Potential of stock tomato to realize skin brightening by inhibiting melanin transfer activity

Abstract

The invention discloses the potential of original tomato to realize skin lightening by inhibiting melanin transfer activity. It is discussed that carotenoid and flavonoid compounds in the breeder seed white tomato (white tomato extract) have effects on skin appearance and safety. The systems and methods include cytotoxicity assessment, damage reduction analysis, melanin estimation, PAR-2 activity assessment, and GSH level estimation. Experimental results show that the white tomato extract can effectively reduce melanin transfer, improve the GSH level and promote skin cell regeneration under the condition of not generating cytotoxic effect. Optimal doses of the white tomato extract have been confirmed, showing significant recovery from injury and reduced LDH release. In addition, the reduction of PAR-2 activity indicates that melanin transfer is inhibited. The present disclosure highlights the potential of colorless carotenoids and flavonoids from the stock tomato in skin care interventions. In general, white tomato extracts are configured for skin lightening and cell regeneration, thereby providing a safe and effective solution for improving skin color and appearance.

Description

Technical Field

[0001] Embodiments of the present invention generally relate to compositions for tomato-based compositions, and more specifically, to methods of making heirloom white tomato-based compositions and related compositions thereof for anti-senescence purposes. Background Art

[0002] The following description includes information that may be helpful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, nor is it an admission that any publication specifically or implicitly referenced is prior art.

[0003] As is well known, skin whitening compositions can be used to whiten the skin to a lighter skin tone or to remove dark spots on the skin. In addition, these skin whitening or skin bleaching compositions are effective because they inhibit the formation of melanin, the pigment that determines the skin color. However, such skin bleaching or skin whitening compositions contain irritating chemicals such as peroxides, glutathione, formaldehyde, etc. that have severe effects on the skin.

[0004] As disclosed in conventional prior art disclosures, the use of hydroquinone inhibits melanocyte activity and helps whiten the skin. However, such less demanding compositions have other disadvantages, namely that hydroquinone is oxidized by air and light. Furthermore, the oxidation products can cause skin irritation and inflammation.

[0005] In addition, the dermatological composition using a paucilamellarliposomal carrier has been disclosed in the prior art disclosures, which helps treat melanoma caused by exposure to sunlight. However, such compositions cannot effectively whiten the skin to a lighter skin color. In addition, some traditional prior art disclosures disclose the use of naturally extracted materials (such as, but not limited to, lemons, oranges, cucumbers, rose hips, etc.) in dermatological compositions. However, such naturally extracted materials limit their usefulness based on age, skin type, and color, and vary greatly among different subjects.

[0006] As disclosed in various prior art disclosures, the use of cosmetic compositions containing carotenoids and polyphenol-containing apple plant extracts contributes to skin whitening. However, these carotenoid-containing compositions are generally highly pigmented and sensitive to light and oxidation, which greatly limits their usefulness.

[0007] Furthermore, the prior art disclosures disclose the use of polyphenolic compounds, primarily resorcinol, which have skin whitening properties. However, these compounds, which are susceptible to oxidation by air and UV, result in color instability, which results in a loss of skin firming efficacy. Additionally, according to related prior art disclosures, extracts obtained from acerola fruit (fruit sofacerola), which contain vitamin C, are incorporated into cosmetics to provide a whitening effect. Thus, the prior art disclosures briefly discuss compositions that can be used for skin whitening. However, the use of elements to increase natural glutathione levels has not yet been introduced to the market.

[0008] Carotenoids, recognized for their nutritional and health importance, serve as precursors to retinoids, accumulating in human skin and providing both health and beauty benefits (Meléndez-Martínez, Manditz et al., 2022, Udensi, Loughman et al., 2022).

[0009] Of note, colorless carotenoids such as phytoene (PT) and phytofluene (PTF) have recently attracted attention for their bioavailability and potential health benefits (Meléndez-Martínez, Manditz, et al., 2022, Biskanaki, Kalofiri et al., 2023, Kiki 2023).

[0010] Colorless carotenoids found in tomatoes, including phytoene, phytoene, lutein, and zeaxanthin, are thought to contribute to antioxidant mechanisms, sparking discussions about their incorporation into nutricosmetics to improve both skin appearance and safety (Etminan, Takkouche, et al. 2004, Canene-Adams, Campbell, et al. 2005, Giovannucci 2005, Engelmann, Clinton, et al. 2011).

[0011] Tomato is recognized as an important source of dietary flavonoids due to its high consumption worldwide. The total flavonoid content of different tomato types varies from 4 mg to 26 mg 100(-1) g FW (Slimestad, Fossen et al., 2008). Tomato contains a large amount of flavonoids, mainly quercetin and quercetin glycosides (isoquercetin, rutin).

[0012] Heirlooms tomatoes, which originated from seeds 50 years ago and have not undergone any hybridization, are well known for their diversity in flavor, color and shape. Heirlooms tomatoes are white tomatoes, and various biological properties have been studied. In this study, the effect of heirlooms tomato powder on melanin transfer inhibition to minimize skin pigmentation levels was investigated. Heirlooms tomato powder is basically a powder rich in colorless carotenoids and flavonoids, which can reduce melanin transfer and increase glutathione levels. To prove this fact, the first strategy was to define the non-cytotoxic dose of tomato powder, and to determine this fact, the MTT assay was used on two cell lines.

[0013] However, due to challenges with detection using wavelengths typical for colored carotenoids, studies have often overlooked colorless carotenoids and flavonoids (Rodriguez-Amaya 2001). Nonetheless, recent studies have explored their antioxidant capacity and isomerism (Melendez-Martinez, Stinco, et al., 2013, Melendez-Martinez, Paulino, et al., 2014, Stinco, Escudero-Gilete, et al., 2016), consistent with the growing interest in dietary compounds that promote skin appearance and safety (Madhere and Simpson, 2010). Tomatoes, which are rich in carotenoids such as lycopene, have been linked to a reduced risk of chronic diseases, with colorless carotenoids such as phytoene and phytoene thought to be biologically active (Etminan, Takkouche, et al., 2004, Canene-Adams, Campbell, et al., 2005, Engelmann, Clinton, et al., 2011). Glutathione, a potent antioxidant, has become a popular skin lightening agent, despite controversy regarding its efficacy and safety (Exner, Wessner, et al., 2000, Sonthalia, Daulatabad, et al., 2016).

[0014] This study highlights the importance of reduced glutathione (GSH) and introduces traditional white tomatoes rich in carotenoids and flavonoids as a unique source that has the potential to provide antioxidant benefits to cells (Khachik, Carvalho et al., 2002).

[0015] Since flavonoids are considered as potential health-promoting substances, efforts have been made to produce tomatoes with standardized levels of flavonoids and other compounds (Stewart, Bozonnet et al., 2000, Slimestad and Verheul, 2005), or to increase the concentration of quercetin and quercetin 3-rutinoside (rutin) in particular (Slimestad, Fossen et al., 2008). Nutricosmetics, bridging the food, nutrition, health and cosmetic fields, consider colorless carotenoids and flavonoids as promising components to promote "beauty from within", which is consistent with the growing interest in dietary compounds that enhance skin appearance and safety (Madhere and Simpson, 2010).

[0016] The emerging paradigm of photoprotection through dietary intervention emphasizes the key role that plant components, particularly carotenoids and flavonoids, play in protecting plants from excessive light and preventing UV damage in humans (Stahl and Sies 2007, Kiefer, Weibel et al. 2010). Tomatoes, consumed and recognized worldwide for their flavonoid content, including quercetin and quercetin glycosides, are currently undergoing efforts to standardize and increase flavonoid concentrations (Crozier, Lean et al. 1997, Stewart, Bozonnet et al. 2000, Slimestad and Verheul 2005, Slimestad, Fossen et al. 2008).

[0017] Tomatoes, known for their carotenoid content such as lycopene, have been linked to a reduced risk of chronic diseases, with colorless carotenoids such as phytoene and phytoene being postulated to have bioactive properties. Glutathione, a potent antioxidant with anti-melanogenic properties, has become a popular skin lightening agent, but controversy remains regarding its efficacy and safety (Exner, Wessner et al., 2000, Sonthalia, Daulatabad et al., 2016).

[0018] Skin melanocytes produce melanin, which acts as a natural barrier against ultraviolet (UV) light and reduces DNA damage in skin cells (Berens, Van Den Bossche et al., 2005). However, abnormal accumulation of melanin can lead to pigmentation disorders, such as melanoma (Buitrago, Hardre et al., 2016), age-related freckles, post-inflammatory pigmentation and melasma (Ashraf, Rafiq et al., 2015). Therefore, controlling pigmentation of human skin remains a top priority for the cosmetic and pharmaceutical industries (Kim, Hwang et al., 2015). It is found that many cosmetic products contain skin whitening ingredients such as vitamin C, hydroquinone, arbutin, azelaic acid, retinoic acid and kojic acid (Hakozaki, Minwalla et al., 2002, Matos, Varela et al., 2015), although the skin whitening ingredients may have negative side effects such as dermatitis, skin irritation and melanocyte death. Therefore, the research on organic color-reducing compounds has been accelerated (Wang, Chang et al., 2017). However, more research is needed to determine the precise process by which these drugs prevent the synthesis and transport of melanin. The transport of melanin to epidermal cells is the only process that causes pigmentation, so blocking this transfer can effectively prevent pigmentation (Ma, Zhao et al., 2010).

[0019] Given the beneficial effects associated with the significant and unique blend of colorless carotenoids and flavonoids in original white tomatoes, the current research effort explored the potential of carotenoids and flavonoids derived from original white tomatoes (TomesOral) in promoting skin lightening and cell regeneration by inhibiting melanin transfer to the epidermis and increasing reduced glutathione (GSH).

[0020] Therefore, there is a recognized need for an alternative effective natural combination of flavonoid and carotenoid preparations with minimal adverse side effects and such an alternative effective natural combination would be very beneficial, effective for skin lightening and suitable for the cosmetic industry. Therefore, there remains a need in the art for innovative, novel and efficient methods for preparing heirloom white tomato based compositions that achieve skin lightening potential by inhibiting melanin transfer activity. Summary of the invention

[0021] The inventive summary does not necessarily disclose all features necessary to define the invention: the invention may lie in a sub-combination of the disclosed features.

[0022] Tomesoral White Tomato is a unique tomato powder derived from heirloom tomatoes, which originated from seeds of plants at least 50 years ago and are recognized for their diverse color, shape and flavor attributes. These tomatoes are rich in phytoene, phytoene, lutein and zeaxanthin obtained from the fruits of organic heirloom white tomatoes. The tomato fruits are carefully sampled and rigorously washed to remove contamination. Subsequently, they are freeze-dried.

[0023] A method for skin lightening and cell regeneration comprises applying an effective amount of a white tomato extract, which is a composition derived from an original white tomato; wherein the composition contains carotenoids and flavonoids that inhibit melanin transfer activity and promote skin cell regeneration.

[0024] The composition effectively reduces melanin transfer, increases GSH levels in the skin, promotes skin cell regeneration, and exhibits no cytotoxic effects.

[0025] The optimal dosage of white tomato extract was identified to show significant recovery from skin damage.

[0026] The composition further reduced LDH release (indicating reduced cell damage) and decreased PAR-2 ​​activity (indicating inhibition of melanin transfer).

[0027] The carotenoids in the white tomato extract are colorless, wherein the composition is specifically derived from an original white tomato.

[0028] The composition is formulated to improve skin tone and appearance.

[0029] In another embodiment, a skin care system for improving skin tone and appearance includes white tomato extract, a formulation derived from pure white tomatoes containing colorless carotenoids and flavonoids; a cytotoxicity assessment module; a damage reduction analysis module; a melanin estimation module; a PAR-2 ​​activity assessment module; and a GSH level estimation module, wherein the system is configured to reduce melanin transfer, increase GSH levels, and promote skin cell regeneration without producing cytotoxic effects.

[0030] The cytotoxicity assessment module evaluates the safety of the composition.

[0031] The damage reduction analysis module assesses recovery from skin damage.

[0032] The melanin estimation module measures the melanin level in the skin.

[0033] The PAR-2 ​​activity assessment module measures the inhibition of melanin transfer.

[0034] The GSH level estimation module measures the increase in GSH level in the skin.

[0035] The composition reduces LDH release, indicating reduced cellular damage.

[0036] The system is further configured for skin lightening and cell regeneration, thereby providing a safe and effective solution for improving skin tone and appearance. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] The accompanying drawings are included to provide a further understanding of the present disclosure and are incorporated in and constitute a part of this specification. The accompanying drawings illustrate exemplary embodiments of the present disclosure and together with the description serve to explain the principles of the present disclosure.

[0038] The accompanying drawings are for illustration purposes only and thus are not intended to limit the present disclosure, and in the accompanying drawings:

[0039] Figure 1 A and C) and B and D) show a bar graph and a line graph, respectively, showing the relative viability percentage of tomato powder at different concentrations according to one embodiment of the present invention.

[0040] Figure 2 : A) Graph shows the level of LDH release in cells after A-375 and HaCaT treatment. Values ​​are obtained as mean ± SEM, and * indicates significant differences (p < 0.05) in cell viability and LDH levels between treated groups and untreated controls.

[0041] Figure 3 : The figure shows the melanin level in cells after treatment according to an embodiment of the present invention.

[0042] Figure 4 The graph shows PAR-2 ​​activity in cells after treatment according to an embodiment of the present invention.

[0043] Figure 5 : The figure shows the GSH level in cells after treatment according to one embodiment of the present invention. DETAILED DESCRIPTION

[0044] The following is a detailed description of the embodiments of the present disclosure illustrated in the accompanying drawings. The embodiments are shown in detail in order to clearly convey the present disclosure. However, the amount of detail provided is not intended to limit the intended variations of the embodiments; on the contrary, the intention is to cover all modifications, equivalent forms, and alternative forms that fall within the spirit and scope of the present disclosure.

[0045] In the following description, numerous specific details are set forth in order to provide a thorough understanding of embodiments of the present invention. It will be apparent to one skilled in the art that embodiments of the present invention may be practiced without some of these specific details.

[0046] A skin care system, method and composition for improving skin tone and appearance, comprising a white tomato extract, a formulation derived from an original white tomato containing colorless carotenoids and flavonoids; a cytotoxicity assessment module; a damage reduction analysis module; a melanin estimation module; a PAR-2 ​​activity assessment module; and a GSH level estimation module, wherein the system is configured to reduce melanin transfer, increase GSH levels, and promote skin cell regeneration without producing cytotoxic effects. method Preparation of tomato fruit powder

[0047] Tomesoral White Tomato is a unique tomato powder derived from stock tomatoes, which originated from seeds of plants at least 50 years ago and are recognized for their diverse color, shape and flavor attributes. These tomatoes are rich in phytoene, phytoene, lutein and zeaxanthin obtained from the fruits of organic stock white tomatoes. The tomato fruits have been carefully sampled and rigorously cleaned to remove contamination. Subsequently, they were subjected to the freeze-drying method outlined by Gaware in 2010, resulting in the production of a powdered form (Gaware, Sutar et al., 2010). Cell line culture

[0048] A-375 melanoma cells and HaCaT cells were seeded onto culture dishes and cultured in Dulbecco's Modified Eagle Medium-High Glucose (DMEM-HG) supplemented with antibiotics and 10% fetal bovine serum (FBS) until reaching the subculturing phase, at which time the cells were used for subsequent experiments. Preparation of stock solutions and dilutions

[0049] Tomato fruit powder stock solution was prepared by dissolving it in distilled water. After mixing, it was passed through a syringe filter and then different dilutions (100ug / ml, 200ug / ml, 500ug / ml, 1000ug / ml and 2000ug / ml) were prepared in normal culture medium (DMEM). Treatment of cell lines with tomato fruit powder

[0050] Second generation A-375 melanoma cell line and HaCaT cell line were seeded into 96-well plates to assess cell proliferation. The cells were treated with specific concentrations of tomato fruit powder for 24 hours. Subsequently, the cells were subjected to a cell viability test and the collected culture medium was used for subsequent experimental analysis. Cell viability assay

[0051] Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test (MTT test) performed on cells cultured in 96-well plates. Cells were placed in different concentrations of fruit powder for 24 hours. After the treatment phase, 25 μl of MTT solution was introduced, and after another 2 to 3 hours, the resulting purple crystals were dissolved by treatment with a detergent such as 10% SDS (sodium dodecyl sulfate) solution. Subsequently, after 3 hours of incubation, absorbance measurements at a wavelength of 570 nm were obtained. LDH release assay

[0052] A-375 melanoma cell line and HaCaT cell line were treated with 6mM H2O2 and then exposed to different concentrations of white tomato extract for 24 hours. Subsequently, the activity of lactate dehydrogenase (LDH) in the treated A-375 cell line and HaCaT cell line was measured in the supernatant of all experimental groups according to the protocol provided by AMP Diagnostics Austria. For the LDH test, 5μl of culture medium samples from each experimental condition were used. Specifically, 5μl of culture medium from each group was mixed with 100μl of experimental working reagent in a 96-well plate, followed by a 5-minute incubation period. The absorbance was then measured using a spectrophotometer at a wavelength of 340nm. Co-culture of cell lines

[0053] In a six-well plate, 2.5 × 10 5 Two days later, A-375 melanoma cells (melanocytes) (5×10 4 ) was added to each well containing keratinocytes. The cells were seeded onto culture dishes and cultured in Dulbecco's Modified Eagle's Medium-High Glucose (DMEM-HG) supplemented with antibiotics and 10% fetal bovine serum (FBS) until 70% confluence was reached, at which point the cells were used for subsequent experiments. Keratinocytes and melanocytes at the same cell density were cultured separately as controls. The cells were treated with tomato fruit powder at an optimal concentration (130 μg / ml) for 48 hours.

[0054] The following groupings were made: Estimation of melanin content

[0055] In 6-well culture plates, different groups of cells were planted in 2ml of culture medium, and the cells were attached overnight in an incubator. White tomato extract (130 μg / ml) at the optimal concentration was applied to cells in MEM for 48 hours. The cells were then dissociated by 0.25% trypsin ethylenediaminetetraacetic acid solution (digested for 2 minutes, then washed to separate melanocytes, and then digested for 8 minutes for keratinocyte digestion, only for co-cultivation), centrifuged at 1200 × g for 15 minutes at 4 ° C. The supernatant was then removed. 200 μl 1N NaOH was used to lyse the cells, and a pipette was used to repeatedly draw to homogenize them. In order to analyze, the crude cell extract was transferred to a 96-well plate in triplicate. Relative melanin content was determined by the absorbance at 405 nm using a microplate reader. Estimation of protease activated receptor 2 (PAR-2) by ELISA

[0056] Solid phase sandwich enzyme-linked immunosorbent assay (ELISA) was performed to assess the activity of protease activated receptor 2 (PAR-2) using a modified version of a previously described protocol (Wajid, Naseem et al., 2015). Briefly, 96-well plates (Corning, USA) were first coated with rabbit polyclonal anti-PAR-2 ​​antibody (Santa Cruz Biotechnology, USA) and incubated for 120 minutes. After washing three times with tris buffered saline (TBS), blocking was performed with 1% bovine serum albumin (BSA). Subsequently, 100 μl of culture medium from both cell groups was added to each well and incubated for 60 minutes. After incubation, the samples were aspirated and the wells were rinsed three times with TBS containing Tween-20 (TBS-T). The plate was then incubated overnight for 90 minutes with HRP-conjugated donkey anti-rabbit secondary antibody (Santa Cruz Biotechnology, USA). After another round of washing, 100 μl of tetramethylbenzidine (TMB) colorimetric solution (Invitrogen Inc., USA) was added to initiate the reaction. The reaction was terminated by adding 0.18 M sulfuric acid, and absorbance was measured at a wavelength of 450 nm. Estimation of GSH

[0057] Glutathione (GSH) levels in all previously listed groups containing stock meal were assessed by analyzing samples from the different experimental groups. GSH concentrations were quantified using a GSH assay kit according to the manufacturer's protocol (Sigma-Aldrich). Statistical analysis:

[0058] Data from experimental groups are presented as mean ± standard error of the mean (SEM) of three replicates. Statistical comparisons between group means were performed using one-way analysis of variance (ANOVA), followed by Bonferroni's test to determine differences between specific groups. Statistical significance was determined by a p value of less than 0.05. Cytotoxicity of original tomato powder and its optimal dosage

[0059] The MTT assay, known for its reliability and simplicity in assessing cytotoxicity, was performed. Figure 1 The relationship between the percentage (%) of cytotoxicity and different concentrations of stock powder in A-375 melanoma cell line and HaCaT cells is shown. It was observed that most tomato powder concentrations did not induce significant cytotoxic effects in both cell lines. Based on these findings, the optimal dose (OC) for A-375 melanoma cell line was determined to be 130 μg / ml and the optimal dose for HaCaT cells was 150 μg / ml, as shown in Figure 2. Figure 1 A to Figure 1 As shown in D.

[0060] Figure 1 A and Figure 1 The bar graph in C shows the relative value of absorbance after MTT assay. Figure 1 B and Figure 1 D. The line graph shows the relative viability percentage of different concentrations of tomato powder, while the boxes represent the OC values ​​of tomato powder. Values ​​are mean ± SEM, (p < 0.05). The damage in the treated group was reduced

[0061] A-375 cells and HaCaT cells were further treated with 6 mM H2O2 for 3 hours, and then the damaged cells were treated with the same dose of stock tomato as used in the previous experiment. When the cells were treated with H2O2, cell viability was significantly reduced, and a significant increase in viability was also observed after tomato powder treatment, showing significant recovery from injury.

[0062] To examine the level of damage and recovery, the LDH assay was used to measure the cytoplasmic enzyme lactate dehydrogenase (LDH) produced by the cells. When cells are damaged or stressed, LDH is released in the cytoplasm. Our results revealed that the levels of LDH released in the A-375 and HaCaT cell-treated groups were lower, even at lower doses, compared to injured cells.

[0063] Based on these LDH release study results and cell viability results, it was determined that the effective tomato powder dose range for recovery from H2O2-induced stress was 100 ug / ml to 2000 ug / ml, with OC approximately 150 ug / ml ( Figure 2 A and Figure 2 B).

[0064] Figure 2 : A) Graph shows the level of LDH release in cells after A-375 and HaCaT treatment. Values ​​are obtained as mean ± SEM, and * indicates significant differences (p < 0.05) in cell viability and LDH levels between treated groups and untreated controls. Reduction of melanin in cells treated with white tomato extract

[0065] Melanin is mainly located in the basal layer of the human epidermis (also known as the basal layer). In this layer, melanin is present in both melanocytes (MCs) responsible for melanin synthesis and adjacent keratinocytes (KCs) that receive and utilize melanin. The results showed that the amount of melanin was reduced in the A-375-white tomato extract group, demonstrating the anti-melanogenic effect of the white tomato extract. Neither group of HaCaT showed a large amount of melanin. However, despite the removal of melanocytes from the culture before melanin estimation, the co-culture - A375 + The HaCaT group still showed a large amount of melanin. In addition, melanin synthesis occurred in melanocytes, and in cells co-cultured with A-375 melanocytes, the transfer of melanin to keratinocytes was evident, explaining the expression of melanin in keratinocytes. In contrast, when these co-cultured cells were treated with white tomato extract and melanocytes were digested after treatment, a significant decrease in the amount of melanin was detected, indicating that the transfer of melanin from melanocytes to keratinocytes was inhibited, as shown in Figure 2. Figure 3 shown.

[0066] also, Figure 3 A graph showing melanin levels in cells after treatment is shown. Values ​​are obtained as mean ± SEM, and * indicates that there is a significant difference in melanin levels between the treated group and the untreated control group (p < 0.05). Activity of protease-activated receptor 2 (PAR-2) in cells treated with white tomato extract

[0067] PAR-2 ​​is involved in various physiological functions, including growth, development, mitosis, injury response, and skin pigmentation. In keratinocytes, PAR-2 ​​regulates growth and differentiation, and its inhibitory effect is beneficial to enhance the growth, differentiation, and regeneration of keratinocytes. In addition, PAR-2 ​​is involved in the phagocytosis of melanin and can trigger the intracellular Ca 2+ Activity. Studies have shown that Ca 2+ There is a relationship between activity and transfer of melanin. The results showed that low percentage activity of PAR-2 ​​in HaCaT cells showed inhibition of melanin transfer, which would further inhibit skin pigmentation. Figure 4 In the co-cultured cells treated with white tomato extract, the percentage of PAR-2 ​​activity from melanocytes to keratinocytes was significantly decreased.

[0068] also, Figure 4 The graph shows PAR-2 ​​activity in cells after treatment. Values ​​are obtained as mean ± SEM, and * indicates that there is a significant difference in PAR-2 ​​activity levels between the treated group and the untreated control (p < 0.05). GSH increased in the treated groups

[0069] Intracellular GSH levels in co-cultured A-375 and HaCaT cells were measured before and after treatment with stock tomato powder (n=5 replicates). GSH levels were significantly increased in cells treated with tomato powder.

[0070] Figure 5 : The graph shows GSH levels in cells after treatment. Values ​​are mean ± SEM, and * indicates that there is a significant difference in GSH levels between the treated group and the untreated control (p < 0.05).

[0071] Figure 5 The graph shows GSH levels in cells after treatment. Values ​​are mean ± SEM, and * indicates that there is a significant difference in GSH levels between the treated group and the untreated control (p < 0.05).

[0072] The present invention discusses that the original tomato powder rich in carotenoids and flavonoids can be a potent agent to inhibit melanin transfer and enhance glutathione production, thereby helping to improve skin tone and obtain long-term skin care benefits. The optimal dosage range for effective treatment without cytotoxic effects was identified and significant recovery from injury, reduced LDH release, decreased PAR-2 ​​activity, inhibition of melanin transfer and increased GSH levels were shown after treatment with tomato powder.

[0073] In conclusion, the present study on the potential of colorless carotenoids and flavonoids from heirloom white tomatoes for skin cell regeneration highlights the promise of skin care intervention. The study confirmed the non-cytotoxic effects of heirloom white tomato powder and demonstrated its efficacy in mitigating melanin transfer, reducing oxidative stress and promoting skin cell lightening.

Claims

1. A method for skin lightening and cell regeneration, comprising applying an effective amount of a white tomato extract, the white tomato extract being a composition derived from an original species of white tomato; wherein the composition contains carotenoids and flavonoids that inhibit melanin transfer activity and promote skin cell regeneration.

2. The method of claim 1, wherein: The composition is effective in reducing melanin transfer.

3. The method of claim 1, wherein: The composition increases GSH levels in the skin.

4. The method of claim 1, wherein: The composition promotes skin cell regeneration.

5. The method of claim 1, wherein: The composition showed no cytotoxic effects.

6. The method of claim 1, wherein: The optimal dosage of white tomato extract was identified to show significant recovery from skin damage.

7. The method of claim 1, wherein: The composition reduces LDH release, indicating reduced cellular damage.

8. The method of claim 1, wherein: The composition reduces PAR-2 ​​activity, indicating that melanin transfer is inhibited.

9. The method of claim 1, wherein: The carotenoids in white tomato extract are colorless.

10. The method of claim 1, wherein: The composition is specifically derived from heirloom white tomatoes.

11. The method of claim 1 configured to improve skin tone and appearance.

12. A skin care system for improving skin tone and appearance, comprising: White Tomato Extract, a formula derived from heirloom white tomatoes that contains colorless carotenoids and flavonoids; Cytotoxicity assessment module; Damage reduction analysis module; Melanin estimation module; PAR-2 ​​activity assessment module; as well as The GSH level estimation module, wherein the skin care system is configured to reduce melanin transfer, increase GSH levels, and promote skin cell regeneration without producing cytotoxic effects.

13. The skin care system of claim 12, wherein: The cytotoxicity assessment module describes the safety of the composition.

14. The skin care system of claim 12, wherein: The damage reduction analysis module assesses recovery from skin damage.

15. The skin care system of claim 12, wherein: The melanin estimation module measures the melanin level in the skin.

16. The skin care system of claim 12, wherein: The PAR-2 ​​activity assessment module measures the inhibition of melanin transfer.

17. The skin care system of claim 12, wherein: The GSH level estimation module measures an increase in the GSH level in the skin.

18. The skin care system of claim 12, wherein: The composition reduces LDH release, indicating reduced cellular damage.

19. The skin care system of claim 12, wherein: The composition showed no cytotoxic effects.

20. The skin care system of claim 12, configured for skin lightening and cell regeneration, thereby providing a safe and effective solution for improving skin tone and appearance.

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