Polat, Heybet Kerem; Turanlı, Yasin; Kurt, Nihat; Gözcü, Sefa; Tunçel, Emre; Bacanak, Hilal; Aytekin, Eren; Ünal, Sedat

Since the early 1960s, nanotechnology has been a critical area of science, allowing for the development of sophisticated nanomaterials. Nanofibers, one of the most widely used nanotechnological drug delivery systems, have emerged as a highly versatile platform within modern pharmaceutical sciences. By combining various polymers with active herbal ingredients, these systems mimic the natural extracellular matrix and provide improved functions such as a high surface area-to-volume ratio, drug targeting, and controlled drug release. Preclinical studies demonstrate that phytochemical-containing nanofibers have improved therapeutic profiles, including higher anti-inflammatory, antioxidant, antimicrobial, and antineoplastic effects. In various biomedical applications, such as promoting tissue engineering (such as bone and nerve regeneration), limiting tumor growth, and accelerating wound closure, it was shown that nanofibers may overcome the physicochemical limitations of herbal drugs such as low solubility and bioavailability. Despite promising preclinical study results, there are significant obstacles in the way of the commercialization of herbal drug-loaded nanofibers. The difficult standardization of multi-component herbal extracts, stabilization drawbacks, and the absence of scalable industrial manufacturing equipment that can maintain repeatable characterization of nanofibers are some of the major obstacles, in addition to a lack of clinical trials. To navigate the clinical translation of nanofibers, interdisciplinary collaboration regarding quality control, safety, and regulatory pathways is strictly necessary due to the case-by-case review approach utilized by regulatory bodies such as the US Food and Drug Administration (FDA) and European Medicines Agency (EMA). This review offers a comprehensive overview of research on herbal drug-loaded nanofibers and contributes a novel perspective with regulatory and clinical translational insights.

Güven, Zühal Bayrakçeken; Türkmenoğlu, Burçin

BACKGROUND: Vaccinium species are characterised by high nutritional value and rich phytochemical composition of their fruits and are therefore used as food and in folk medicine. The aim of this study was to investigate the potential of fruits and leaves of the Vaccinium arctostaphylos L. and Vaccinium myrtillus L. for use as cosmeceuticals and against hyperpigmentation-related skin diseases. The antityrosinase and melanogenesis effects were determined by inhibition of mushroom tyrosinase enzyme, inhibition of tyrosinase enzyme in α-MSH (melanocyte-stimulating hormone)-induced B16F10 cells and melanin inhibition. RESULTS: The ethyl acetate sub-extract from the fruits of V. arctostaphylos showed very strong tyrosinase enzyme inhibition (median inhibitory concentration (IC50) = 9.71 μg mL−1), which was approximately as high as that of the standard compound, and also showed significant melanin inhibition and antityrosinase effect compared to kojic acid. The ethyl acetate extracts from the fruits and leaves of V. arctostaphylos showed antioxidant effect as strong as the standard compounds gallic acid and Trolox. The phytochemical contents of fruit and leaf extracts were analysed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). ADMET (absorption, distribution, metabolism, excretion, and toxicity) profiles of the compounds were predicted and their likeness to drug candidates were analysed. Finally, molecular docking analysis was performed with the crystal structures of different targets determined for compounds (rutin, protocatechuic acid, gentisic acid, catechin, chlorogenic acid, caffeic acid, p-coumaric acid, taxifolin, salicylic acid, 4-hydroxybenzoic acid, luteolin-7-glycoside, quercetin). CONCLUSION: The fruit extracts of V. arctostaphylos in particular appear to be a potential and promising source for the cosmeceutical and pharmaceutical industry due to their content of active ingredients and strong biological effect.

Akman, Tugrul Cagri; Simsek, Samed; Aksit, Zeynep; Aksit, Huseyin; Aydin, Ali; Adem, Şevki; Tufekci, Ali Riza; Yıldız, İlyas

BACKGROUND: Ebenus macrophylla Jaub. & Spach (EM), an endemic Turkish plant, is a rich source of bioactive compounds with potential therapeutic applications. This study aimed to analyze the phytochemistry of its leaf, stalk, and root extracts and evaluate their antioxidant, antimicrobial, enzyme inhibitory, and antiproliferative effects. RESULTS: Liquid chromatography–electrospray ionization–tandem mass spectrometry (LC-ESI-MS/MS) analysis identified 39 phenolic compounds. Among the major phenolics identified, hesperidin (in the leaf and stalk), isoquercitrin (in the leaf), catechin (in all parts), and vanillic acid (in the root) were particularly abundant. The leaf extract showed the strongest antioxidant activity (2,2-Diphenyl-1-picrylhydrazyl (DPPH) Half maximal inhibitory concentration (IC50): 14.08 μg mL−1) and the highest phenolic content (62.27 mg GAE g−1), which is attributed to its high levels of hesperidin and shikimic acid. Antimicrobial assays showed significant activity by the leaf extract against Clostridium perfringens and Pseudomonas aeruginosa (Minimum Inhibitory Concentration (MIC): 31.3 μg mL−1). In enzyme inhibition studies, the leaf extract showed stronger effects than standard inhibitors on α-glucosidase (IC50: 1.56 μg mL−1−1) and tyrosinase (IC50: 18.01 μg mL−1). Molecular docking analyses confirmed that hesperidin forms multiple stabilizing interactions with α-glucosidase and blocks substrate access. Antiproliferative studies showed that the root extract was significantly active against A549, A2780, and HeLa cancer cell lines (Growth Inhibition 50% (GI50): 1.00–3.22 μg mL−1) and minimal toxicity toward normal BEAS-2B cells. CONCLUSION: The distribution of phenolic compounds detected in different plant parts is the main factor determining the strong antioxidant, antimicrobial, antidiabetic, and antiproliferative potential of EM. This study demonstrates that the species is a pharmaceutically promising natural bioactive source and provides the scientific basis for further in vivo research.

Güven, Zühal Bayrakçeken; Karadayı, Mehmet; Yilmaz, Mustafa Abdullah; Tugay, Osman; Cakir, Oguz

BACKGROUND: Food additives are widely used to extend the shelf life of foods and maintain their quality. In this study, the potential of Prunus laurocerasus and Prunus cerasifera fruit pits (endocarp and seed) as food additives was investigated in terms of cytotoxicity, antigenotoxicity, antioxidant activity, enzymatic anti-browning, and urease enzyme inhibition, and their phytochemical contents were comprehensively analyzed. RESULTS: While the extracts showed low cytotoxicity against the healthy L929 cell line, they exhibited a concentration-dependent increase in selective cytotoxicity against A549, CaCo-2, Hep3B, MCF-7, and PC-3 cells. Furthermore, the genotoxic safety of the extracts was assessed in both prokaryotic and eukaryotic model test systems. The extracts showed no cytogenotoxic or mutagenic effects in the Allium cepa and Ames/Salmonella assays. P. cerasifera endocarp ethyl acetate extract exhibited the highest antioxidant activity (DPPH, CUPRAC, and TEAC: 576.24, 402.42, and 680.14 mg Trolox g−1 extract, respectively) and strong enzyme inhibition, with tyrosinase (IC50: 10.46 ± 1.48 μg mL−1) nearly comparable to kojic acid and potent urease inhibition (IC50: 28.60 ± 2.08 μg mL−1). Phytochemical analysis of the extracts was performed using liquid chromatography–tandem mass spectrometry, with epicatechin (10.722 mg g−1 extract) detected in the highest amount in the endocarp of P. laurocerasus and quinic acid (11.679 mg g−1 extract) detected in the endocarp of P. cerasifera. CONCLUSION: Overall, according to the conducted cytotoxicity and genotoxicity assays, P. laurocerasus and P. cerasifera fruit pits were found to be safe, rich in bioactive phytochemicals, and exhibited strong antioxidant and enzyme inhibitory activities, highlighting their potential as sustainable functional ingredients or natural food additives.

Aydın, Bilge; Ertuğrul, Muhammed Sait; Aydın, İsmail Çağrı; Uğan, Rüstem Anıl; Özkaraca, Mustafa; Yuca, Hafize; Özbek, Hilal; Kazaz, Cavit; Okkay, Ufuk; Karakaya, Songül

BACKGROUND: Various Vicia species belonging to the Fabaceae family are known to be used in different parts of the world both in human nutrition and as animal feed, and also have pharmacological potential in the prevention and treatment of various diseases. With this perspective, in the present study, the possible antiparkinsonian effects of extracts prepared from Vicia canescens subsp. variegata were investigated in cell culture and on 6-hydroxydopamine (6-OHDA)-induced Parkinson's model rats. The effects of the plant on the pathophysiology of Parkinson's disease were evaluated by biochemical and histopathological analyses; in addition, the structures of the compounds isolated from the active extracts were elucidated and the in vitro antioxidant and antiparkinsonian activities of these compounds were determined. RESULTS: Methanol and n-butanol extracts prepared from the aerial parts of V. canescens subsp. variegata showed significant antiparkinsonian effects both in cell culture and in a rat model. Histopathological and biochemical analyses were also parallel to in vivo experiments. As a result of the isolation studies, six flavonol glycosides were obtained from the plant, and their antioxidant capacities as well as antiparkinsonian effects were subsequently evaluated. The highest biological effect was observed in quercetin-3-O-α-l-rhamnopyranoside (quercitrin, VCB-3). CONCLUSION: In the light of the studies, the antiparkinsonian effects of V. canescens subsp. variegata was evaluated. It was seen that they prevented neurodegeneration and the oxidative stress that occurred, and the most effective compounds were determined. Two new compounds named viciacoside A and B have been added to the literature as a result of this research.

Akman, Tuğrul Çağrı; Şimşek, Samed; Akşit, Zeynep; Akşit, Hüseyin; Aydin, Ali; Tüfekçi, Ali Riza; Adem, Sevki; Yilmaz, Mustafa Abdullah

BACKGROUND: The objective of this study is to investigate the antiproliferative, antioxidant, antimicrobial, and enzyme activity capacities and phytochemical compositions of Thymus pectinatus (TP), Thymus convolutus (TC), which are endemic to Türkiye. Quantitative analysis of phenolic compounds in the extracts was conducted using liquid chromatography–tandem mass spectrometry, targeting 53 phenolic compounds. RESULTS: Rosmarinic acid, quinic acid, and cynaroside were identified as the major compounds, exhibiting quantitative variation in both extracts. The extracts had a high total phenolic content, with 113.57 ± 0.58 mg gallic acid equivalents (GAE)/g extract for TP and 130.52 ± 1.05 mg GAE/g extract for TC. Furthermore, although both extracts exhibited high total flavonoid content; the TP extract (75.12 ± 1.65 mg quercitin equivalents (QE)/g extract) displayed a higher flavonoid content than the TC extract (30.24 ± 0.74 mg QE/g extract) did. The extracts had a promising antiproliferative effect on C6, HeLa, and HT29 cancer cell lines with a less cytotoxic effect (10.5–14.2%) against normal cells. Both extracts exhibited very potent inhibitory activity against the xanthine oxidase enzyme, with half-maximal inhibitory concentration values of respectively 2.07 ± 0.03 μg mL−1 and 2.76 ± 0.06 μg mL−1 and moderate activity against tyrosinase and α-glucosidase. Docking simulations proved that rosmarinic acid and cynaroside, the major components of the extracts, were the most potent inhibitors of xanthine oxidase. According to antimicrobial activity results, the TC extract exhibited moderate activity against Staphylococcus aureus, and the TP extract had strong activity against both Enterococcus faecium and S. aureus. CONCLUSION: These findings emphasize the beneficial effects of the two endemic Thymus species on human health and suggest their potential use as plant-derived bioactive agents.

Gözcü, Sefa; Akşit, Zeynep; Şimşek, Samed; Kandemir, Ali; Aydın, Ali; Yılmaz, Mustafa Abdullah; Akşit, Hüseyin

BACKGROUND: Ferulago setifolia K. Koch (Apiaceae) has been the subject of this study, aiming to comprehensively determine its phenolic fingerprint and evaluate its various biological activities. The liquid chromatography–tandem mass spectrometry analysis of the 70% methanol extract of F. setifolia (FS) revealed the presence of 23 phytochemicals, among which chlorogenic acid, quinic acid, kaempferol-3-O-glucoside, and quercetin-3-O-glucoside were identified as the major phenolics in the extract. RESULTS: The biological screening included examinations of antioxidant, antibacterial, antiproliferative, and cytotoxic activities. The FS extract displayed moderate 2,2-diphenyl-1-picrylhydrazyl radical scavenging and ferric-reducing capacity, indicating moderate antioxidant activity. Furthermore, FS exhibited significant antiproliferative effects on cancer cells while showing low cytotoxicity on normal cells. The antibacterial activity findings revealed that FS demonstrated potent activity against Pseudomonas aeruginosa, Bacillus cereus, Staphylococcus aureus, and Escherichia coli. CONCLUSION: The findings of this study suggest that the methanolic extract of FS holds promise as a potential source of biologically active compounds. It can be utilized for the development of pharmaceutical formulations, thanks to its significant antiproliferative and antibacterial activities. Additionally, FS can serve as a valuable source of chlorogenic acid for industrial applications.

Arslan, Betul; Xiong, Youling L; Soyer, Ayla

BACKGROUND: The influence of protein hydrolysate produced from bovine liver protein hydrolysate (LPH) by enzymatic hydrolysis, using Alcalase/Protamex (1:1), on lipid dispersions was investigated. LPH production was optimized to maximize the antioxidant activity (at 45, 50, and 55 °C for 12, 18, and 24 h). Different concentrations of LPHs (1, 3, and 5 mg/g) were added to emulsions and to liposomes. Lipid oxidation level and particle size of the lipid dispersions were monitored for 14 days of storage at 25 °C. RESULTS: Radical scavenging activity and reducing power were the highest at 45 °C after 24 h of hydrolysis. Electrophoresis pattern showed that the antioxidant activity was arising from the peptides with molecular weight around 10 kDa. Lipid oxidation occurred more rapidly in samples without LPH during storage. In emulsions, lower thiobarbituric acid-reactive substance and conjugated diene values were measured with increasing concentrations of LPH at day 14. Accordingly, particle size of the samples containing 5 mg/g of LPH was smaller than those of other groups. Phase separation was observed only in lecithin emulsion without LPH at day 14. The use of LPH in liposome limited the lipid oxidation and maintained the size of the particles independently from the concentration. CONCLUSION: This study highlights the potential applications of animal by-products as natural antioxidants in complex food systems. The results demonstrate that LPH, particularly when hydrolyzed at optimized conditions, can effectively inhibit lipid oxidation. The findings suggest that biphasic systems incorporating LPH have promising prospects for enhancing the stability and quality of food products.

Gözcü, Sefa; Polat, Heybet Kerem; Gültekin, Yakup; Ünal, Sedat; Karakuyu, Nasıf Fatih; Şafak, Esra Köngül; Doğan, Osman; Pezik, Esra; Haydar, Muhammet Kerim; Aytekin, Eren

BACKGROUND: Allergic conjunctivitis is one of the most common eye disorders. Different drugs are used for its treatment. Hesperidin is an active substance isolated from Citrus sinensis L. (Rutaceae) fruit peels, with known anti-inflammatory activity but low solubility. It was complexed with cyclodextrin and encapsulated in situ gel to extend its duration in the eye. RESULTS: The optimized formulation comprised 1% hesperidin, 1.5% hydroxyethyl cellulose, and 16% poloxamer 407. The viscosity at 25 °C was 492 ± 82 cP, and at 35 °C it was 8875 ± 248 cP, the pH was 7.01 ± 0.03, gelation temperature was 34 ± 1.3 °C, and gelation time was 33 ± 1.2 s. There was a 66% in vitro release in the initial 2 h, with a burst effect. A lipoxygenase (LOX) inhibition test determined that hesperidin was active at high doses on leukotyrens seen in the body in allergic diseases. In cell-culture studies, the hesperidin cyclodextrin complex loaded in situ gel, BRN9-CD (poloxamer 16%, hydroxy ethyl cellulose (HEC) 1.5%), enhanced cell viability in comparison with the hesperidin solution. It was determined that BRN9-CD did not cause any irritation in the ocular tissues in the Draize test. CONCLUSION: The findings of this study demonstrate the potential of the in situ gel formulation of hesperidin in terms of ease of application and residence time on the ocular surface. Due to its notable LOX inhibition activity and positive outcomes in the in vivo Draize test, it appears promising for incorporation into pharmaceutical formulations.

Pérez-Navarro, José; Hermosín-Gutiérrez, Isidro; Gómez-Alonso, Sergio; Kurt-Celebi, Aynur; Colak, Nesrin; Akpınar, Erdal; Hayirlioglu-Ayaz, Sema; Ayaz, Faik A

BACKGROUND: ‘Karaerik’ is a novel table grape (Vitis vinifera L.) native to Turkey and widely cultivated in areas bordering the city of Erzincan. Because of the demonstrated beneficial effects on human health of the grape phenolic composition, the aim of this work was to conduct a detailed profiling of non-anthocyanin phenolic fractions from different grape tissues of the ‘Karaerik’ table grape. Both qualitative and quantitative characterization of phenolic compounds were achieved using high-performance liquid chromatography–diode array detection–electrospray ionization–tandem mass spectrometry. Total phenolic content and oxygen radical absorbance capacity were also determined to evaluate the antioxidant properties of this table grape. RESULTS: A high number of non-anthocyanin phenolic compounds was identified in ‘Karaerik’ table grape skins and seeds, including 11 flavonols, six hydroxycinnamic acid derivatives, two stilbenes, several monomeric and dimeric flavan-3-ols and proanthocyanidins. Quercetin-type derivatives dominated the flavonol profile of grape skins, followed by myricetin type. Tartaric acid esters of three acids (caffeic, coumaric and ferulic acids) were the main hydroxycinnamic acid derivatives in this cultivar. Qualitative and quantitative differences were observed in flavan-3-ol composition among the grape tissues. Proanthocyanidins were the most abundant class of phenolic compounds in ‘Karaerik’ grapes, being mainly located in seeds. Higher antioxidant capacity values were determined in grape seeds, in correlation with the total phenolic content. CONCLUSION: These results provide useful information for a better understanding of phenolic antioxidants from the ‘Karaerik’ table grape and will contribute to promoting the varietal identity and health-related properties of this fruit.