Introduction
The cosmetics industry is seeing increased demand for high-performance products, beauty foods, fermented cosmetics, and natural ingredient formulations. This trend aligns with a growing focus on healthier lifestyles and enhanced beauty. In response, there’s active development in natural formulations, cellular research, and clinical trials. Popular ingredients like green tea catechins, isoflavones, and curcumin are favored for their skin benefits, with ongoing research exploring new natural ingredients to further improve cosmetics (Lee, 2007).
There is a growing emphasis on eco-friendly wellness trends that prioritize natural ingredients over synthetic ones, driving significant research and development of new materials. Key initiatives include producing cosmetic raw materials from enzymes derived from natural sources and utilizing fermentation techniques with microorganisms for bioconversion, all aimed at promoting sustainability and eco-friendliness in product development (Pérez-Rivero & López-Gómez, 2023).
The fermented cosmetics market in South Korea is growing rapidly, leveraging traditional fermentation techniques from food processing to extract specific components with minimal nutrient loss. These components are then incorporated into cosmetics to improve absorption and effectiveness. Commonly used ingredients include plum, honey, red ginseng, polyphenols, yeast, soybean, moringa leaf, probiotics, and Bacillus-fermented extracts. Meanwhile, industries like food quality enhancement and pharmaceuticals employ microbial protein-degrading enzymes such as Actinidin, Ficin, Bromelain, and Papain— found in fruits like kiwi, pear, pineapple, papaya, and fig— for purposes like meat tenderization. Despite the large scale of the domestic cosmetics market in Korea, it lags behind the pharmaceutical industry in adopting advanced technologies, with more than half of its raw materials being imported. As a result, there is a strong need for ongoing research and development in domestic cosmetic raw materials through biotechnology (Chen et al., 2018).
It plays a vital role in activating the immune system and facilitating wound healing, focusing on the damaged area and utilizing advanced defense mechanisms to restore it to its normal state. This process is crucial in the occurrence of various diseases (Strbo et al., 2014). Mast cells, when activated by external invaders during inflammatory responses, produce a range of inflammatory mediators that create defense mechanisms to eliminate antigens effectively. However, if these mediators continue to increase, they can lead to chronic inflammatory skin conditions. Therefore, it is essential to suppress the inflammatory response that generates these mediators (Lundberg, 2000). Antiinflammatory agents that suppress the excessive activity of inflammatory mediators have been extensively studied for treating chronic inflammatory skin conditions. However, research on skin side effects and resistance remains limited, highlighting the need for substantial efforts to develop natural substances with anti-inflammatory properties that can effectively address various inflammatory conditions (Ginwala et al., 2019).
As a result, ongoing research efforts, both domestically and internationally, are focused on discovering materials through the bioconversion of natural and plantbased sources. However, studies on bioconverting plant materials like the bark of Picrasma quassioides using plant-derived enzymes from pineapple and kiwi remain limited. Additionally, research on natural substances with anti-inflammatory effects, which could address various inflammatory conditions, is also underexplored. Therefore, this study aims to investigate the enhanced anti-inflammatory properties of bioconverted Picrasma quassioides bark using enzymes derived from pineapple and kiwi.
Picrasma quassioides, widely found in Korea, Taiwan, Japan, China, and India, belongs to the dicotyledonous plant order Sapindales and the family Simaroubaceae. It is a deciduous hardwood tree that grows up to about 10 meters in height, with leaves less than 10 cm long, arranged in feather-like clusters of 10-14 leaflets. The tree produces greenish-yellow flowers, approximately 5-8 mm in diameter, that typically bloom separately as male and female flowers in June. Male flowers have around five divided stamens, while female flowers contain about five pistils. Known for its resilience to dryness and cold, Picrasma quassioides has dense, hard wood commonly referred to as “bitterwood”, Traditionally, it has been used to treat tonsillitis, pharyngitis, and eczema (Mohd Jamil et al., 2020). Traditionally, the woody parts of Picrasma quassioides are harvested, dried, and then commonly used. In China, the stems and root bark from Picrasma quassioides, known as “high tree bark”, are believed to have effects similar to those of bitterwood. Picrasma quassioides contains carbohydrates, fat, and ash, with notably higher levels of protein and fat compared to other plants. The main components of Picrasma quassioides include quassinoids, tirucallanes, ionones, and alkaloids. The fruits of Picrasma quassioides are rich in compounds such as arbutin, phlorin, koaburaside, syringin, citrusin B, cnidioside B, flavaprenin 7,4-diglucoside, phenylpropanoids, and phenolic compounds (Yoshikawa et al., 1995). The Simaroubaceae family, to which Picrasma quassioides belongs, is known for its anticancer activity and immunemodulating effects, as well as its anti-inflammatory, antioxidant, anti-hypertensive, and detoxifying properties.
Bioconversion is a technology that uses microorganisms, enzymes, and biological agents to transform precursor substances into desired products or enhance specific components. This process includes bioprocessing, biocatalysis, and biosynthesis, aiming to improve ingredient efficacy and convert prodrugs into active drugs with better bioavailability (Bae et al., 2004). At its core, bioconversion leverages the functions of living organisms to create new bioproducts or replace traditional chemical processes with biological alternatives. It’s an innovative and environmentally friendly technology designed for modern needs (Willke & Vorlop, 2004).
Traditional fermentation has long relied on simple raw materials to create products, but bioconversion takes a different approach. By harnessing the power of microorganisms and enzymes, bioconversion transforms precursor substances into entirely new compounds. This advanced and energy-efficient technology has the potential to revolutionize industries such as pharmaceuticals, cosmetics, and food development, leading to more effective and innovative solutions.
In this study, we focus on Picrasma quassioides, exploring whether bioconversion can unlock its potential as a functional cosmetic ingredient. Through this process, we aim to discover new ways to enhance the effectiveness of cosmetics, pushing the boundaries of what’s possible in the field.
Methods
1. Picrasma quassioides, kiwi, pineapple
The Picrasma quassioides used in the experiment was bought in dried form from an herbal market in Jecheon, Chungbuk. To eliminate foreign substances and impurities, the bark was washed three times with cold water, then dried with hot air at 60℃, and ground into a powder using a mixer. Pineapple (from the Philippines) and kiwi (from Jeju Island), which were used as sources of plant enzymes, were purchased from a major domestic store. The skins of the pineapple and kiwi were removed, cut into suitable sizes, ground with a mixer, and then homogenized for use. The peony root bark extract used as a control (from Jecheon, Chungbuk) was prepared using the same method as Picrasma quassioides, with the exception of pineapple and kiwi.
2. Reagents and equipments
The extraction process utilized a HEATING MANTLE (USA, MS-DMB605) and a LAB STIRRER (USA, MS5060). The Sandwich ELISA Kit, obtained from Cell Signaling Technology (USA), targeted the following proteins: p-p38 alpha (Thr180/Tyr182), p-Akt-1 (Ser473), p-SAPK/JNK (Thr183/Tyr185), p-MEK1/2 (Ser217/221), p-Stat3 (Tyr705), p-NF-κB, and p-IκB-α (Ser32). Fetal bovine serum (FBS) from Sigma (USA), DMEM medium from Sigma (USA), and Hygromycin B from Roche (Germany) were used. Additional materials included Antibiotic-Antimycotic from Sigma (USA), MTS samples from Promega using the CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay kit (Cat. NO. G5421), and filter paper from Whatman International Ltd., Maidstone (England). All other reagents were of premium or first-grade quality.
3. Bioconversion conditions
Preparation of Picrasma quassioides Bark enzyme reaction mixture (B-P.Q): The enzyme reaction mixture was prepared by blending 100 g of pulverized Picrasma quassioides bark powder with 10-100 g of pulverized kiwi (1-10% in water) and 10-100 g of pulverized pineapple (1-10% in water). NF-κB inhibition experiments were conducted to determine the optimal reaction ratio and duration for these ingredients. It was found that the ideal conditions for biological transformation were 10% Picrasma quassioides bark extract combined with 2% kiwi and 2% pineapple for 7 days. The reaction mixture was then filtered and freeze-dried to obtain B-P.Q for further use. For the Picrasma quassioides extract (P.Q), which included the same proportions of Picrasma quassioides bark, kiwi, and pineapple, the mixture was not incubated for 7 days. Instead, the filtrate was directly frozen and freeze-dried.
4. Cell culture conditions
The NIH 3T3 fibroblast cell line, derived from mice and obtained from Signosis, was seeded in 75T flasks and cultured at 37℃ with 5% CO2. The culture medium consisted of Dulbecco’s Modified Eagle’s Medium (DMEM; Sigma, USA) supplemented with 10% Fetal Bovine Serum (FBS, Sigma, USA), 1% Antibiotic-Antimycotic (Sigma, USA), and 1% Growth Supplement. The medium was changed every 3 days. Cells were passaged when they reached approximately 80-90% confluency, and experiments were performed using cells that had undergone at least one passage to ensure stability.
5. MTS assay
The NIH 3T3 cell line was seeded into 96-well plates at a density of 4×104 cells/mL in 200 μL per well and incubated at 37℃ with 5% CO2 for 24 hours to stabilize. After stabilization, samples were treated with concentrations ranging from 1 mg/mL to 5 μg/mL for 72 hours (3 days) at 37℃ under 5% CO2. PBS, used in the same volume as the samples, served as the control. The MTS assay was conducted using the CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay Kit (Cat. No. G5421) from Promega. MTS solution mixed with PMS solution was added to each well at 20 μL per well and incubated for 1 hour at 37℃ with 5% CO2. Absorbance was then measured at 492 nm using an ELISA reader (Molecular Devices, USA).
6. Measurement of NF-κB
We used the NF-κB Luciferase reporter NIH 3T3 stable cell line from Signosis. The cells were cultured in DMEM media with Hygromycin (100 μg/mL, Sigma) at 37℃ with 5% CO2 to maintain vector integrity. Cells were counted to a density of 5×105 cells/mL, and 100 μL were seeded into 96-well plates and incubated for 24 hours. The samples were then treated as specified. After 12 hours, cells were stimulated with 20 μg/mL TNF-α and 10 μg/mL IL-1β, and harvested 8 hours later. To measure NF-κB activity, cell culture supernatants were removed, and cells were lysed with lysis buffer to extract intracellular Luciferase. Luciferase activity was assessed using a Luminoskan Ascent luminometer (Thermo Electron, USA).
7. Measurement of NF-κB-related signaling proteins activity
The mouse-derived fibroblast cell line 3T3 was used for measuring NF-κB-related cytokines. Cells were seeded at a density of 5×105 cells/mL in 96-well plates, with 100 μL per well, and incubated for 24 hours. After this period, samples were treated. TNF-α (20 μg/mL) and IL-1β (10 ng/mL) were added to induce NF-κB and related signaling pathways. NF-κB-related cytokines were measured at 5, 10, 20, 40, and 80 minutes using a Sandwich ELISA kit (Cell Signaling, USA), as detailed in Table 1, to assess the activity of phosphorylated signaling proteins. The experiment was conducted following the instructions provided in the manual.
8. CAM (Chorioallantoic Membrane) Assay
To assess the ability to inhibit angiogenesis, we used fertilized eggs (Pulmuone, Korea) purchased from a local agricultural cooperative in Wonju, all less than two days post-hatch. A fertilized egg is identified by visible blood vessels and a heartbeat when a window is created. After acquisition, the eggs were incubated at 37℃ with 80-90% humidity. Three days later, a small hole was made at the narrow end of the eggshell membrane, and 4-5 mL of albumin was withdrawn using a 5 mL syringe. The puncture site was sealed to prevent infection and drying, and the eggs were re-incubated with the hole facing downward. Four days later, a 2-3 cm diameter window was created at the air sac end of the egg. Only those eggs confirmed as fertilized were sealed with cellophane tape and re-incubated. Five days after this, when the CAM (chorioallantoic membrane) region had developed and was approximately 2-5 mm in diameter, 5 μg samples were applied to quartered Thermanox coverslips and air-dried overnight in a clean bench to avoid contamination. The tape covering the window was removed, the CAM region was identified, and the Thermanox coverslip with the sample was placed on the CAM region using tweezers. The window was then resealed with cellophane tape. After seven days, the tape was removed, and 1 mL of Intralipose (fat emulsion) was drawn into a syringe, air bubbles were removed, and it was injected just below the CAM to highlight blood vessels against a white background. Angiogenesis inhibition was observed and documented with close-up photographs using a digital camera (Auerbach et al., 2003).
9. Tube Formation (HUVECs Differentiation) Assay
Human umbilical vein endothelial cells (HUVECs) used in the experiment were obtained from MCTT Co., Ltd. The primary cultured cells were grown in plates coated with 0.1% gelatin, using endothelial cell medium (Innoprot, ES) supplemented with 3 ng/mL βFGF, 5 units/mL heparin, and 20% FBS, and were maintained under 5% CO2 at 37℃. The tube formation assay, which assesses HUVEC differentiation, followed a modified method based on Grant et al. (Grant, Kinsella et al. 1992). BD MatrigelTM Basement Membrane Matrix (BD Biosciences, Cat. No. 354230) was applied to a 24-well plate, 100 μL per well, and allowed to coat at 37℃ for 30 minutes. The endothelial cells were then mixed with endothelial cell medium (Innoprot, ES) containing 5% FBS, 2 ng/mL βFGF, and 2 ng/mL EGF, and 450 μL of this mixture was added to each well to achieve a density of 3.6×104 cells per well. The B-P.Q ingredient was added to achieve a final concentration of 50 μg/mL, while the control group received medium containing only 5% FBS. The cells were incubated at 37℃ with 5% CO2 for 18-24 hours, and tube formation was observed under a microscope (Olympus IX53, 100x magnification).
10. HUVECs Cell Migration Inhibition Assay
This experiment used a modified method based on Jang et al (Jiang et al., 2017). HUVECs were seeded at a density of 1×105 cells per well in a 6-well plate and incubated at 37℃ with 5% CO2 for 15 hours to allow stabilization. After 15 hours, a scratch was made on the cell monolayer using a yellow tip. The cells were then washed with PBS and replaced with fresh medium. The control group was provided with medium containing 5% FBS, while the other groups received 180 μL of medium containing 5% FBS, 2 ng/mL βFGF, and 2 ng/mL EGF. Each sample was treated by adding 20 μL to achieve the desired concentration. The cells were incubated at 37℃ with 5% CO2 for 9 hours, then stained with 0.2% crystal violet solution. The extent of cell migration was observed and photographed under a microscope (Olympus IX53, 100x magnification).
Result
1. Cell cytotoxicity and NF-κB inhibition
To evaluate the effects of bioconverted Picrasma quassioides (B-P.Q) extract, untreated Picrasma quassioides extract (P.Q), and peony root bark extract (BOT)—which is recognized for its potent anti-inflammatory properties (Ekiert et al., 2022)—on NIH 3T3 fibroblast cell viability, we treated the cells with these extracts at various concentrations and assessed their viability. We tested B-P. Q and P.Q at concentrations of 50 and 500 μg/mL, and BOT at 50 μg/mL, with none showing cytotoxic effects. Therefore, further experiments were conducted using each extract at 50 μg/mL.
Based on the optimal response ratio identified from NF-κB activity inhibition experiments (which involved adding 2% kiwi and 2% pineapple to 10% Picrasma quassioides bark extract), we investigated the ideal reaction time. The extracts were reacted for 0, 1, 3, 7, and 14 days, and each mixture was tested on 3T3 cells to assess changes in NF-κB activity. As shown in Figure 1, the mixture reacted with sophora bark extract for 7 days demonstrated the highest NF-κB activity inhibition, surpassing even the peony root bark extract (BOT), which is recognized for its anti-inflammatory effects through NF-κB signaling pathway inhibition. Consequently, the optimal conditions for biological transformation were determined to be 10% Picrasma quassioides bark extract combined with 2% kiwi and 2% pineapple for 7 days. The resulting products were then filtered and freeze-dried (B-P.Q) for further use.
2. Measurement of NF-κB related signaling Protein Activation
To verify the anti-inflammatory effects of bioconverted Picrasma quassioides extract (B-P.Q), we measured the activation levels of NF-κB-related signaling proteins, such as p-p38, p-Akt-1, SAPK/JNK, and MEK1/2, which are involved in the inflammatory response. Inflammation was induced by treating with TNF-α and IL-1β. Preliminary experiments identified the peak of each signal at specific times, and the inhibition rates of each substance were confirmed. Changes in the activation of NF-κB-related signaling proteins were measured using the Sandwich ELISA kit (Cell Signaling, USA) according to the manual specified in the materials and methods section.
1) p-p38
p-p38 is a type of p38 MAP kinase with four isoforms: p38α, β, γ (ERK6, SAPK3), and δ (SAPK4). It is known to be activated by various intracellular stresses, including inflammatory cytokines, UV radiation, LPS, and growth factors (Raingeaud et al., 1995). Maximum activation of p-p38 is observed about 20 minutes after treating cells with TNF-α and IL-1β. Using this time frame, the effectiveness of each substance was assessed, as illustrated in Figure 2A. The group was treated only with TNF-α and IL-1β, using PBS as a substitute for the sample (positive control, P.C.) showed a 16.05-fold increase in p-p38 activation compared to the control group (N.C.). BOT reduced activation by 10.53% compared to the treatment group, while P.Q., without enzyme treatment, reduced it by 24.54%. B-P.Q., treated with the enzyme, achieved a 27.24% reduction, with significant differences between the enzyme-treated and untreated groups (p<0.05). This suggests that B-P.Q. may offer superior efficacy in reducing inflammatory responses, which is crucial for minimizing redness and irritation in cosmetic applications.
2) p-Akt-1
Akt, which influences IKK-α, β, and γ, is phosphorylated into p-Akt and plays a key role in activating NF-κB (Kim & Chung, 2002). The peak activation of p-Akt-1 occurs around 40 minutes after cells are exposed to TNF-α and IL-1β. At this time point, the efficacy of each substance was assessed, as shown in Figure 2B. The treatment group (P.C.) demonstrated a 2.57-fold increase in p-Akt-1 activation compared to the control group (N.C.). BOT reduced p-Akt-1 activation by 25.42% compared to the treatment group, while P.Q., which was not enzyme-treated, showed a 20.73% reduction. B-P.Q., treated with the enzyme, achieved a 31.28% reduction in activation. However, the difference in efficacy between enzyme-treated and untreated groups was not statistically significant. From a cosmetic perspective, the ability to reduce p-Akt-1 activation can contribute to improved skin appearance by potentially mitigating inflammatory responses and supporting overall skin health.
3) p-SAPK/JNK
SAPK/JNK, activated by environmental stress and various stimuli, plays a role in the activation of NF-κB and AP-1 (Chen et al., 2021). It operates through mechanisms triggered by stresses such as UV, mediated by MAPKs like ERK and JNK (Yang et al., 2018).
Cells exposed to TNF-α and IL-1β exhibit peak activation of p-SAPK/JNK at 20 minutes post-stimulation. Figure 2C displays the effectiveness of each substance at this time point. The treatment group (P.C.) showed a 17.26-fold increase in p-SAPK/JNK activation compared to the control group (N.C.). BOT reduced p-SAPK/JNK activation by 41.25% relative to the treatment group, while P.Q., which was not enzyme-treated, decreased activation by 36.62%. B-P.Q., which underwent enzyme treatment, achieved a 37.76% reduction. However, the difference in effectiveness between enzyme-treated and untreated groups was not statistically significant. From a cosmetic perspective, these results highlight the potential of these substances in modulating inflammatory pathways, which is crucial for improving skin conditions characterized by redness and irritation.
4) p-MEK1/2
MEK1 and MEK2, also known as MAPK or Erk kinases, are pivotal in signaling pathways that regulate cell growth and differentiation. Activated by various growth factors and cytokines, MEK1/2 play a crucial role in processes triggered by membrane depolarization and calcium influx (Alessi et al., 1994).
Cells exposed to TNF-α and IL-1β reach peak activation of p-MEK1/2 at 5 minutes after stimulation. Figure 2D illustrates the efficacy of each substance at this time point. The treatment group (P.C.) showed a 3.96-fold increase in p-MEK1/2 activation compared to the control group (N.C.). BOT decreased p-MEK1/2 activation by 15.01% relative to the treatment group, while P.Q., which was not enzyme-treated, reduced activation by 14.09%. B-P.Q., treated with the enzyme, resulted in a 9.44% reduction. Although B-P. Q.’s inhibitory effect was not prominent at the 5-minute mark, significant inhibition was observed from 10 to 80 minutes. From a cosmetic perspective, this indicates that B-P.Q may offer a delayed yet effective reduction in signaling pathways associated with inflammation, which is crucial for addressing issues such as skin redness and irritation.
5) p-iκB
TNF-α and IL-1β can trigger the phosphorylation of IκB (inhibitor of NF-κB), leading to its degradation. This event is a crucial step in the NF-κB signaling pathway, as phosphorylated IκB allows NF-κB to move into the nucleus, where it activates genes involved in inflammation, immune responses, and cell survival (Sivamaruthi et al., 2023). Therefore, inhibiting the phosphorylation of IκB effectively means blocking the NF-κB signaling pathway.
Cells exposed to TNF-α and IL-1β reached peak activation of p-IκB at 5 minutes after stimulation. Figure 3A demonstrates how each substance performed at this critical time point. The treatment group (P.C.) showed a 16.66-fold increase in activation compared to the control group (N.C.). The BOT reduced this activation by 20.10% relative to the treatment group, while P.Q., without enzyme treatment, achieved a 28.68% reduction. Enzyme-treated B-P.Q. achieved a 42.91% reduction, although the difference in effectiveness between enzyme-treated and untreated groups did not reach statistical significance. This data suggests that B-P.Q. may have notable potential for reducing inflammatory responses related to skin conditions, which is significant for cosmetic applications targeting redness and inflammation.
6) p-Stat3
Stat3 is a key signaling protein frequently elevated in tumor sites, highlighting its role as an oncogene and anti-apoptotic factor. Beyond its involvement in oncogene regulation, Stat3 is also crucial for cellular defense against primary inflammatory responses (Yu et al., 2009). In the NF-κB signaling context, inhibiting Stat3 is considered an important strategy for managing inflammatory responses and targeting chronic skin conditions like psoriasis (Andrés et al., 2013). In this experimental model (Figure 3B), while no significant changes were observed across the groups, B-P.Q. generally showed lower values compared to others.
Other cytokines and transcription factors:
NF-κB is made up of five protein types: the p50 subunit family (p50, p52) and the p65 subunit family (p65, c-Rel, RelB). These proteins can form either homodimers (such as p50-p50 or p65-p65) or heterodimers (such as p50-p65 or p50-Rel). Typically, NF-κB is composed of p50-p65 heterodimers. Therefore, changes in NF-κB activity are more influenced by the location of p65 rather than its quantity. As shown in Figure 3C, the amount of p65 remained consistent, indicating no significant changes. Similarly, Figures 3D show that the levels of Akt-1 was stable throughout the experimental period. This series of results suggests that the NF-κB-related experiments were conducted accurately, providing reliable insights into their potential cosmetic applications.
3. Chorioallantoic membrane assay results
The chorioallantoic membrane (CAM), which rapidly develops new blood vessels during the embryonic development of fertilized eggs, is utilized in CAM assays to evaluate angiogenesis inhibition. In this widely used method, the CAM acts as the site for assessing new blood vessel formation. To confirm normal vascular conditions, a cover slip is placed on the CAM without any substances, and the area is observed after 3 days (Ribatti, 2008). Substances that inhibit new blood vessel formation in the CAM area are reported to have potential as treatments for chronic skin inflammation. From a cosmetic perspective, reducing inflammation is highly important (Auerbach et al., 2003). Recent studies have confirmed the antiangiogenic effects of seven natural substances, including garlic, ginseng, and rosemary, while other substances like turmeric have been reported to promote blood vessel formation (Kim et al., 2008). These compounds are used in cosmetics for their important roles as anti-inflammatory agents and for reducing redness. Research into the role of natural substances in inhibiting new blood vessel formation is ongoing. In this study, we used CAM assays to compare the anti-angiogenic effects of bioconverted B-P.Q and non-bioconverted P.Q extracts to assess their potential for treating chronic inflammatory skin conditions. Both extracts were tested at a concentration of 50 μg/mL, similar to the conditions used in NF-κB-related signaling protein assays. As shown in Figure 4, while P.Q inhibited angiogenesis compared to the control group treated with PBS, B-P.Q demonstrated a significantly greater suppression of new blood vessel formation. This confirms the potent antiangiogenic effect of B-P.Q.. The anti-angiogenic effect is closely linked to reducing skin redness, making it highly significant from a cosmetic perspective.
4. Tube formation inhibition assay results
Endothelial cells (HUVECs) proliferate and grow invasively to form inflammatory tissues, differentiate into endothelial cells, and ultimately form blood vessels (Kocherova et al., 2019). To evaluate the anti-inflammatory effects of various substances, we performed a tube formation assay to assess endothelial cell differentiation. HUVECs were treated with angiogenic factors βFGF and EGF, and B-P.Q. was applied at a concentration of 50 μg/mL under identical conditions. As depicted in Figure 5, the bioconverted Picrasma quassioides extract (B-P.Q.) effectively inhibited tube formation. This finding confirms B-P.Q.’s anti-angiogenic properties, which are highly relevant for reducing redness and irritation in cosmetic formulations.
5. HUVECs cell migration inhibition assay results
HUVECs, which are derived from umbilical cord veins, play a key role in migrating to sites where new blood vessels form or existing vessels are damaged. Inhibiting their migration can be beneficial for treating chronic inflammatory skin conditions like psoriasis and atopic dermatitis(García-Caballero et al., 2011), as these diseases often involve excessive angiogenesis. Therefore, controlling angiogenesis and cell migration is crucial for managing such conditions.
To investigate the potential benefits of bioconverted Picrasma quassioides extract (B-P.Q.), we conducted an experiment to assess its effect on endothelial cell migration. Using human umbilical vein endothelial cells (HUVECs) stimulated with βFGF (2 ng/mL) and EGF (2 ng/mL), we treated the cells with B-P.Q. at a concentration of 50 μg/mL. The results, illustrated in Figure 6, showed that B-P. Q. effectively inhibited the migration of these cells, which is a crucial step in new blood vessel formation. This finding suggests that B-P.Q. could be highly beneficial in cosmetics for reducing excessive blood vessel formation, which is often associated with redness and irritation. By incorporating B-P.Q. into skincare formulations, it may improve the appearance of chronic inflammatory conditions, making it a valuable ingredient for enhancing skin health and reducing unwanted redness.
Discussion
The study provides valuable insights into the potential applications of bioconverted Picrasma quassioides extract (B-P.Q.) for managing skin inflammation and improving skin conditions.
Key Points for Cosmetic Applications:
1. Anti-Inflammatory Effects: NF-κB-Related Signaling Proteins: The study reveals that B-P.Q. significantly reduces the activation of key inflammatory signaling proteins such as p-p38, p-Akt-1, p-SAPK/JNK, and p-IκB compared to other treatments. In cosmetics, reducing these inflammatory markers can help minimize skin redness and irritation, leading to a more even and calm complexion (Seo et al., 2014). While both BOT and P.Q. show some anti-inflammatory effects, B-P.Q. generally demonstrates superior efficacy. This suggests that B-P.Q. might be more effective in formulations aimed at reducing inflammation and redness. Anti-inflammatory effects are not limited to this study. For instance, traditional Chinese medicines that inhibit p38 also have melanin production suppression capabilities (Tsang et al., 2012). Additionally, inhibiting p-SAPK/JNK is highly effective in combating wrinkles. For instance, JNK/SAPK signaling is crucial for the efficient reprogramming of human fibroblasts (Neganova et al., 2016). This suggests that anti-inflammatory effects can be utilized to address various skin issues.
2. Anti-Angiogenic Properties: B-P.Q. shows a greater suppression of new blood vessel formation compared to P.Q., indicating a strong anti-angiogenic effect. This is particularly relevant for reducing visible redness and inflammation in cosmetic products, as excessive blood vessel formation can exacerbate these issues (Seo et al., 2014). B-P.Q. inhibits tube formation and cell migration in endothelial cells, which is beneficial for managing chronic inflammatory skin conditions like psoriasis and atopic dermatitis. By controlling angiogenesis and cell migration, B-P.Q. could help in reducing the appearance of redness and irritation in cosmetic applications. Genistein was the most potent inhibitor of angiogenesis among the isoflavone compounds tested (Su et al., 2005). Genistein suppresses wrinkles and inflammation caused by UV exposure (Tang et al., 2022), so these effects are closely interconnected.
Practical Implications for Cosmetic Formulations:
1. Redness and Irritation Control: Incorporating B-P. Q. into cosmetic products could provide anti-inflammatory benefits, helping to reduce skin redness and irritation. This would be valuable in products designed for sensitive skin or those targeting conditions like rosacea. Chronic inflammation, a hallmark of rosacea, can cause a variety of symptoms such as flushing, telangiectasia, inflammatory papules and pustules, and ocular manifestations. Therefore, managing rosacea is critically important from an aesthetic perspective (Paiva-Santos et al., 2023).
2. Overall Skin Health: The ability of B-P.Q. to modulate inflammatory pathways and inhibit angiogenesis suggests it could support overall skin health, making it a promising ingredient for skincare lines focused on calming and protecting the skin. Hesperidin is a citrus flavanone glycoside with strong anti-inflammatory properties that inhibits UVB-induced angiogenesis in the skin (Kim et al., 2021). And acts as a promising skincare bioactive (Rodrigues & Pintado, 2024). Hesperidin can counteract UV effects by reducing epidermal thickening, regulating angiogenesisrelated factors (such as PI3K/Akt, VEGF, HIF-1α, and MMPs), modulating apoptotic proteins triggered by oxidative stress, and affecting immune cells and inflammatory cytokines, which helps decrease inflammation.
3. Potential for New Products: The data supports the development of new cosmetic products featuring B-P.Q. as a key ingredient. These products could be marketed for their advanced anti-inflammatory and anti-angiogenic properties, potentially differentiating them from existing offerings in the market. Plants with anti-inflammatory and anti-angiogenic properties sometimes have skin whitening effects (Jung et al., 2013). Plants with both anti-inflammatory and anti-angiogenic properties not only contribute to skin whitening but are also valuable for overall skin health. Unlike simple anti-inflammatory agents, angiogenesis inhibitors can significantly enhance skin beauty by preventing the formation of new blood vessels that contribute to chronic, inflammatory skin conditions (Lee et al., 2021). Since such conditions negatively impact skin appearance, these ingredients are likely to play a key role in improving aesthetic outcomes and promoting a healthier, more beautiful complexion.
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