Search Results (684)

This study assessed the impact of biofertilisers on primary and secondary (specialised) metabolites in six lettuce cultivars (‘Kiribati’, ‘Murai’, ‘Aquino’, ‘Gaugin’, ‘Aleppo’, and ‘Carmesi’) grown in anthropogenic soil during an autumn greenhouse experiment. Four treatments were tested: control (no fertilisation), effective microorganisms (EM), Trichoderma, and their combination. Red cultivars showed higher total antioxidant capacity (TAC) and total phenolic content (TPC), with red Lollo ‘Carmesi’ having the highest TAC, TPC, carotenoids, total soluble solids, sweetness index, and sugars. Red Oak ‘Murai’ exhibited the highest chlorophyll b and total chlorophyll, while green cultivars ‘Kiribati’ and ‘Aquino’ excelled in chlorophyll a and overall taste, respectively. Biofertilisers did not affect TAC or most chlorophyll types but increased TPC (EM by 18.6% and combined treatment by 19.6%) and chlorophyll a (EM by 28.6% and Trichoderma by 23.8%). Combined fertilisers improved taste with reduced glucose and fructose content and sweetness index, though sucrose remained unchanged compared to the control. Major organic acids (malic, citric, and tartaric) were most abundant in ‘Murai’ and ‘Kiribati’, unaffected by treatments. Phenolics content peaked in ‘Murai’ and ‘Carmesi’, characterised by chicoric and chlorogenic acid, caffeoylquinic acid glucoside, and flavonoids (quercetin derivatives, kaempferol); biofertilisers mainly influenced gallic acid, while kaempferol was affected by all biofertilisers and increased in the range of 12.5–25%. The key sesquiterpene lactones identified were lactucin, lactucopicrin-15-oxalate, and 11β,13-dihydrolactucin. The content of sesquiterpene lactones analysed in this study increased significantly, especially with EM treatment (14.7–185.7%) and combined fertilisers (12.5–128.6%), highlighting the lactone-rich cultivars ‘Carmesi’ and ‘Gaugin’. Red cultivars ‘Carmesi’ and ‘Murai’ exhibited the most favourable phytochemical profiles, suitable for cultivation and processing of quality-based products. In contrast, the green cultivar ‘Aquino’ received the highest sensory scores, delivering the most appealing overall taste despite its lower metabolite content. EM treatment and combined fertilisers are recommended for increasing chlorophyll a, myo-inositol, TSS, propionic acid, TPC, kaempferol, and major lactones under greenhouse autumn conditions. Full article

Waste orange peels (WOP) are a major orange processing residue, and they may be a rich source of precious bioactive polyphenols. Amongst the various WOP constituents, hesperidin holds a prominent position as the most abundant polyphenolic metabolite, with proven biological properties. The current work was performed to provide detailed information on the effect of various acid catalysts to assist hesperidin recovery, using an ethanol organosolv treatment. The treatment developed was first examined by comparing inorganic (HCl) and natural organic (oxalic, citric) acids for their influence on process performance, extraction kinetics, and severity. Following this, optimization was accomplished through response surface methodology, and the extracts produced were investigated with respect to their polyphenolic composition and antioxidant characteristics. The HCl-catalyzed treatment, carried out with 70% ethanol/2% HCl, was proven the most efficacious, giving a total polyphenol yield of 30.7 mg gallic acid equivalents per g of dry mass, and it was shown that the treatment yield was related to severity, obeying a power model. Liquid chromatography–tandem mass spectrometry analysis of the extract generated under optimized conditions (170 min, 80 °C) revealed that hesperidin was extensively hydrolyzed into hesperetin 7-O-glucoside and aglycone (hesperetin). Such an effect was very limited with the oxalic acid-catalyzed treatment, whereas citric acid did not affect the original polyphenolic composition. Overall, the HCl-catalyzed treatment was of significantly higher performance, providing a total flavanone yield of 21.22 mg per g dry mass. The results of this investigation may be of value in adjusting treatment settings for (i) increased flavonoid recovery from WOP and (ii) producing extracts enriched in hesperidin and/or its hydrolysis derivatives. Such practical recommendations may assist the establishment of WOP valorization processes in an integrated biorefinery prospect. Full article

Red walnuts have been widely studied because of their strong antioxidant activity and ornamental value. However, research on the mechanism of anthocyanin biosynthesis in walnuts remains in the initial stage. The regulatory mechanism of TT2-type R2R3-MYB transcription factors in anthocyanin biosynthesis in walnuts is also unclear. Therefore, this study used ‘D2-1’ and ‘Jinghong 1’ walnuts as plant materials. The testa of ‘Jinghong 1’ was red, and its anthocyanin content was significantly higher than that of ‘D2-1’, mainly composed of cyanidin-3-O-arabinoside, cyanidin-3-O-galactoside, and cyanidin-3-O-glucoside. Differentially expressed genes between ‘D2-1’ and ‘Jinghong 1’ testa were enriched in phenylpropanoid biosynthesis and flavonoid biosynthesis. Next, this study identified a TT2-type R2R3-MYB transcription factor JrMYB1L, which was involved in regulating the anthocyanin biosynthesis in the testa of ‘Jinghong 1’. The overexpression of JrMYB1L could promote anthocyanin accumulation in walnut leaves and activate the expression of JrCHS, JrCHI, JrF3H, JrDFR, JrANS, JrUFGT, JrLAR, and JrANR. In addition, yeast two-hybrid results proved that JrMYB1L, JrbHLH42, and JrWD40 proteins could interact with each other. The results of yeast one-hybrid and dual-luciferase assays indicated that JrMYB1L could activate the expression of JrCHS and JrUFGT by binding to their promoters. Based on the above results, this study proposed a possible regulatory mechanism. JrMYB1L activated the expression of JrCHS and JrUFGT in the form of JrMYB1L-JrbHLH42-JrWD40 complex, thereby promoting anthocyanin accumulation in the testa of ‘Jinghong 1’. In summary, this study lays a theoretical foundation for revealing the regulatory mechanism of anthocyanin biosynthesis in red walnut and contributes to the breeding of new varieties of red walnuts with more edible and ornamental value. Full article

The purpose of this study was to present a chemical analysis of the metabolome of cell aggregates of the tree fern Cyathea smithii (J.D. Hooker) cell suspension culture. The LC/MS and GC/MS techniques were used for identification of metabolites. The kinetics of fresh weight, dry weight, and ash content showed 3.5-fold increases during 15-day-long culture. The analysis demonstrated high metabolic activity of cultured cells. In total, 160 metabolites from primary and secondary metabolism and almost 2000 compounds of unknown identity were identified. Three flavonoids—the chalcone isookanin [(2S)-2-(3,4-dihydroxyphenyl)-7,8-dihydroxy-2,3-dihydrochromen-4-one], a methoxy derivative of the flavone gardenin B (5-Hydroxy-2-(4-methoxyphenyl)-6,7,8-trimethoxy-4H-1-benzopyran-4-one), and the isoflavone tectoridin (4′,5-Dihydro-6-methoxy-7-(O-glucoside)isoflavone)—had not been previously detected in the cell culture of C. smithii. Principal component analysis revealed five distinct groups of samples; groups 4 and 5 showed the greatest similarity and corresponded to cultures on days 12 and 15, respectively. The number of differentiating compounds was 75, indicated by a heatmap showing positive and negative correlations between the days of culture. The studies described in this paper are crucial for further identification of metabolites and establishing the relationship between the metabolic composition of tree fern cells in culture and their biological activity, assessed by physiological parameters. By determining the relationship between the chemical composition of cells and their growth from culture initiation to senescence, we will provide a more complete picture of the potential for environmental factors to regulate this relationship. Based on previous studies, environmental stimuli such as electromagnetic fields or light of different wavelengths can result in altered growth physiology and cell mass, as well as metabolite diversification and accumulation. The research results presented in this paper provide a foundation for further studies aimed at predicting and regulating the productivity of C. smithii cells in suspension culture and elucidating the significance of tree fern-derived metabolic products in human cell biology, particularly in thyroid cells. Full article

Horseradish (Armoracia rusticana L.) root (HRP) and leaf (HLP) pomaces, by-products of juice production by cold-pressing, were analyzed as a novel potential source of natural antioxidants. Chromatography analysis (UHPLC Q-ToF MS) of the bioactive compounds of pomaces was performed along with spectrophotometric determination of total phenolic content (TPC), total flavonoid content (TFC), total phenolic acid (hydroxycinnamic) content (TPAC), and antioxidant capacity (via 1,1-diphenyl-2-picrylhydrazyl (DPPH^•) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic-acid) (ABTS^•+) radicals’ scavenging activity and ferric reducing antioxidant power (FRAP)). The concentrations of TPC, TFC, and TPAC differed among the pomaces, significantly favoring HLP. However, both horseradish pomaces (HRP and HLP) contained a considerable amount of various phenolics, with kaempferol and its glucosides dominating. In addition, they exhibit pronounced antioxidant activity, which is confirmed by all three methods used (DPPH, ABTS, and FRAP). These results highlight the potential of valorizing horseradish processing waste as a natural, reliable source of health-promoting bioactive compounds and functional ingredients in food products, thereby fortifying food, preventing oxidation, and prolonging shelf-life. In addition, this study supports endeavors to reduce food waste by providing new insights into the valorization of horseradish pomace, thus contributing to sustainable development and environmental protection. Full article

Polygonum cuspidatum is a well-known and versatile medicinal plant. In Taiwan, P. cuspidatum is typically found in central mountainous regions. Once acclimated, it can also thrive in flat areas, where it is known as flatland P. cuspidatum. Flatland P. cuspidatum has several advantages over alpine P. cuspidatum; for example, flatland P. cuspidatum grows faster and has larger leaves. This study enhanced the functionality of different parts of flatland P. cuspidatum (flowers, leaves, and rhizomes) by using microwave-assisted extraction (MAE) technology and Box–Behnken response surface methodology. Experiments revealed that the combination of MAE parameters that yielded optimal results was influenced by which plant part was used as input material. Regarding whitening activity, the extracts were ranked as follows: leaf > rhizome > flower. Leaf extracts had higher total flavonoid content, and rhizome extracts had higher total phenolic content. Regarding antiaging activity, the extracts were ranked as follows: rhizome > leaf > flower. The rankings for antimicrobial activity were as follows: leaf > rhizome > flower. Regarding anti-inflammatory activity, the extracts were ranked as follows: flower > rhizome > leaf. The rhizome extract exhibited slight cytotoxicity. UHPLC-UV-Q-TOF-HRMS/MS analysis identified 27, 34, and 37 bioactive compounds in the leaf, rhizome, and flower extracts, respectively. Given the relatively low pharmacological activity observed in the MAE-optimized flower extract, fermentation with Aspergillus oryzae was employed to enhance its efficacy. This process significantly enhanced the extract’s pharmacological properties, including its whitening, antiaging, and antimicrobial properties. Increased levels of 3-O-caffeoylquinic acid, decursin, quercetin, quercitrin, kaempferol, resveratrol, epicatechin gallate, resveratrol-3-O-D-(2-galloyl)-glucopyranoside, resveratrol-4′-O-β-D-glucoside, apigenin, emodin-8-O-(6′-O-malonyl)-glucoside, physcion, emodin, and torachrysone in the fermented flower extract likely contributed to its enhanced pharmacological activities. The results of this study indicate that the newly developed flatland P. cuspidatum extracts can be considered viable substitutes for alpine P. cuspidatum extracts. Full article

This study employed wild sea buckthorn (Hippophae rhamnoides L.) fruits harvested in Qinghai Province as experimental material. Following compositional analysis of their flavonoids, the antibacterial efficacy and mechanistic pathways of flavonoids in sea buckthorn against Helicobacter pylori (H. pylori) were systematically examined through in vitro and animal model experiments. The results showed that the main flavonoids in sea buckthorn were rutin, quercetin-3-O-glucoside, quercetin, catechin, isorhamnetin, kaempferol-3-O-glucoside, kaempferol and epicatechin. Among them, quercetin, isorhamnetin and kaempferol had strong inhibitory activity against H. pylori. In vitro, sea buckthorn flavonoids inhibited the growth of H. pylori through multiple mechanisms: inducing morphological transformation from rod-shaped to spherical bacteria, promoting cell shrinkage and rupture, disrupting the cell membrane to cause leakage of intracellular macromolecules, increasing membrane permeability, and suppressing urease activity. Sea buckthorn flavonoids exert therapeutic effects on H. pylori-infected mice through multiple mechanisms, including the alleviation of gastric mucosal inflammation via the Nuclear Factor KappaB (NF-κB) signaling pathway, the down-regulation of gastrin-17 (GAS17) to suppress gastric acid production, and the up-regulation of epidermal growth factor (EGF) expression to promote gastric mucosal repair and modulate the composition of gastric microbiota. This study systematically elucidated the anti-H. pylori activity and antibacterial mechanisms of flavonoids derived from sea buckthorn fruits, providing a theoretical basis for the screening of natural antibacterial agents from this plant source. Full article

The pomegranate, a widely consumed fruit, produces large quantities of waste, mainly from its peel. Pomegranate peels (PPs) contain high amounts of antioxidant compounds, such as polyphenols, flavonoids, and anthocyanins, which can be isolated from them and used for the benefit of humans and the environment. In the present work, a study of recovery of these compounds by ultrasound-assisted extraction (UAE) was carried out, whose parameters were optimized. The optimal results were a total polyphenol content of 195.55 mg gallic acid equivalents/g, total flavonoid content of 74.78 mg rutin equivalents/g, total anthocyanin content of 992.87 μg cyanidin 3-O-glucoside equivalents/g, and ascorbic acid content of 15.68 mg/g, while the antioxidant activity determined through ferric-reducing antioxidant power and DPPH assays was 2366.89 and 1755.17 μmol ascorbic acid equivalents/g, respectively. In parallel, an artificial intelligence (AI)-based framework was developed to model and predict antioxidant and phytochemical responses from UAE parameters. Six machine learning models were implemented on the experimental dataset, with the Random Forest (RF) regressor consistently achieving the best predictive accuracy. Partial dependence analysis revealed ethanol concentration as the dominant factor influencing outcomes, while ultrasonic power and extraction time exerted comparatively minor effects. Although dataset size limited model generalizability, the RF model reproduced experimental outcomes within experimental variability, underscoring its suitability for predictive extraction optimization. These findings demonstrate the complementary role of machine learning in accelerating antioxidant compound recovery research and its potential to guide future industrial-scale applications of AI-assisted extraction. Full article

This research investigates the use of apple and sour cherry pomace to fortify biscuits, aiming both to improve their nutritional profile and to support the sustainable reuse of fruit processing by-products. Apple and sour cherry pomace, known for their high content of bioactive compounds, were added to biscuit formulations at inclusion levels of 5%, 10%, and 15%. Enrichment notably boosted the concentration of health-promoting constituents. Biscuits containing 15% sour cherry pomace recorded the highest amounts of polyphenols (475.16 mg gallic acid equivalents/100 g), flavonoids (204.10 mg catechin equivalents/100 g), and anthocyanins (28.58 mg cyanidin-3-glucoside equivalents/100 g). In contrast, biscuits fortified with 15% apple pomace displayed stronger antiradical activity (30.80%) and higher sugar content (46.31 g glucose equivalents/100 g) than their sour cherry counterparts. FTIR spectroscopy confirmed the presence of characteristic vibrations associated with these bioactive compounds in both the pomace and the enriched biscuits. Overall, the results show that incorporating apple and sour cherry pomace is a practical way to create functional biscuits with enhanced nutritional qualities while promoting the sustainable use of fruit industry residues. Full article

Tomatoes are among the most consumed vegetables within the Mediterranean diet, recognized as one of the healthiest dietary patterns in the world. This study evaluated the effects of four fertilization treatments on the antioxidant and metabolic profiles of four local tomato varieties: Cornabel (V1), Cuban Pepper (V2), Corno Andino (V3), and Roli Rosa (V4). Treatment 1 (T1) used 1.28 kg/m² of commercial woody compost (C/N ratio 13), while Treatment 2 (T2) served as the control, initially without fertilization. Treatments 3 and 4 (T3, T4) incorporated ramial chipped wood (RCW) at 15 kg/m² and 7.5 kg/m², respectively, without tillage. Each treatment × variety combination included four biological replicates (n = 4). Untargeted metabolomic profiling via UHPLC-QToF and statistical analyses identified 163 compounds, 37 of which showed significant varietal differences (p < 0.05). The flavonoid eriodictyol was more abundant in the Pebroter varieties (V1, V2, V3), whereas luteolin-8-O-glucoside (orientin) predominated in V4, likely due to differential expression of biosynthetic genes. RCW (T3) yielded higher levels of antioxidant compounds like phenolic acids, and flavonoids compared to compost and control; however, no dose–response was observed using different doses of RCW (T3 and T4). These findings demonstrate that RCW may modulate the antioxidant metabolite profile of tomatoes, highlighting its potential as a sustainable fertilization strategy for enhancing antioxidant compounds. Full article

Forsythia suspensa (Thunb.) Vahl, a pharmacopoeial medicinal plant, is valued for its therapeutic efficacy in heat-clearing detoxification, dispelling wind-heat, and promoting blood circulation to resolve stasis. Flavonoids, ubiquitous secondary metabolites in F. suspensa, are critically linked to pharmacological activities and exhibit diverse biological functions. To elucidate the chemotypic divergence and ecological drivers of its bioactive compounds, we conducted flavonoid metabolomic profiling across ten wild populations F. suspensa using UPLC-MS/MS. Results revealed significant inter-population variation in all twenty-nine flavonoid metabolites analyzed. Notably, Notably, Cinchonain Ic was significantly enriched in the JX population, Flavanomarein in the LT population, and Desmethylxanthohumol in the HX population. Association analysis with environmental variables further indicated that Sulfuretin, Apigenin-5-O-glucoside, and Flavanomarein were positively correlated with multiple precipitation-related variables (bio12-Annual Precipitation, bio14-Precipitation of Driest Month, bio17-Precipitation of Driest Month, and bio19-Precipitation of Coldest Quarter), whereas Vicenin 2 was negatively correlated with bio12, bio17, and bio19. Homoplantaginin showed a positive correlation with bio4 (Temperature Seasonality) and bio7 (Temperature Annual Range). Loureirin B was positively correlated with elevation but negatively correlated with high-temperature variables (bio5-Max Temperature of Warmest Month, bio8-Mean Temperature of Wettest Quarter, and bio10-Mean Temperature of Warmest Quarter). 5-Demethoxynobiletin was positively associated with both precipitation (bio12, bio17, bio19) and temperature variables (bio1-Annual Mean Temperature, bio6-Min Temperature of Coldest Month, bio9-Mean Temperature of Driest Quarter and bio11-Mean Temperature of Coldest Quarter). Cinchonain Ic was positively correlated with bio2 (Mean Monthly Temperature Range), and Oroxin A was negatively correlated with elevation. These findings demonstrated that flavonoids accumulation in F. suspensa was predominantly influenced by temperature heterogeneity, with precipitation serving as a secondary factor, while latitude and elevation play only limited roles. This study systematically investigates the divergence and environmental drivers of flavonoids in F. suspensa populations, clarifies the molecular ecological basis of its adaptation to environmental heterogeneity, and provides valuable insights for leveraging ecological factors to enhance medicinal potential, ultimately supporting targeted breeding and optimized field management strategies. Full article

Dental caries is a major global health concern, with Streptococcus mutans playing a key role in biofilm formation and acid production, which lead to enamel demineralization. Natural products, particularly plant-derived extracts, offer promising alternatives to conventional antibacterial agents. This study aimed to analyze the chemical composition of Ferula varia 70% ethanol extract (FVE) and evaluate its potential to inhibit biofilm formation by S. mutans. The aerial parts of F. varia were extracted with 70% ethanol and analyzed using LC-UV-ESI-MS/MS to determine the chemical profile. The anti-biofilm activity of FVE was evaluated using a crystal violet assay against S. mutans. Phytochemical analysis identified 14 compounds, including major phenolic acids (e.g., chlorogenic acid, gallic acid) and flavonoids (e.g., isoquercitrin, isorhamnetin-3-O-glucoside). FVE exhibited significant, dose-dependent inhibition of S. mutans biofilm formation. Importantly, the FVE concentration of 5 mg/mL inhibited S. mutans biofilm development by 100%. The potent antibiofilm activity of FVE against S. mutans is likely due to the synergistic action of its rich content of phenolic acids and flavonoids, which possess known anti-virulence properties. These findings support the potential use of FVE as a natural ingredient in oral hygiene products to prevent dental plaque and caries. Full article

This study examines the antidiabetic potential of Rubus ulmifolius, a shrub traditionally used for medicinal and nutritional purposes. The aim was to assess the fruit extract’s inhibitory effect on α-amylase and α-glucosidase, therapeutic targets in type 2 diabetes (T2D), given their role in carbohydrate digestion. Considering the role of islet amyloid polypeptide (IAPP) aggregation in pancreatic β-cell dysfunction in T2D, the extract’s effect on inhibiting this process was also evaluated. Cytocompatibility and antioxidant effects were tested in Caco-2 cells, while caspase-1 activity was measured to evaluate anti-inflammatory potential. Phytochemical analysis of R. ulmifolius fruits revealed various phenolic compounds, with anthocyanin as the most abundant one. The cyanidin-3-O-glucoside accounted for 86% of all anthocyanins. Among flavonoids, the most represented ones were quercetin-HMG-glucoside and a kaempferol derivative, while ellagic acid glucuronide was the predominant ellagitannin. The extract showed significantly higher α-glucosidase inhibition with an IC₅₀ value of 2.8 µg/mL, 32 times more effective than acarbose, and it markedly inhibited IAPP aggregation in a dose-dependent manner. It demonstrated antioxidant activity in cellular and acellular systems, without cytotoxicity. Caspase-1 activity was also reduced in intestinal cells. These findings support R. ulmifolius fruits as a promising functional food for managing T2D and preserving pancreatic function. Full article

Red grapes are recognized as valuable sources of phenolic compounds with nutritional and technological importance. Anthocyanins strongly influence the color, stability, and antioxidant activity of wines, thereby contributing to both quality and potential health effects. In this study, berries of nine red grapevine cultivars (Alicante Bouschet, Burgund Mare, Busuioacă de Bohotin, Cabernet Franc, Cabernet Sauvignon, Cadarcă, Malbec, Sangiovese, and Syrah) were examined for their anthocyanin composition, total phenolic and flavonoid content, physicochemical parameters, and fatty acid profiles. Anthocyanins were characterized using High Performance Liquid Chromatography coupled with Mass Spectrometry (HPLC-MS), total polyphenols and flavonoids were quantified spectrophotometrically, and fatty acids were determined by Gas Chromatography coupled with Mass Spectrometry (GC-MS). Substantial variability was observed across cultivars for the analyzed traits, with nine anthocyanins identified (total levels ranging from 70.79 ± 13.84 to 335.75 ± 87.62 mg malvidin-3-O-glucoside equivalents (MGE) per 100 g fresh weight (FW). Total phenolics ranged from 107.51 ± 11.11 to 432.13 ± 42.91 mg gallic acid equivalents (GAE) per 100 g FW, and flavonoids from 34.23 ± 11.45 to 162.51 ± 39.63 mg catechin equivalents (CE) per 100 g FW. Ten fatty acids were identified, with linoleic acid being the most abundant. Alicante Bouschet and Burgund Mare showed the highest levels of total anthocyanins, polyphenols, and flavonoids, while Cabernet Franc, Cabernet Sauvignon, and Sangiovese exhibited the richest profiles of polyunsaturated fatty acids, together highlighting their potential as valuable sources of bioactive and nutritional compounds for functional food applications. Cabernet Franc and Sangiovese, characterized by higher titratable acidity and balanced pH, showed favorable traits for producing stable, high-quality wines. Analysis of the data further grouped the cultivars based on their chemical and lipid profiles. Overall, these findings show the notable biochemical differences among the red grapevine cultivars and their potential uses in food and wine production. Full article

A corneal abrasion results from the disruption or loss of cells in the corneal epithelium. If inadequately treated, it can compromise visual clarity. The wound healing process of a corneal abrasion involves epithelial migration, proliferation and adhesion. Clitoria ternatea flower extract (CTE) is rich in flavonoids, anthocyanins and other bioactive compounds. It has antioxidant, anti-inflammatory and wound-healing properties. This study explores the potential of CTE to be used as a natural supplement to improve corneal wound healing. Phytochemical profiling via LC–MS identified a total of 51 distinct bioactive constituents. The anthocyanin content, quantified in terms of cyanidin-3-glucoside equivalent, was quantified at 33.06 mg per gram of extract. The extract exhibited 33.8% DPPH radical scavenging activity and a total polyphenol content equivalent to 24.14 mg/g gallic acid. Human telomerase-immortalized corneal epithelial (hTCEpi) cells maintained in keratinocyte basal medium were utilized to determine cytotoxicity and wound-healing effects. The optimal extract concentration of 0.08 mg/mL, quantified via MTT assay, resulting in enhanced cell viability. Scratch assays demonstrated a higher percentage of wound closure in the CTE-treated group at 6 and 12 h relative to the untreated group, with statistical significance (p < 0.05). The gene expressions of CK3 and Cx43, quantified via qRT-PCR, showed no significant differences between groups. However, within the CTE-treated group, CK3 expression increased at 12 h relative to 0 h and 6 h, and Cx43 expression rose significantly at 12 h compared with 0 h (p < 0.05). Immunofluorescence confirmed positive protein expression of both markers. These findings suggest that CTE possesses potent antioxidant properties and promotes corneal epithelial wound healing through upregulation of CK3 and Cx43 in vitro. Full article