This work shows the synthesis of a polyvinylpyrrolidone (PVP) hydrogel by heat-activated polymerization and explores the production of hydrogels with an open porous network by lyophilisation to allow the three-dimensional culture of human oral mucosa stem cells (hOMSCs). The swollen hydrogel showed a storage modulus similar to oral mucosa and elastic solid rheological behaviour without sol transition. A comprehensive characterization of porosity by scanning electron microscopy, mercury intrusion porosimetry and nano-computed tomography (with spatial resolution below 1 μm) showed that lyophilisation resulted in the heterogeneous incorporation of closed oval-like pores in the hydrogel with broad size distribution (5 to 180 μm, d₅₀ = 65 μm). Human oral mucosa biopsies were used to isolate hOMSCs, expressing typical markers of mesenchymal stem cells in more than 95% of the cell population. Direct contact cytotoxicity assay demonstrated that PVP hydrogel have no negative effect on cell metabolic activity, allowing the culture of hOMSCs with normal fusiform morphology. Pore connectivity should be improved in future to allow cell growth in the bulk of the PVP hydrogel.

Tissue engineering scaffolds attempt to mimic the stem cell environment by creating different biophysical and chemical signals. On the other hand, stem cells are able to sense these characteristics and change their destiny. Scientists try to explain these phenomena through scaffold design and in vitro experiments, but the mechanisms implicated remain unclear. Moreover, environment-cell interactions are a key process to get organs and tissues arrangement. Therefore, this chapter deals with the mechanical signals and mechanism involved in cell behaviour through scaffolds as a strategy in tissue engineering.

Vat photopolymerization is an additive manufacturing technique that utilizes photosensitive resins to fabricate 3D polymeric objects with high precision. However, these objects often lack mechanical strength. This study investigated the strengthening of a resin based on epoxidized soybean oil acrylate, specifically designed for vat photopolymerization, by the in situ formation of hydroxyapatite nanocrystals. First, a stable alpha tricalcium phosphate (α-TCP)-resin feedstock mixture was developed (~30 vol.% α-TCP), which proved suitable for fabricating monoliths as well as complex triply periodic minimal surface (gyroid, diamond, and Schwarz) porous structures through vat photopolymerization. The results demonstrated that the incorporation of α-TCP particles led to a significant mechanical improvement of the resin. Second, post-printing hydrothermal treatments were utilized to transform the α-TCP particles into hydroxyapatite crystals within the resin. It was observed that the space between hydroxyapatite crystals within the composites was occupied by the cured resin, resulting in a more compact, stronger, and mechanically more reliable material than the porous hydroxyapatite produced by the hydrolysis of α-TCP mixed with water. Moreover, water absorption during the hydrothermal treatments caused the plasticization of the cured resin. As a consequence, the hydroxyapatite-resin composites displayed slightly lower mechanical properties compared to the as-printed α-TCP-resin composite.