Current in vitro tissue models struggle to recapitulate the structural, vascular, and mechanical complexity of human tissues, limiting their physiological relevance for disease modelling and preclinical testing. Self-organising three-dimensional cultures such as spheroids and organoids capture key aspects of cellular organisation and differentiation, but they commonly lack controlled geometry, perfusable vasculature, and reproducible mechanical microenvironments. Conversely, biofabrication strategies, such as three-dimensional (3D) bioprinting and organ-on-chip (OoC) microfluidic devices, offer spatial control, integrated perfusion, and dynamic mechanical stimulation, yet often fall short in recapitulating the full cellular diversity and self-organisation of native tissues. Notably, emerging hybrid approaches that embed self-organising biological units (e.g., organoids and spheroids) into engineered scaffolds or microfluidic platforms combine biological relevance, architectural fidelity, and functional control. Advances in bioink chemistry, sacrificial-printing vascularisation, and chip–organoid interfaces now enable perfusable, multicompartment tissues suitable for disease modelling and preclinical testing. This review highlights the most recent (2020–2025) progress in organoid vascularisation, bioprinting strategies for prevascularised constructs, and OoC integration, outlining remaining challenges and emphasising priorities for next-generation hybrid cellular and tissue models.

The evolutionarily conserved Notch signalling pathway regulates the fate, proliferation and differentiation of cells in most developing organs, thus affecting their morphogenesis and function. Here, we investigated the role of the Notch2 receptor in the generation and function of epithelial cells of the continuously erupting rodent incisors. We used transgenic Notch1-Cre^ERT2/+;Rosa26^mT/mG and Notch2-Cre^ERT2/+;Rosa26^mT/mG mice to compare the contribution of Notch1- and Notch2-expressing cells and their progeny in the generation of the different epithelial cell populations. Furthermore, we examined if the dental epithelium organisation and enamel structure are affected in early postnatal incisors of Keratin14^Cre/+;Notch2^fl/fl mice using immunofluorescent staining, gene expression analysis, microcomputed tomography and scanning electron microscopy. Our results showed that Notch2 deletion resulted in smaller incisors with disorganised dental epithelium and defective enamel. Delayed eruption was correlated with alterations in the proliferative and differentiation status of epithelial stem cells in the cervical loop area of the incisors. Similar results were obtained with in vitro studies, where inhibition of the Notch signalling by the CB103 blocker recapitulated the in vivo phenotype. In conclusion, this study demonstrates for the first time the importance of Notch2 in epithelial cell fate acquisition, dental epithelium organisation and enamel structure in rodent incisors.

Biodegradable polymeric scaffolds incorporating bioactive mineral phases are a promising approach for dentin-pulp tissue engineering. Although poly(lactic-co-glycolic acid) (PLGA) microparticles have been widely used as scaffolds due to their biocompatibility and tunable degradation profile, their inherent bioactivity is limited. Furthermore, fluorapatite (FAP), a fluoride-substituted apatite ceramic, exhibits enhanced chemical stability and mineral-related properties that may be useful for regenerative biomaterial design. In this study, we investigated the effect of nano-FAP functionalization on the physicochemical properties of porous PLGA microscaffolds and their interaction with human dental pulp stem cells (hDPSCs). Porous PLGA microscaffolds were fabricated using a double-emulsion solvent evaporation method and subsequently functionalized with FAP suspensions ranging from 0.1 to 5 mg/mL (0.01–0.5% w/v). The scaffolds were evaluated using scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), electrical conductivity measurements, cell viability assays, and immunofluorescence. Lower and intermediate FAP concentrations-maintained surface pore accessibility and supported hDPSC viability, whereas the highest concentration (5 mg/mL; 0.5% w/v) reduced visible surface pore size and showed less favorable cellular responses. The 2.5 mg/mL (0.25% w/v) FAP condition provided the most favorable overall balance among the evaluated physicochemical and biological parameters. These preliminary in vitro findings support further investigation of FAP-functionalized PLGA microscaffolds in advanced three-dimensional and in vivo models.

Rodent incisors exhibit pronounced labial–lingual asymmetry, with enamel deposited exclusively on the labial surface and cementum on the lingual surface. This asymmetry arises early in development as the initially symmetrical dental epithelium rotates and reorients along the jaw axis. Here, we investigated the cellular and molecular mechanisms underlying this process. Quantitative cell-cycle analyses and transcriptomic profiling before and during epithelial rotation revealed accelerated proliferation, shorter cell-cycle duration, and enrichment of DNA replication and mitotic genes within the labial epithelium compared with the lingual epithelium. The evolutionarily conserved Notch signalling pathway, a regulator of cell proliferation and fate, was asymmetrically active during this period, with Notch2 expression restricted to the labial domain. Lineage tracing demonstrated that Notch2-expressing cells and their progeny preferentially contribute to the enamel-producing compartment. Epithelial-specific deletion of Notch2 produced no obvious early morphological phenotype and induced upregulation of other Notch pathway components. By contrast, pharmacological Notch inhibition reduced epithelial asymmetry and altered S-phase progression, and decreased Ccnd1 and Cdk6 expression. Together, these findings implicate Notch signalling in regional cell-cycle dynamics during early mouse incisor asymmetry, while suggesting that other Notch components may compensate for Notch2 loss.