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Nutraceuticals are increasingly investigated for their capacity to modulate oxidative and inflammatory stress, yet preclinical testing still relies largely on immortalized cell lines or animal models that poorly recapitulate human epithelial complexity. To address this gap, we developed an integrated platform based on patient-derived colon organoids generated from non-tumoral mucosa and maintained under proliferative or differentiation conditions to model distinct epithelial states. The system combines millifluidic measurement of individual organoid mass, density, and diameter with bulk RNA sequencing and digital PCR profiling to enable multiparametric characterization. Transcriptional analysis revealed state-specific gene programs and shifts in epithelial and immune-related pathways, while biophysical measurements captured structural remodeling. In this pilot validation, a defined oxidative insult followed by nutraceutical treatment elicited coordinated transcriptional and phenotypic responses. This integrated approach provides a scalable and physiologically relevant framework for functional nutraceutical profiling and mechanistic studies of epithelial stress responses.

Rationale: Obesity-associated metabolic diseases such as metabolic dysfunction-associated steatotic liver disease (MASLD) and type 2 diabetes mellitus (T2DM) are increasing rapidly, necessitating physiologically relevant in vitro models of the liver-pancreas axis. While recent multi-organoid systems have advanced inter-organ modeling, many systems still fall short of replicating the complex and directional metabolic interactions required to accurately reflect disease progression. This is partly due to inherent limitations such as reliance on passive diffusion for metabolic exchange and the use of shared or non-compartmentalized media, which restrict tissue-specific functions and fail to mimic in vivo-like physiological gradients.Methods: To overcome these limitations, we developed a multi-organoid device (MOD) that incorporates convective flow and physically separates liver and pancreatic organoids in distinct media environments. To evaluate the effectiveness of this co-culture system, we assessed metabolic transport using FITC-dextran and examined pancreatic and liver organoid function by measuring insulin and albumin secretion respectively. The effectiveness of the MOD in modeling MASLD-induced T2DM was further validated through functional assays and transcriptomic analysis.Results: The MOD successfully recapitulated key pathological features of MASLD-induced T2DM. Convective flow significantly enhanced directional transport of glucose and other metabolic molecules compared to passive diffusion, as validated by simulation and diffusion assays. Media separation preserved organoid function, increasing insulin and albumin secretion by 1.8- and 1.6-fold, respectively, compared with the non-separated group. Importantly, the device achieved rapid glucose regulation following glucose stimulation, with normoglycemia restored within 2 hours closely mimicking physiological glucose regulation not previously attainable in existing systems. Under MASLD conditions, the platform further revealed that liver-derived Fetuin-A was associated with β-cell apoptosis in pancreatic organoids.Conclusion: This MOD effectively models the pathophysiological cascade linking MASLD and T2DM by integrating organ-specific environments, convective flow, and multi-organ crosstalk. It offers a robust and biologically relevant tool for mechanistic studies of metabolic diseases and provides a promising platform for preclinical drug screening and therapeutic development.

AbstractAimsP-glycoprotein (P-gp), an efflux transporter with diverse compound effects, is a vital part of cardiac function. To determine if the selective substrate tracer [18F]MC225 also functions in cardiac P-gp, micro-engineered heart tissues (µ-EHTs) utilizing human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes were used. This model offers advantages in potentially reducing animal experiments and allowing direct evaluation on human cells. However, its adoption in nuclear medicine remains very limited. This study aims to evaluate [18F]MC225 as a measurement method for cardiac P-gp function using a heart-on-chip model.Methods and resultsµ-EHTs were treated with the P-gp inhibitor Tariquidar (200 nM for 30 min) or the P-gp inducer Doxorubicin (1 µM for 24 h) and incubated with [18F]MC225 (1 MBq/mL for 30 min). First, we identified and confirmed the expression of P-gp in the µ-EHTs using immunofluorescent staining, which showed an increase of P-gp expression after Doxorubicin treatment. According to γ-counter measurements, Tariquidar-treated tissues exhibited a higher uptake (117.5 ± 33.67%, n = 24) (P = 0.035) than the control, compared to Doxorubicin-treated tissues which exhibited a lower uptake (63.97 ± 21.89%, n = 20) (P < 0.001) compared to its controls. Autoradiography visualized radioactive distribution in each µ-EHT and confirmed the γ-counter measurements.Conclusion[18F]MC225 effectively evaluates and measures cardiac P-gp function in µ-EHTs on the heart-on-chip platform. This research sets the stage for future studies using P-gp function to evaluate the efficacy and safety of novel cardiovascular drugs using µ-EHTs. Graphical Abstract Graphical Abstract