Ewa Barg

Background/Objectives: Cannabidiol (CBD) exhibits cytotoxic activity against cancer cells, but its poor aqueous solubility may limit its pharmaceutical application. This study investigated Santa Barbara Amorphous No. 15 (SBA-15) mesoporous silica as a CBD carrier and evaluated its biological effects in A549 and HCC827 non-small cell lung cancer cell lines using 2D and 3D culture models. Methods: SBA-15 was loaded with 40 or 50 wt% CBD by melt infiltration and characterized by powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC), and transmission electron microscopy (TEM). Free and SBA-15-loaded CBD were evaluated using the PrestoBlue assay in 2D cultures, live/dead fluorescence imaging in 3D spheroids, and YO-PRO-1/PI flow cytometry. Preliminary short-term survival was assessed in Galleria mellonella larvae at a single dose, with five larvae per group. Results: The mesoporous structure of SBA-15 was preserved after CBD loading. At 40 wt%, CBD was predominantly amorphous and confined within the pores, whereas at 50 wt%, partial crystallization outside the silica matrix was observed. The PrestoBlue results indicated that A549 cells were more sensitive than HCC827 cells to the CBD-containing formulations. SBA-15 alone did not reduce cell viability. Both cell lines were less sensitive under 3D conditions. Flow cytometry demonstrated increased membrane permeabilization, with limited YO-PRO-1-positive/PI-negative populations. No reduction in larval survival was observed under the tested conditions. Conclusions: CBD loading affected its physical state within SBA-15 but did not substantially alter its cytotoxic activity in 2D cultures. Whether SBA-15 improves CBD dissolution remains to be determined. Biological responses depended on the cell line and culture dimensionality. Unloaded SBA-15 did not markedly reduce cellular metabolic activity, and no larval mortality was observed at the single tested dose. Cancer selectivity could not be assessed because non-malignant lung cells were not included.

Background/Objectives: Ovarian cancer remains one of the deadliest gynecological malignancies due to late diagnosis, frequent recurrence, and the development of resistance to platinum-based chemotherapy. Naphthyridines constitute a diverse class of heterocyclic compounds with promising anticancer properties. The aim of this study was to evaluate the anticancer activity of three structurally related 2,7-naphthyridine derivatives (Naph-1, Naph-2, and Naph-3), identify the most active compound, and investigate its mechanism of action in ovarian cancer models. Methods: Cytotoxic activity was evaluated using the PrestoBlue assay in human cancer cell lines IGROV-1, MDAH-2774, SKOV-3, K562, MCF-7, and A549 and a non-cancerous CHO-K1 cell line. The most active derivative, Naph-3, was subsequently characterized in the two most sensitive ovarian cancer models (IGROV-1 and MDAH-2774) by evaluating its effects on 3D spheroid viability, cell proliferation, migration, apoptosis, and cell cycle progression. Electrochemical impedance spectroscopy was used as a label-free method for real-time monitoring of treatment-induced cellular responses. Cisplatin served as the reference compound. Results: Among the tested derivatives, Naph-3 demonstrated the strongest anticancer activity, with the highest sensitivity observed in ovarian cancer cell lines IGROV-1 (IC₅₀ 30.31 µM) and MDAH-2774 (IC₅₀ 53.68 µM). Naph-3 inhibited cell proliferation and migration, induced G1-phase cell cycle arrest, and promoted apoptotic cell death in both IGROV-1 and MDAH-2774 cells. In 3D spheroid models, Naph-3 retained significant activity in IGROV-1 spheroids, whereas MDAH-2774 spheroids displayed pronounced resistance to treatment. Although Naph-3 was less potent than cisplatin, it exhibited greater selectivity toward cancer cells, with lower toxicity against non-cancerous CHO-K1 cells. Impedance spectroscopy in a wide frequency range provided complementary kinetic information, supporting the interpretation that Naph-3 exerts a predominantly cytostatic effect, whereas cisplatin induces a more pronounced cytotoxic response. Conclusions: Naph-3 emerged as the most promising derivative among the tested compounds and exhibited multifaceted anticancer activity in IGROV-1 and MDAH-2774 ovarian cancer models. Its favorable selectivity profile, combined with antiproliferative, pro-apoptotic, and anti-migratory properties, highlights its potential as a promising compound for the development of novel therapeutic strategies against ovarian cancer.

Introduction and aim: Down syndrome is the most common chromosomal disorder characterized by a wide spectrum of clinical symptoms such as immune system dysregulation and co-occurring metabolic disorders, including an increased risk of cardiovascular disease. Therefore, the aim of this study was to evaluate the immunometabolic interactions in children adolescents and adults with Down syndrome (DS) and to compare selected inflammatory and metabolic parameters with those observed in their siblings. Materials and methods: The study included n= 63 individuals who were divided into two groups: group with DS n=42 (mean age: 14.2 ± 6.6) and control group (CG) n=21 (mean age: 15.6 ± 6.9). In addition, patients in both groups were also divided according to age ≤ 18 years and >18 years of age. Carbohydrate-lipid and immunological profiles were analyzed using spectrophotometric and immunoenzymatic methods. Statistical analysis was performed using R studio software. Results: In the DS group ≤ 18 years significantly higher obesity rates, i.e., Ponderal Mass Index (TMI), were observed (p=0.04), which was also associated with statistically significantly higher level of non-HDL (p=0.02) and apoB (p=0.04). Among lipid parameters, apolipoprotein A demonstrated relatively high diagnostic utility (AUC = 0.818, sens%=60.0, spec%=95.2). Significantly lower cytokine levels were observed in the DS group for IL-10 (p=0.006), IL-13 (p<0.01), and IL-22 (p=0.002). The highest diagnostic utility among the assessed cytokines was demonstrated for IL-5 (AUC = 0.814, sens%=71.10, spec%=88.1). Conclusion: The analyses conducted indicate significant differences between the studied groups of patients with Down syndrome and the control group. The presence of an additional copy of 21 the chromosome leads to changes in the immune system, influences the heterogeneous cytokine profile, and consequently may increase the development of metabolic disorders.

Background: Advanced therapy medicinal products (ATMPs) require strict control of critical process parameters (CPPs) to ensure manufacturing efficiency. The relative impact of donor systemic factors, such as vitamin D status, versus technical process parameters on dental pulp-derived stem cell (DPSC) production remains unclear. Methods: In this prospective observational study, 250 adults undergoing extraction of impacted mandibular third molars were included. Dental pulp was processed under a standardized SOP using different preparation methods and enzyme conditions. Primary endpoints were serum 25(OH)D concentration and cell yield; secondary endpoints included number of passages and cryovials. Results: Mean 25(OH)D concentration was 30.1 ± 14.5 ng/mL and was higher in supplemented individuals (38.2 ± 14.0 vs. 25.6 ± 12.7 ng/mL; p < 0.0001) but was not associated with cell yield (ρ = 0.14, p = 0.168) or passages (ρ = 0.07, p = 0.406). In contrast, process parameters showed strong effects: scissor preparation resulted in a substantially higher yield than mechanical methods (median 5.00 vs. 1.00 million cells; p = 3.6 × 10⁻¹³), and type II collagenase was independently associated with a higher yield (+2.04 million cells; p = 0.026). The number of passages was the strongest predictor of yield (β = 2.28 million per passage; p < 10⁻²⁶). Post-thaw viability remained high (mean 90.1% and range 81–98%). Conclusions: Manufacturing efficiency of DPSCs is primarily determined by critical process parameters, particularly preparation method, enzyme selection, and passage control, whereas donor vitamin D status did not significantly influence outcomes under the studied SOP. These findings highlight process standardization as the key driver of reproducible ATMP manufacturing.

Background/Objectives: Metformin (Met) is a potent antidiabetic drug that also exhibits anticancer, antioxidant, and organ-protective properties. Luteolin (Lut), a naturally occurring flavonoid found in many plant species, possesses anticancer, antioxidant, and anti-inflammatory effects. Since both compounds affect cellular metabolism and oxidative balance, the analysis of metabolites produced in living cells provides insight into biochemical alterations occurring in cancer cells and enables monitoring of treatment response. Methods: In this study, Met (1–20 mM) and Lut (1–100 µM) were tested in vitro, both individually and in combination, to evaluate their effects on cell viability, free radical levels, and metabolite profile alterations in cancer and normal cell lines (MDA-MB-231, SW620, and V79). Cell viability was assessed using the MTT assay at two time points (24 h and 48 h), while reactive oxygen species (ROS) levels were measured after hydrogen peroxide stimulation (100 µM H₂O₂) using the DCF-DA assay. Metabolomic changes induced by Met and Lut were analyzed by ¹H NMR spectroscopy. Results: The analysis showed that Lut reduced the viability of MDA-MB-231 cells at both time points, whereas Met decreased viability only after prolonged incubation. Met did not inhibit the proliferation of SW620 colorectal cancer cells, while Lut reduced viability at higher concentrations (100 µM after 24 h, and 50–100 µM after 48 h). Conclusions: The combination of metformin and luteolin demonstrated additive effects in reducing cell viability and oxidative stress compared with single-compound treatments. Normal V79 fibroblasts responded to both Met and Lut, individually and in combination. Both compounds exhibited moderate antioxidant properties in cells exposed to 100 µM H₂O₂. Lut (25 µM) reduced free radical levels in MDA-MB-231 cells, whereas Met (2.5 mM) did so in SW620 cells. The combination of both compounds increased ROS levels in SW620 cells subjected to oxidative stress. Overall, co-treatment with metformin and luteolin altered metabolic pathways and induced changes in intra- and extracellular metabolite levels across all tested cell lines. The observed additive effects suggest that the combined use of metformin and luteolin may enhance anticancer and antioxidant responses, warranting further in vivo studies to confirm these interactions.

Lung cancer is the leading cause of cancer mortality worldwide, with a poor prognosis driven by late diagnosis, systemic toxicity of existing therapies, and rapid development of multidrug resistance (MDR) to agents such as paclitaxel and cisplatin. MDR arises through multiple mechanisms, including overexpression of efflux transporters, alterations in apoptotic pathways, and tumour microenvironment-mediated resistance. The application of nanotechnology offers a potential solution to the aforementioned challenges by facilitating the enhancement of drug solubility, stability, bioavailability, and tumour-specific delivery. Additionally, it facilitates the co-loading of agents, thereby enabling the attainment of synergistic effects. Halloysite nanotubes (HNTs) are naturally occurring aluminosilicate nanocarriers with unique dual-surface chemistry, allowing hydrophobic drug encapsulation in the positively charged lumen and functionalisation of the negatively charged outer surface with targeting ligands or MDR modulators. This architecture supports dual-delivery strategies, enabling simultaneous administration of phytocannabinoids and chemotherapeutics or efflux pump inhibitors to enhance intracellular retention and cytotoxicity in resistant tumour cells. HNTs offer additional advantages over conventional nanocarriers, including mechanical and chemical stability and low production cost. Phytocannabinoids such as cannabidiol (CBD) and cannabigerol (CBG) show multitarget anticancer activity in lung cancer models, including apoptosis induction, proliferation inhibition, and oxidative stress modulation. However, poor solubility, instability, and extensive first-pass metabolism have limited their clinical use. Encapsulation in HNTs can overcome these barriers, protect against degradation, and enable controlled, tumour-targeted release. This review examined the therapeutic potential of HNT-based phytocannabinoid delivery systems in the treatment of lung cancer, with an emphasis on improving therapeutic selectivity, which represents a promising direction for more effective and patient-friendly treatments for lung cancer.

Background: Colorectal cancer is one of the leading and most lethal neoplasms. Standard chemotherapy is ineffective, especially in metastatic cancer, and does not target cancer stem cells. A promising approach to improve cancer treatment is the combination therapy of standard cytostatic drugs with natural compounds. Several plant-derived compounds have been proven to possess anticancer properties, including the induction of apoptosis and inhibition of cancer invasion. This study was focused on investigating in vitro the combination of camptothecin (CPT) with celastrol (CEL) or resveratrol (RSV) as a potential strategy to target metastatic (LOVO) and stem-like (LOVO/DX) colon cancer cells. Methods: The genotoxic effects that drive cancer cells into death-inducing pathways and the ability to inhibit the migratory properties of cancer cells were evaluated. The γH2AX+ assay and Fast-Halo Assay (FHA) were used to evaluate genotoxic effects, the annexin-V apoptosis assay to rate the level of apoptosis, and the scratch test to assess antimigratory capacity. Results: The results showed that both combinations CPT-CEL and CPT-RSV improve general genotoxicity of CPT alone on metastatic cells and CSCs. However, the assessment of specific double-stranded breaks (DSBs) indicated a better efficacy of the CPT-CEL combination on LOVO cells and CPT-RSV in LOVO/DX cells. Interestingly, the combinations CPT-CEL and CPT-RSV did not improve the pro-apoptotic effect of CPT alone, with both LOVO and LOVO/DX cells suggesting activation of different cell death mechanisms. Furthermore, it was found that the combinations of CPT-CEL and CPT-RSV improve the inhibitory effect of camptothecin on cell migration. Conclusions: These findings suggest the potential utility of combining camptothecin with celastrol or resveratrol in the treatment of colon cancer, including more aggressive forms of the disease. So far, no studies evaluating the effects of combinations of these compounds have been published in the available medical databases.

Background: This study investigates the impact of environmental pollution on the quality and viability of dental stem cells (DSCs) from impacted third molars. By comparing DSCs from patients in industrial areas with high air pollution and those from non-industrial regions, the research assesses the adverse effects of heavy metals on stem cell proliferation. Methods: Impacted lower third molars were collected from 28 patients—10 from industrial and 18 from non-industrial areas. Patients were divided into two age groups: 18–27 years and 28–38 years old. Dental pulp was extracted under sterile conditions, and DSCs were isolated and cultured. Heavy metal concentrations in dental tissues were measured using atomic absorption/emission spectrometry. Results: The study found significantly higher concentrations of copper and lead in the dental tissues of patients in industrial areas. Cell viability was lower in samples from these areas, with a statistically significant difference in average doubling time and the number of cells obtained after the first passage. There was no significant impact of gender on heavy metal content, except for higher iron levels in men. Conclusions: Exposure to industrial pollutants negatively affects the viability and proliferation of DSCs, but there are no differences in differentiation in the osteogenic medium regarding cell mineralization. These studies highlight the importance of environmental factors for oral health, suggesting that residents of polluted areas may face greater difficulties in dental and regenerative treatments. Further research is needed to develop strategies to mitigate the effects and improve clinical outcomes for affected populations.

Phytocannabinoids, compounds found in Cannabis sativa L., are used in oncology and palliative care to reduce the adverse reactions of standard therapies. Cancer patients use formulations of Cannabis sativa L. to manage the anxiety, pain, and nausea associated with cancer treatment, and there is growing evidence that some of them may exhibit anticancer properties. In this study, we tested the anticancer potential of selected cannabinoids CBD (cannabidiol) and its quinone derivative CBD-HQ (cannabidiol hydroquinone), CBG (cannabigerol) and its acid derivative CBG-A (cannabigerolic acid), as well as a combination of CBD+CBG on the colon cancer cell line SW-620. The MTT assay was used to determine the cannabinoids’ ability to induce colon cancer cell death. All cannabinoids were cytotoxic at the lowest concentration (3 μg/mL). The half maximal inhibitory concentration (IC50) ranged from 3.90 to 8.24 μg/mL, depending on the substance. Cytotoxicity was confirmed in a 3D spheroidal cell culture with calcein and propidium iodide staining. The amount of intracellular reactive oxygen species (ROS) was examined using a DCF-DA assay. CBG showed the lowest antioxidant activity of all the cannabinoids tested. The level of intracellular ROS decreased only by 0.7–18%. However, CBG-A induced the strongest reduction in ROS level by 31–39%. Our results suggest that cannabinoids represent an interesting research direction with great implementation potential. These preliminary results represent the beginning of research into the potential of these substances for anticancer treatment and underscore the potential for further research.

Colorectal cancers are one of the leading cancers worldwide and are known for their high potential for metastasis and resistance to therapy. The aim of this study was to investigate the effect of various combination therapies of irinotecan with melatonin, wogonin, and celastrol on drug-sensitive colon cancer cells (LOVO cell line) and doxorubicin-resistant colon cancer stem-like cells (LOVO/DX cell subline). Melatonin is a hormone synthesized in the pineal gland and is responsible for circadian rhythm. Wogonin and celastrol are natural compounds previously used in traditional Chinese medicine. Selected substances have immunomodulatory properties and anti-cancer potential. First, MTT and flow cytometric annexin-V apoptosis assays were performed to determine the cytotoxic effect and the induction of apoptosis. Then, the potential to inhibit cell migration was evaluated using a scratch test, and spheroid growth was measured. The results showed important cytotoxic effects of the drug combinations on both LOVO and LOVO/DX cells. All tested substances caused an increase in the percentage of apoptotic cells in the LOVO cell line and necrotic cells in the LOVO/DX cell subline. The strongest effect on the induction of cancer cell death was observed for the combination of irinotecan with celastrol (1.25 µM) or wogonin (50 µM) and for the combination of melatonin (2000 µM) with celastrol (1.25 µM) or wogonin (50 µM). Statistically significant improvements in the effect of combined therapy were found for the irinotecan (20 µM) and celastrol (1.25 µM) combination and irinotecan (20 µM) with wogonin (25 µM) in LOVO/DX cells. Minor additive effects of combined therapy were observed in LOVO cells. Inhibition of cell migration was seen in LOVO cells for all tested compounds, while only irinotecan (20 µM) and celastrol (1.25 µM) were able to inhibit LOVO/DX cell migration. Compared with single-drug therapy, a statistically significant inhibitory effect on cell migration was found for combinations of melatonin (2000 µM) with wogonin (25 µM) in LOVO/DX cells and irinotecan (5 µM) or melatonin (2000 µM) with wogonin (25 µM) in LOVO cells. Our research shows that adding melatonin, wogonin, or celastrol to standard irinotecan therapy may potentiate the anti-cancer effects of irinotecan alone in colon cancer treatment. Celastrol seems to have the greatest supporting therapy effect, especially for the treatment of aggressive types of colon cancer, by targeting cancer stem-like cells.