CN110227057A - A kind of hair gel cream of hair regeneration and preparation method thereof - Google Patents
A kind of hair gel cream of hair regeneration and preparation method thereof Download PDFInfo
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- CN110227057A CN110227057A CN201910497603.XA CN201910497603A CN110227057A CN 110227057 A CN110227057 A CN 110227057A CN 201910497603 A CN201910497603 A CN 201910497603A CN 110227057 A CN110227057 A CN 110227057A
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Abstract
The present invention provides a kind of hair regeneration hair gel creams, with the weight of the hair regeneration hair gel cream for 100%, the component including following parts by weight: 0.3%-3% acrylate analog copolymer；0.5%-5% glycine betaine；1%-10% hyaluronic acid；0.05%-1% sodium hydroxide；2%-10% pleiotrophic factor solution；Surplus is deionized water；Wherein, the pleiotrophic factor solution is modified pleiotrophic factor solution.Modified pleiotrophic factor solution can induce the proliferation and activation of growth factor signal in people's hair follicle dermal papilla cell, the expression of a variety of growth factors is also stimulated simultaneously, improve the accumulating level of albumen in people's hair follicle dermal papilla cell, promote the proliferation of people's hair follicle dermal papilla cell, further promotes the generation of wool fibre and the development of hair follicle；In addition, strengthening the recovery dynatron effect of stem cell present invention adds other complexing agents, it is more advantageous to pleiotrophic factor and is contacted with skin of head, preferably hair growth and alopecia areata problem.
Description
Technical field
The present invention relates to stem cell and its alopeciaing therapeutic field more particularly to the hair gel creams and its preparation of a kind of hair regeneration
Method.
Background technique
The hair of health is the important factor in order of people's external image, and alopecia areata caused by alopecia, line of gaining fame and fortune move back, are bald
Various lives are caused to perplex Deng to people.
Hair can be divided into four papilla, hair follicle, hair root and hair shaft parts, the physiological characteristic and function of hair from bottom to top
Depend primarily on scalp epidermis papilla below, hair follicle and sebaceous glands etc..Under normal condition, hair also has the growth week of oneself
Phase can occur nature physiological and fall off when growing into certain length.But when people is overstrain one's nerves or carries out dyeing and perm to hair
Afterwards, it causes that abnormal alopecia will be caused when hair follicle, papilla impaired or smegma exception, including gene, hormone, outer
The reason of a series of complex such as wound and iatrogenic event, will cause alopecia areata, bald etc..
In all alopecia cases, the ratio of male pattern baldness has risen to 71%.Currently have for hair loss patient several
Therapeutic choice, such as using oral perhaps external drug or progress operative treatment.However, drug therapy can only be of short duration alleviation
Symptom, hair can start shedding off again after patient discontinues medication.It is a kind of using self single hair follicle and Follicular Unit Transplantation
Reliable therapeutic method of surgery, but the quantity of hair follicle donor is limited, and hair transplantation survival rate is low.Therefore, alopecia
It treats and is badly in need of some new treatment means.
Hair follicle be it is a kind of can undergo growth, decline, suspend mode cyclic process regenerative system, it is by corium and epidermis two
Part forms.Hair cycle and regeneration be periodical reconstructed tissue complex process, it be related to growth factor, cell factor,
Hormone, adhesion molecule and relevant enzyme.MSC is mescenchymal stem cell, has been widely used in regenerative medicine.More and more bodies
Outer and In vivo study shows that the secretion of MSC has extensive biological function, and MSC secretion many in regeneration and repair process has
Growth factor, they are capable of the hair inducibility of Assisted Activation DP cell.Dermal papilla cell (DPC) derives from corium
Mesenchymal cell has very important effect in terms of control growth cycle of hair and hair follicle formation.Some researchers think,
The dysfunction of hair papilla cell makes hair follicle cycle is unbalance to lead to alopecia.Studies have found that stem cell can activate wnt albumen
Signal transduction, and then enhance the activity of DPC, enable hair follicle to be converted to active anagen phase rank from static stand-down
Section, and promote hair regeneration under stable state and respond depilation stress, achieve the effect that hair regeneration.In the research of forefathers,
Successful preparation condition culture medium MSC-CM.Hair cycle is accelerated using MSC-CM, it is related to increase internal hair induction
The expression of gene, and restore the hair inducibility that DP cell is destroyed.But the microenvironment of internal inflammation will affect stem cell
Effect play, keep stem cell induced efficiency lower, hair regeneration can not be promoted well, the purpose of clinical treatment is not achieved.
Summary of the invention
The purpose of the present invention is to provide a kind of hair regeneration hair gel creams, it is intended to it is easy to solve anti-hair loss products in the prior art
It is influenced by internal inflammatory microenvironment, the bad problem of anticreep effect.
For achieving the above object, The technical solution adopted by the invention is as follows:
A kind of hair regeneration hair gel cream, with the weight of the hair regeneration hair gel cream for 100%, including following parts by weight
Component:
Wherein, the pleiotrophic factor solution is modified pleiotrophic factor solution.
A kind of and preparation method of hair regeneration hair gel cream comprising following steps:
Each component is weighed respectively according to above-mentioned hair regeneration hair gel cream；
The acrylate analog copolymer, the glycine betaine, the hyaluronic acid are added to the water mixing, heats, obtains
First chamber；
The sodium hydroxide is added in first chamber, is cooled to 35-45 DEG C, obtains second chamber；
The modified pleiotrophic factor solution is added in the second chamber, is uniformly mixed so as to obtain hair regeneration hair gel cream.
Compared with prior art, a kind of hair regeneration hair gel cream that the present invention is prepared, main component includes propylene
Sour lipid copolymer, glycine betaine, hyaluronic acid, sodium hydroxide, pleiotrophic factor solution and water；Wherein, wherein the pleiotrophic factor
Solution is modified pleiotrophic factor solution.Modified pleiotrophic factor solution can induce growth factor in people's hair follicle dermal papilla cell to believe
Number proliferation and activation, while the expression of a variety of growth factors can also be stimulated, improve albumen in people's hair follicle dermal papilla cell
Accumulating level, promote people's hair follicle dermal papilla cell proliferation, further promote wool fibre generation and hair follicle development；This
Outside, present invention adds other complexing agents, including acrylate analog copolymer, glycine betaine, hyaluronic acid, sodium hydroxide, water
Equal substances, are made hair gel cream, strengthen the recovery dynatron effect of stem cell, be more advantageous to pleiotrophic factor contacts with skin of head, multiple-effect
The factor after scalp enters deep skin, be on the one hand can the stimulating growth factor effect, promote generation and the hair follicle of wool fibre
Development, on the other hand, the inhibitor that can be used as inflammatory reaction inhibit the inflammation at papilla and hair follicle position, reach hair regeneration
Purpose, preferably hair growth and alopecia areata problem.
In addition, a kind of preparation method of hair regeneration hair gel cream is additionally provided, the preparation side of above-mentioned hair regeneration hair gel cream
Method only needs each component carrying out mixing treatment according to the proportion, and the method which uses distributed rendering can make
Each component is uniformly dispersed, while carrying out heating mixing to different component using different temperatures, is able to maintain each component good
Performance while so that each component is can be realized above-mentioned beneficial effect, is incubated for its stability.In addition, the preparation method simple process,
Condition is controllable, and equipment requirement is low, prepares convenient and efficient, can be used for industrialized production.
Detailed description of the invention
Fig. 1 is the increment phenomenon of various concentration pleiotrophic factor solution processing hHFDC provided in an embodiment of the present invention.
Fig. 2 a is that the hair loss patient hair amount before hair gel cream is smeared is not used；Fig. 2 b is to have used to be not added with pleiotrophic factor solution
Hair gel cream smear 30 days after hair amount；Fig. 2 c is that the hair loss patient hair amount before hair gel cream is smeared is not used；Fig. 2 d is to use
Add hair amount of the hair gel cream smearing of pleiotrophic factor solution after 30 days.
Specific embodiment
To keep the purpose, technical solution and technical effect of the embodiment of the present invention clearer, the present invention will be implemented below
Technical solution in example is clearly and completely described, it is clear that and described embodiments are some of the embodiments of the present invention, and
The embodiment being not all of.In conjunction with the embodiment in the present invention, those of ordinary skill in the art are not making creative work
Under the premise of every other embodiment obtained, shall fall within the protection scope of the present invention.
In the description of the present invention, it is to be understood that, term " first ", " second " are used for description purposes only, and cannot
It is interpreted as indication or suggestion relative importance or implicitly indicates the quantity of indicated technical characteristic.Define as a result, " the
One ", the feature of " second " can explicitly or implicitly include one or more of the features.In the description of the present invention,
The meaning of " plurality " is two or more, unless otherwise specifically defined.
Present example provides a kind of hair regeneration hair gel cream, with the weight of the hair regeneration hair gel cream for 100%, packet
Include the component of following parts by weight:
Wherein, the pleiotrophic factor solution is modified pleiotrophic factor solution.
Specifically, with the weight of the hair regeneration hair gel cream for 100%, the additive amount of the pleiotrophic factor solution is
2%-10%.If additive amount is too low, effective component pleiotrophic factor content is too low in hair regeneration hair gel cream, then patient used
Cheng Zhong, can not well the regeneration of few hair the problem of；If adding too much, effective component mistake in hair regeneration hair gel cream
Height can impact the stability of hair follicle system, while can also play inhibiting effect, and people's hair follicle dermal papilla cell is inhibited to increase
It grows, further suppresses the generation of wool fibre and the development of hair follicle, hair loss state can not be treated, can not effectively solve hair
The problem of regeneration.
Specifically, the pleiotrophic factor solution is modified pleiotrophic factor solution.Preferably, the modified pleiotrophic factor solution
It is the supernatant being prepared by mustard seed low-kappa number umbilical cord mesenchymal stem cells cell monolayer.Sinapic acid, also known as sinapsis alba acid,
3,5- dimethoxy-4 's-hydroxycinnamic acid is a kind of naturally occurring hydroxycinnamic acid of small molecule, and hydroxycinnamic acid, which belongs to, to be had
The phenolic acid class of bioactivity is contained mainly in various herbal materials, including veterinary antibiotics, herbal medicine and high bran cereal.Modification is more
Effect factor solutions are the supernatants being prepared by mustard seed low-kappa number umbilical cord mesenchymal stem cells cell monolayer, and sinapic acid can be with
The proliferation and activation of growth factor signal in people's hair follicle dermal papilla cell are induced, while a variety of growth factors can also be stimulated
Expression, the cell factor of the pretreated stem cell secretion of sinapic acid can be improved the accumulation of albumen in people's hair follicle dermal papilla cell
Level promotes the proliferation of people's hair follicle dermal papilla cell, further promotes the generation of wool fibre and the development of hair follicle.
Preferably, in the modified pleiotrophic factor solution, the concentration of the modified pleiotrophic factor is 20-100mg/mL.
In modified pleiotrophic factor solution, the concentration of modified pleiotrophic factor is lower than 20mg/mL, since concentration is too low, active constituent content
Very little, proliferative induction and activation can not be carried out to growth factor signal in people's hair follicle dermal papilla cell, while can not also stimulated
The expression of a variety of growth factors can not further promote the generation of wool fibre and the development of hair follicle, can not effectively solve hair again
Raw problem；If the concentration of pleiotrophic factor is too high, being higher than 100mg/mL, then effective component is excessively high in modified pleiotrophic factor solution,
Excessive effective component is easy to influence the stability of hair follicle system, then effective component is excessively high in hair regeneration hair gel cream, can be to hair
The stability of bag system impacts, while can also play inhibiting effect, and the expression quantity of growth factor is made to will receive inhibition, into one
The accumulating level for inhibiting to step albumen in people's hair follicle dermal papilla cell, can not treat hair loss state, can not be effective
Solve the problems, such as hair regeneration.
In a preferred embodiment of the invention, in modified pleiotrophic factor solution, the concentration of the sinapic acid is 80-120 μM.
If the additive amount for adding the sinapic acid is very few, it is unable to fully pre-process umbilical cord mesenchymal stem cells, can not activates
Cell factor in stem cell can not stimulate the expression of a variety of growth factors, can not further promote generation and the hair of wool fibre
The development of capsule；If the excessive concentration of the sinapic acid, growth factor expression excess will lead to, in people's hair follicle dermal papilla cell
The accumulating level of albumen is excessive, and excessive effective component is easy to influence the stability of hair follicle system, while can also play inhibition and make
With making one the inducing action of the proliferation of growth factor signal and activation in hair follicle dermal papilla cell and receive inhibition, make to take off
Heat condition is unable to get improvement, can not make hair regeneration.In a preferred embodiment of the invention, the additive amount that sinapic acid is added
It is 100 μM.
Specifically, also added in the hair gel cream acrylic acid esters co-polymer be with acrylate (with methyl acrylate,
Based on ethyl ester, butyl ester and methyl methacrylate) it is the general name of polymer that raw material is generated through copolyreaction.Preferably, this hair
The bright cross-linked copolymer for selecting acrylic acid and VP, adding this substance makes hair gel cream have certain hydrophily, it is easier to smear.?
In the preferred embodiment of the present invention, with the weight of the hair regeneration hair gel cream for 100%, the additive amount of acrylate analog copolymer
For 0.3%-3%.
Specifically, also added glycine betaine in the hair gel cream, glycine betaine is a kind of alkaloid, has strong hygroscopicity
It can, it can be ensured that the hair gel cream being prepared is paste form, is easy to use.In a preferred embodiment of the invention, with the hair
The weight of hair regeneration hair gel cream is 100%, and the additive amount of glycine betaine is 0.5%-5%.
Specifically, also added hyaluronic acid in the hair gel cream, the Nutrition and Metabolism of hair follicle skin can be improved, make hair follicle
Skin is tender, while moisturizing is again good skin penetration enhancer, can further improve hair follicle dermal papilla cell
Absorption to pleiotrophic factor solution activates growth factor, promotes the generation of wool fibre and the development of hair follicle.Of the invention preferred real
It applies in example, with the weight of the hair regeneration hair gel cream for 100%, the additive amount of hyaluronic acid is 1%-10%.
Specifically, also added sodium hydroxide in the hair gel cream, sodium hydroxide solution is alkaline solution, can be accelerated each
The dissolution of substance keeps hair gel cream quality fine and smooth, easy to use.In a preferred embodiment of the invention, with the hair regeneration gel
The weight of cream is 100%, and the additive amount of sodium hydroxide is 0.05%-1%.
The application is determined by the adding proportion to other additives, it is ensured that other substances and pleiotrophic factor solution
Being formed has a certain proportion of hair gel cream, acts on each substance collaboration, strengthens the recovery dynatron effect of stem cell, be on the one hand
Can the stimulating growth factor effect, promote generation and the hair follicle development of wool fibre, on the other hand, can be used as the suppression of inflammatory reaction
Preparation inhibits the inflammation at papilla and hair follicle position, achievees the purpose that hair regeneration, preferably hair growth and alopecia areata problem.
To sum up, a kind of hair regeneration hair gel cream that the present invention is prepared, main component include acrylics copolymerization
Object, glycine betaine, hyaluronic acid, sodium hydroxide, pleiotrophic factor solution and water；Wherein, the pleiotrophic factor solution is modified multiple-effect
Factor solutions.Modified pleiotrophic factor solution can induce the proliferation of growth factor signal in people's hair follicle dermal papilla cell and swash
It is living, while the expression of a variety of growth factors can also be stimulated, the accumulating level of albumen in people's hair follicle dermal papilla cell is improved, is promoted
Into the proliferation of people's hair follicle dermal papilla cell, further promote the generation of wool fibre and the development of hair follicle；In addition, the present invention is added
The substances such as other complexing agents, including acrylate analog copolymer, glycine betaine, hyaluronic acid, sodium hydroxide, water, are made Gel
Curry cream strengthens the recovery dynatron effect of stem cell, is more advantageous to pleiotrophic factor and contacts with skin of head, pleiotrophic factor enters through scalp
After deep skin, be on the one hand can the stimulating growth factor effect, promote generation and the hair follicle development of wool fibre, on the other hand,
The inhibitor that can be used as inflammatory reaction inhibits the inflammation at papilla and hair follicle position, achievees the purpose that hair regeneration, preferably
Hair growth and alopecia areata problem.
Correspondingly, the embodiment of the invention also provides a kind of above-mentioned preparation methods of hair regeneration hair gel cream.This method packet
Include following steps:
G01. each component is weighed respectively according to above-mentioned hair regeneration hair gel cream；
G02. the acrylate analog copolymer, the glycine betaine, the hyaluronic acid are added to the water mixing, are heated to
80-90 DEG C, obtain first chamber；
G03. the sodium hydroxide is added in first chamber, is cooled to 35-45 DEG C, obtains second chamber；
G04. the pleiotrophic factor solution is added in the second chamber, is uniformly mixed so as to obtain hair regeneration hair gel cream.
Specifically, being used for hair regeneration hair gel cream each component preferred content and type institute for example above in above-mentioned steps G01
It states, in order to save length, details are not described herein.
In above-mentioned steps G02, the acrylate analog copolymer, the glycine betaine, the hyaluronic acid are added to the water
It mixes, mixes as conventional processing mode, such as stir, as long as each component is uniformly mixed.Specifically, heating stepses
In, heating temperature be 80~90 DEG C, if heating temperature is too low, the acrylate analog copolymer, glycine betaine, hyaluronic acid without
Method is completely dissolved into water, if heating temperature is excessively high, will lead to the wasting of resources, can also make to react danger coefficient rising.It is preferred that
, the heating time is 15~25 minutes.It is the acrylate analog copolymer, glycine betaine, transparent if heating time is too short
Matter acid can not be completely dissolved into water, and reaction not exclusively, if heating time is too long, will affect each material composition.
Specifically, the sodium hydroxide is added in first chamber, is cooled to 35-45 DEG C in above-mentioned steps G03,
Obtain second chamber；Specifically, the sodium hydroxide is added in first chamber, since sodium hydroxide is highly basic, it is added
In water, dissolution meeting heat release rises solution system temperature.If pleiotrophic factor solution under the high temperature conditions, is added, then it can make multiple-effect
Factor solutions inactivation does not have the proliferation of growth factor signal and the effect of activation in induction people's hair follicle dermal papilla cell, together
When can not also stimulate the expression of a variety of growth factors therefore after sodium hydroxide is added, to be cooled to 35-45 DEG C, further it is excellent
Choosing, will above-mentioned first combination cooling to 40 DEG C, 40 DEG C of optimum temperatures acted on for pleiotrophic factor.
Specifically, the pleiotrophic factor solution is added in the second chamber, mixes in above-mentioned steps G04
To hair regeneration hair gel cream.The pleiotrophic factor solution is added as modified pleiotrophic factor solution, is uniformly mixed so as to obtain hair regeneration gel
Cream.Mixing is conventional processing mode, such as stirring, as long as each component is uniformly mixed.
Preferably, in the hair regeneration hair gel cream, the modified pleiotrophic factor solution the preparation method is as follows:
S01. umbilical cord mesenchymal stem cells are provided to be cultivated to obtain the first cultivating system；
S02. sinapic acid is added in first cultivating system to be pre-processed, obtains the second cultivating system；
S03. second cultivating system digested, neutralized, washed, be centrifuged, be concentrated collection obtain supernatant multiple-effect
Factor solutions.
Specifically, in above-mentioned steps S01, umbilical cord mesenchymal stem cells is provided and are cultivated to obtain the first cultivating system.
Umbilical cord mesenchymal stem cells (Mesenchymal Stem Cells, MSCs), which refer to, is present in one of neonatal umbilical cord tissue
Versatile stem cell, differentiation potential with higher can be broken up to multiple directions, can be divided into many kinds of histocytes,
It is the aquicultural stem cell of pleiotrophic factor that the application, which selects umbilical cord mesenchymal stem cells, can promote cell Proliferation, simultaneously
Since the immune performance of umbilical cord mesenchymal stem cells is strong, have one using the pleiotrophic factor solution that the cell culture obtains
Surely the effect for inhibiting inflammation, reinforces the anti-inflammatory performance of pleiotrophic factor solution.Specifically, the umbilical cord mesenchyma of the offer is dry thin
Born of the same parents' the preparation method is as follows:
S11., umbilical cord mesenchymal stem cells primary cell is provided；
S12. the culture solution of the umbilical cord mesenchymal stem cells primary cell is replaced, culture to cell reaches 70% plocoid
State is digested, is neutralized, is washed, and umbilical cord mesenchymal stem cells suspension is obtained；
S13. the umbilical cord mesenchymal stem cells suspension is centrifuged, obtains umbilical cord mesenchymal stem cells；
S14. the umbilical cord mesenchymal stem cells are cultivated, obtains a large amount of umbilical cord mesenchymal stem cells, and at -80 DEG C
Freezen protective.
Specifically, above-mentioned steps are operated in superclean bench, in superclean bench in above-mentioned steps S11
In operated, it is preferred that umbilical cord is rinsed using the physiological saline containing 2 times of 100U/mL penicillin and streptomysin, is removed
Umbilical cord blood remained on surface, the Wal Tong Shi glue on umbilical cord is removed, and by Wal Tong Shi glue be cut into 1 cubic millimeter with
Under bulk, be spread evenly across on culture dish, be slow added into serum-free medium and cultivated.Preferably, by Wall Tong Shi
It is relatively cut into 1 cubic millimeter of bulk below, it is more uniform when its purpose is to be coated with Wal Tong Shi glue, simultaneously
And make in cell cultivation process, good growing space is provided for cell, keeps the growth of cell unrestricted, on the other hand
Ensure that the quantity of cell culture is enough.If obtained blocky size is greater than 1 cubic millimeter, it will lead to and cultivated in culture dish
The cell quantity arrived is very little, wastes culture medium resource.In a preferred embodiment of the invention, in the block-like China of the even spread
Serum-free medium is added in the culture dish of your Tong Shi glue to be cultivated, the serum-free medium of the addition is 10mL, according to training
Support ware in even spread blocky Wal Tong Shi glue quantity, be added 10mL serum-free medium can be provided for it is enough
Nutriment promotes cell growth；It is further preferred that the incubator condition cultivated is 37 DEG C, volume fraction is
Umbilical cord mesenchymal stem cells primary cell can be obtained under above-mentioned condition of culture in 5% carbon dioxide, saturated humidity incubator.
Specifically, in above-mentioned steps S12, after the umbilical cord mesenchymal stem cells primitive cell culture 7 days, replacement
The culture solution of the umbilical cord mesenchymal stem cells primary cell, it is preferred that the culture solution is serum-free medium, replacement culture
Liquid is that purpose is to provide enough nutrition for cell growth, promotes cell fusion.After replacing culture solution, culture to cell reaches
70% Fusion Strain, when the Fusion Strain of cell reaches 70%, cell is in state appropriate and is digested, if degrees of fusion
Excessively high, then bad generation digestion reaction, digestion effect is bad, then will affect subsequent cell culture, meanwhile, if degrees of fusion is excessively high,
Cell will form competition each other, compete nutritional ingredient and the growing space of culture medium；If degrees of fusion is too low, digest
Cell quantity is very few, will affect subsequent cell and collects.Specifically, after culture reaches 70% Fusion Strain to cell, to it
It digested, neutralized, further obtain umbilical cord mesenchymal stem cells suspension.Preferably, the digestion is first to remove in cell
Serum-free medium, then add digestive juice and digested.In a preferred embodiment of the invention, the digestive juice selects tryptose
Enzyme-sodium ethylene diamine tetracetate (EDTA) is digested, and still more preferably, uses 0.25% and 0.1% trypsase-respectively
Sodium ethylene diamine tetracetate (EDTA) solution digests 1 minute respectively, shrinks umbilical cord mesenchymal stem cells, makes between cell and cell
It is separated from each other and becomes individual cells.After digestion reaction, the serum free medium supernatant previously removed is added, neutralizes tryptose
Enzyme-EDTA solution, and gently blown and beaten using pipette, obtain umbilical cord mesenchymal stem cells suspension.
Specifically, the umbilical cord mesenchymal stem cells suspension is centrifuged, umbilical cord is obtained in above-mentioned S13 step
Mescenchymal stem cell.Preferably, it is 1000rpm that the method for the centrifugation, which is setting revolving speed, and centrifugation time is 8 minutes, in removal
Clear liquid rejoins Serum-free complete medium and umbilical cord mesenchymal stem cells precipitating is resuspended, obtains umbilical cord mesenchymal stem cells.
Specifically, cultivate the umbilical cord mesenchymal stem cells in above-mentioned S14 step, obtain between a large amount of umbilical cords
Mesenchymal stem cells.Preferably, in the expansion incubation, secondary culture is carried out on the culture dish of 10cm.It is excellent in the present invention
It selects in embodiment, the umbilical cord mesenchymal stem cells for selecting third generation culture to obtain, in its logarithm growth stage, according to the method described above
It digested, neutralized, washed, collect cell, using cells frozen storing liquid gravity treatment umbilical cord mesenchymal stem cells, be sufficiently mixed uniformly,
It is sub-packed in cryopreservation tube, cryopreservation tube is placed in the freezing storing box equipped with isopropanol, in -80 DEG C of freezen protectives according to every pipe 1mL.It is excellent
Choosing, the cells frozen storing liquid is the mixed solution of 90% fetal calf serum and 10% dimethyl sulfoxide.
In above-mentioned S01 step, it is preferred that the umbilical cord mesenchymal stem cells of offer are the third that above-mentioned culture obtains
For umbilical cord mesenchymal stem cells.Under normal conditions, the umbilical cord mesenchymal stem cells of people activity at second and third generation is strongest,
After algebra increases to a certain extent (such as 30 Dai Yihou), cell activity can be substantially reduced；It is preferred that being preferably implemented in the present invention
In example, tested using third generation umbilical cord mesenchymal stem cells.The third generation umbilical cord mesenchymal stem cells are cultivated,
The first obtained cultivating system be 40%~50% cell monolayer, when first cultivating system be 40%~50% single layer
Cell can be such that the sinapic acid of subsequent addition is preferably pre-processed, preferably activation stem cell, make stem cell secretion cell
The factor enhances the repairing activity of stem cell.
Specifically, sinapic acid is added in first cultivating system and is pre-processed, obtains in above-mentioned S02 step
Second cultivating system；In a preferred embodiment of the invention, sinapic acid is added in first cultivating system to carry out stem cell
Pretreatment, is continuously cultivated, and incubation time is 46~50 hours, and in the second obtained cultivating system, cell monolayer fusion reaches
To 75%-80%.If incubation time is too short, the cell differentiation cultivated is too low, and cell quantity is less, after being unfavorable for
The effective component of continuous reaction, the modification pleiotrophic factor solution being centrifuged is inadequate；If incubation time is too long, obtained cell
Differentiation degree is excessively high, and cell is easy to be at war with to nutriment, space size in a limited space, make thin to grow
Born of the same parents' growing way is bad, meanwhile, incubation time is too long, is unfavorable for cell and carries out subsequent processing.
Specifically, being digested, neutralizing, being washed by second mixture in above-mentioned steps S03, it is centrifuged, receipts is concentrated
Collection obtains pleiotrophic factor solution.Specifically, removing the medium supernatant of second mixture, digestive juice is added and disappears
Change.Preferably, the digestive juice is selected from one kind or several of trypsin solution, sodium ethylene diamine tetracetate solution, collagenase solution
Kind.In a preferred embodiment of the invention, selected digestive juice is trypsin-EDTA solutions.Still more preferably, it uses respectively
0.25% and 0.1% trypsin-EDTA sodium (EDTA) solution digests 1 minute respectively, keeps umbilical cord mesenchyma dry
Cellular contraction, making to be separated from each other between cell and cell becomes individual cells.After digestion reaction, add previously removed without blood
Clear medium supernatant is neutralized trypsin-EDTA solutions, and is gently blown and beaten and washed using pipette, is obtained between umbilical cord
Mesenchymal stem cell suspension.Further, centrifugal treating is carried out to umbilical cord mesenchymal stem cells suspension obtained above, collects training
Obtained supernatant is supported, in a preferred embodiment of the invention, in the centrifugation reaction process, the revolving speed of centrifugation is 2000rpm, from
The heart time is 5 minutes.If the revolving speed of centrifugation is too high, cell will lead in centrifugal process, due to excessively high centrifugal force, make thin
The activity of born of the same parents is affected；If centrifugation revolving speed it is too low, cell body can not be precipitated, obtained supernatant i.e. multiple-effect because
The residual that a large amount of cell bodies are had in sub- solution influences subsequent use.Centrifugation time is set as 5 minutes, if centrifugation time is too long,
Influence reaction efficiency；If centrifugation time is too short, cell body can not be precipitated, the i.e. modified pleiotrophic factor of obtained supernatant
The residual that a large amount of cell bodies are had in solution influences subsequent use.The i.e. modified pleiotrophic factor of the supernatant being collected by centrifugation is molten
Liquid.Specifically, further the modification pleiotrophic factor solution being collected by centrifugation is concentrated, modified pleiotrophic factor solution is improved
Concentration, it is preferred that method that the present invention is concentrated is still more preferably, selected using the method being concentrated by ultrafiltration
The molecular size range of ultrafiltration membrane is 5KDa, is mainly rejected by solute, and hydrone passes through film, further will modified multiple-effect because
Sub- solution is concentrated 10 times, is utilized the mescenchymal stem cell conditioned medium supernatant of mustard seed acid active, i.e., modified multiple-effect because
Sub- solution.The concentration for improving modified pleiotrophic factor solution can induce growth factor signal in people's hair follicle dermal papilla cell
Proliferation and activation, while the expression of a variety of growth factors can also be stimulated, improve albumen in people's hair follicle dermal papilla cell
Accumulating level promotes the proliferation of people's hair follicle dermal papilla cell, further promotes the generation of wool fibre and the development of hair follicle, into one
Walk hair growth and alopecia areata.
To sum up, the preparation method of above-mentioned hair regeneration hair gel cream only needs each component carrying out mixing treatment i.e. according to the proportion
Can, the method which uses distributed rendering can make each component be uniformly dispersed, while using different temperatures to different groups
Divide and carry out heating mixing, each component is made to be able to maintain good performance, while so that each component is can be realized above-mentioned beneficial effect,
It is incubated for its stability.In addition, the preparation method simple process, condition is controllable, and equipment requirement is low, prepares convenient and efficient, can be used for
Industrialized production.
Now by taking hair regeneration hair gel cream each component content and preparation method as an example, the present invention is carried out further specifically
It is bright.
Embodiment 1
Hair regeneration hair gel cream each component content and preparation method thereof.Wherein, hair regeneration hair gel cream each component is included
Described in weight ratio ingredient table 1 as follows.Wherein, pleiotrophic factor solution the preparation method is as follows: selection third generation umbilical cord between fill
When matter stem cell is cultivated to obtain the cell monolayer containing 40%, 80 μM of sinapinic acid solution is added, is further cultured for 48 hours, makes
Cell monolayer fusion reaches 75%, then is digested, neutralized, washed, and being centrifuged, collection is concentrated to obtain medium supernatant is multiple-effect
Factor solutions.And measure in the precipitating that centrifugation obtains, the quantity of living cells umbilical cord mesenchymal stem cells is about 5 × 105A/mL,
Ensure that sinapic acid being capable of successful activation umbilical cord mesenchymal stem cells.
The preparation method of hair regeneration hair gel cream: weighing required substance according to the composition and ratio of above-mentioned each ingredient respectively, will
0.5% acrylate analog copolymer, 1% glycine betaine, 2% hyaluronic acid are added to the water mixing, are heated to 80 DEG C, heat 15 points
Clock dissolves above-mentioned three kinds of substances；It is cooled to 35 DEG C again, 0.1% sodium hydroxide, 4% pleiotrophic factor solution is added, is uniformly mixed
Obtain hair regeneration hair gel cream.
Embodiment 2
Hair regeneration hair gel cream each component content and preparation method thereof.Wherein, hair regeneration hair gel cream each component is included
Described in weight ratio ingredient table 1 as follows.Wherein, pleiotrophic factor solution the preparation method is as follows: selection third generation umbilical cord between fill
When matter stem cell is cultivated to obtain the cell monolayer containing 45%, 100 μM of sinapinic acid solution is added, is further cultured for 48 hours,
So that cell monolayer fusion is reached 80%, then digested, neutralized, washed, it is more for being centrifuged, collection is concentrated to obtain medium supernatant
Imitate factor solutions.And measure in the precipitating that centrifugation obtains, the quantity of living cells umbilical cord mesenchymal stem cells is about 6 × 105A/
ML, it is ensured that sinapic acid being capable of successful activation umbilical cord mesenchymal stem cells.
The preparation method of hair regeneration hair gel cream: weighing required substance according to the composition and ratio of above-mentioned each ingredient respectively, will
1% acrylate analog copolymer, 2% glycine betaine, 4% hyaluronic acid are added to the water mixing, are heated to 85 DEG C, heat 20 minutes,
Dissolve above-mentioned three kinds of substances；It is cooled to 40 DEG C again, 0.5% sodium hydroxide, 6% pleiotrophic factor solution is added, is uniformly mixed
To hair regeneration hair gel cream.
Embodiment 3
Hair regeneration hair gel cream each component content and preparation method thereof.Wherein, hair regeneration hair gel cream each component is included
Described in weight ratio ingredient table 1 as follows.Wherein, pleiotrophic factor solution the preparation method is as follows: selection third generation umbilical cord between fill
When matter stem cell is cultivated to obtain the cell monolayer containing 50%, 120 μM of sinapinic acid solution is added, is further cultured for 48 hours,
So that cell monolayer fusion is reached 80%, then digested, neutralized, washed, it is more for being centrifuged, collection is concentrated to obtain medium supernatant
Imitate factor solutions.And measure in the precipitating that centrifugation obtains, the quantity of living cells umbilical cord mesenchymal stem cells is about 7 × 105A/
ML, it is ensured that sinapic acid being capable of successful activation umbilical cord mesenchymal stem cells.
The preparation method of hair regeneration hair gel cream: weighing required substance according to the composition and ratio of above-mentioned each ingredient respectively, will
2% acrylate analog copolymer, 3% glycine betaine, 6% hyaluronic acid are added to the water mixing, are heated to 85 DEG C, heat 20 minutes,
Dissolve above-mentioned three kinds of substances；It is cooled to 40 DEG C again, 0.8% sodium hydroxide, 8% pleiotrophic factor solution is added, is uniformly mixed
To hair regeneration hair gel cream.
Comparative example
Hair regeneration hair gel cream each component content and preparation method thereof.Wherein, hair regeneration hair gel cream each component is included
Described in weight ratio ingredient table 1 as follows.Wherein, pleiotrophic factor solution the preparation method is as follows: selection third generation umbilical cord between fill
When matter stem cell is cultivated to obtain the cell monolayer containing 50%, it is added without sinapinic acid solution, is further cultured for 48 hours, makes single layer
Cell fusion reaches 80%, then is digested, neutralized, washed, and being centrifuged, collection is concentrated to obtain medium supernatant is pleiotrophic factor
Solution.And measure in the precipitating that centrifugation obtains, the quantity of living cells umbilical cord mesenchymal stem cells is about 6 × 105A/mL, it is ensured that
Sinapic acid being capable of successful activation umbilical cord mesenchymal stem cells.
The preparation method of hair regeneration hair gel cream: weighing required substance according to the composition and ratio of above-mentioned each ingredient respectively, will
1% acrylate analog copolymer, 2% glycine betaine, 4% hyaluronic acid are added to the water mixing, are heated to 85 DEG C, heat 20 minutes,
Dissolve above-mentioned three kinds of substances；It is cooled to 40 DEG C again, 0.5% sodium hydroxide is added, 6% pleiotrophic factor solution is uniformly mixed
To hair regeneration hair gel cream.
Measurement is analyzed to the characteristic further progress of the above-mentioned pleiotrophic factor solution being prepared and hair regeneration hair gel cream.
Analysis determining method is as follows:
The pleiotrophic factor liquid and people's hair follicle dermal papilla cell being prepared respectively with Examples 1 to 3 and comparative example
(hHFDC) 72h is co-cultured, 3- [4,5- dimethyl -2- thiazolyl] -2,5- diphenyl -2H- tetrazolium bromide (MTT) is then used
Reagent further cultivates 3h, then removes medium, the formaldehyde crystal generated with dimethyl sulfoxide (DMSO) dissolution MTT.With light splitting
Photometer measurement wavelength is the absorbance of 570nm, detects the increment quantity of hHFDC, with this to analyze the spy of pleiotrophic factor solution
Property.
Hair regeneration hair gel cream performance evaluation method for measuring: 20 hair loss patients are randomly selected, are divided into 2 groups, control group
(hair gel cream is not added with pleiotrophic factor group) and test group (hair gel cream is added to pleiotrophic factor group), every group of 10 people are sooner or later each to smear
At hair gel cream to alopecia, average one square centimeter of alopecia area smears the hair gel cream applying amount of soya bean size, the 30th
It when carry out follow-up and acquiring head photo.
To pleiotrophic factor solution property analysis measurement result such as Fig. 1, as shown in Figure 1, activation hair regeneration multiple-effect because
Sub- liquid has been obviously promoted the increment of hHFDC, and competence for added value is related with pleiotrophic factor solution concentration, it can be clearly seen that, with
The activity of the increase of the concentration of pleiotrophic factor solution, cell increases；And in comparative example, since the pleiotrophic factor of its preparation is molten
The preparation of liquid is not added sinapinic acid solution and is pre-processed, therefore obtained pleiotrophic factor solution cannot promote the increasing of hHFDC
Value.
To result such as Fig. 2 of the performance evaluation of hair regeneration hair gel cream measurement, firstly, the hair loss patient of 10 bit test groups,
There is no adverse events, meanwhile, wherein 8 micro- patients reflect that depilation phenomenon has improvement, can see by acquiring photo comparison
As shown in Fig. 2, the patient that Fig. 2 (a) is control group does not smear the hair situation before hair gel cream, Fig. 2 (b) is the photo of acquisition
The patient of control group has smeared the hair situation after the hair gel cream without addition pleiotrophic factor solution, and Fig. 2 (c) is test group
Patient does not have the hair situation before hair gel cream, and Fig. 2 (d) is that the patient of test group has smeared the Gel for being added to pleiotrophic factor solution
Hair situation after curry cream.It can thus be seen that using be added to pleiotrophic factor solution hair gel cream and be not added with multiple-effect because
The hair gel cream group of son is compared, and the hair loss patient hair for being added to the test group of the hair gel cream of pleiotrophic factor solution obviously becomes close, is taken off
Hair is improved, and the hair loss state of the hair loss patient for the control group of hair gel cream for being not added with pleiotrophic factor solution is used not obtain
To improvement, it can be seen that, the hair gel cream of what the application was prepared be added to pleiotrophic factor solution has significantly to alopecia is improved
Effect.
The hair regeneration hair gel cream each component parts by weight of 1 Examples 1 to 3 of table match
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (9)
1. a kind of hair regeneration hair gel cream, which is characterized in that with the weight of the hair regeneration hair gel cream for 100%, including such as
The component of lower parts by weight:
Wherein, the pleiotrophic factor solution is modified pleiotrophic factor solution.
2. hair regeneration hair gel cream as described in claim 1, which is characterized in that the modified pleiotrophic factor solution is by mustard seed
The supernatant that low-kappa number umbilical cord mesenchymal stem cells cell monolayer is prepared.
3. hair regeneration hair gel cream as described in claim 1 or 2 is any, which is characterized in that the modified pleiotrophic factor solution
In, the concentration of modified pleiotrophic factor is 20-100mg/mL.
4. hair regeneration hair gel cream as claimed in claim 2, which is characterized in that described in the modified pleiotrophic factor solution
The concentration of sinapic acid is 80-120 μM.
5. a kind of preparation method of hair regeneration hair gel cream comprising following steps:
Each component is weighed respectively according to any hair regeneration hair gel cream of the claims 1~4；
The acrylate analog copolymer, the glycine betaine, the hyaluronic acid are added to the water mixing, are heated to 80-90 DEG C,
Obtain first chamber；
The sodium hydroxide is added in first chamber, is cooled to 35-45 DEG C, obtains second chamber；
The modified pleiotrophic factor solution is added in the second chamber, is uniformly mixed so as to obtain hair regeneration hair gel cream.
6. the preparation method of hair regeneration hair gel cream as claimed in claim 5, which is characterized in that the modified pleiotrophic factor is molten
Liquid the preparation method is as follows:
Umbilical cord mesenchymal stem cells are provided to be cultivated to obtain the first cultivating system；
Sinapic acid is added in first cultivating system to be pre-processed, the second cultivating system is obtained；
Second cultivating system is digested, neutralized, is washed, is centrifuged, collection is concentrated is obtained the modified pleiotrophic factor of supernatant
Solution.
7. the preparation method of hair regeneration hair gel cream as claimed in claim 5, which is characterized in that first cultivating system
In, the cell monolayer content cultivated is 40%~50%.
8. the preparation method of hair regeneration hair gel cream as claimed in claim 5, which is characterized in that second cultivating system
In, the cell monolayer content cultivated is 75%~80%.
9. the preparation method of hair regeneration hair gel cream as claimed in claim 5, which is characterized in that in the concentration step, adopt
With the method for ultrafiltration concentration.
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Application publication date: 20190913 |