In this paper the first complete phytochemical study of Stachys sylvatica (Lamiaceae) aerial parts, collected in Tuscan-Emilian Apennine, is reported. Twenty-three known compounds were isolated from the chloroform, chloroform-methanol 9:1, and methanol extracts; they were subsequently characterized by means of one- and two-dimensional nuclear magnetic resonance spectroscopy (1D- and 2D-NMR) and mass spectrometry as six simple phenolic derivatives (chlorogenic acid (1), caffeic acid (2), p-coumaric acid methyl ester (3), niduloic acid (4), hydroxytyrosol (5), tyrosol (6)), one glycosylated aliphatic alcohol (ebracteatoside B (7)), six glycosylated phenylpropanoids (decaffeoyl verbascoside (8), lavandulifolioside (9), betanyoside F (10), verbascoside (11), isoacteoside (12), cystanoside D (13)), four flavonoids (stachyspinoside (14), chrysoeriol 7-O-β-d-glucopyranoside (15), naringenin 7-O-(6''-E+Z-p-coumaroyl)-β-d-glucopyranoside (16), apigenin 7-O-(6''-E+Z-p-coumaroyl)-β-d-glucopyranoside (17)), three lignans (syringaresinol 4'-O-β-d-glucopyranoside (18), pinoresinol 4-O-β-d-glucopyranoside (19), 2-episesaminol 2-O-β-d-glucopyranoside (20)), and three ionones (loliolide (21), vomifoliol (22), (3S,5R,6R,7E,9R)-3,5,6,9-tetrahydroxy-7-megastigmene (23)). In addition, a further phenolic derivative (ethyl caffeate (b)) and four carboxylic acids (azelaic acid (a), trihydroxyoctadecadienoic acid (c), trihydroxyoctadecenoic acid (d), and dihydroxydodecadienoic acid (e)) have been tentatively identified by ultra-high performance liquid chromatography coupled to an electrospray ionization source high-resolution Orbitrap mass spectrometer (UHPLC-ESI-HR-Orbitrap/MS). Chemosystematic relationships between S. sylvatica and other related species were highlighted. Graphical Abstract

Saffron, branded as Repron^®, is effective in slowing the progression of several neurodegenerative diseases. Its production, however, requires specific cultivation techniques and procedures that, together with low yields, make it expensive. To address this challenge, hydroponic cultivation has been adopted. Previous studies have shown that hydroponically cultivated saffron and Repron^® share comparable chemical compositions and neuroprotective effects under oxidative stress conditions. In this study, we evaluated the protective properties of extracts derived from Crocus sativus L. waste, compared with those of saffron derived from stigmas. Human retinal pigment epithelium (ARPE-19) cells were pre-treated with extracts of various plant waste fractions before being subjected to three stress conditions: H₂O₂-induced oxidative stress (500 μM, 3 h), lipopolysaccharide (LPS; 0.25 mg/mL, 24 h), and hyperglycemia (25 mM glucose, 96 h). Saffron Repron^® served as a positive control. The results revealed that the extract derived from C. sativus waste had superior protective effects against oxidative stress and inflammation by preserving the state of the mitochondria and tight junctions (ZO-1); conversely, the tepal extract alone was more effective under hyperglycemic conditions by also modulating acrolein levels. These results suggest that different plant fractions contain bioactive compounds with specific protective actions, which together lead to increased cell survival.

Background/Objectives: Natural polysaccharides have many bio-pharmacological effects, which make them compounds with potential in healthcare. Limnospira platensis (Spirulina), a well-known blue–green cyanobacterium with relevance in the market of nutraceuticals, produces polysaccharides with recognized antioxidant and anti-inflammatory activities. Noteworthy, the growth of the cyanobacterium biomass may be obtained in a more sustainable manner under mixotrophic conditions. In the present study, we compared the antioxidant and anti-inflammatory effects of polysaccharide-enriched extracts from the cyanobacterium cultured under autotrophism (Auto−P extract) or mixotrophism (Mixo−P extract); this latter was realized using medium added with brewery wastewater (BWW). Methods and Results: Non-cellular investigation showed a better antioxidant profile for Mixo−P extract in the OH radical scavenging assay and a similar activity between the extracts in ABTS and ferrous chelation assays. The antioxidant protective activity of L. platensis extracts investigated on HaCat cells in the range of 0.3–10 μg/mL (not cytotoxic concentrations), against hydrogen peroxide (H₂O₂, 600 μM)-induced damage, revealed a similar activity by the two extracts. When tested against the inflammatory stimuli with lipopolysaccharide (LPS, 10 μg/mL) or tumor necrosis factor-α (TNF-α, 10 ng/mL), both Auto−P and Mixo−P showed an ability to prevent the effects of the inflammatory agents on cell viability and on interleukin-1β (IL-1β) and interleukin-6 (IL-6) release, with a slightly greater potency by Mixo−P extract. Conclusions: In conclusion, our data suggest the possible use of L. platensis polysaccharide-enriched extracts in biological-made pharmaceuticals for skin disorders or in cosmeceuticals. In addition, this study demonstrates that mixotrophic cultivation of L. platensis may be an alternative and sustainable way for biotechnological applications of the cyanobacterium biomass.

Bioprinting has emerged as a transformative technology in biofabrication, enabling the precise spatial arrangement of biomaterials, living cells, and bioactive factors to generate functional three-dimensional biological constructs. Recent advances are redefining the scope and impact of this field through innovations in both materials and methodologies. Multi-material and multiscale printing strategies are enhancing the ability to replicate the hierarchical architecture and functional gradients of native tissues, while the valorization of waste-derived biomaterials for bioink formulation is introducing sustainable solutions without compromising performance. The integration of bioprinting with organ-on-a-chip systems is providing highly sophisticated in vitro models for disease research and drug discovery, and in situ bioprinting techniques are opening new possibilities for direct, patient-specific tissue repair. Parallel to these developments, four-dimensional bioprinting introduces the dimension of time, allowing printed constructs to change shape, properties, or function in response to environmental stimuli. The application of artificial intelligence in process monitoring and quality control is improving reproducibility, predictive accuracy, and manufacturing efficiency, thus paving the way for standardized production. Looking ahead, the emerging concept of five-dimensional bioprinting— integrating spatial, temporal, and functional control—suggests a paradigm shift in the design and manufacturing of living systems. Collectively, these advances are broadening the technological capabilities of biofabrication and accelerating the translation of bioprinting from experimental settings toward transformative clinical and industrial applications. This review synthesizes current progress while outlining the opportunities and challenges that will shape the next generation of bioprinting technologies.

Tendon tissue engineering requires scaffolds with an appropriate structure that can be cultured under cyclic strain in order to replicate both mechanical cues and biological commitments. This study presents a hybrid scaffold combining 3D-printed auxetic polycaprolactone (PCL) with electrospun gelatin-(3-Glycidyloxypropyl)trimethoxysilane (GPTMS) functionalized with collagen microfibers suitable for the purpose of tenogenic commitment and in vitro modelling. The auxetic core was fabricated via pneumatic melt extrusion at 1000 mbar and printing speed of 200 mm/min, achieving a re-entrant angle of 60.92 ± 2.1° and strut thickness of 350 ± 30 μm with high geometric fidelity. Mechanical characterization revealed that PCL scaffolds exhibited an elastic modulus of 51.97 ± 6.19 MPa and ultimate tensile strain of 6.75 ± 0.59%. Integration of electrospun gelatin-GPTMS without collagen increased the elastic modulus to 66.54 ± 7.16 MPa while reducing ultimate tensile strain to 4.14 ± 1.27%. Incorporation of 1 mg/mL and 2 mg/mL μCollaFibR™ significantly enhanced ultimate tensile strain to 21.70 ± 0.20% and 19.94 ± 0.20%, while decreasing elastic modulus to 47.65 ± 1.33 MPa and 39.83 ± 1.85 MPa. Field Emission-Scanning Electron Microscopy confirmed hierarchical architecture with collagen microfibers (1-5 μm diameter) interspersed throughout the electrospun gelatin matrix. Human tendon stem/progenitor cells seeded at 1 × 10⁵ cells/cm² maintained viability exceeding 85% at day 3 across all formulations, exhibiting elongated morphology and alignment along nanofiber axes. The developed multimaterial platform successfully integrates auxetic mechanics with extracellular matrix-mimetic topographies, offering a biomimetic scaffold structure for subsequent in vitro modelling of tenogenic events.

Existing in vivo models for retinal pathologies are costly, time-consuming, and subject to regulatory challenges, promoting interest in advanced in vitro models that replicate the blood-retinal barrier. Challenges related to vascularization and structural multiscale interactions hinder the development of in vitro models, which are useful to study nutrient and drug exchange between the retina and systemic circulation. To address these issues, we developed a blood-retinal barrier in vitro model for studying retinal pathophysiology. It features an electrospun scaffold (Bruch’s membrane) placed between a culture chamber (vitreous humor) and a bioinspired microfluidic network (choroidal vasculature). The network, modelled from indocyanine green angiography of the human retina, is laser-engraved into a 15 mm PDMS disc with channels 70–800 μm wide. PLGA and gelatin electrospun scaffold were investigated to mimic Bruch’s membrane. The scaffold is fixed to the microfluidic network, assembled into a culture chamber, and integrated within a bioreactor. A peristaltic pump ensures a consistent flow in the channels. Biological validation included ARPE-19 cells cultured on Transwell, PLGA, or gelatin membranes, HUVECs within membrane-coupled microchannels, and dual-layer constructs. Cells were examined separately and together, allowing evaluation of epithelial, endothelial, and construct responses. All configurations showed progressive increases in metabolic activity and limited LDH release over 28 days. Cell-seeded constructs reduced FITC–dextran transport relative to acellular controls, with stable values between days 21 and 28. Blank-corrected TEER likewise increased during ARPE-19 culture and plateaued by day 21. PLGA constructs showed the greatest electrical and solute-transport resistance, although acellular controls confirmed a substantial material contribution. Confocal microscopy demonstrated HUVEC attachment and F-actin organization along the microchannels, while ARPE-19 monolayers exhibited continuous junction-associated ZO-1 staining that remained stable between days 21 and 28. Overall, the platform supported long-term epithelial–endothelial culture and provided a stable, perfusable system for studying oBRB transport and cellular interactions.