WO2021045712A1 - Use of plant exosomes for enhancing viability and hair growth capacity of skin cells - Google Patents
Use of plant exosomes for enhancing viability and hair growth capacity of skin cells Download PDFInfo
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- WO2021045712A1 WO2021045712A1 PCT/TR2020/050796 TR2020050796W WO2021045712A1 WO 2021045712 A1 WO2021045712 A1 WO 2021045712A1 TR 2020050796 W TR2020050796 W TR 2020050796W WO 2021045712 A1 WO2021045712 A1 WO 2021045712A1
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- 210000001808 exosome Anatomy 0.000 title claims abstract description 72
- 230000003779 hair growth Effects 0.000 title claims abstract description 27
- 210000004927 skin cell Anatomy 0.000 title claims abstract description 26
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8962—Allium, e.g. garden onion, leek, garlic or chives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
- A61K8/14—Liposomes; Vesicles
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
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- A—HUMAN NECESSITIES
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q7/00—Preparations for affecting hair growth
Definitions
- the present invention relates to use of plant exosomes as a product having the effect of enhancing viability and, by stimulating the hair follicles, enhancing hair growth in skin cells.
- Mammalian cells include small vesicular structures called exosomes. When these exosomes are isolated from healthy cells, they can be used to restore the cells, which are damaged or under a certain stress and which cannot complete their self regeneration, back to their earlier healthy state [1] Recently, use of the stem cells in cell based therapies is also one of the most promising studies. It is asserted that use of the exosomes released from these cells will have positive effects on wound healing and hair growth/strengthening in clinical application [2].
- the vesicles are small sacs which are involved in the transport and storage of substances within the cell and are separated by at least one lipid bilayer from the cytoplasm fluid.
- Exosomes are vesicles, which are released by many organisms from prokaryotes to high eukaryotes and plants, and which contain lipid bilayer vesicles of different sizes [3]. The importance of these vesicles lies behind the capacity of transferring information to the other cells in order to influence the cell function. Signal transfer via exosomes is carried out by means of biomolecules in many different categories consisting of proteins, lipids, nucleic acid and sugars [4].
- Intense hair loss is a problem which can be encountered both in men and women. There are many reasons of this problem depending on hormones, age, stress and mineral deficiency [6]. Although there is no study conducted on cells regarding the viability and hair growth stimulation effects of the plant exosomes related to skin and hair, the characterization of the plant exosomes and their effects on cancer cells have begun to be studied [7].
- the European patent application document no. EP3354257 discloses a composition which comprises plant extract-derived extracellular vesicles and is uses for improving skin and preventing hair loss.
- the said application discloses about the effects of the vesicles obtained by passing the filtered plant juice through different centrifuges.
- Exosome isolation method involves isolation via ultracentrifuge technique which “is not considered pure” in the literature.
- WO2017078641A1 and WO2017057881A1 are also documents known in the art related to use of exosomes in promoting hair growth.
- the objective of the invention is to enable to revitalize and strengthen unhealthy skin and hair that have lost their vitality, to stop hair loss, to provide faster hair growth by stimulating hair follicles, and to provide a regulating effect in improving the effects such as wrinkles and age spots caused by aging skin cells.
- Figure 1 is a graphical representation of the evaluation of the effect of administration of the exosomes obtained from Garlic to skin cells at different concentrations for 24 hours on cell viability using MTS test.
- Figure 2 is a graphical representation of the evaluation of the effect of administration of the exosomes obtained from Pineapple to skin cells at different concentrations for 24 hours on cell viability using MTS test.
- Figure 3. is a graphical representation of the evaluation of the effect of administration of the exosomes obtained from Pomegranate to skin cells at different concentrations for 24 hours on cell viability using MTS test.
- Figure 4. is a graphical representation of the evaluation of the effect of administration of Garlic-Pineapple-Pomegranate exosomes at non toxic concentrations for 24 hours on cell viability using MTS test
- Figure 5 is a graphical representation of evaluation of administration of the mixtures obtained from plant exosomes and stem cell exosome to skin cells on TGF-B 1 gene expression levels
- Figure 6. is a graphical representation of evaluation of administration of the mixtures obtained from plant exosomes and stem cell exosome to skin cells on WNT5a gene expression levels.
- Figure 7 is a graphical representation of evaluation of administration of the mixtures obtained from plant exosomes and stem cell exosome to skin cells on Shh gene expression levels.
- the present invention relates to use of plant exosomes as a product having the effect of enhancing viability and, by stimulating the hair follicles, enhancing hair growth in skin cells. Pineapple, garlic and pomegranate are preferred as the plants from which the exosomes are isolated.
- plant-based exosomes are used for this purpose.
- the effects of the plant exosomes of the present invention can vary according to the plant from which the exosome is isolated. While these can be the entire plant, fruit, leaf, seed and root, they may also be differentiated tissues like the plant’s culture medium, stem cell, waste material, shell or phloem.
- the plant exosomes can be isolated by many methods such as isolation by two phase separation, graduated centrifuge, ultrafiltration, chromatographic methods, polymer based isolation and isolation by microbeads.
- exosomes are isolated from Pineapple, Pomegranate and Garlic.
- the two phase liquid system is utilized for isolation of exosomes from plant lysate, which system comprises the steps of removing the large size particles and impurities resulting from plant disintegration by centrifugation performed between 2,000 g and 10,000 g for 5-20 minutes filtering to remove the particles sized 220 nanometers and above, and separating the obtained homogeneous exosome-protein mixture.
- Exosomes are cleared of nonexosomal proteins, cellular fats and other impurities by utilizing the chemical tendency of the PEG phase to the proteins and of the DEX (dextran) phase to the phospholipid structured membranes in the two-phase liquid system.
- the scope of the invention involves the effect of not only promoting hair growth, but also the vitality and rejuvenation of skin cells. Accordingly, the scope of the invention involves mixing the plant exosomes into a solution as various personal care products such as creams and shampoos.
- plant exosomes of the present invention have effects of enhancing viability and promoting hair growth on skin and hair cells indicates that they can serve as regulators to revitalize and strengthen unhealthy skin and hair where the skin and hair cells have lost their vitality, to stop hair loss, to provide faster hair growth and to improve the effects such as wrinkles and age spots caused by aging skin cells.
- Effective doses of the plant exosomes preferred within the scope of the invention are as follows. Furthermore, as these exosomes can be used in different combinations, a range of use has been determined. These exosomes can be used alone or in combination of 2 or 3. The preferred ratios of composition in a triple combination are as follows:
- plant exosomes extracted from these plants can be used with other plant- extracted exosomes.
- the plant exosome mixture that is used enhances regeneration of the hair follicles in the skin cells.
- the plant exosome mixture enables regrowth of hair as the dormant hair follicle becomes active since the mixture provides an increase in the TGFB 1 gene.
- a two-phase fluid system is preferred to be used for separating the homogeneous exosome-protein mixture obtained as a result of the preparation steps.
- Exosomes are cleared of nonexosomal proteins, cellular fats and other impurities by utilizing the chemical tendency of the PEG phase to the proteins and the DEX phase to the phospholipid structured membranes in the two-phase liquid system.
- the DEX phase formed by means of the concentrations of the polymers that are used in the solution separate the exosomes.
- the isolated exosomes were marked by the surface markers CD9, CD63 and HSP70 antibodies which are carried by the exosomes and the exosomes carrying these markers were measured by flow cytometry device. At the same time, the dimensions of the isolated exosomes were measured by means of Zeta Sizer device, and the results are as seen in Figures 1 - 3.
- DMEM Dulbecco's modified Eagle's medium
- PSA Biological Industries, Beit Haemek, Israel
- Cell viability was measured by using 3-(4,5-di-methyl-thiazol-2-yl)-5-(3-carboxy- methoxy-phenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium (MTS)-method (CellTiter96 AqueousOne Solution; Promega, Southampton, UK).
- IOmI MTS solution was added onto the cells within a IOOmI medium and they were incubated at 37°C in dark for 2 hours. After the incubation process, cell viability was observed by performing absorbance measurement via ELISA plate reader (Biotek, Winooski, VT) device at 490 nm wavelength.
- RNAs were isolated and cDNA was synthesized from the cells that were seeded in 6-well culture plates (Coming Glasswork, Coming, NY) at 50,000 cells/well in Dulbecco's modified Eagle's medium (DMEM). The synthesized cDNAs were mixed with primers in Fermentas Maxima SYBR Green mixture product such that the final volume will be 20ml and the expression levels of the genes were analyzed by using BIO-RAD device.
- the doses of the plant exosomes required to induce hair growth were determined according to the results of toxicity and cell viability tests shown in Figure 1-4. In these figures, the doses that enhance skin cell viability at a maximum level are determined.
- Figure 5-8 the effects of the plant exosome application on the expression levels of the genes of sonichedgehog (Shh) [8] that is used in determining the hair follicle in the skin cell, WNT5a [9] that increases the regeneration of the hair follicle, and TGFB1 [10] that is responsible for the growth of hair follicles, were examined, and the plant exosome mixture that is used has been shown to be effective on these genes.
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Abstract
The present invention relates to use of plant exosomes as a product having the effect of enhancing viability and, by stimulating the hair follicles, enhancing hair growth in skin cells. The objective of the invention is to enable to revitalize and strengthen unhealthy skin and hair that have lost their vitality, to stop hair loss, to provide faster hair growth by stimulating hair follicles, and to provide a regulating effect in improving the effects such as wrinkles and age spots caused by aging skin cells.
Description
DESCRIPTION
USE OF PLANT EXOSOMES FOR ENHANCING VIABILITY AND HAIR GROWTH CAPACITY OF SKIN CELLS
Field of the Invention
The present invention relates to use of plant exosomes as a product having the effect of enhancing viability and, by stimulating the hair follicles, enhancing hair growth in skin cells.
Background of the Invention
Mammalian cells include small vesicular structures called exosomes. When these exosomes are isolated from healthy cells, they can be used to restore the cells, which are damaged or under a certain stress and which cannot complete their self regeneration, back to their earlier healthy state [1] Recently, use of the stem cells in cell based therapies is also one of the most promising studies. It is asserted that use of the exosomes released from these cells will have positive effects on wound healing and hair growth/strengthening in clinical application [2].
The vesicles are small sacs which are involved in the transport and storage of substances within the cell and are separated by at least one lipid bilayer from the cytoplasm fluid. Exosomes are vesicles, which are released by many organisms from prokaryotes to high eukaryotes and plants, and which contain lipid bilayer vesicles of different sizes [3]. The importance of these vesicles lies behind the capacity of transferring information to the other cells in order to influence the cell function. Signal transfer via exosomes is carried out by means of biomolecules in many different categories consisting of proteins, lipids, nucleic acid and sugars [4].
Functional interactions of extracellular vesicles with cells were first found in 1982 upon determining experimentally that vesicles isolated from seminal plasma increase sperm motility [5]. From this point on, studies have been conducted in
many different tissues until today on the developments related to the molecular mechanism of vesicles and bringing the issues left in the dark into light.
Intense hair loss is a problem which can be encountered both in men and women. There are many reasons of this problem depending on hormones, age, stress and mineral deficiency [6]. Although there is no study conducted on cells regarding the viability and hair growth stimulation effects of the plant exosomes related to skin and hair, the characterization of the plant exosomes and their effects on cancer cells have begun to be studied [7].
The European patent application document no. EP3354257, an application in the state of the art, discloses a composition which comprises plant extract-derived extracellular vesicles and is uses for improving skin and preventing hair loss. The said application discloses about the effects of the vesicles obtained by passing the filtered plant juice through different centrifuges. Exosome isolation method involves isolation via ultracentrifuge technique which “is not considered pure” in the literature.
The United States patent application document no. US2018256488, an application in the state of the art, discloses a composition for preventing hair loss and promoting hair growth, containing ginseng-derived exosome-like vesicles.
The documents numbered EP2629782A1, W02017023690A1,
WO2017078641A1 and WO2017057881A1 are also documents known in the art related to use of exosomes in promoting hair growth.
The problems in the applications in the prior art can be listed as follows:
• Many substances used as hair growth promoters are chemicals and have toxic effects.
• Obtaining the exosomes used as hair growth promoters from animal cells and use thereof are not ethically appropriate.
• The side effects resulting due to the fact that the products claiming to promote hair growth and skin rejuvenation are chemicals extend to heart diseases and renal failure.
• The high doses used for an effective result are toxic and irritant to the skin.
Summary of the Invention
The objective of the invention is to enable to revitalize and strengthen unhealthy skin and hair that have lost their vitality, to stop hair loss, to provide faster hair growth by stimulating hair follicles, and to provide a regulating effect in improving the effects such as wrinkles and age spots caused by aging skin cells.
Detailed Description of the Invention
“Use of plant exosomes for enhancing viability and hair growth capacity of skin cells” developed to fulfill the objectives of the present invention is illustrated in the accompanying figures, in which:
Figure 1. is a graphical representation of the evaluation of the effect of administration of the exosomes obtained from Garlic to skin cells at different concentrations for 24 hours on cell viability using MTS test.
Figure 2. is a graphical representation of the evaluation of the effect of administration of the exosomes obtained from Pineapple to skin cells at different concentrations for 24 hours on cell viability using MTS test.
Figure 3. is a graphical representation of the evaluation of the effect of administration of the exosomes obtained from Pomegranate to skin cells at different concentrations for 24 hours on cell viability using MTS test.
Figure 4. is a graphical representation of the evaluation of the effect of administration of Garlic-Pineapple-Pomegranate exosomes at non toxic concentrations for 24 hours on cell viability using MTS test
Figure 5. is a graphical representation of evaluation of administration of the mixtures obtained from plant exosomes and stem cell exosome to skin cells on TGF-B 1 gene expression levels
Figure 6. is a graphical representation of evaluation of administration of the mixtures obtained from plant exosomes and stem cell exosome to skin cells on WNT5a gene expression levels.
Figure 7. is a graphical representation of evaluation of administration of the mixtures obtained from plant exosomes and stem cell exosome to skin cells on Shh gene expression levels.
The present invention relates to use of plant exosomes as a product having the effect of enhancing viability and, by stimulating the hair follicles, enhancing hair growth in skin cells. Pineapple, garlic and pomegranate are preferred as the plants from which the exosomes are isolated.
Within the scope of the invention, it is aimed to enhance the viability and hair growth capacity of skin cells, and plant-based exosomes are used for this purpose. The effects of the plant exosomes of the present invention can vary according to the plant from which the exosome is isolated. While these can be the entire plant, fruit, leaf, seed and root, they may also be differentiated tissues like the plant’s culture medium, stem cell, waste material, shell or phloem. The plant exosomes can be isolated by many methods such as isolation by two phase separation, graduated centrifuge, ultrafiltration, chromatographic methods, polymer based isolation and isolation by microbeads.
Among the different applications listed above for isolating exosomes from plants, isolation with two-phase liquid system which provides the purest exosome isolation is preferred within the scope of the invention. Within the scope of the
invention, the exosomes are isolated from Pineapple, Pomegranate and Garlic. The two phase liquid system is utilized for isolation of exosomes from plant lysate, which system comprises the steps of removing the large size particles and impurities resulting from plant disintegration by centrifugation performed between 2,000 g and 10,000 g for 5-20 minutes filtering to remove the particles sized 220 nanometers and above, and separating the obtained homogeneous exosome-protein mixture. Exosomes are cleared of nonexosomal proteins, cellular fats and other impurities by utilizing the chemical tendency of the PEG phase to the proteins and of the DEX (dextran) phase to the phospholipid structured membranes in the two-phase liquid system.
Within the scope of the invention, not only the effect of one plant exosome but the use of specific concentrations of 3 plants is discussed. It is observed that the effects of the plant exosomes when used alone are not as high as those when they are used in the combination. In other words, no plant exosome is sufficient for promoting hair growth and skin vitality. Moreover, the scope of the invention involves the effect of not only promoting hair growth, but also the vitality and rejuvenation of skin cells. Accordingly, the scope of the invention involves mixing the plant exosomes into a solution as various personal care products such as creams and shampoos.
The fact that plant exosomes of the present invention have effects of enhancing viability and promoting hair growth on skin and hair cells indicates that they can serve as regulators to revitalize and strengthen unhealthy skin and hair where the skin and hair cells have lost their vitality, to stop hair loss, to provide faster hair growth and to improve the effects such as wrinkles and age spots caused by aging skin cells.
Effective doses of the plant exosomes preferred within the scope of the invention are as follows. Furthermore, as these exosomes can be used in different combinations, a range of use has been determined. These exosomes can be used
alone or in combination of 2 or 3. The preferred ratios of composition in a triple combination are as follows:
Pineapple: 1 milligram - 1 nanogram Garlic: 10 milligrams - 1 nanogram Pomegranate 1 milligram - 1 nanogram
In addition, it is contemplated and discussed within the scope of the invention that the plant exosomes extracted from these plants can be used with other plant- extracted exosomes.
The advantages of using the plant exosomes of the present invention to enhance the viability and hair growth capacity of skin cells can be listed as follows:
• The plant exosome mixture that is used enhances regeneration of the hair follicles in the skin cells.
• Enhances viability and regeneration of the skin cells.
• Induces hair growth in hair follicles.
• Plays a role in eliminating wrinkles on the skin and recuperation of aging skin as it enhances cell viability.
• The dose specified in this mixture has a substantial effect in eliminating signs of aging, wrinkles and age spots.
• Enables enhancing viability of hair cells and a faster and healthier hair growth by stimulating the hair follicles.
• At the same time, the plant exosome mixture enables regrowth of hair as the dormant hair follicle becomes active since the mixture provides an increase in the TGFB 1 gene.
• The fact that the product is plant-based increases its usability.
EXPERIMENTAL STUDIES A two-phase fluid system is preferred to be used for separating the homogeneous exosome-protein mixture obtained as a result of the preparation steps. Exosomes
are cleared of nonexosomal proteins, cellular fats and other impurities by utilizing the chemical tendency of the PEG phase to the proteins and the DEX phase to the phospholipid structured membranes in the two-phase liquid system. The DEX phase formed by means of the concentrations of the polymers that are used in the solution separate the exosomes. The isolated exosomes were marked by the surface markers CD9, CD63 and HSP70 antibodies which are carried by the exosomes and the exosomes carrying these markers were measured by flow cytometry device. At the same time, the dimensions of the isolated exosomes were measured by means of Zeta Sizer device, and the results are as seen in Figures 1 - 3.
A toxicity analysis was performed to determine the effects of the plant exosomes on viability of skin cells. After the cells were seeded in 96- well culture plates (Corning Glasswork, Coming, NY) at 5000 cells/well in Dulbecco's modified Eagle's medium (DMEM) containing 10% exosome-depleted fetal bovine serum (Invitrogen) and 1% PSA (Biological Industries, Beit Haemek, Israel) in the culture medium, the viability levels of the cells were measured on day 1, 2 and 3. Cell viability was measured by using 3-(4,5-di-methyl-thiazol-2-yl)-5-(3-carboxy- methoxy-phenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium (MTS)-method (CellTiter96 AqueousOne Solution; Promega, Southampton, UK). IOmI MTS solution was added onto the cells within a IOOmI medium and they were incubated at 37°C in dark for 2 hours. After the incubation process, cell viability was observed by performing absorbance measurement via ELISA plate reader (Biotek, Winooski, VT) device at 490 nm wavelength.
At the same time, expression levels of the genes related to cell viability and hair growth were measured in order to determine the effect of the exosomes on promoting viability and hair-growth. Cultured cells may lose their own properties and acquire new properties. These properties may be both in morphological level and gene expression level. Real Time PCR method was applied to observe the changes in gene expression level. Total RNAs were isolated and cDNA was
synthesized from the cells that were seeded in 6-well culture plates (Coming Glasswork, Coming, NY) at 50,000 cells/well in Dulbecco's modified Eagle's medium (DMEM). The synthesized cDNAs were mixed with primers in Fermentas Maxima SYBR Green mixture product such that the final volume will be 20ml and the expression levels of the genes were analyzed by using BIO-RAD device.
As can be seen in the figures as well, the doses of the plant exosomes required to induce hair growth were determined according to the results of toxicity and cell viability tests shown in Figure 1-4. In these figures, the doses that enhance skin cell viability at a maximum level are determined. At the same time, in Figure 5-8, the effects of the plant exosome application on the expression levels of the genes of sonichedgehog (Shh) [8] that is used in determining the hair follicle in the skin cell, WNT5a [9] that increases the regeneration of the hair follicle, and TGFB1 [10] that is responsible for the growth of hair follicles, were examined, and the plant exosome mixture that is used has been shown to be effective on these genes.
REFERENCES
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[5]. Stegmayr, B. and G. Ronquist (1982). "Promotive effect on human sperm progressive motility by prostasomes." Urol Res 10(5): 253-257.
[6]. Norwood, O. T. (1975). Male pattern baldness: classification and incidence. Southern medical journal , <58(11), 1359-1365.
[7]. Raimondo S, Naselli F, Fontana S, et al. Citrus Union -derived nanovesicles inhibit cancer cell proliferation and suppress CML xenograft growth by inducing TRAIL-mediated cell death. Oncotarget. 2015;6(23): 19514- 19527.
[8]. Paladini, Rudolph D., et al. "Modulation of hair growth with small molecule agonists of the hedgehog signaling pathway." Journal of investigative dermatology 125.4 (2005): 638-646.
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[10]. Mori, Osamu, Hiroshi Hachisuka, and Yoichiro Sasai. "Effects of transforming growth factor bΐ in the hair cycle." The Journal of dermatology 23.2 (1996): 89-94.
Claims
1. Plant exosomes used for their effects of enhancing viability, and, by stimulating the hair follicles, enhancing hair growth in skin cells.
2. Plant exosomes according to Claim 1 characterized in that they are isolated from single, double or triple combinations selected from a group comprising pineapple, garlic and pomegranate.
3. Plant exosomes according to Claim 2 characterized by a triple combination comprising 1 milligram - 1 nanogram of plant exosomes isolated from pineapple plant, 10 micrograms - 1 nanogram of plant exosomes isolated from garlic plant, and 1 milligram - 1 nanogram of plant exosomes isolated from pomegranate plant.
4. Plant exosomes according to Claim 1, characterized in that they are isolated from the entire plant, fruit, leaf, seed, root, as well as the differentiated tissues of the plant such as the culture medium, stem cell, waste material, shell or phloem.
5. Plant exosomes according to any one of the preceding claims, characterized in that they are isolated by an isolation method selected from a group comprising the isolation methods of two phase separation, graduated centrifuge, ultrafiltration, chromatographic methods, polymer based isolation and isolation by microbeads.
6. Plant exosomes according to Claim 5 characterized in that they are isolated by the two phase liquid system isolation method.
7. Plant exosomes according to any one of the preceding claims characterized in that they are mixed into a solution as personal care products.
8. Two-phase liquid system, which is performed for isolation of the plant exosomes according to Claim 7 from plant lysate, comprising the steps of -removing the large size particles and impurities resulting from plant disintegration by centrifugation performed between 2,000 g and 10,000 g for 5-20 minutes,
-filtering to remove the particles sized 220 nanometers and above, and -separating the obtained homogeneous exo some-protein mixture.
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| US17/639,614 US20220331230A1 (en) | 2019-09-02 | 2020-09-02 | Use of plant exosomes for enhancing viability and hair growth capacity of skin cells |
| EP20860753.1A EP4025232A4 (en) | 2019-09-02 | 2020-09-02 | Use of plant exosomes for enhancing viability and hair growth capacity of skin cells |
| CN202080061501.4A CN114401729A (en) | 2019-09-02 | 2020-09-02 | Use of plant exosomes for enhancing skin cell viability and hair growth capacity |
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| TR2019/13215A TR201913215A2 (en) | 2019-09-02 | 2019-09-02 | USE OF HERBAL EXOSOMES TO INCREASE THE VITALITY OF SKIN CELLS AND THEIR HAIR REMOVAL CAPACITY |
| TR2019/13215 | 2019-09-02 |
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| CN113846046A (en) * | 2021-10-22 | 2021-12-28 | 科丝美诗(中国)化妆品有限公司 | Dendrobium-derived exosome-like vesicle, preparation method thereof and application thereof in alopecia improvement |
| KR102534286B1 (en) * | 2022-10-07 | 2023-05-26 | (주)3일애 | Cosmetic composition containing aloe vera-derived exosomes and its manufacutring method |
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| CN116036196A (en) * | 2022-12-29 | 2023-05-02 | 广州远想生物科技股份有限公司 | Anti-hair loss hair-nourishing essence and preparation method thereof |
| CN116942847B (en) * | 2023-08-30 | 2024-04-09 | 深圳市华健生物技术有限公司 | Drug-loaded plant exosome targeting hair follicle, and preparation method and application thereof |
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| KR102475127B1 (en) * | 2015-09-30 | 2022-12-07 | (주)아모레퍼시픽 | Composition comprising ginseng derived exosome like vesicles for preventing hair loss or promoting hair growth |
| US11911713B2 (en) * | 2018-02-15 | 2024-02-27 | Yeditepe Universitesi | Exosome isolation method by two phase fluid system |
| CN113226474A (en) * | 2018-09-06 | 2021-08-06 | 耶迪特普大学 | Use of plant exosomes for displaying a modulating effect on cells of the immune system |
| TR201911667A2 (en) * | 2019-08-01 | 2021-02-22 | Univ Yeditepe | Plant exosomes used for the treatment of diseases caused by protein deposits formed in cells. |
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| US20130216596A1 (en) * | 2010-09-21 | 2013-08-22 | Lipotec, S.A. | Nanocapsules containing microemulsions |
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| WO2017052271A1 (en) * | 2015-09-25 | 2017-03-30 | ㈜프로스테믹스 | Composition for preventing or treating hair loss |
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| CN113846046A (en) * | 2021-10-22 | 2021-12-28 | 科丝美诗(中国)化妆品有限公司 | Dendrobium-derived exosome-like vesicle, preparation method thereof and application thereof in alopecia improvement |
| KR102534286B1 (en) * | 2022-10-07 | 2023-05-26 | (주)3일애 | Cosmetic composition containing aloe vera-derived exosomes and its manufacutring method |
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| TR201913215A2 (en) | 2021-03-22 |
| EP4025232A1 (en) | 2022-07-13 |
| US20220331230A1 (en) | 2022-10-20 |
| EP4025232A4 (en) | 2024-02-07 |
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