Electrostimulation of a 3D in vitro skin model to activate wound healing

The aim of the work is to propose a methodology for the stimulation of a 3D in vitro skin model to activate wound healing. The presented work is in the frame of the national research project, CronXCov, “Checking the CHRONIC to prevent COVID-19”, devoted to understand how physiologic and inflamed skin on chip 3D models evolve upon a range of physical (e.g., electrical, mechanical, optical) stimulations, over time. Thanks to the 3D modelling, using Next Generation Sequencing and the network medicine frame of analysis to process the data, we will systematically characterize the effects of the applied stimuli, offering new insight for the exploitation of wound healing

Development of in vitro 3D culture system to mimic lung cancer tissue

A 3D culture system based on a photocurable matrix has been developed. The aim is to create a 3D printable platform mimicking lung cancer tissue, to study tumor microenvironment evolution, in terms of structural (architecture) and molecular (signalling) components

3D printable acrylate polydimethylsiloxane resins for cell culture and drug testing

Nowadays, most of the microfluidic devices for biological applications are fabricated with only few wellestablished materials. Among these, polydimethylsiloxane (PDMS) is the most used and known. However, it has many limitations, like the operator dependent and time-consuming manufacturing technique and the high molecule retention. TEGORad or Acrylate PDMS is an acrylate polydimethylsiloxane copolymer that can be 3D printed through Digital Light Processing (DLP), a technology that can boast reduction of waste products and the possibility of low cost and rapid manufacturing of complex components. Here, we developed 3D printed Acrylate PDMS-based devices for cell culture and drug testing. Our in vitro study shows that Acrylate PDMS can sustain cell growth of lung and skin epithelium, both of great interest for in vitro drug testing, without causing any genotoxic effect. Moreover, flow experiments with a druglike solution (Rhodamine 6G) show that Acrylate PDMS drug retention is negligible unlike the high signal shown by PDMS. In conclusion, the study demonstrates that this acrylate resin can be an excellent alternative to PDMS to design stretchable platforms for cell culture and drug testing

3D printed microfluidic devices for cell culture

A successful application of the 3D printed materials in the biomedical field requires extensive studies to ensure their biocompatibility at every step of the process. Here, different components suitable for cell applications, including a microfluidic device, were 3D printed using common resins and a deep analysis of their biocompatibility and post printed protocols was conducted

A METHODOLOGICAL APPROACH TO INTERPRET AND COMPARE THE VISCOELASTIC BEHAVIOR OF BIOLOGICAL TISSUES AND HYDROGELS

Cell behavior is strongly influenced by the physical properties of the microenvironment and complex mechanotransduction mechanisms are involved in cell and tissue development, homeostasis and even pathologies. Thus, when developing materials mimicking the extracellular matrix of healthy or pathological tissues their mechanical features should be closely considered. In this context, nanoindentation is a powerful technique for mechanically characterizing biomaterials and hydrogels at the cell-length scale, however, standardized experimental protocols and data analysis techniques are lacking. Here, we propose a methodological approach for quantitatively analyzing and comparing the time-dependent mechanical responses of different samples. As an explanatory study, stress-relaxation nanoindentation tests were performed on human and pig lung samples and on hydrogels in order to quantify and compare their viscoelastic properties

A programmable culture platform for stimulation and in situ sensing of lung epithelial cells

A programmable dynamic cell culture chamber compatible with a standard multi-well plate was designed and characterized. The system is integrated with an array of OECT biosensors, in view of an in-situ monitoring of the dynamic cultures

Visible light-induced crosslinking of unmodified gelatin with PEGDA for DLP-3D printable hydrogels

The possibility to 3D shape hydrogels is attracting an enormous interest in the biomedical field both for their application as scaffold or for the design of new medical hydrogels. Digital light processing (DLP) printing can create layer-by-layer models with high resolution and printing speed. Herein, a hybrid natural-synthetic hydrogel is propsed using cold-water fish gelatin as innovative co-initiating species of a camphorquinone photo-initiator instead of the traditional aliphatic or aromatic amines, for the crosslinking of PEGDA monomer. Such system allows to chemically bound gelatin to the PEGDA monomer without any previous modification and leads to the production of DLP-3D printable hydrogels. The real-time photorheological measurements showed that Gelatin behave as a binder during photopolymerization and ATR-FTIR spectroscopy proved that gelatin segments were chemically incorporated within PEGDA network. Mechanical and biological properties were improved by increasing gelatin content. Furthermore, the 3D digital light processing of this material leads to the creation of precise and rapidly printed structures that are biocompatible and able to support cell viability and proliferation. Considering those features, the proposed hydrogel may be a promising candidate in 3D-printed devices for cell culture

Therapeutic Silencing of miR-214 Inhibits Tumor Progression in Multiple Mouse Models

We previously demonstrated that miR-214 is upregulated in malignant melanomas and triple-negative breast tumors and promotes metastatic dissemination by affecting a complex pathway including the anti-metastatic miR-148b. Importantly, tumor dissemination could be reduced by blocking miR-214 function or increasing miR-148b expression or by simultaneous interventions. Based on this evidence, with the intent to explore the role of miR-214 as a target for therapy, we evaluated the capability of new chemically modified anti-miR-214, R97/R98, to inhibit miR-214 coordinated metastatic traits. Relevantly, when melanoma or breast cancer cells were transfected with R97/R98, anti-miR-214 reduced miR-214 expression and impaired transendothelial migration were observed. Noteworthy, when the same cells were injected in the tail vein of mice, cell extravasation and metastatic nodule formation in lungs were strongly reduced. Thus, suggesting that R97/R98 anti-miR-214 oligonucleotides were able to inhibit tumor cell escaping through the endothelium. More importantly, when R97/R98 anti-miR-214 compounds were systemically delivered to mice carrying melanomas or breast or neuroendocrine pancreatic cancers, a reduced number of circulating tumor cells and lung or lymph node metastasis formation were detected. Similar results were also obtained when AAV8-miR-214 sponges were used in neuroendocrine pancreatic tumors. Based on this evidence, we propose miR-214 as a promising target for anti-metastatic therapies