Background/Aim: Among various types of brain tumors, glioblastoma is the most malignant and highly aggressive brain tumor that possesses a high resistance against anticancer drugs. To understand the underlined mechanisms of tumor drug resistance, a new and more effective research approach is required. The three dimensional (3D) in vitro cell culture models could be a potential approach to study cancer features and biology, as well as screen for anti-cancer agents due to the close mimicry of the 3D tumor microenvironments. Materials and Methods: With our developed 3D alginate scaffolds, Ilumina RNA-sequencing was used to transcriptomically analyze and compare the gene expression profiles between glioblastoma cells in traditional 2-dimensional (2D) monolayer and in 3D Ca-alginate scaffolds at day 14. To verify the reliability and accuracy of Illumina RNA-Sequencing data, ATP-binding cassette transporter genes were chosen for quantitative real-time polymerase chain reaction) verification. Results: The results showed that 7,411 and 3,915 genes of the 3D glioblastoma were up-regulated and down-regulated, respectively, compared with the 2D-cultured glioblastoma. Furthermore, the Kyoto Encyclopaedia of Genes and Genomes pathway analysis revealed that genes related to the cell cycle and DNA replication were enriched in the group of down-regulated gene. On the other hand, the genes involved in mitogen-activated protein kinase signaling, autophagy, drug metabolism through cytochrome P450, and ATP-binding cassette transporter were found in the up-regulated gene collection. Conclusion: 3D glioblastoma tumoroids might potentially serve as a powerful platform for exploring glioblastoma biology. They can also be valuable in anti-glioblastoma drug screening, as well as the identification of novel molecular targets in clinical treatment of human glioblastoma.

This study aimed to profile the metabolites, piperine content, and biological activities (antioxidant, anti-inflammatory, anticancer, antibacterial, and apoptotic cell) of 15 samples of Piper nigrum L. from various geographic regions of Thailand. UHPLC-QTOF-MS was applied to establish alkaloid profiles of the P. nigrum L. samples. PCA and HCA analysis showed that the samples from the PN-1, PN-2, and PN-11 sources differed significantly from those of other sources in terms of compound type and alkaloid concentration profile. Comparative analysis based on a validated HPLC-DAD method was used for the standardization of 15 varieties of P. nigrum L. with piperine as a standard phytomarker which revealed that PN-4 source possessed a high content of piperine. PN-2 showed the highest antioxidant capacity (DPPH activity) while PN-10, PN-12, and PN-15 showed the highest FRAP activity. PN-11 source showed high inhibition of anti-inflammatory and anticancer properties. Furthermore, PN-3, and PN-4 also showed the highest both anti-inflammatory and antibacterial activity. Almost samples have been shown to induce late apoptosis and necrosis in SW480 cell line. This study will also be useful for the development of quality control parameters through the standardization of varieties of P. nigrum L. as raw materials in traditional medicine formulations. HIGHLIGHTS Metabolite profiling of the nigrum L. varieties from 15 different geographical locations in Thailand Quality control parameter for standardization based on marker analysis by HPLC-DAD using piperine as the marker compound Comparisons of metabolites among 15 different cultivations by Principal Component Analysis (PCA) and hierarchical cluster analysis (HCA) Biological activity of the nigrum L. varieties from 15 different geographical locations in Thailand GRAPHICAL ABSTRACT

Background: Abutilon indicum (L.) Sweet (Malvaceae) is a traditional medicinal plant known for its antidiabetic properties in Ayurveda and other health systems. Aims: This study aims to profile metabolites in Abutilon indicum (L.) Sweet extracts (AI) and elucidate their antidiabetic mechanisms through bioinformatics and experimental methods. Study Design: The ethanolic (AIE) and aqueous (AIA) extracts were evaluated for their inhibitory effects on α ‐glucosidase and α ‐amylase, as well as their impact on glucose metabolism in 3T3‐L1 adipocytes and L6 skeletal muscle cells. AIE was characterized via HPLC‐DAD‐QTOF‐MS, with network pharmacology and molecular docking analyses used to explore molecular targets. Methods: In vitro assays were performed to assess enzyme inhibition, and cell line studies HPLC‐DAD‐QTOF‐MS were utilized for compound characterization. Network pharmacology and molecular docking were conducted to reveal underlying antidiabetic mechanisms. Results: LC‐MS‐QTOF analysis identified gallic acid, stigmasterol, and riboflavin as abundant compounds. The AIE exhibited significant α ‐glucosidase (IC ₅₀ = 74.15 ± 1.61 μg/mL) and α ‐amylase inhibition (IC ₅₀ = 13.41 ± 0.71 μg/mL). Moreover, it enhanced glucose consumption in 3T3‐L1 cells (IC ₅₀ = 6.25 μg/mL) and promoted glucose uptake in L6 myotubes. Network pharmacology analyses highlighted the PI3K–Akt signaling pathway’s role in facilitating glucose transport. Conclusion: The phytochemicals in AIE may contribute significantly to its antidiabetic effects, particularly through the modulation of glucose transport via the PI3K–Akt pathway. Future studies should focus on the preclinical development of safe herbal formulations utilizing these mechanisms for effective diabetes management.

Centella asiatica (L.) Urb. (Apiaceae) is widely incorporated into dermatological and wound-healing formulations, where dechlorophylization is commonly used to improve extract stability, although its biological consequences are not fully understood. In this study, an ethanolic extract of C. asiatica leaves (CAE) was compared with a dechlorophylized C. asiatica extract (DCAE) in terms of cytotoxicity, wound-healing, anti-inflammatory, and antimicrobial activities, complemented by network pharmacology analysis. DCAE exhibited improved cytocompatibility, remaining non-cytotoxic up to 100 µg/mL, whereas CAE showed cytotoxic effects above 25 µg/mL. In contrast, CAE demonstrated superior wound-healing performance, reducing the residual wound area to 21.77% compared with 36.81% for DCAE and inducing higher Ki67 expression (1.94-fold versus 1.58-fold). These findings suggest that chlorophyll-associated constituents may contribute to keratinocyte proliferation and re-epithelialization. Although DCAE was enriched in pentacyclic triterpenes, including asiatic acid, asiaticoside, madecassic acid, and madecassoside, it showed stronger anti-inflammatory effects by more effectively suppressing CXCL10, IL-6, TNF-α, and IL-8 expression. Both extracts displayed weak antibacterial activity but moderate antifungal effects against Candida albicans, likely mediated through membrane disruption. Network pharmacology further indicated that wound-healing activity is associated with the regulation of COL1A1 and VEGFA through quercetin, kaempferol, and asiatic acid by way of the PI3K-Akt and MAPK signaling pathways, whereas anti-inflammatory effects involve modulation of PTGS2 and TNF through the NF-κB and IL-17 pathways. Overall, DCAE enhances cytocompatibility and anti-inflammatory activity, while CAE retains stronger regenerative capacity. These findings support the rational design of tailored formulations for specific dermatological applications, such as promoting wound repair or alleviating chronic skin inflammation.