• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    The growth factor concentrate is intended for the cosmetic treatment of a subject in need thereof; it comprises decapsulated growth factors derived from plasma rich in blood platelets and can be preserved over a prolonged period thanks to the method of the invention. The invention also relates to an anti-aging cosmetic composition comprising said growth factor concentrate, as well as a skin rejuvenation method using such a composition.
    公开号:CH708629B1
    申请号:CH01399/14
    申请日:2014-09-17
    公开日:2016-12-30
    发明作者:Yip Chan Kin；Man Tam Wing
    申请人:Bestop Group Holdings Ltd；
    IPC主号:
    专利说明:

    Field of the invention
    The present invention relates to a growth factor concentrate and the use thereof, particularly the use in a cosmetic treatment.
    Context of the invention
    [0002] Growth factors are a group of naturally occurring proteins or steroid hormones that are capable of stimulating cell growth, proliferation, and cell differentiation. Growth factors are important for regulating different cellular processes and play an important role in maintaining a healthy skin structure and function.
    [0003] Certain types of cells, such as the keratinocytes constituting the epidermis and the dermis of the skin, can secrete growth factors. However, the concentration of growth factors can be decreased in a way related to age. Since the skin has to be fertilized by growth factors, theoretically some cosmetic products or current cosmetic treatments apply growth factors to the skin surface, in which the growth factors are extracted from the subject requiring them. The cosmetic effects that are expected by the application of growth factors to the skin surface include, but are not limited to: the reduction of fine lines, expression lines and wrinkles following the synthesis of new collagen; reduction of dark spots and pigmentation; improvement of density, smoothness and firmness; and reducing an uneven skin texture and tone.
    [0004] Platelet rich plasma (PRP) extracted from the subject requiring this is used for treatment with a growth factor. However, the PRP must be used freshly, and can not be stored for a long time. The average shelf life of platelets is typically only 5 to 9 days after extraction. In skin care products, growth factors would be used repeatedly, and possibly for long periods of time. A normal cosmetic treatment of growth factors requires several treatments covering a period of 3 to 6 months. Typically, each time before receiving the treatment, the subject requiring it must undergo a venipuncture and wait at least 30 minutes before extracting the PRP. Some products use additives such as a preservative or other chemical to extend the shelf life of growth factors.
    In addition, the absorption of growth factors applied to the surface of the skin may not be as predictable because of the blockage of the epidermis. In addition, the PRP treatment known in the art increases the number of platelets surrounding the skin cells, but only a small amount of growth factors can be released from the platelets at a slow rate.
    It is necessary to provide growth factors without additive and stable self-administered by the subject in need, having a high released concentration and a longer shelf life.
    Summary of the invention
    Given the above context, it is an object of the present invention to describe a growth factor concentrate for cosmetic treatment of a subject in need thereof, the growth factor concentrate comprising derived decapsulated growth factors. platelet rich plasma.
    In an exemplary embodiment of the present invention, the platelet rich plasma is extracted from the subject.
    In another exemplary embodiment of the present invention, the growth factor concentrate is preserved by the steps of:<tb> (a) <SEP> extracting a sample of whole blood from the subject, the platelet-rich plasma with the growth factors being contained therein;<tb> b) <SEP> obtaining the platelet-rich plasma from the blood sample;<tb> c) <SEP> stabilization of platelet rich plasma;<tb> d) <SEP> decapsulation of growth factors of stabilized platelet rich plasma;<tb> e) SEP filtration of decapsulated growth factors;<tb> f) <SEP> Lyophilization of the filtered growth factors to obtain the growth factor concentrate; and<tb> g) <SEP> preservation of freeze-dried growth factor concentrate.
    The growth factor concentrate is stored for an extended period of time.
    In another exemplary embodiment of the present invention, the decapsulation step further comprises the step of freezing and thawing the platelet-rich plasma. In another exemplary embodiment, the platelet rich plasma is frozen at a temperature between -50 ° C and -60 ° C for at least 30 minutes; in another further exemplary embodiment, the platelet-rich plasma is thawed at a temperature of 37 ° C for 10 minutes. In an exemplary embodiment, the decapsulated growth factors are filtered through a cellulose acetate membrane in the filtration step. In another additional exemplary embodiment, the lyophilization step further comprises the steps of: f1. freezing growth factors filtered at a temperature between -50 ° C and -60 ° C for at least 3 hours; and f2. lyophilization of the frozen growth factors at a temperature between -40 ° C and -60 ° C and at a pressure of about 0.18 millibar for at least 10 hours. In another exemplary embodiment, the lyophilized growth factor concentrate is stored at a temperature in the range of 2 ° C to 10 ° C in the storage step.
    In another exemplary embodiment of the present invention, the growth factor concentrate is a cryo-crystallized growth factor concentrate.
    In another further exemplary embodiment of the present invention, the growth factor concentrate has a shelf life of at least 22 months.
    According to another aspect of the present invention, an anti-aging cosmetic composition for a subject in need thereof is disclosed, the anti-aging cosmetic composition comprising an effective amount of the growth factor concentrate as described above.
    In another aspect of the present invention, a non-invasive method for stimulating skin rejuvenation in a subject in need thereof is described, the method comprising topically applying a cosmetic composition comprising an effective amount of the growth factor as described above.
    In a further aspect of the present invention, a method for rejuvenating the skin of a subject in need thereof is described, the method comprising the steps of:<tb> a) <SEP> cleaning the target area of the subject's skin;<tb> b) <SEP> applying a probe from a radiofrequency generating device to the cleaned skin to create channels in the cleansed skin; and<tb> c) <SEP> applying a cosmetic composition comprising an effective amount of the growth factor concentrate to the cleansed skin of step b so that the cosmetic composition can penetrate the deeper layers of the cleansed skin through the channels.
    Brief description of the figures
    [0017]<tb> Fig. 1 <SEP> represents the standard deviation of contamination of extracted growth factor concentrate and fresh platelets in a safety study of the growth factor concentrate described in the present invention.<tb> Fig. "SEP" represents the density of growth factors released under the epidermis of the subject requiring the growth factor concentrate applied by the process described in the present invention and the fresh growth factor applied by the injection.<tb> Fig. <SEP> represents the total number of cells after treatment of the growth factor concentrate described in the present invention and a typical treatment using PRP.<tb> Fig. <SEP> represents the results of studies of the application of the growth factor concentrate described in the present invention.
    Detailed Description of the Preferred Embodiments
    In the present context and in the claims, "comprising" means comprising the following elements but without excluding others.
    Preparation of the growth factor concentrate
    1. Collection of PRP
    A sample of 30 ml to 60 ml of whole blood is extracted from the subject by venipuncture or any other suitable means and stored in a sodium citrate blood collection tube or other suitable container to facilitate transportation.
    The PRP layer is separated from the blood cell layer by centrifugation at 3000 r / min or any other suitable means. Growth factors are now concentrated in the separate PRP solution.
    2. Stabilization of the PRP layer
    A buffering agent is added to the separate PRP layer to stabilize the pH value of the PRP in a desired range to avoid unwanted fluctuations in pH value at the extreme temperatures. Such undesirable fluctuations in pH value may affect the efficiency of growth factors in subsequent steps including a temperature change.
    The buffering agent that can be added to the PRP solution as a lyoprotectant is Tris buffer.
    3. Decapsulation of growth factors
    Platelets are prone to apoptosis at a low storage temperature which can release inhibitors and destroy the growth factors present therein. On the other hand, as mentioned above, the rate of uptake of growth factors by the skin cell may be low, since only a small amount of growth factors are released from the platelets during processing. In the context of the invention, the inventors have planned to decapsulate growth factors from platelets since the decapsulated growth factors tend to be less vulnerable to inhibitors and easier to absorb by the cutaneous cell.
    There are several methods that can release (decapsulate) growth factors from platelets, including, but not limited to, freeze-thaw, mechanical lysis, liquid homogenization, sonication and manual grinding. . A freeze-thaw process is used in one embodiment to decapsulate growth factors from the stabilized PRP solution.
    The stabilized PRP solution was frozen at -50 ° C to -60 ° C for 30 minutes, and then thawed in a 37 ° C water bath for 10 minutes. The above freeze-thaw cycle is repeated 3 more times. By means of freezing and thawing, the formation of ice crystals mentioned above in the platelets increases in volume and finally breaks the membrane of the platelets; therefore, growth factors are released from the platelets and a decapsulated growth factor solution is then obtained.
    By decapsulating growth factors from platelets, the retention period of growth factors can be greatly prolonged and the absorption rate can also be increased.
    4. Filtration of the decapsulated growth factor
    In order to reduce the amount of pathogens and other contaminants such as bacteria, erythrocytes or cell fragments that can affect the sterility of the solution and the lifetime of the growth factors, the factor solution of Decapsulated growth is filtered on a 0.2 μm cellulose acetate membrane. As described in FIG. 1, the filtered growth factor solution clearly has a lower standard deviation of contamination under the same conditions as that of fresh platelets, which shows that the filtered growth factor solution is safer than fresh platelets.
    The filtered growth factor solution is then dispensed into vials of serum which have been sealed with rubber stoppers and frozen. In one embodiment, these vials are frozen at -50 ° C to -60 ° C for at least 3 hours to prepare for freeze drying.
    5. Freeze
    Lyophilization, also called freeze drying, works by freezing the material and then reducing the surrounding pressure to allow the frozen water in the material is directly sublimated from the solid phase to the gas phase. This process is used in the preservation of perishable material or to make the material more suitable for transport.
    The vials containing the filtered solution of growth factor are removed from storage; in one embodiment of the present invention, the storage temperature is -55 ° C. The freely covered vials are then placed in a freeze-dryer. In one embodiment, the lyophilization process is maintained at a temperature between -40 ° C and -60 ° C and at a pressure of about 0.18 millibar for at least ten hours until growth factors are carefully dried. The growth factor concentrate is obtained in powder form (also called cryo-crystallized growth factor, C-GF).
    6. Storage
    After visual inspection of the flasks to ensure sufficient dryness of the powder, the flasks are closed hermetically in the air and removed from the lyophilizer at a temperature, in one embodiment, from 2 ° C to 10 ° C. ° C with humidity below 70%, vials can be stored for up to six months. In another embodiment of the present invention, the growth factor concentrate prepared by the above steps and stored in a typical household refrigerator can then have a shelf life of up to 22 months.
    Administration of the final product with a growth factor concentrate
    The growth factor concentrate prepared and preserved from the above steps can be used in the cosmetic treatment of the subject requiring this. In one embodiment, the growth factor concentrate may be applied as a serum to rehydration treatment areas with normal 1: 1 solute or administered as a powder directly to open wet wounds or ulcers.
    The results of the administration of the growth factor concentrate of the present invention in the cosmetic treatment are shown in FIGS. 2, 3 and 4. Fig. 2 shows that there is a 10-fold significant proliferation of cell number in the application of the growth factor concentrate of the present invention directly to the surface of the skin as compared to that in the application of fresh growth factor by injection. This shows that by applying the growth factor concentrate of the present invention directly to the skin surface, the area of skin to be treated would have more growth factors released than by applying fresh growth factor by injection.
    FIG. 3 presents the results of a total cell count study after the conventional growth factor treatment and the growth factor concentrate of the present invention. It has been observed that cell numbers increase using both types of growth factors. However, after 20 days of treatment, the number of cells using the growth factor concentrate of the present invention is 1.5 times that obtained using a conventional growth factor.
    FIG. 4 shows the results of actual clinical examples before and after treatment using the growth factor concentrate of the present invention. It shows that the growth factor concentrate of the present invention could reduce nasolabial / ocular folds, enlarged pores and bags under the eyes; reduce dark spots and pigmentation; and improve density, smoothness and firmness.
    Application on the surface of the skin
    In order to improve the absorption rate of the growth factor concentrate during the treatment of the subject requiring it directly on the skin surface, the bipolar radiofrequency (RF) technology is used in one embodiment of the invention. present invention. Radiofrequency technology as a rejuvenation method is partially based on remodeling of the skin after a controlled lesion and could be used for laser skin resurfacing, dermabrasion and deep chemical exfoliation.
    An embodiment of the present invention relates to a method comprising the steps of:<tb> a) <SEP> cleaning the target area of the subject's skin;<tb> b) <SEP> cleansing skin cleaned with a probe of a radio frequency device; and<tb> c) <SEP> applying a cosmetic composition comprising an effective amount of the growth factor concentrate of the present invention to the massaged skin.
    In the same embodiment, an RF energy of 10 J / cm <3> with 1.7 MHz is adapted. By massaging the skin before applying the growth factor concentrate, the growth factor concentrate can then easily pass through the epidermis by permeation and the concentration thereof under the epidermis would be increased.
    [0039] Exemplary embodiments of the present invention are thus fully described. Although the description refers to particular embodiments, it will be apparent to those skilled in the art that the present invention may be practiced with variation of these specific details. Therefore, this invention should not be considered as limited to the embodiments described herein.
    For example, the present invention is applicable to various types of growth factors including and not limited to platelet derived growth factor (PDGF), transforming growth factor beta (TGF-B), insulin-like growth factor (IGF). -1), platelet factor 4 (PF-4), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), hepatocyte growth factor (HGF), bone morphogenetic protein (BMP), and growth factor fibroblasts (FGF).
    权利要求:
    Claims (13)
    [1]
    A growth factor concentrate for the cosmetic treatment of a subject said growth factor concentrate comprising decapsulated growth factors derived from platelet rich plasma.
    [2]
    2. Growth factor concentrate according to claim 1, as it can be obtained and stored for an extended period of time by the steps of:a) extracting a whole blood sample from a subject;b) obtaining said platelet-rich plasma from said blood sample;c) stabilizing said platelet rich plasma;d) decapping growth factors from the platelets of said stabilized platelet rich plasma;e) filtering said decapsulated growth factors;f) lyophilizing said filtered growth factors; andg) preserving said freeze-dried concentrate of growth factor.
    [3]
    The growth factor concentrate of claim 2 wherein said decapping step may further comprise the step of freezing and thawing said platelet rich plasma.
    [4]
    The growth factor concentrate according to claim 3 wherein, in said freezing step, said platelet-rich plasma can be frozen at a temperature between -50 ° C and -60 ° C for 30 minutes.
    [5]
    A growth factor concentrate according to claim 3 wherein, in said thawing step, said platelet-rich plasma can be thawed at a temperature of 37 ° C for 10 minutes.
    [6]
    The growth factor concentrate of claim 2 wherein, in said filtering step, said decapsulated growth factors can be filtered through a cellulose acetate membrane.
    [7]
    The growth factor concentrate of claim 2 wherein said lyophilization step may further comprise the steps of:f1) freezing said filtered growth factors at a temperature between -50 ° C and -60 ° C for at least 3 hours; and(f2) lyophilizing said frozen growth factors at a temperature between -40 ° C and -60 ° C and at a pressure of about 0.18 millibar for at least 10 hours.
    [8]
    The growth factor concentrate according to claim 2 wherein in said preservation step, said freeze-dried growth factors can be stored at a temperature in the range of 2 ° C to 10 ° C.
    [9]
    The growth factor concentrate according to claim 1, said growth factor concentrate being a cryo-crystallized growth factor concentrate.
    [10]
    The growth factor concentrate according to claim 1, said growth factor concentrate having a shelf life of at least 22 months.
    [11]
    An anti-aging cosmetic composition comprising an effective amount of said growth factor concentrate of any one of claims 1 to 10.
    [12]
    A non-invasive cosmetic method of stimulating skin rejuvenation in a subject comprising topically applying a cosmetic composition comprising an effective amount of said growth factor concentrate of any one of claims 1 to 10.
    [13]
    13. A cosmetic process for rejuvenating the skin of a subject, comprising the steps of:a) cleaning the target area of the subject's skin;b) massage of the cleaned skin with a probe of a radiofrequency apparatus; andc) applying a cosmetic composition comprising an effective amount of said growth factor concentrate of any one of claims 1 to 10 to the massaged skin.
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    同族专利:
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    法律状态:
    2015-08-14| NV| New agent|Representative=s name: ING. MARCO ZARDI C/O M. ZARDI AND CO. S.A., CH |
    优先权:
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    HK13110684.5A|HK1194912A2|2013-09-17|2013-09-17|Growth factor concentrate and the use thereof|
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