one's woes can be selected like this containing the cosmetics of composition that can confirm to reduce Galectin-7.
In the following embodiments, in the composition being coupled to cosmetics, as can be confirmed the material reducing galactose agglutinin generation by above-mentioned screening, galactose, Salix alba Extract, dog rose extract can be illustrated.
Assigning to implement this test about the one-tenth be coupled in cosmetics, as long as accumulate the information that obtains in advance, when the cosmetics of the people of coarse skin of selecting to pour out one's woes, by contrasting with the Galectin-7 amount of skin, suitable cosmetics can be selected.
Below, be described about specific embodiment, but the present invention is not limited to following embodiment.
Embodiment
1. the mensuration of the expression of the Galectin-7 of people's different parts
< experimenter >
From having the cheek of 35 men and women (comprising the people with atopic dermatitis (AD) past medical history) of healthy skin, inner forearm portion, buttocks collected specimens.
< cuticula sample extraction method, protein quantification >
Stick cuticula test pieces (ASAHIBIOMED company) gather cuticula at the cheek of experimenter, arm, buttocks.(Tissue Protein Extraction Reagent Thermo Scientific system (product#78510) carries out the extraction of cuticula protein to use the T-PER Extraction buffer of 500 μ l to grind method (add tested material in a reservoir, method that beaded glass that diameter is about 2mm and extract carry out mechanical shaking extraction) from cuticula test pieces with pearl.The albumen quality of each sample is measured by Pierce BCA protein Assay Kit (Thermo Scientific#23225).Be determined as the reaction reagent liquid adding 200 μ l Pierce BCA protein Assay Kit in cuticula extracting solution of protein described in 10 μ l, after hatching 30 minutes at 60 DEG C, measure the absorbance at 562nm place.Meanwhile, with BSA production standard curve, calculate albumen quality from the value of absorbance.
The mensuration > of the Galectin-7 in < cuticula
The ELISA kit (galectin-7:DY1339E) of R & D systems is similarly used by the cuticula extracting solution of protein extracted from adhesive tape to measure the expression of Galectin-7.First, the concentration of being specified by the immobilized antibody kit becoming initial antibody in the 96 orifice plate #3590 of costar is spent the night at 20 DEG C and is carried out immobilization.Then, place at 37 DEG C with 1%BSA and carry out closed culture plate in 1 hour, and after cleaning culture plate with 0.01%Tween-PBS, the standard items and the sample that each 100 μ l are become typical curve carry out application of sample, hatch 2 hours at 25 DEG C.After cleaning culture plate with 0.01%Tween-PBS, 2 antibody kit prescribed concentration are hatched 2 hours at 25 DEG C.And then, after cleaning culture plate with 0.01%Tween-PBS, Streptavidin-HRP kit prescribed concentration is hatched 30 minutes at 25 DEG C.Finally, after cleaning culture plate with 0.01%Tween-PBS, TMB solution (the Promega as chromogenic substrate is added according to 100 μ l/well, G7431)), after the time that kit is specified, add 100 μ l stop buffers (0.5M sulfuric acid), the absorbance measuring 450nm place carrys out calculation expression amount.
< result >
Measurement result as shown in Figure 1.
As shown in Figure 1, can confirm daily suffering compared with the usual absent variable arm of the coarse skin of the cheek of environmental stimuli and what is called, buttocks (hip), the expression of Galectin-7 is significantly high.This test findings shows that the Galectin-7 as coarse skin index is useful.
2. the change of the Galectin-7 expression in artificial coarse skin model
The women 14 with healthy skin implements test as experimenter.
The coarse skin modelling > that < is undertaken by SDS
Put 2 pressing quantity of surfactant (lauryl sodium sulfate, hereinafter referred to as SDS) 10% solution in left forearm inside portion and be coated with 100 times back and forth.Spray hot water (35 DEG C, 1L) and rinse surfactant.In addition, hot water (35 DEG C, 1L) is also sprayed in contrast in right forearm inside portion.This operation carries out 4 times in 1 day, and 1 operation repeats with the interval state of more than 1 hour.Aforesaid operations continues the coarse skin model of manual manufacture over 2 days.
< cutin gathers >
Gather 4 respectively at depth direction and do not implement the cuticula that SDS cleaned and implemented the various piece that SDS cleans.Gather the cuticular date within the 4th, 8,12 day, gathering before cleaning with after cleaning.
The mensuration > of < cuticula sample extraction, protein quantification, Galectin-7
Measure in the same manner as the method for above-mentioned 1..
< result >
Indivedual measurement results of experimenter as shown in Figure 2.Can confirm that coarse skin makes the measurement result processing rear 8th ~ 12 days Galectin-7s and significantly rises.
The coarse skin modelling > that < is undertaken by acetone
The women 3 with healthy skin implements test as experimenter.
The glass funnel (diameter 3cm) of upper and lower opening is fixed with rubber strip, and acetone (WAKO is superfine)/ether (WAKO is superfine) equivalent mixed liquor pipettor is injected 2ml in funnel, put both arms process on a shaker 20 minutes.
Then, after the acetone/diethyl ether mixed liquor in dropping pipet removing funnel, inject tap water, and put both arms process 5 minutes (ungrease treatment) on a shaker.Second day, carry out same operation in a same part and again carry out ungrease treatment, make dry skin.In addition, the position of water treatment will only be used in contrast.
< cutin gathers >
4 cuticula do not implemented acetone treatment and implement acetone treatment position are gathered respectively at depth direction.Gathering the date is gather for the 1st, 5,8,12,16,20 day after acetone treatment the previous day and acetone treatment.
The mensuration > of < cuticula sample extraction, protein quantification, Galectin-7
Operate in the same manner as the test method of above-mentioned 1.
< result >
Indivedual measurement results of experimenter as shown in Figure 3.Can confirm that coarse skin makes the measurement result processing rear 8th ~ 12 days Galectin-7s and significantly rises.
Can confirm the generation with coarse skin from the result of above coarse skin modelling test, Galectin-7 also rises.
3. the mensuration of the Galectin-7 of the people of healthy skin
< experimenter >
Gather 608 cheek samples with men and women's (comprising the people with atopic dermatitis past medical history) of healthy skin and confirm the distribution of Galectin-7.In addition, about the wherein experimenter of 35, measure as coarse skin in the past evaluation index through skin water loss (TEWL), and confirm and the measurement result of Galectin-7 with or without correlationship.
< cuticula sample extraction method, protein quantification >
1. same with above-mentioned, stick cuticula test pieces (ASAHIBIOMED company) at the cheek of experimenter and gather cuticula.
The mensuration > of the Galectin-7 in < cuticula
1. same with above-mentioned, used by the cuticula extracting solution of protein extracted from adhesive tape the ELISA kit (galectin-7:DY1339E) of R & D systems to measure the expression of galectin-7.
< is through skin water loss (TEWL (transepidermal water loss) value >
TEWL uses the Delfin of Keystone company to be used for by the mean value measuring 2 times respectively resolving.
< result >
The Galectin-7 measurement result of 608 is as shown in table 1 and Fig. 4.
Table 1
According to table 1, Fig. 4, within the Galectin-7 of the cheek skin of the people of 80% healthy skin belongs to 435pg/ μ g.In addition, listening to investigation and can confirm from experimenter, has the people of atopic dermatitis past medical history, self-consciousness to the Galectin-7 being easy to coarse etc. the people of skin and most demonstrating 500pg/ more than μ g.Therefore, such as can using the index of the measured value 500pg/ μ g of Galectin-7 as coarse skin.
In addition, with the correlativity of TEWL as shown in Figure 5.
Can confirm that Galectin-7 and TEWL exist certain correlationship.
Useful as the index of coarse skin from the mensuration of the known Galectin-7 of above test findings.And then, judge to detect and examine with TEWL the potential coarse skin do not measured.
4. the test of the generation depressant of the Galectin-7 of end user's dimensional culture epidermis model and the mensuration through skin water loss (TEWL) of people's dimensional culture epidermis model
1. improve skin barrier function or suppress the selection through the medicament of skin water loss
(1) cell
End user's dimensional culture epidermis model.
As people's dimensional culture epidermis model, LabCyte EPI-MODEL24 pass (EPI model 24, Japan Tissue Engineering Co., Ltd. system) is used to carry out the inhibit feature of the Galectin-7 in the cell of appraiser's dimensional culture epidermis model.
About people's dimensional culture epidermis model, change to fluid nutrient medium from gel entrapment culture and carry out nutrient culture media exchange, at 37 DEG C, 5%CO
2spend the night in incubator and carry out front cultivation.And, people's dimensional culture epidermis model as on physiology, functionally reappear people sink conditions external model and be utilized.
(2) test specimen
Select 5 kinds of compositions of Salix alba Extract, dog rose extract, galactose, lactoferrin, Cer NS and ceramide 5 potpourri.
(3) generation of Galectin-7 reduces test and TEWL mensuration 1 (Salix alba Extract, dog rose extract)
Tested substance is added according to the concentration shown in table 2 in the cultivation cup that people's dimensional culture epidermis model is equipped with, and at 37 DEG C, 5%CO
2cultivate 24 hours in incubator.
Table 2
| Sample | Composition |
| Contrast | 1%1,3 butylene glycols (BG)/phosphate buffer (PBS) |
| White willow 1 | 73.26 μ g/ml Salix alba Extract (solid constituent), 1%BG/PBS |
| White willow 2 | 24.42 μ g/ml Salix alba Extract (solid constituent), 1%BG/PBS |
| Dog rose 1 | The 345.9 micro-extract of μ g/ml dog roses (solid constituent), 1%BG/PBS |
| Dog rose 2 | 115.3 μ g/ml dogs rose extract (solid constituent), 1%BG/PBS |
White willow 1, white willow 2 use the Dermaveil preparation naturally vigorous and graceful processed of a Japanese ball.The naturally vigorous and graceful Dermaveil processed of Japan's one ball is the water/BG solution of white willow bark extract.Dog rose 1, dog rose 2 use the naturally vigorous and graceful Pharcolex rose B processed of Japan one or nine to prepare.The naturally vigorous and graceful Pharcolex rose B processed of Japan's one ball is the water/BG solution of dog rosaceous fruit extract.
Dimensional culture epidermis model is cleaned with PBS (-) after cultivation completes.Then, the concentration with 0.03% adds SDS (lauryl sodium sulfate, Wako), cleans after 4 hours with PBS (-).
After cleaning with PBS (-), same tested material is measured 2 times by the portable moisture evapotranspirometer (VapoMeter, Delfin system) having a joint through skin water loss (TEWL) Franz pond type cultivated on cup, mean value is calculated as TEWL value.TEWL measurement result as shown in Figure 7.Measuring number of times is 3 times, shows standard deviation in figure.
The people epidermal tissue of people's dimensional culture epidermis model is moved in sample hose (TM-6262.0ml capacity), adds 1 ZB-50 (zirconia
) with 350 μ L cell pyrolysis liquids, use pearl mill formula cell breaking plant with 2,800rpm homogenate 120 second, obtain homogenate solution.This homogenate solution is carried out centrifuging with 4,400G, and the polycarbonate membrane making to be mixed into, insoluble protein become sediment and are separated.Supernatant is moved into dark plate, measures Galectin-7 concentration and protein content.
And the mensuration of Galectin-7 uses Human Galectin-7DuoSet Economy Pack (R & D Systems Inc.) and measures 3 times, and result mean value ± standard deviation represents (Fig. 8).
Any one in Salix alba Extract, dog rose extract all can improve people's dimensional culture epidermis model coarse skin, reduce TEWL, reduce the generation of Galectin-7 simultaneously.
(4) generation of galactose agglutinin reduces test and TEWL mensuration 2 (galactoses)
As described above, galactose to be added in people's dimensional culture epidermis model and to measure Galectin-7.The galactose sample that test uses is added on adjustment in nutrient solution and becomes the concentration shown in table 3.
Table 3
| Sample | Composition |
| Contrast | Phosphate buffer (PBS) |
| Galactose 1 | 1% galactose/PBS |
| Galactose 2 | 0.3% galactose/PBS |
TEWL measures and uses Franz pond type similarly to carry out with the portable moisture evapotranspirometer (VapoMeter, Delfin system) of joint.Result as shown in Figure 9.
The mensuration of Galectin-7 similarly uses Human Galectin-7DuoSet Economy Pack (R & D Systems Inc.) and measures 3 times, and result mean value ± standard deviation represents (Figure 10).
Galactose suppresses the generation of the Galectin-7 of people's dimensional culture epidermis model.
(5) generation of Galectin-7 reduces test 3 (lactoferrin)
Lactoferrin S (a Japanese ball naturally vigorous and graceful company limited system) as commercial cosmetic products contain raw material is diluted to 20 times or 100 times with PBS, adds to as described above in people's dimensional culture epidermis model and to measure Galectin-7.
And, using 20 times of dilutions as lactoferrin 1, using 100 times of dilutions as lactoferrin 2, be presented in following result.
The mensuration of Galectin-7 uses Human Galectin-7DuoSet Economy Pack (R & D Systems Inc.) and measures 3 times in the same manner as the test of described (3), and result mean value ± standard deviation represents (Figure 11).
As shown in figure 11, lactoferrin does not suppress the generation of the Galectin-7 of people's dimensional culture epidermis model.
(6) generation of galactose agglutinin reduces test and TEWL mensuration 4 (Cer NS and ceramide 5 potpourris)
Using as the Cer NS of commercial cosmetic products contain raw material and ceramide 5 mixed in equal amounts, be diluted to 4000 times to prepare the mixed solution of Cer NS and ceramide 5 with 0.5% hydrolecithin.Amount to containing 250 μ g/ml Cer NSs and ceramide 5 in this mixed solution.As described above, this mixed solution is added in people's dimensional culture epidermis model to make Cer NS and ceramide 5 become 12.5 μ g/ml respectively.In addition, cultivate in the people's dimensional culture epidermis model after similarly adding SDS load to.Measure TEWL after cultivation completes, then measure the Galectin-7 amount in people's dimensional culture epidermis model.
TEWL measures and uses Franz pond type similarly to carry out with the portable moisture evapotranspirometer (VapoMeter, Delfin system) of joint.Result as shown in figure 11.
The mensuration of Galectin-7 similarly uses Human Galectin-7DuoSet Economy Pack (R & D Systems Inc.) and measures 2 times, and result mean value represents (Figure 12).
Potpourri respectively containing 12.5 μ g/ml Cer NSs and ceramide 5 inhibits the TEWL of people's dimensional culture epidermis model rise and suppress the generation of Galectin-7.
Can distinguish from above test findings, dog rose, white willow, galactose, Cer NS and ceramide 5 potpourri can reduce the generation of the Galectin-7 of skin.Therefore, the cosmetics containing mentioned component can select the cosmetics as coarse skin.
Claims (5)
1. select a method for cosmetics, it is characterized in that,
When selecting cosmetics to coarse skin, Galectin-7 expression is high when measuring the expression of Galectin-7 of keratoderma, select the cosmetics having coordinated the composition with the effect reducing Galectin-7.
2. the method for selection cosmetics according to claim 1, is characterized in that,
The method measuring the expression of the Galectin-7 of keratoderma is undertaken by enzyme-linked immunosorbent assay.
3., according to the system of selection of cosmetics according to claim 1 or claim 2, it is characterized in that,
The composition with the effect reducing Galectin-7 is the composition selected by following method, even if tested substance contact people dimensional culture epidermis model, the amount of the Galectin-7 in described model cell is fewer than the amount of the Galectin-7 do not contacted in the described model cell of tested substance, be judged as that tested substance has the effect reducing Galectin-7.
4., according to the system of selection of cosmetics according to claim 1 or claim 2, it is characterized in that,
Having the composition of effect reducing Galectin-7 is any one in the potpourri of Salix alba Extract, dog rose extract, galactose, Cer NS and ceramide 5.
5. a method, is characterized in that,
What mensuration information and the Galectin-7 in cosmetics of the Galectin-7 amount of contrast client skin suppressed composition contains information, selects the cosmetics of the galactose agglutinin suppression composition containing the Galectin-7 concentration being suitable for client.
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| CN114187972A (en) * | 2021-12-10 | 2022-03-15 | 华东理工大学 | Calculation simulation method for screening functional ingredients of cosmetics |
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| JP2001163794A (en) * | 1999-12-03 | 2001-06-19 | Shiseido Co Ltd | Promoter for production of hyaluronic acid and preparation for external use for skin |
| JP2006104098A (en) * | 2004-10-04 | 2006-04-20 | Ichimaru Pharcos Co Ltd | Hyaluronidase activity inhibitor |
| JP2006117612A (en) * | 2004-10-25 | 2006-05-11 | Ichimaru Pharcos Co Ltd | Active oxygen eliminating agent |
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| JP5970184B2 (en) * | 2011-12-26 | 2016-08-17 | 株式会社 資生堂 | Evaluation method of cosmetics |
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| CN105734009A (en) * | 2016-04-01 | 2016-07-06 | 陕西博溪生物科技有限公司 | In-vitro recombined human skin epidermis model and preparation method and application thereof |
| CN105734009B (en) * | 2016-04-01 | 2020-05-19 | 广东博溪生物科技有限公司 | In-vitro recombinant human skin epidermis model and preparation method and application thereof |
| CN114187972A (en) * | 2021-12-10 | 2022-03-15 | 华东理工大学 | Calculation simulation method for screening functional ingredients of cosmetics |
| CN114187972B (en) * | 2021-12-10 | 2024-02-09 | 华东理工大学 | Calculation simulation method for screening functional components of cosmetics |
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