BACKGROUND Chemoresistance significantly limits the therapeutic efficacy of neoadjuvant chemotherapy (NACT) in advanced gastric cancer (AGC). There is an urgent need to identify robust biomarkers predictive of NACT response and to elucidate the molecular mechanisms that drive resistance. In this study, we systematically assess whether intercellular adhesion molecule 2 (ICAM2 ) predicts NACT response in patients with AGC and delineate its mechanistic role in chemoresistance. AIM To investigate the predictive significance and mechanistic role of ICAM2 in mediating 5-fluorouracil (5-FU) resistance in gastric cancer (GC). METHODS Real-time PCR, Western blotting, enzyme-linked immunosorbent assay, and immunohistochemistry were conducted to assess alterations in ICAM2 expression between 5-FU-sensitive and -resistant GC cells as well as in AGC patient samples. Cytotoxicity assays, colony formation, flow cytometry, analyses of apoptosis-related proteins, and xenograft experiments were employed to elucidate the role of ICAM2 in mediating chemoresistance. The mechanism underlying ICAM2 -mediated chemoresistance was further explored through RNA sequencing (RNA-seq), nuclear-cytosolic fractionation, co-immunoprecipitation, luciferase reporter, and chromatin immunoprecipitation assays. RESULTS Low ICAM2 expression correlated significantly with poor NACT response, advanced tumor stage, worse differentiation, and reduced overall survival and disease-free survival in AGC patients. Pre-NACT serum ICAM2 demonstrated high predictive accuracy (area under the curve = 0.876) in discriminating chemotherapy responders from non-responders. Mechanistically, ICAM2 knockdown conferred 5-FU resistance through two intertwined processes: Inhibition of caspase-dependent apoptosis and promotion of immunosuppressive M2 macrophage polarization within the tumor microenvironment. At the molecular level, loss of ICAM2 activated the TGF-β/Smad pathway, leading to transcription factor SP1-mediated pleiotrophin (PTN) upregulation. Elevated PTN further enhanced GC cell survival and may contribute to M2 macrophage polarization, thereby amplifying chemoresistance. Importantly, targeted inhibition of TGF-β signaling reversed ICAM2-associated chemoresistance in both cell culture and xenograft models. CONCLUSION Our study highlights the clinical impact of ICAM2 downregulation predicting poor outcome and NACT response in AGC patients, and reveals a novel ICAM2/TGF-β/Smad/SP1/PTN signaling mediating 5-FU resistance in GC.

Volatile organic compounds (VOCs) are prevalent in both indoor and outdoor environments and have been linked to health effects. This study aimed to assess VOC-induced effects on the respiratory epithelium using an invitro human bronchial epithelial air-liquid interface (ALI) model. A human bronchial epithelial cell line, 16HBE, was cultured at ALI and exposed to relevant concentrations of two representative VOCs, acrolein or formic acid, and matched filtered air (control) in a CelTox exposure system for two hours to replicate an acute inhalation exposure. Cells were allowed to recover for 24 h before cell lysate and culture media were collected for analysis. Cytotoxicity, based on LDH activity, significantly increased at the highest doses tested for both VOCs. A dose-dependent increase in barrier permeability was observed for confluent cells exposed to acrolein and formic acid. Acrolein and formic acid exposure induced IL-8, TNFα, and HMOX-1 expression, genes indicative of proinflammatory signaling and oxidative stress, respectively. Formic acid, but not acrolein, exposure also increased expression of PINK1, a gene indicative of mitophagy. Benchmark concentration (BMC) modeling of invitro acrolein data yielded a BMCL (benchmark concentration lower confidence limit) that substantiates the stringency of OSHA's 8-hour permissible exposure limit (PEL). In contrast, BMC modeling of invitro formic acid data produced BMCLs below existing regulatory exposure thresholds. Collectively, these findings demonstrate that this model is a plausible invitro tool to investigate VOC-induced effects on the airway and supports its utility in VOC safety evaluation.

Posttranscriptional regulation is particularly prominent during the maternal-to-zygotic transition (MZT), a developmental phase during which a large proportion of maternally provided mRNAs is repressed and cleared from metazoan embryos. RNA-binding proteins (RBPs) are key components of the posttranscriptional regulatory machinery. We show that the ORB2 RBP, the Drosophila ortholog of the human cytoplasmic polyadenylation element binding protein (hCPEB) 2-4 protein subfamily, binds to hundreds of maternally provided, rare-codon-enriched mRNAs in early embryos and that ORB2 targets are translationally repressed and unstable during the MZT. We identify a U-rich motif enriched in ORB2 targets' 3'UTRs and show that this motif confers ORB2 binding and repression to a luciferase reporter mRNA in S2 tissue culture cells. When tethered to a luciferase reporter, ORB2 and hCPEB2 (but not ORB and hCPEB1) repress translation and the C-terminal Zinc-binding ("ZZ") domain of ORB2 is necessary and sufficient for repression. ORB2 interacts with a suite of posttranscriptional regulators in early embryos; a subset of these interactions is lost upon deletion of the ZZ domain, notably with the Cup repressive complex. ORB2 targets significantly overlap with those previously identified for the repressive RBP, Smaug (SMG). Analysis of the early embryo's translatome in the presence or absence of the endogenous ZZ domain shows that mRNAs bound by ORB2 but not by SMG move onto polysomes upon ZZ domain deletion, whereas cobound transcripts do not, consistent with coregulation of the latter set of transcripts by both RBPs. Our results assign a function to the ZZ domain and position ORB2 in the posttranscriptional network that regulates maternal transcripts during the Drosophila MZT.

Investigation of toxicological profile and possible side effects of engineered nanomaterials (ENMs) is of high importance. Historically, two-dimensional (2D) cell culture was used to study the toxicity of the ENMs, but due to their inability to simulate in vivo cell behavior, three-dimensional (3D) cell culture systems have been developed. Nanotoxicity studies initiate with in vitro experiments and continue with in vivo studies, which are very challenging and sometimes accompanied by conflicting data due to the in vitro-in vivo gap. Thus, scientists are turning their attention to microfabrication techniques and engineered systems "called organ-on-a-chips", which act as an intermediate between in vivo and in vitro systems. The present account tries to review the classical study models and suitably cover the emerging 3D culture models including scaffold-free and scaffold-based 3D cell cultures, 3D co-culture with direct contact and without cell-cell contact methods as well as microfluidic-based tissue chips and organoids. Overall, this review aims to give readers a better insight about the ENMs' toxicology and fill the gaps between the knowledge and practical techniques. Hopefully, the presented information will resolve the issues of 2D in vitro cultures and display the clinically relevant responses to the concerns of therapeutic ENMs.

Rising numbers of organ failures have intensified the demand for high-performance biomaterials to support the development of bioartificial organs and advanced bioreactors. Hollow fiber membranes (HFMs) are particularly well-suited for such applications, including bioartificial kidney, liver, and 3D cell culture systems, due to their unique architecture and functional versatility. In this study, we engineered HFMs by blending amphiphilic Pluronic F127 (PF127) with poly(ether sulfone) (PES), aiming to enhance both separation efficiency and cellular attachment and proliferation. Physicochemical characterization revealed that PF127 incorporation resulted in a concentric, porous membrane structure with significantly improved porosity as compared to that of plain PES HFMs. Biocompatibility was assessed using human embryonic kidney (HEK293) and hepatocellular carcinoma liver (HepG2) cell lines. Confocal microscopy, MTT cell viability assays, flow-cytometry-based live/dead assays, and calcein AM/propidium iodide staining demonstrated that PF127/PES HFMs strongly support the attachment and proliferation of viable cells. The attached cells exhibited high metabolic activity and formed three-dimensional spheroids, indicating the bioactive influence of PF127. Hemocompatibility evaluation by hemolysis and terminal complement complex (SC5b9) showed that the HFMs fabricated were hemocompatible, suggesting a diminished inflammatory response. Additionally, separation performance evaluation demonstrated a high ultrafiltration coefficient, highest for 2.5 PF127 (173.83 ± 7.31 mL m-2 h-1 mmHg-1) and efficient removal of a broad range of uremic toxins, including urea, creatinine, macroglobulin analogs, and protein-bound toxins such as indoxyl sulfate. Collectively, the enhanced cytocompatibility with kidney and liver cells, hemocompatibility, and separation capability of PF127/PES HFMs make them promising scaffolds for bioartificial kidney and liver applications.

Compared with positive control MRTX849, the synthesized compounds 7g, 7p, 7q, 7r, 7v, and 7y displayed stronger antiproliferative activities against H358 cells with IC50 values of < 1 nM (3D cell culture) and comparable inhibitory potency against KRASG12C. Among them, 7q (MH5) exhibited satisfactory cellular selectivity, moderate pharmacokinetic characters, and good anticancer effects on pancreatic, colorectal cancer xenograft in vivo. Meaningfully, 7q combined with Nrf2 inhibitor ML385 or PARP7 inhibitor RBN-2397 greatly enhanced the sensitivity of 7q against lung cells (H1373) in vivo. Furthermore, combination therapy of 7q with pan-USP inhibitor PR-619 obtained a statistically significant synergistic inhibition of H1373 cell growth in vitro and in vivo. Our findings indicate that 7q may be a promising drug candidate for the treatment of cancers harboring the KRASG12C mutation, and the results of the combination regimen established a pharmacological foundation for addressing drug resistance.

Is the endometrial microbiome altered in women who fail to get pregnant after ART and do microbial-derived metabolites influence endometrial cellular mechanisms important for embryo implantation? The endometrial microbiome in women who fail to get pregnant after ART is more diverse and has fewer lactobacilli species than the endometrial microbiomes of women who become pregnant; the short-chain fatty acid butyrate, a common metabolite found in the presence of increased microbial diversity, diminishes endometrial epithelial barrier function and increases the expression of inflammatory markers. Shifts in the endometrial microbial community structure have been linked to fertility and pregnancy complications although the underlying mechanisms are poorly understood. Microbial metabolites at other mucosal surfaces, such as the gut, act as important modulators of immune and barrier function, particularly in epithelial cells. Effects of changes in local bacterial microbial populations on fertility, and how their metabolites might influence endometrial cell function have not been explored. In this prospective longitudinal study of ART outcomes, 29 nulliparous women with unexplained infertility were recruited between October 2016 and February 2018. Endometrial tissue samples were taken for microbiome analysis and endometrial transcriptomics prior to ART. For primary cell culture studies, endometrial biopsies were obtained from fertile women of reproductive age undergoing laparoscopic surgical investigation between February 2021 and September 2023. In vitro models of implantation were established using endometrial cell lines and primary endometrial stromal cells. Microbiome 16S sequencing analysis was performed on bacterial DNA isolated from endometrial biopsies and correlated with receptivity markers. Endometrial RNA sequencing data from women undergoing ART were used to analyse differential gene expression of receptivity and decidualization markers in women who had a positive or negative ART cycle outcome. In vitro models, using both established endometrial cell lines and primary human endometrial epithelial cells and stromal cells, were developed to investigate the effects of microbial-derived metabolites. An in vitro model of peri-implantation was used to test the effect of butyrate on endometrial epithelial receptivity and stromal cell decidualization. Endometrial microbiome 16S sequencing revealed a lower abundance of Lactobacillus spp. and significantly higher abundance of pathogenic species such as Prevotella spp. and Corynebacterium spp. in women who did not become pregnant after ART. Endometrial microbiota from women who had positive ART outcomes showed significantly lower diversity indices. Intriguingly, analysis of endometrial RNA sequencing data from women with unexplained infertility undergoing ART showed that negative ART outcomes were associated with higher levels of some receptivity and decidualization markers in their endometrial tissue. Butyrate, but not lactate or acetate, also increased some markers of epithelial receptivity and stromal decidualization. Butyrate exposure also activated defence mechanisms in cultured endometrial epithelial cells by inducing expression of antimicrobial peptide(s) and inflammation markers, as well as impairing the barrier integrity of endometrial epithelial cell monolayers. The RNA-seq data used for the study can be found in GEO database, GEO ID GSE144895. The data for the 16S sequencing can be accessed in SRA BioProject number PRJNA1338067. Limitations of our study include the cohort size and technical challenges that precluded absolute butyrate measurement in endometrial tissue biopsies. Biopsy collection from women undergoing gynaecological investigation varied in menstrual cycle staging and fertility diagnoses, which may contribute to the variability between responses obtained from in vitro stimulations. The transferred embryos were not genetically tested, but were all of good or top quality. Our findings indicate that the endometrial microbiome is altered in women who fail to become pregnant after ART, and that the microbial-derived metabolite butyrate can induce inflammation and impair endometrial epithelial barrier function and drive increased gene expression levels of markers for epithelial receptivity and stromal decidualization in in vitro models of peri-implantation. Endometrial microbial dysbiosis and higher expression of receptivity markers were found in women who failed to establish pregnancy post-ART. Negative ART outcomes in this cohort were found to correlate with the presence of a wider, more diverse microbial community that includes Prevotella spp., which is among the butyrate-producing bacteria. Further investigation of the microbial metabolome in healthy endometrium would help clarify the physiological role of butyrate and other bacterial metabolites in endometrial function. This research was supported by the Grant for Fertility Innovation from Merck KGaA, grant award number 15692. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. N/A.

Background/Objectives: Indocyanine green (ICG) is an FDA-approved, near-infrared fluorescent dye widely used for tumor imaging. This study aimed to evaluate the photodynamic efficacy and selectivity of ICG as a photosensitizer in photodynamic therapy (PDT) against MCF-7 breast cancer cells in 2D monolayers and 3D collagen-embedded cell cultures that simulate ECM diffusion, and to confirm direct generation of singlet oxygen (1O2) as the primary cytotoxic species. Methods: MCF-7 breast adenocarcinoma cells and HMEC normal mammary epithelial cells were cultured in 2D monolayers, with MCF-7 cells additionally grown in 3D collagen type I matrices to mimic tumor environments. Cells were incubated with 50 µM ICG for 30 min, washed, and irradiated with a 780 nm diode laser at 39.8 mW/cm2. Cell viability was quantified using the Muse® Count &amp; Viability assay at multiple time points, while ICG uptake and penetration were assessed via flow cytometry, fluorescence microscopy, and confocal imaging. Direct 1O2 production was measured through its characteristic 1270 nm phosphorescence using time-resolved near-infrared spectrometry. Results: ICG-PDT reduced MCF-7 viability to 58.3 ± 7.4% in 2D cultures (41.7% cell kill, p &lt; 0.0001) and 70.2 ± 10.7% in 3D cultures (29.8% cell kill, p = 0.0002). In contrast, normal HMECs maintained 91.0 ± 1.3% viability (only 9% reduction, p = 0.08), resulting in a therapeutic index of approximately 4.6. IC50 values in 2D MCF-7 cultures decreased over time from 51.4 ± 3.0 µM at 24 h to 27.3 ± 3.0 µM at 72 h. ICG uptake was higher in 2D (78%) than in 3D (65%) MCF-7 cultures, with diffusion in 3D collagen exhibiting linear depth-dependent penetration. Notably, the singlet-oxygen phosphorescence signal, though weak and requiring highly sensitive detectors, provided direct evidence of efficient 1O2 generation. Conclusions: ICG as a photosensitizer in photodynamic therapy using clinically compatible parameters is highly cytotoxic to MCF-7 breast cancer cells while largely sparing HMECs in 2D cell culture. Direct spectroscopic evidence confirms efficient 1O2 generation, which contributes significantly to the cytotoxicity. The reduced efficacy in 3D versus 2D models highlights the importance of penetration barriers also present in solid tumors. These results support further preclinical and clinical investigation of ICG as a dual imaging-and-therapy (theragnostic) agent for selective photodynamic treatment of breast cancer.

Electrospun polymeric patch integrated with an electrochemical biosensor for real-time superoxide monitoring in cell culture models of chronic wounds.

Metformin, the common anti-hyperglycemic agent, is emerging with pharmacological significance as an effective anti-cancer modulator. Its efficacy as an anti-cancer modulator is reported in pre-clinical and clinical studies. Therefore, an attempt was made to identify the possible invitro anti-cancer molecular mechanisms studied on breast cancer (BC) cell lines. An advanced literature search was conducted in the PubMed database using search terms "Metformin, Cell culture, Breast neoplasms." Different anti-cancer molecular mechanisms induced by metformin (MET) identified in cell culture studies are presented in this paper. It was identified that MET induces molecular pathways that exert anti-cancer effects when treated on BC cells. Inhibition of oxidative phosphorylation, adenosine monophosphate-activated protein kinase mediated anticancer effects, anti-proliferation and inhibition of cell migration, alteration of tumor micro-environment, synergetic effects with conventional chemotherapies and other potential molecules, induced apoptosis and ferroptosis were mainly identified as MET-induced pathways that affect BC cells. Metformin induces diverse anti-cancer biochemical pathways through which it exhibits a potential to be used as an anti-cancer therapeutic in BC.

An alternative model for HEV infection in the HepaRG cell line.

Role of Cell Culture Scaffold Stiffness on Sonoporation Efficiency.

Organoid-driven nanomedicine platform development.

Experimental production of synthetic infectious porcine circovirus 2b particles in swine testicular cells.

Expression of kynurenine aminotransferase-2 in different mouse brain-derived cells: A comprehensive study in cell cultures.

Differential effects of type I, III, and V collagens on mammary epithelial development in vitro.

Perfluorohexanoic acid (PFHxA) induces cytochrome P450 expression and enzymatic activity in well-differentiated primary human bronchial epithelial (HBE) cells.

Chemical composition, antiproliferative, and cell death potential of crude and digested jabuticaba (Myrciaria cauliflora) leaf extracts in HepG2 human hepatocellular carcinoma cells in 2D and 3D culture: a bioprospecting approach.

A novel microbubble delivery platform with high payload of paclitaxel upon focused ultrasound for enhanced glioblastoma treatment.

In this study, different shape memory polyurethane (SMPU)-based electrospun nanofibers containing Fe3O4 magnetic nanoparticles (MNPs) were prepared. The shape-memory behavior of these scaffolds enables minimally invasive applications within biological systems, while their magnetic properties enhance the growth, proliferation, and differentiation of the bone cells. SMPU was synthesized via a two-step pre-polymerization method. The effects of MNPs on hydrogen bonding, crystallinity, thermal properties, hydrophilicity, water absorption, and mechanical and shape memory properties of SMPU were investigated. The results revealed that MNPs restricted the hydrogen bonds formation and promoted microphase separation in SMPU hard and soft segments. Moreover, a reduction in the degree of crystallinity of oft segments in the SMPU nanocomposites was observed by the addition of MNPs. Scanning electron microscopy was employed to determine the average diameters and size distributions of the SMPU nanofibers. In addition, the results showed that the prepared electrospun nanofibrous mats have adequate mechanical and shape memory properties for practical biomedical applications. Bioactivity studies indicated that the presence of MNPs could enhance the In-vitro cell cultivation of MG63 bone cells on the nanofibers. These results indicated that the prepared electrospun nanofibers could be utilized as a potential candidate for shape memory-assisted smart wound healing applications.