Breast cancer remains the most common malignancy in women, yet, many patients fail to achieve full remission despite significant advancements. This is largely due to tumour heterogeneity and the limitations of current experimental models in accurately replicating the complexity of in vivo tumour environment. In this study, we present a compartmentalised alginate hydrogel platform as an innovative in vitro tool for three-dimensional breast cancer cell culture. To mimic the heterogeneity of tumour tissues, we developed a core–shell structure (3.5% alginate core and 2% alginate shell) that mimic the stiffer, denser internal tumour matrix. The human triple-negative breast cancer cell line (MDA-MB-231) was embedded in core–shell alginate gels to assess viability, proliferation and hypoxic activity. Over one week, good cells proliferation and viability was observed, especially in the softer shell. Interestingly, cells within the stiffer core were more positive to hypoxic marker expression (HIF-1α) than those embedded in the shell, confirming the presence of a hypoxic niche, as observed in vivo. When cultured in the MIVO^® milli fluidic organ-on-chip resembling the physiological fluid flow conditions, cancer cells viability became comparable between core and shell hydrogel area, emphasising the importance of the fluid flow in nutrients diffusion within three-dimensional matrixes. Cisplatin chemotherapy treatment further highlighted these differences: under static conditions, cancer cell death was prominent in the softer shell, whereas cells in the stiffer core remained resistant to cisplatin. Conversely, drug diffusion was more homogeneous in the core–shell structured treated in the organ-on-chip, leading to a uniform reduction in cell viability. These findings suggest that integrating a compartmentalised core–shell cell laden alginate model with the millifluidic organ on chip offers a more physiologically relevant experimental approach to deepening cancer cell behaviour and drug response.

In this work, immune modulation of the monocyte-derived dendritic cells was studied by in vitro cell culture and organ-on-chip models. Self-assembled mannosylated peptide nanofibers were prepared and integrated with the MUC1 TnThr mimetic Tn-MIM to stimulate DCs’ maturation and differentiation. Man-PA/Tn-MIM nanofibers displayed remarkable cell-penetrating capabilities, also allowing the correct presentation of the antigen mimetic with a corresponding induction of lymphocytes, iNKT and NK cells recruitment. The Tn-MIM integrated nanofibrers were also found to direct immune response towards Th1 phenotype which is responsible for the production of cellular and humoral immunity. Organ-on-chip tests confirmed the cytotoxicity of lymphocytes stimulated with DC exposed to Man-PA/Tn-MIM. These results will help in the development of immunotherapies avoiding complications due to heterogeneous antigen display and to the use of adjuvants.

Fasting-mimicking diets have emerged as promising adjuvant strategies to enhance chemotherapy efficacy. However, conventional preclinical models do not adequately reproduce the microenvironmental and transport-related conditions associated with nutrient modulation, limiting mechanistic investigation of fasting-related therapeutic responses. Here, we developed a nutrient-responsive cancer-on-chip (CoC) platform integrating 3D human breast cancer models with systemic-like cisplatin administration under defined glucose conditions (high, low, no glucose). Complete glucose deprivation markedly reduced metabolic activity and cell viability, while mild reduction was largely tolerated, reflecting tumor metabolic adaptability. Tumor models were exposed to glucose modulation and cisplatin, with or without 24 h glucose-free pre-conditioning, in static and dynamic 3D systems. Static cultures displayed limited treatment response, whereas CoC exhibited significantly increased cell death, highlighting the limited predictive capacity of conventional models. Glucose-free pre-conditioning enhanced chemotherapy sensitivity, while fasting initiated concurrently with treatment produced minimal impact. Glucose reintroduction after glucose-free pre-conditioning impaired metabolic recovery, revealing a transient vulnerability window. Importantly, in healthy fibroblasts, glucose deprivation reduced metabolic activity without markedly affecting viability. A qualitative comparison with published clinical data from fasting-mimicking diet trials indicated that dynamic CoC models reproduced several treatment-related response trends more consistently than static systems. These findings suggest the potential of nutrient-responsive CoC systems as complementary tools for investigating context-dependent responses to fasting-associated therapeutic strategies.

Metastasis to the bone significantly impacts patient morbidity and mortality for multiple cancers. The lack of in vitro models able to recapitulate bone fo