Biology subjects

Cidonio, G.

Publications and source records attributed to Cidonio, G..

6 recordsLinked to original sources

bioRxiv2025

3D bone printing via primed differentiation of stem cells with ultrasound (3DBonUS)

Bone disorders and skeletal defects represent a significant clinical challenge. Tissue engineering and regenerative medicine (TERM) strategies using 3D bioprinting have emerged as promising alternatives, but still limited by the inability of directing stem cell differentiation in a controlled and reproducible manner. Advancing beyond this, we are proposing 3D bone printing via primed differentiation of stem cells with ultrasound (referred to here as 3DBonUS). This approach synergistically integrates low-intensity pulsed ultrasound (LIPUS) with a microfluidic-assisted 3D bioprinting platform enabling the biophysical stimulation of human bone marrow stromal cells (HBMSCs) during extrusion, promoting osteogenic differentiation without the need for post-fabrication treatments. Moreover, the incorporation of microbubbles enhanced the effects of LIPUS by amplifying mechanical signals at the cellular level. 3DBonUS was found to significantly upregulate key osteogenic markers (RUNX-2, ALP, COL1A1, BMP-2, OCN, OPN) as confirmed by immunofluorescence and RT-qPCR analysis. Furthermore, the LIPUS-treated constructs showed a significant increase in alkaline phosphatase activity and calcium deposition, indicating enhanced mineralisation. The 3DBonUS strategy represents a new modality in skeletal biofabrication, harnessing targeted minimally-invasive mechanical stimulation, with potential for manufacturing scalability and clinical application. Future studies will aim to validate 3DBonUS in vivo to assess the ultimate regenerative potential with enhanced osteogenic properties.

bioengineering↗

bioRxiv2025

Aqueous two-phase bioinks for discrete packing and compartmentalisation of 3D bioprinted cells

The unparalleled ability of aqueous two-phase systems (ATPS) to reproduce microscale cellular and biomaterial compartmentalisation to selectively modulate cell behaviour and functionality is ideal for tissue engineering and regenerative medicine (TERM) purposes. Herein, we introduce new ATPS biomaterial inks for 3D bioprinting of water-in-water (W/W) emulsions, enabling precise cellular crowding for tissue regeneration in vitro and ex vivo. Gelatin methacryloyl (GelMA) was hierarchically dispersed in an alginic acid phase depending on sodium chloride (NaCl) concentration (0-36 g/L). Emulsion droplet size (12.8{+/-}2.6 {micro}m to 52.4{+/-}11.4 {micro}m) influenced degradation and spatial cell localisation (A549, C2C12, MG63). A microfluidic-assisted 3D bioprinting approach allowed fine-tuning of fibre structure adjusting ATPS deposition by modulating flow rates and printing speed. Rheological properties supported the findings of the two-phase partitioning, aiding the selection of the ATPS ink formulation for functional cell-laden construct fabrication. Encapsulation of C2C12 cells revealed enhanced cytoskeletal remodelling at higher salt concentrations. Increased GelMA phase promoted human bone marrow stromal cells (HBMSCs) crowding, mineral deposition and skeletal differentiation. In ovo studies demonstrated degradation control and vascular infiltration via salt modulation. Altogether, ATPS bioinks offer a versatile platform for the assembling of complex, hierarchical tissues with microscale precision, expanding biofabrication strategies for TERM applications.

bioengineering↗

bioRxiv2025

Engineering a microfluidic-assisted 3D bioprinting approach for the hierarchical control deposition and compartmentalisation of graded bioinks

The advent of 3D bioprinting has revolutionised tissue engineering and regenerative medicine (TERM). Today, tissues of single cell type can be printed with extreme resolution and printing fidelity. However, the ultimate functionality of the desired tissue is limited, due to the absence of a multicellular population and diversity in micro-environment distribution. Currently, 3D bioprinting technologies are facing challenges in delivering multiple cells and biomaterials in a controlled fashion. The use of interchangeable syringe-based systems has often favoured the delamination between interfaces, greatly limiting the fabrication of interconnected tissue constructs. Microfluidic-assisted 3D bioprinting platforms have been found capable of rescuing the fabrication of tissue interfaces, but often fails to guarantee printing fidelity, cell density control and compatimentalisation. Herein, we present the convergence of microfluidic and 3D bioprinting platforms into a new deposition system capable of harnessing a microfluidic printhead for the continuous rapid fabrication of interconnected functional tissues. The use of flow-focusing and passive mixer printhead modules allowed for the rapid and dynamic modulation of fibre diameter and material composition, respectively. Cells were compartmentalised into discrete three-dimensional layers with defined density patterns, confirming the punctual control of the presented microfluidic platform in arranging cells and materials in 3D. In ovo and in vivo studies demonstrated the functionality of 3D bioprinted constructs with patterned vascular endothelial growth factor (VEGF) and transforming growth factor-{beta}1 (TGF-{beta}1), respectively. This, in turn, facilitated the simulation of diverse cellular environments and proliferation pathways within a single construct, which is currently unachievable with conventional 3D bioprinting techniques, offering new opportunities for the fabrication of functionally graded materials and physiologically-relevant skeletal tissue substitutes.

bioengineering↗

bioRxiv2024

Chronic mycobacteria infection triggers macrophage senescence

Chronic infections with intracellular pathogens such as Mycobacterium abscessus (Mab) pose significant health challenges due to their capacity to persist within host cells and evade immune responses. This study investigates the cellular responses to chronic Mab infection in macrophages, particularly focusing on cellular senescence. Using an in vitro model of chronic infection in murine alveolar-like macrophages, we found that Mab induces a senescent phenotype characterised by decreased proliferation, altered morphology, DNA damage signalling activation, and upregulation of senescence markers such as p21 and SA-{beta}-galactosidase. Intriguingly, senescent macrophages secreted pro-inflammatory cytokines, consistent with a senescence-associated secretory phenotype (SASP), which promoted secondary senescence in neighbouring uninfected cells. This paracrine transmission of senescence underscores a potentially deleterious effect of Mab-induced SASP on tissue microenvironments, fostering a pro-inflammatory niche that may contribute to pathogen persistence. These findings highlight Mab-induced senescence as a key factor in chronic infection pathology, suggesting that targeting senescent cells and SASP-related pathways could enhance treatment outcomes in chronic bacterial infections.

microbiology↗

bioRxiv2023

Human bone tissue-derived ECM hydrogels: Controlling physicochemical, biochemical, and biological properties through processing parameters

Decellularized tissues offer significant potential as biological materials for tissue regeneration due to their ability to preserve the complex compositions and architecture of the native extracellular matrix (ECM). While the use of decellularized ECM hydrogels from bovine and porcine bone tissues has been extensively studied, the evaluation and derivation of decellularized matrices from human bone tissue remain largely unexplored. The objective of this study was to investigate how the physiochemical and biological properties of ECM hydrogels derived from human bone ECM could be controlled by manipulating bone powder size (45-250 m, 250-1000 m, and 1000-2000 m) and ECM composition through modulation of digestion time (3, 5, and 7 days). The current studies demonstrate that a reduction in material bone powder size and an increase in ECM digestion time resulted in enhanced protein concentrations in the ECM hydrogels, accompanied by the presence of a diverse array of proteins. Furthermore, these adjustments in the physicochemical properties generated improved gelation strength of the hydrogels. The evaluation of human bone marrow-derived stromal cells (HBMSCs) cultured on ECM hydrogels derived from 45-250 m bone powder, treated for 7 days, demonstrated enhanced osteogenic differentiation compared to hydrogels derived from both larger bone powders and collagen gels. In conclusion, this study highlights the significant promise of human bone ECM hydrogels as biologically active materials for bone regeneration. The ability to manipulate digestion time and bone powder size enables the generation of hydrogels with enhanced release of ECM proteins and appropriate gelation and rheological properties, offering new opportunities for application in bone tissue engineering.

bioengineering↗

bioRxiv2023

Biofabrication of nanocomposite-based scaffolds containing human bone extracellular matrix for the differentiation of skeletal stem and progenitor cells

Autograft or metal implants are routinely used in skeletal repair but can fail to provide a long-term clinical resolution, emphasising the need for a functional biomimetic tissue engineering alternative. An attractive sustainable opportunity for tissue regeneration would be the application of human bone waste tissue for the synthesis of a material ink for 3D bioprinting of skeletal tissue. The use of human bone extracellular matrix (bone-ECM) offers an exciting potential for the development of an appropriate micro-environment for human bone marrow stromal cells (HBMSCs) to proliferate and differentiate along the osteogenic lineage. Extrusion-based deposition was mediated by the blending of human bone-ECM (B) with nanoclay (L, Laponite(R)) and alginate (A) polymer, to engineer a novel material ink (LAB). The inclusion of nanofiller and polymeric material increased the rheological, printability, and drug retention properties and, critically, the preservation of HBMSCs viability upon printing. The composite human bone-ECM-based 3D constructs containing vascular endothelial growth factor (VEGF) enhanced vascularisation following implantation in an ex vivo chick chorioallantoic membrane (CAM) model. Addition of bone morphogenetic protein-2 (BMP-2) with HBMSCs further enhanced vascularisation together with mineralisation after only 7 days. The current study demonstrates the synergistic combination of nanoclay with biomimetic materials, (alginate and bone-ECM) to support the formation of osteogenic tissue both in vitro and ex vivo and offers a promising novel 3D bioprinting approach to personalised skeletal tissue repair. Graphical AbstractEngineering nanoclay-based bone ECM novel bioink for bone regeneration. Human bone trabecular tissue was demineralised, decellularised and blended with nanoclay (Laponite(R)) and alginate after digestion. The resulting ink was investigated for printability following rheological and filament fusion investigation. The microstructural arrangement of the blends was examined together with viability and functionality of bioprinted HBMSCs. Finally, the ability of the novel blend to support drug release ex vivo in a CAM model was determined confirming the potential of the bone ECM ink to support bone formation. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=191 SRC="FIGDIR/small/536074v1_ufig1.gif" ALT="Figure 1"> View larger version (64K): org.highwire.dtl.DTLVardef@b9dc13org.highwire.dtl.DTLVardef@1f6f0dorg.highwire.dtl.DTLVardef@1b3c098org.highwire.dtl.DTLVardef@295192_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗