CN109568191A - A kind of hair growth accelerating agent and preparation method thereof - Google Patents
A kind of hair growth accelerating agent and preparation method thereof Download PDFInfo
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Abstract
The embodiment of the present invention provides a kind of hair growth accelerating agent and preparation method thereof, is related to cosmetic technical field, can be not only used for the treatment of clinically alopecia seborrheica, scalds dye damage to alopecia areata, scalp and all has good therapeutic effect.The hair growth accelerating agent includes: that nourishing shines Essence and freeze-dried powder is shone in nourishing, the nourishing is shone Essence and is formulated by functional component with mass fraction, it is that fat mesenchymal stem cell (adipose tissue-derived stromal cells, ADSCs) conditioned medium makes that freeze-dried powder is shone in the nourishing.
Description
Technical field
The present invention relates to cosmetic technical fields more particularly to a kind of hair growth accelerating agent and preparation method thereof.
Background technique
As social competition's increase and environment are worsening, the trend risen increasingly is presented in the generation of alopecia.The whole world is big
Puzzlement there are about 50% adult man by alopecia." Chinese alopecia census of population " statistics indicate that, China's alopecia crowd about 3
Hundred million, wherein male pattern baldness number about 2.3 hundred million；The young and the middle aged is many after 80s even after 90s at " main force " in alopecia crowd
Student be added to alopecia ranks, alopecia more rejuvenation.Newest report display, pressure have become one line city 25-45 of China
Year male pattern baldness arch-criminal.Allegro work, the life of high pressure, overweight study can all aggravate alopecia status.
In so huge alopecia group, alopecia seborrheica crowd accounts for major part.Male sex hormone is to lead to seborrheica take off
The main reason for hair.Internal testosterone is changed into protona under the catalytic action of 5α-reductase, on protona and hair follicle
Androgen receptor combines active highest, and the androgen receptor after activation causes the activation of downstream signaling pathway to eventually lead to alopecia.
It is mainly both at home and abroad Western medicine for the drug of hair growth, such as minoxidil, the expansible head after scalp absorbs
Veins beneath the skin improves the microcirculation of perifollicolar, reduces perifollicolar inflammation cellular infiltration, so that the hair follicle of atrophy be made to increase simultaneously
The growth for promoting hair, makes fine hair on birds or animals hair become terminal hair；In addition, Finasteride be it is a kind of can inhibit 5α-reductase activity to prevent
The drug of male sex hormone synthesis.But minoxidil and Finasteride have a better effect primarily directed to androgenetic alopecia, have
Certain limitation, and long-term administration is wanted, side effect can be generated to human body.It is the product of Chinese medicine that, which also there are many main components in the country,
But since its ingredient is single, do not go to solve the problems, such as from the angle that hair follicle physiological is grown, so effect is not significant.It is close several
Year the hair transplant technology that comes into vogue and clinical treatment alopecia a kind of directly effective mode, but for alopecia area compared with
Greatly, the patient of oligotrichosis, can not meet demand.
Go deep into to hair follicle microstructure and hair regeneration mechanism study, it has been found that hair regeneration by hair follicle from
The comprehensive regulation of body stem cell and perifollicolar stem cell microenvironment.Stem cell is that one kind has self-replacation, self-renewing
Initial cell has the ability to Various Tissues cell differentiation, in vivo can be by secreting a variety of regulatory factors to adjusting group
Knit the normal physiological metabolism of organ.Therefore, the reconstituted product that exploitation is directed to the stem cell microenvironment reparation of hair follicle position, which becomes, to be controlled
Treat the development trend of alopecia agent.
Chinese patent CN200510048870.7 is disclosed using made of low passage human dermal papilla cells conditioned medium
The ability that there is freeze-dried powder induction hair follicle to be formed has apparent curative effect in treatment alopecia areata.But in the case of cultivating in vitro, the
6-9 no longer shows aggregative behavior for later cell, and the ability that induction hair follicle is formed also gradually weakens and disappears, so even if at
Function separates hair papilla cell, can not also maintain the functional status of hair papilla cell for a long time under cultivation conditions.And due to people's
Hair follicle sample is not easy to obtain, and hair papilla cell position is hidden, and structures surrounding is complicated, a kind of high there is presently no being formed
Effect, the succinct method for being separately cultured hair papilla cell, are unfavorable for scale cell culture.
Chinese patent CN201510792991 is disclosed using made of adipose-derived stem cells culture solution and ancillary drug
Treatment of the hair restorer for alopecia easily loses activity since stem cell medium is not easy to store, if in production hair restorer
Make its loss of activity in the process, just fall flat, and directly use culture solution, is unfavorable for clinical expansion.
In conclusion domestic Chinese patent drug and import Western medicine do not go to solve from hair follicle stem cells and hair follicle microenvironment angle
Alopecia problem, without fundamentally improving alopecia problem and promoting hair restoration.Though the operation of surgery hair transplant can quickly improve baldness
Bad vision, reach certain cosmetic result, but it is expensive to perform the operation, and needs repeatedly transplanting can be only achieved mostly ideal
Effect.The hair-growing effects of hair papilla cell culture solution are significant, but hair papilla cell materials are not easy, are separately cultured difficulty.Directly
It is easy to lose activity using stem cell medium, therapeutic effect is not achieved.So people still research and develop a kind of cost in craving
The good hair growth accelerating product of low, easy to use and therapeutic effect.
Summary of the invention
The embodiment of the invention provides a kind of hair growth accelerating agent and preparation method thereof, can be not only used for clinically seborrheica
The treatment of alopecia scalds dye damage to alopecia areata, scalp and all has good therapeutic effect.
Technical problem solved by the invention is realized using following technical scheme: a kind of nourish shines Essence combination taste
The preparation method and application for shining the hair growth accelerating agent of freeze-dried powder is supported, selects fatty group from a wealth of sources and being easily isolated stem cell
It knits, collects cell culture fluid after ex vivo stem cell separation, identification, expansion culture and conditional stimulus, through culture solution concentration, freeze
Dry, quality inspection obtains nourishing and shines freeze-dried powder；Cacumen Platycladi extract, kuh-seng root extract, Angelica roots, Roots of Panax Notoginseng are mentioned
It takes object, resveratrol to use glycerol and water as solvent, obtains nourishing after dissolution and shine Essence；Essence dissolution is shone with nourishing
Freeze-dried powder is shone in nourishing, and as hair growth accelerating agent can be used for clinically men and women's alopecia seborrheica, alopecia areata, scalp and scald dye damage etc.
The treatment of scalp problems.
Present invention application nanometer micropin and electroporation promote to seep instrument as physical enhancement method, instruct making for hair growth accelerating agent
With.
The preparation method and application for the hair growth accelerating agent that Essence combines nourishing to shine freeze-dried powder, step are shone in a kind of nourishing
It is rapid as follows:
Step 1: the preparation for shining Essence is nourished
Take 3-8 parts of Cacumen Platycladi extract, 2-7 parts of kuh-seng root extract, 1-6 parts of Angelica roots, Radix Notoginseng root extract 1-6
Part, it 1-6 parts of resveratrol, uses glycerol and water as solvent, dissolves above-mentioned various Effect raw materials, make to nourish to shine in Essence to plant
The final concentration of 5-10mg/ml of object functional component mixture.
In the present invention, the Thuja extract can activate hair follicle, moreover it is possible to which activation of melanocyte enhances the work of tyrosinase
Property, have the function of dark hair.Angelica extract has promoting blood circulation effect.Shrubby sophora extract plays the role of antagonism androgen.Radix Notoginseng mentions
Object is taken to have trichogenous effect.Scientific research in recent years shows that resveratrol plays the role of promoting hair regeneration.
Step 2: the preparation for shining freeze-dried powder is nourished
(1) collection of ADSCs conditioned medium: to ADSCs carry out conditional stimulus, after continue to cultivate, collect culture arrive third day
ADSCs conditioned medium, be stored in 4 DEG C, conditioned medium dispensed in time in next day and is stored in -20 DEG C.Validity period is 6
A month.
(2) the removal of impurities concentration of conditioned medium: the conditioned medium being collected into is carried out using ultrafiltration concentration technology first
Removal of impurities, primary concentration and secondary concentration, finally obtain secondary concentration liquid, as bioactie agent freeze-dried powder stoste.
(3) filtration sterilization of bioactie agent freeze-dried powder stoste: using miillpore filter to bioactie agent freeze-dried powder
Stoste is filtered degerming.
(4) total protein concentration measures: being surveyed using BCA method to bioactie agent freeze-dried powder stoste total protein concentration
It is fixed.Protein concentration should be 0.5-2.0 mg/mL.
(5) pH value measures: being measured using pH meter to bioactie agent freeze-dried powder stoste pH value.Freeze-dried powder stoste pH
It should be 6.5 ~ 8.0.
(6) add freeze drying protectant: using the total protein concentration finally measured as standard, then plus corresponding proportion mannitol,
The mixture of trehalose and Dextran 40 is as freeze drying protectant.
(7) freeze-drying of bioactie agent freeze-dried powder stoste: filtered liquid is dispensed, and is freezed after packing dry
It is dry, it is spare to obtain freeze-dried powder storage at normal temperature.
In the present invention, the conditional stimulus can be vitamin C, cell factor, hypoxemia, in tetra- kinds of methods of low dosage UVB
It is one or more.Conditional stimulus can make cell that stress reaction occur, and secrete more growth factors.
In the present invention, the ADSCs conditioned medium contains cytokine profiles, such as VEGF, PDGF, SCF, TGF,
HGF, IGF etc. can adjust the microenvironment of hair follicle growth, activate hair papilla cell, and hair follicle is made to enter growth period, promote hair
Regeneration.
In the present invention, the filtering with microporous membrane aperture is 0.22-0.45 μm, it is therefore an objective to which concentrate filtration sterilization retains
The small cytokines albumen of 14KB-44KB, removes most unrelated protein in freeze-dried powder stoste.
In the present invention, the freeze drying protectant, it is therefore an objective to the protein in freeze-dried powder stoste be protected to exempt from freeze-drying process
By low temperature damage.The ratio that added mannitol accounts for freeze-dried powder stoste is 10%-15%；Trehalose is 0.6%-1%；Dextran 40
It is 0.4%-1%.
In the present invention, (6), (7) step are intended to aseptically carry out.
Step 3: the use of hair restorer
(1) preparation of hair growth accelerating agent: being nourished to shine Essence dissolution and nourish with one and shine freeze-dried powder, sufficiently shaken up, spare.
(2) cleaning disinfection: hair loss patient scalp is cleaned with physiological saline first, then is carried out disinfection with Iodophor.
(3) micropin imports: hair growth accelerating agent uniformly being dripped in patient's scalp alopecia position, carries out importing behaviour using micropin instrument
Make.Each patient needs 30-40min with micropin importing process.
(4) massage promotees to seep: the remaining importing liquid of scalp reuses rush infiltration instrument and imports in scalp after the completion of micropin import operation
Position is massaged, and to promote to import the absorption of liquid, massages 10-15min.After the completion of massage, on hospital gauze wiping hair
Remaining importing liquid promotees to seep massage operation completion after the importing liquid of scalp sites fully absorbs.
In the present invention, the physiological saline be medical 0.9% NaCl injection；The Iodophor is that medical grade sterilizes Iodophor.
In the present invention, the micropin instrument is 36 needle nanometer micropins.Microneedle cutaneous mechanism is to utilize microneedle skin
The cuticula of epidermis enters skin certain depth, forms the minim channel vertical with skin, and drug can effectively be promoted by the channel
Into transdermal penetration and in the accumulation of privileged site.Micron-sized hole, hole are formed in skin surface after micropin insertion and removal
Gap can pass through in order to pharmaceutical preparation, thus the local action in skin or the capillary absorbance by skin.
In the present invention, it is electroporation introducing apparatus that the rush, which seeps instrument, the keratoderma shape under the effect of high voltage pulse electric current
At of short duration reversible aqueous duct, increase the permeability of drug.
In the present invention, micropin technology and physical enhancement technical tie-up are entered into skin table using that can further speed up drug
The speed of chrotoplast increases the percutaneous absorbtion effect of drug, and skin does not have apparent excitement and feeling of pain.
Beneficial effects of the present invention:
(1) ingredient: plant extracts contained by Essence is shone in nourishing, itself has effects that educate hair, shines jelly in conjunction with nourishing
Cytokine component abundant in dry powder can not only make scalp microenvironment restore normal, moreover it is possible to activate hair follicle stem cells, reach
Hair tonic purpose.
(2) technique: freeze-dried powder is shone using the nourishing being concentrated by ultrafiltration and prepared by freeze drying process, can be cell factor
Activity is maximumlly saved, and this dosage form facilitates the storage and popularization and use of product.
(3) application method: hair growth accelerating agent combination nanometer micropin and electroporation of the invention promotees to seep the use of instrument, can be maximum
The absorption of the guarantee hair restorer of limit, and not damaged scalp.
(4) act on: the hair growth accelerating agent in the present invention can be not only used for the treatment of clinically alopecia seborrheica, to spot
Bald, scalp scalds dye damage and all has good therapeutic effect.
Detailed description of the invention
Attached drawing 1 is to nourish to shine Essence outside drawing in the agent of this hair growth accelerating；
Attached drawing 2 is to nourish to shine freeze-dried powder outside drawing in the agent of this hair growth accelerating；
Attached drawing 3 is that two male baldness patients use schematic diagram before hair growth accelerating agent of the present invention；
Attached drawing 4 is that two male baldness patients use schematic diagram after hair growth accelerating agent 3 months of the present invention.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in the embodiment of the present invention
Technical solution is clearly and completely described, it is clear that described embodiments are some of the embodiments of the present invention, rather than complete
The embodiment in portion.Based on the embodiments of the present invention, those of ordinary skill in the art are without making creative work
Every other embodiment obtained, shall fall within the protection scope of the present invention.
The embodiment of the present invention provides a kind of preparation method of hair growth accelerating agent, specifically includes:
Step 1: the preparation for shining Essence is nourished
Take 8 parts of Cacumen Platycladi extract, 5 parts of kuh-seng root extract, 3 parts of Angelica roots, 3 parts of Radix Notoginseng root extract, white black false hellebore
It 2 parts of alcohol, uses glycerol and water as solvent, dissolves above-mentioned various Effect raw materials, make to nourish and shine plant functional component in Essence
The final concentration of 5-10mg/ml of mixture.
Such as Fig. 1, Essence outside drawing is shone to nourish in the agent of this hair growth accelerating, is weak yellow liquid.
Step 2: the preparation for shining freeze-dried powder is nourished
(1) collection of ADSCs conditioned medium: to ADSCs carry out conditional stimulus, after continue to cultivate, collect culture arrive third day
ADSCs conditioned medium, be stored in 4 DEG C, conditioned medium dispensed in time in next day and is stored in -20 DEG C.Validity period is 6
A month.
(2) the removal of impurities concentration of conditioned medium: the conditioned medium being collected into is carried out using ultrafiltration concentration technology first
Removal of impurities, primary concentration and secondary concentration, finally obtain secondary concentration liquid, as bioactie agent freeze-dried powder stoste.
(3) filtration sterilization of bioactie agent freeze-dried powder stoste: using miillpore filter to bioactie agent freeze-dried powder
Stoste is filtered degerming.
(4) total protein concentration measures: being surveyed using BCA method to bioactie agent freeze-dried powder stoste total protein concentration
It is fixed.Protein concentration should be 0.85-2.0 mg/mL.
(5) pH value measures: being measured using pH meter to bioactie agent freeze-dried powder stoste pH value.Freeze-dried powder stoste pH
It should be 6.5 ~ 8.0.
(6) add freeze drying protectant: using the total protein concentration finally measured as standard, then plus corresponding proportion mannitol,
The mixture of trehalose and Dextran 40 is as freeze drying protectant.
(7) freeze-drying of bioactie agent freeze-dried powder stoste: filtered liquid is dispensed, and is freezed after packing dry
It is dry, obtain nourishing that shine freeze-dried powder storage at normal temperature spare.
Such as Fig. 2, freeze-dried powder outside drawing is shone to nourish in the agent of this hair growth accelerating, is white powder.
Step 3: the use of hair growth accelerating agent
(1) preparation of hair growth accelerating agent: being nourished to shine Essence dissolution and nourish with one and shine freeze-dried powder, sufficiently shaken up, spare.
(2) cleaning disinfection: hair loss patient scalp is cleaned with physiological saline first, then is carried out disinfection with Iodophor.
(3) micropin imports: hair growth accelerating agent uniformly being dripped in patient's scalp alopecia position, carries out importing behaviour using micropin instrument
Make.Each patient needs 30min with micropin importing process.
(4) massage promotees to seep: the remaining importing liquid of scalp reuses rush infiltration instrument and imports in scalp after the completion of micropin import operation
Position is massaged, and to promote to import the absorption of liquid, massages 15min.After the completion of massage, remained on hospital gauze wiping hair
Importing liquid, after the importing liquid of scalp sites fully absorbs, promote seep massage operation complete.
Step 4: clinical application
The present invention picks out the alopecia seborrheica male work that 2 ages are respectively 24,32 years old after identifying by biological safety
Patient is used for hair restorer, in the case where patient's informed consent, according to the application method of hair restorer, at patient's alopecia.
Regular observation of taking pictures, observed hair-growing effects after 3 months.
Hair loss patient one, male, a large amount of alopecias suddenly before 24 years old, 3 years, multiple herbal treatment are invalid.
Hair loss patient two, male, 32 years old, heavy because working, psychological pressure was big, suffers from alopecia more than 2 years.
Every hair loss patient does in the set time weekly and nourishes the use for shining nourishing liquid.
It is that two male baldness patients use schematic diagram before hair growth accelerating agent of the present invention such as Fig. 3；Fig. 4 is that two males are bald
It sends out patient and uses schematic diagram after hair growth accelerating agent 3 months of the present invention.
In the alopecia seborrheica patient of 2 hair growth accelerating agent treatment, 2 people have significant curative effect.It is combining micropin and is promoting to seep
After hair restorer hair growth patient 2 weeks of the present invention of instrument application, former alopecia position scalp color can be clearly visible and turn black, scalp
Fat secretion is also reduced, and is illustrated that the hair follicle in patient's alopecia area gradually activates, is initially entered growth period.It uses hair restorer one month
Afterwards, alopecia area villous is grown, and color is partially yellow, uniform fold.The freeze-dried powder combination plant extracts of ADSCs conditioned medium exists
Pure Chinese medicine compound or Western medicine preparation are substantially better than on treatment alopecia seborrheica.We to use promote seep instrument and be not used promote seep
The patient of instrument compares, and discovery, which promotees infiltration instrument, can quickly promote absorption of patient's scalp to hair restorer, and mechanism is obvious.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (5)
1. a kind of hair growth accelerating agent, which is characterized in that the hair growth accelerating agent includes: that nourishing shines Essence and jelly is shone in nourishing
Dry powder, the nourishing shine what Essence was formulated by functional component with mass fraction, and it is rouge that freeze-dried powder is shone in the nourishing
The production of fat mescenchymal stem cell (adipose tissue-derived stromal cells, ADSCs) conditioned medium.
2. a kind of preparation method of hair growth accelerating agent, which is characterized in that the method is used to prepare as described in claim 1 stop
Hair shed and generating hair agent, the preparation method include following operating procedure:
Step 1: the preparation for shining Essence is nourished
Take 3-8 parts of Cacumen Platycladi extract, 2-7 parts of kuh-seng root extract, 1-6 parts of Angelica roots, Radix Notoginseng root extract 1-6
Part, it 1-6 parts of resveratrol, uses glycerol and water as solvent, dissolves above-mentioned various Effect raw materials, make to nourish to shine in Essence to plant
The final concentration of 5-10mg/ml of object functional component mixture；
Step 2: the preparation for shining freeze-dried powder is nourished
(1) collection of ADSCs conditioned medium: to ADSCs carry out conditional stimulus, after continue to cultivate, collect culture arrive third day
ADSCs conditioned medium, be stored in 4 DEG C, conditioned medium dispensed in time in next day and is stored in -20 DEG C,
Validity period is 6 months；
(2) the removal of impurities concentration of conditioned medium: cleaned first using ultrafiltration concentration technology to the conditioned medium being collected into,
Primary concentration and secondary concentration, finally obtain secondary concentration liquid, as bioactie agent freeze-dried powder stoste；
(3) filtration sterilization of bioactie agent freeze-dried powder stoste: using miillpore filter to bioactie agent freeze-dried powder stoste
It is filtered degerming；
(4) total protein concentration measures: being measured using BCA method to bioactie agent freeze-dried powder stoste total protein concentration, egg
White concentration should be 0.5-2.0 mg/mL；
(5) pH value measures: being measured using pH meter to bioactie agent freeze-dried powder stoste pH value, freeze-dried powder stoste pH should be
6.5~8.0；
(6) add freeze drying protectant: using the total protein concentration finally measured as standard, then plus corresponding proportion mannitol, seaweed
The mixture of sugar and Dextran 40 is as freeze drying protectant；
(7) freeze-drying of bioactie agent freeze-dried powder stoste: filtered liquid is dispensed, and is freeze-dried after packing,
It is spare to obtain freeze-dried powder storage at normal temperature.
3. the preparation method of hair growth accelerating agent according to claim 2, which is characterized in that the method also includes:
(1) preparation of hair growth accelerating agent: being nourished to shine Essence dissolution and nourish with one and shine freeze-dried powder, sufficiently shaken up, spare；
(2) cleaning disinfection: hair loss patient scalp is cleaned with physiological saline first, then is carried out disinfection with Iodophor；
(3) micropin imports: hair growth accelerating agent is uniformly dripped in patient's scalp alopecia position, carries out import operation using micropin instrument,
Each patient needs 30-40min with micropin importing process；
(4) massage promotees to seep: the remaining importing liquid of scalp reuses rush infiltration instrument in scalp imported parts after the completion of micropin import operation
It is massaged, to promote to import the absorption of liquid, massages 10-15min；
After the completion of massage, remaining importing liquid on hair is wiped with hospital gauze, after the importing liquid of scalp sites fully absorbs,
Promote to seep massage operation completion.
4. the preparation method of hair growth accelerating agent according to claim 2 or 3, which is characterized in that the condition of the ADSCs is pierced
Swash, one of tetra- kinds of vitamin C, cell factor, hypoxemia, low dosage UVB methods can be used or a variety of is handled.
5. the preparation method of hair growth accelerating agent according to claim 2 or 3, which is characterized in that micropin, which imports, uses 36 needles
Nanometer micropin promotees to seep and seeps instrument using electroporation rush.
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