Search Results (617)

Background: Cyclodextrin (CD) polymers have attracted increasing attention due to their favourable drug delivery properties and broad pharmaceutical applicability. While the bioavailability and biological behaviour of native cyclodextrins have been extensively investigated, considerably less information is available regarding modified cyclodextrin polymers. Therefore, the present study aimed to investigate the permeation and cellular uptake of an epichlorohydrin-crosslinked β-cyclodextrin polymer using multiple in vitro and ex vivo models. Methods: Fluorescently labelled β-cyclodextrin polymers were applied in all experiments. Membrane permeation studies were performed using an in-line diffusion cell system with membranes of different pore sizes. In vitro transport and cellular uptake were investigated on HaCaT, Caco-2, and TR146 cell monolayers, while ex vivo permeation studies were carried out using skin, buccal, and intestinal tissues. Results: The results demonstrated a strong size-dependent transport behaviour across synthetic membranes. Cell monolayer studies revealed cell-line-dependent differences in polymer intracellular distribution. Lysosomal accumulation was observed in HaCaT and Caco-2 cells, whereas no intracellular accumulation was detected in TR146 cells. These findings suggest differences in polymer permeation among the investigated cell models. Ex vivo studies demonstrated the tissue permeation of cyclodextrin polymers, with marked accumulation within skin layers, indicating predominant dermal retention. Furthermore, strong correlations were identified between the in vitro and ex vivo skin and intestinal models. Conclusions: Overall, the findings demonstrate that β-cyclodextrin polymers exhibit complex, barrier-dependent transport behaviour across different biological models. The observed differences in permeation and intracellular localization suggest that multiple transport processes may contribute to their biological interactions, which provide a foundation for future studies aimed at elucidating the molecular mechanisms governing polymer uptake and permeation. Full article

Excessive ethanol consumption is a cause of gastric ulceration. This study aimed to identify the gastroprotective action of rufigallol, an anthraquinone, against ethanol-induced gastric ulcers and assess its antioxidant and anti-inflammatory properties. Healthy rats were grouped into five groups (n = 8); the control group received sterile saline orally; the ethanol group received ethanol (5 mL/kg) to generate gastric ulcers on the last day of the experiment; the pretreated rufigallol group was administered 10 or 20 mg/kg rufigallol orally for a week before gastric ulcer initiation; and the drug control group received omeprazole (20 mg/kg) for a week with ethanol treatment. The results showed that oral rugalallol significantly reduced gastric ulcers, as indicated by decreased gastric juice volume and increased preventive percentage, gastric pH value, and pepsin activity. Histopathology confirmed a reduction in the gastric ulcer index following rufigallol treatment. Rufigallol pre-treatment significantly increased antioxidant levels (CAT, SOD, and GSH) and decreased MDA levels compared to the ethanol group. Furthermore, rufigallol treatment decreased MPO, pro-inflammatory cytokines, and mediator levels. It also reduced COX-2, IFN-γ, and NLRP3 expression and restored NO and PGE2 levels. In vitro experiments using HT-29 and HaCaT cell lines confirmed that rufigallol reduced cytokine production and exhibited anti-inflammatory activity in response to lipopolysaccharide stimulation. These results suggested that rufigallol administration may provide gastroprotective benefits against ethanol-induced gastric ulcers, potentially by reducing oxidative stress and gastric inflammation. Full article

Candida spp. can cause systemic infections with high mortality in immunocompromised patients. Phytotherapy may be an alternative for adjunctive treatment of fungal infections. This study evaluated the phytochemical profile, cytotoxicity, genotoxicity, and antibiofilm activity of the hydroalcoholic extract of Origanum vulgare L. against Candida albicans, Candida tropicalis, and Candida dubliniensis. The extraction and quantification (flavonoids and phenols) were performed, and its antioxidant activity (DPPH) and the presence of bio-active compounds were investigated using high-performance liquid chromatography with Diode Array Detection (HPLC-DAD). Cytotoxicity and genotoxicity tests were conducted using HaCat cell lines. Antifungal activity on planktonic cultures was evaluated using the standard (CLSI M27-S4). The analysis of the extract on biofilms was verified with different exposure times. The extract demonstrated the presence of bioactive molecules, and antioxidant activity. The cytotoxicity test showed viability >70%. Genotoxicity revealed the presence of a few micronuclei in some dilutions. The Minimum Fungicidal Concentration was obtained for C. albicans and C. tropicalis. In the biofilm analysis, there was a reduction of more than 60% in all Candida spp. species at the 24 h exposure time. The findings suggest that the O. vulgare extract exhibits activity against Candida spp. and shows biocompatibility in human keratinocytes. Full article

Background/Objectives: Cadherin-17 (CDH17, LI-cadherin) is a non-classical cadherin with an atypical structure and unique functions. CDH17 expression is restricted to normal intestinal epithelium. Furthermore, CDH17 functions as an oncoprotein that promotes tumor migration and invasion in colorectal, gastric, and pancreatic cancers. Therefore, CDH17 is an important diagnostic marker and therapeutic target. The CDH17-directed strategies, including monoclonal antibodies (mAbs), bispecific Abs, antibody–drug conjugates (ADCs), and chimeric antigen receptor (CAR) T cells, have been evaluated in preclinical and clinical studies. Therefore, developing mAbs that specifically recognize cell surface-expressing CDH17 is essential for advancing both tumor diagnosis and therapy. Methods: Anti-human CDH17 mAbs (named Ca₁₇Mabs) were developed by immunizing a mouse with CDH17-overexpressed cells and a high-throughput screening using flow cytometry. Results: Among Ca₁₇Mabs, a clone, Ca₁₇Mab-5 (IgG₁, κ) specifically recognized CDH17-overexpressed Chinese hamster ovary-K1 (CHO/CDH17) cells with no detectable cross-reactivity to 21 other CDHs by flow cytometry. Ca₁₇Mab-5 also detected endogenous CDH17 in human colorectal cancer cell lines, COLO201 and COLO205. The apparent dissociation constant (K_D) values of Ca₁₇Mab-5 for CHO/CDH17 and COLO205 were estimated as 1.5 × 10⁻⁸ M and 1.3 × 10⁻⁸ M, respectively. Furthermore, Ca₁₇Mab-5 detected endogenous CDH17 by Western blotting. In immunohistochemistry, Ca₁₇Mab-5 exhibited clear membranous staining in normal colon epithelium, colorectal, gastric, and pancreatic cancers. Conclusions: Ca₁₇Mab-5 is a versatile tool for detecting CDH17 and has potential for tumor diagnosis. Full article

Background/Objectives: Artemisia annua essential oil (EO) possesses diverse biological activities; however, its practical application is limited by volatility, instability, and poor bioavailability. This study aimed to develop chitosan-coated nanostructured lipid carriers (CH-NLCs) for efficient encapsulation and delivery of A. annua EO and to evaluate their physicochemical characteristics and biological performance. Methods: The nanoformulation exhibited favorable physicochemical properties, including a high encapsulation efficiency (85.97 ± 1.30%) and a strongly positive surface charge (approximately +45 mV), indicating good colloidal stability. Structural analyses by SEM, FTIR, and XRD confirmed successful encapsulation of the EO within the nanocarrier matrix. Results: The CH-NLC formulation significantly enhanced larvicidal activity against Aedes aegypti larvae, reducing the LC₅₀ value from 213 ppm for the free EO to 142 ppm. Enhanced antifungal activity was also observed, with 47–56% greater inhibition against Malassezia furfur, Trichophyton mentagrophytes, and Candida albicans compared with the free EO. Furthermore, CH-NLC demonstrated improved cytotoxic activity against skin cancer cell lines, achieving IC₅₀ values of 21.4 ± 1.7 µg/mL and 30.1 ± 1.6 µg/mL against A431 and A375 cells, respectively, while maintaining lower toxicity toward normal HaCaT keratinocytes. Mechanistic investigations revealed enhanced apoptosis and an approximately 3-fold increase in intracellular reactive oxygen species (ROS) levels in treated cancer cells. Conclusions: Collectively, these findings indicate that chitosan-coated nanostructured lipid carriers effectively improve the stability and biological efficacy of A. annua essential oil and represent a promising platform for future biomedical and biocidal applications. Full article

Background: The HPV16 E6 oncoprotein facilitates the ubiquitin-mediated degradation of the tumor suppressor p53, constituting a pivotal mechanism underlying viral immune evasion, cellular immortalization, and ultimately, malignant transformation. This study aimed to develop a reliable detection tool for the HPV16 E6 protein. Methods: Recombinant GST-E6 and E6-His proteins were expressed and purified using a prokaryotic expression system. Female BALB/c mice were immunized with GST-E6, and two hybridoma cell lines (G11A11 and A4) stably secreting anti-HPV16 E6 monoclonal antibodies were generated via hybridoma technology. Antibody pairing experiments identified A4 and G11A11 as suitable for sandwich ELISA. The optimal detection system was established using A4 antibody at 2 μg/mL for coating, HPV16 E6-His at 5 μg/mL as the detection antigen, and G11A11-HRP at a 1:200 dilution as the detection antibody. To validate the reliability, Hacat-HPV16E6 cell lysates were tested in parallel with a commercial ELISA kit. Results: After purification, the titers of both antibodies reached 1:204,800. The lower limit of quantification (LOQ) was 4.79 ng/mL and the limit of detection (LOD) was 3.39 ng/mL. The comparison with the commercial kit showed good consistency, with percentage differences ranging from 20% to 40%, confirming that the established ELISA is reliable for quantitative detection. Conclusions: This study successfully yielded high-titer and highly specific anti-HPV16 E6 monoclonal antibodies and developed a specific double-antibody sandwich ELISA, thereby furnishing a technical foundation for both fundamental research and laboratory-based applications related to HPV16-associated tumors. Full article

Coleus Lour. and Plectranthus L’Hér. (Lamiaceae) species are recognized as promising sources of bioactive compounds for skin health, supported by their traditional use in skin-related conditions; however, scientific validation of these traditional uses remains limited for several species. In this work, the biological activities of eight Coleus and Plectranthus species (C. barbatus, C. cylindraceus, C. grandidentatus, C. hadiensis, C. madagascariensis, P. ambiguus, P. ecklonii, and P. fruticosus) were evaluated to support their ethnomedicinal relevance. Extracts were assessed for general toxicity (Artemia salina model), antimicrobial activity (well diffusion and microdilution methods) and antioxidant capacity (DPPH, TBARS and cellular assays). In addition, anti-inflammatory activity (ability to suppress nitric oxide), cytotoxicity in skin cell lines (HaCaT and HFF-1) and inhibition of skin-related enzymes (elastase, tyrosinase and collagenase) were evaluated. Antibacterial activity was predominantly observed against Gram-positive bacteria, with no general toxicity observed. P. ambiguous and P. ecklonii showed moderate-to-high antioxidant activity, while anti-inflammatory effects were observed for C. hadiensis and C. madagascariensis and P. ambiguous. None of the extracts showed cytotoxicity. Enzyme inhibition assays highlighted C. cylindraceus and P. ecklonii. HPLC-DAD analysis confirmed the presence of rosmarinic acid. Overall, the findings support the traditional use of Coleus and Plectranthus species for skin-related conditions and highlight their potential as sources of bioactive compounds for dermatological and cosmeceutical applications. Full article

Multifunctional drugs represent an emerging strategy for treating complex skin disorders and melanoma. A series of benzothiazole-based hybrids incorporating gallic and syringic acid moieties was synthesized and evaluated as multifunctional agents for skin-related applications. Six hydrazone (GAHYDR1–3) and acyl-hydrazone (GACIN1–3) derivatives were obtained and fully characterized. Hydroxylated compounds showed the strongest antioxidant activity, with GAHYDR1 and GACIN1 displaying low DPPH IC₅₀ values and high FRAP reducing power. UV–Vis studies revealed strong UVA–UVB absorption, with molar extinction coefficients comparable to or exceeding those of PBSA. Photoprotective evaluation showed SPF values up to 10.09 (GACIN2) and broad-spectrum behavior for selected derivatives. Antioxidant activity remained substantially stable over 3 months in solution. Antiproliferative assays against Colo38, A375, and HaCaT cell lines indicated generally low cytotoxicity toward non-tumor cells. Notably, GAHYDR3 exhibited selective activity against A375 melanoma cells (IC₅₀ = 8.75 µM; SI = 8.12). Overall, phenolic substitution emerged as a key determinant of biological activity, highlighting hydroxylated benzothiazole hybrids as promising antioxidant and photoprotective agents, with GAHYDR3 representing a potential lead for anti-melanoma development. Full article

Current treatment options for hair loss remain limited. Therefore, this study compared a botanical extract derived from multiple plants with the pharmaceutical agent minoxidil for topical application. The evaluated parameters included inflammatory cytokines (IL-1β, IL-6, TNF-α), growth factors (TGF-β, VEGF, KGF), and 5α-reductase type II (SRD5A2) expression in the human keratinocyte cell line HaCaT, as measured by ELISA. Both the botanical extract and minoxidil reduced IL-6 levels by 21% and 35%, and TNF-α levels by 13% and 35%, respectively. Treatment with the botanical extract and minoxidil increased VEGF expression by 50% and 85%, and KGF by 16% and 31%, respectively, while reducing SRD5A2 expression by 21% and 28%, respectively. Overall, the results of this in vitro study suggest that the botanical extract exhibits a response pattern similar to that of minoxidil, characterized by the suppression of pro-inflammatory cytokines and SRD5A2, along with enhanced expression of growth factors VEGF and KGF in HaCaT cells. These results provide a promising basis for further in vivo studies. Full article

Natural therapeutic alternatives are increasingly explored in endometriosis, a highly prevalent gynecological disorder with limited therapeutic options. Rosmarinus officinalis (rosemary) has attracted increasing scientific interest due to its biological activity. This study aimed to characterize a hydroethanolic rosemary extract (RE) and evaluate its effects on key cellular processes involved in endometriosis pathophysiology. Major phenolic compounds in RE were quantified by RP-HPLC, and antioxidant activity was assessed using DPPH, ABTS, and FRAP assays. After RE treatment, cell viability (WST-1), migration (wound healing assay), cell cycle distribution (DAPI staining), apoptosis (Annexin V/PI), p21 and cyclin A expression (Western blot), and intracellular ROS levels (DCFH-DA) were evaluated in endometrial stromal (t-HESC, St-T1b) and endometriotic epithelial (12-Z) cells. Phytochemical analysis revealed rosmarinic acid (RA) at 4.2%, while carnosic acid (CA) and carnosol (CS) together accounted for 23.7% of the extract. RE reduced cell viability and cell migration in 12-Z and t-HESC cells (p < 0.05). S-phase accumulation with a concomitant reduction in the G1 phase was observed across all evaluated cell lines (p < 0.05), along with increased p21 and cyclin A expression in stromal cells (p < 0.05). RE induced cell death in both 12-Z (p < 0.05) and St-T1b cells (p < 0.0001). In t-HESC cells, RE reduced both basal and H₂O₂-induced ROS levels (p < 0.01). These findings indicate that RE modulates key mechanisms involved in endometriosis pathophysiology, supporting its multi-target therapeutic potential as a nutraceutical approach for endometriosis management. Full article

The valorization of culinary by-products into functional bioactive resources represents a significant advancement in sustainable biotechnology. This study characterizes an extract derived from “battuto toscano” by-products, a traditional blend of garlic, onion, carrot, and celery trimmings, recovered through circular economy principles. Comprehensive antioxidant profiling was performed alongside biological evaluations on human cell lines and anti-glycation assays. Results from Folin–Ciocalteu, FRAP, and TEAC assays confirmed a high concentration of secondary metabolites with significant scavenging capacity. In vitro testing on primary human fibroblasts and HaCaT keratinocytes revealed a concentration- and time-dependent biological response, with lower concentrations showing better compatibility and transiently enhancing HaCaT metabolic activity. Furthermore, BTE reduced AGE-associated fluorescence in the BSA–glucose model, particularly at 5 mg/mL, supporting its potential anti-glycation activity. These findings establish “battuto toscano” by-products as a reservoir of sustainable biomolecules. This study offers a transformative resource for the pharma/nutraceutical sectors by bridging culinary tradition with biomedical innovation. Full article

The cone berries of Juniperus communis L. are rich in bioactive compounds, but biological properties of extracts are strongly influenced by the solvents used to obtain them. Therefore, this study aimed to evaluate the effect of solvent fractionation on the targeted polyphenolic profile and associated antioxidant, antimicrobial, and anticancer activities of pseudo-fruit extracts. The crude ethanolic extract was subjected to liquid–liquid partitioning to obtain ethyl acetate and n-butanol-soluble fractions, which were characterized by HPLC–MS and FTIR, while total polyphenol content was determined using the Folin–Ciocâlteu method and biological activities were assessed through DPPH, antimicrobial assays, and in vitro cytotoxicity on A375 melanoma and HaCaT keratinocyte cell lines. The ethyl acetate-soluble fraction showed the highest polyphenol content (361.08 ± 17.72 mg chlorogenic acid equivalents/g extract) and was enriched in hyperoside, whereas the n-butanol-soluble fraction contained lower phenolic levels and higher rutoside content; both fractions exhibited antioxidant activity correlated with phenolic content and weak-to-moderate antimicrobial activity, particularly against Streptococcus pyogenes. Cytotoxicity assays revealed a dose-dependent antiproliferative effect, with the ethyl acetate fraction displaying higher activity and greater selectivity toward melanoma cells, confirmed by apoptosis-related morphological changes. These findings demonstrate that solvent polarity plays a critical role in enriching bioactive phytochemicals and support the potential of J. communis fractions as sources of antioxidant and selective anticancer compounds. Full article

Wound healing remains a persistent health problem with no definitive solution. It is crucial to characterize the complex wound healing process and the various growth factors, cytokines, and polypeptides involved. Transforming growth factor beta1 (rhTGF-β1) stimulates different cell types, providing multifunctionality in the wound healing process. Since proteins are sensitive to proteases, drug delivery systems are needed. Developed polymeric carrier systems are as important as the active substance. The carrier systems used in our study aim to contribute to wound healing in addition to the rhTGF-β1. We hypothesized that PLGA nanoparticles embedded in PVA/Chitosan (PVA/Chi) hydrogels could enhance the therapeutic effect of rhTGF-β1. PVA/Chitosan hydrogels were prepared by the freezing/thawing method. Several characterization studies (Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), texture analysis, and cell culture) were performed to investigate the potential of the prepared formulations to enhance the therapeutic effect of rhTGF-β1. Hydrogel formulations reduced the inhibitory effect of rhTGF-β1 on keratinocytes. The H5 hydrogel exhibited a proliferative effect on fibroblast cells, which play a crucial role in wound healing, resulting in a 78.8% increase compared to the control. As the PVA content in the hydrogel formulations increased, bioadhesion and viscosity also increased. Although TGF-β1 inhibited keratinocytes, it induced migration of both NIH-3T3 and HACAT cell lines. The formulations developed exhibit the potential to improve the therapeutic efficacy of rhTGF-β1 in wound healing. A small amount of the protein can have the same therapeutic efficacy and fewer side effects because the developed polymeric carrier systems contribute to the therapeutic efficacy. Full article

Fusarium oxysporum f. sp. lycopersici is one of the most destructive soilborne pathogens affecting tomato production, reducing plant growth and yield and highlighting the need for sustainable management alternatives. Streptomyces spp. are promising microbial candidates due to their ability to combine antifungal activity with plant growth promotion characteristics. The objective of this study was to evaluate the biofertilizer and antifungal potential of Streptomyces spp. in Chonto tomato (Solanum lycopersicum Mill.) under greenhouse conditions. Seventy actinobacterial strains were screened in vitro against F. oxysporum, and eight exhibited significant antagonistic activity. Based on antagonistic activity, enzymatic profile, cytotoxicity, and plant growth-promoting potential, strains 1B260 and 445 were selected for greenhouse assays. Strain 1B260 achieved 43.5% mycelial growth inhibition and showed the highest phosphate-solubilizing capacity (420 µg/mL), while both strains displayed proteolytic and cellulolytic activity, low cytotoxicity in human skin cell lines (HaCaT and HDFa), nitrogen fixation, and ammonia production. In greenhouse assays under non-infected conditions, 1B260 showed the most consistent biofertilizer effect, promoting stem elongation. Under pathogen pressure, strain 445 improved plant performance compared to the infected control. Overall, strains 1B260 and 445 exhibited complementary roles in tomato crop management, highlighting the potential of multifunctional Streptomyces inoculants for sustainable biofertilization and biocontrol strategies. Full article

Cervical cancer remains one of the most lethal cancers affecting women, with infection by high-risk Human Papillomavirus (HR-HPV), especially HPV-16, recognized as a primary cause. Phyllanthus urinaria, a plant that grows in Indonesia, has demonstrated notable both antiviral and anticancer properties. This study aimed to investigate the potential of P. urinaria as both an antiviral and anti-cervical cancer agent. The HPV-16 E6 protein was modeled using homology modelling, with model accuracy verified through torsional angle assessment and identification of conserved regions. Molecular docking was performed to examine E6–p53 interactions. Fraction of n-hexane compounds of P. urinaria were further evaluated for their interaction with E6 by molecular docking and molecular dynamics simulation. Additionally, in vitro cytotoxicity assays were conducted using HaCaT (normal keratinocyte) and HeLa (cervical cancer) cell lines. Compounds from P. urinaria were found to interact with E6 within conserved regions and these interactions were more stable conformationally than those observed for p53. In vitro assay demonstrated that P. urinaria exhibited moderate cytotoxicity against HeLa cells but had limited toxicity toward HaCaT cells. The n-hexane fraction of P. urinaria leaves exhibits anti-cervical cancer activity by inhibiting HPV-16 E6 and eliminating cervical cancer cells. Full article