Bioengineered Nanoparticles Loaded-Hydrogels to Target TNF Alpha in Inflammatory Diseases.

Rheumatoid Arthritis (RA) is an incurable autoimmune disease that promotes the chronic impairment of patients' mobility. For this reason, it is vital to develop therapies that target early inflammatory symptoms and act before permanent articular damage. The present study offers two novel therapies based in advanced drug delivery systems for RA treatment: encapsulated chondroitin sulfate modified poly(amidoamine) dendrimer nanoparticles (NPs) covalently bonded to monoclonal anti-TNF α antibody in both Tyramine-Gellan Gum and Tyramine-Gellan Gum/Silk Fibroin hydrogels. Using pro-inflammatory THP-1 (i.e., human monocytic cell line), the therapy was tested in an inflammation in vitro model under both static and dynamic conditions. Firstly, we demonstrated effective NP-antibody functionalization and TNF-α capture. Upon encapsulation, the NPs were released steadily over 21 days. Moreover, in static conditions, the approaches presented good anti-inflammatory activity over time, enabling the retainment of a high percentage of TNF α. To mimic the physiological conditions of the human body, the hydrogels were evaluated in a dual-chamber bioreactor. Dynamic in vitro studies showed absent cytotoxicity in THP-1 cells and a significant reduction of TNF-α in suspension over 14 days for both hydrogels. Thus, the developed approach showed potential for use as personalized medicine to obtain better therapeutic outcomes and decreased adverse effects.

Synthesis of mussel-inspired polydopamine-gallium nanoparticles for biomedical applications.

Aim: To established a simple, controlled and reproducible method to synthesize gallium (Ga)-coated polydopamine (PDA) nanoparticles (NPs). Materials & methods: PDA NPs were synthesized in alkali medium with posterior Ga shell formation due to ion chelation on the NP surface. Results: The obtained results with energy-dispersive x-ray spectroscopy confirmed the incorporation of Ga on the PDA NP surface. The cytotoxicity of Ga-coated PDA NPs was evaluated in vitro at different concentrations in contact with human adipose-derived stem cells. Further cell analysis also demonstrated the benefit of Ga-coated PDA NPs, which increased the cell proliferation rate compared with noncoated PDA NPs. Conclusion: This study indicated that Ga could work as an appropriate shell for PDA NPs, inducing cell proliferation at the analyzed concentrations.

Decellularized hASCs-derived matrices as biomaterials for 3D in vitro approaches.

Tissue regeneration strategies have been greatly evolving in the last years due to the use of more realistic approaches. These approaches rely in the use of biomaterials for the development of three dimensional (3D) structures that emulate the in vivo microenvironment of different tissues. Recently, extracellular matrices (ECM) secreted by cells have been caught a great deal of attention as an attractive biomaterial for the development 3D structures. In fact, different cells and/or different cellular culture conditions gave rise to different ECM's compositions, which can be used for the development of more physiologically relevant 3D structures. Nevertheless, the recovery of cell-derived ECM requires the use of a proper decellularization protocol. Herein, we report a decellularization protocol to recover the ECM produced by human adipose derived stem cells. This protocol comprises multiple steps (chemical, physical or enzymatic) which are described here in more detail. Furthermore, it is describe the methods that have been used to evaluate the effectiveness of this decellularization protocol. Overall, this protocol enables the production of hASCs-derived matrices that can be further used for the production of more physiologically relevant 3D in vitro models for tissue regeneration strategies.

Entrapped in cage (EiC) scaffolds of 3D-printed polycaprolactone and porous silk fibroin for meniscus tissue engineering.

The meniscus has critical functions in the knee joint kinematics and homeostasis. Injuries of the meniscus are frequent, and the lack of a functional meniscus between the femur and tibial plateau can cause articular cartilage degeneration leading to osteoarthritis development and progression. Regeneration of meniscus tissue has outstanding challenges to be addressed. In the current study, novel Entrapped in cage (EiC) scaffolds of 3D-printed polycaprolactone (PCL) and porous silk fibroin were proposed for meniscus tissue engineering. As confirmed by micro-structural analysis the entrapment of silk fibroin was successful, and all scaffolds had excellent interconnectivity (≥99%). The EiC scaffolds had more favorable micro-structure compared with the PCL cage scaffolds by improving the pore size while keeping the interconnectivity almost the same. When compared with the PCL cage, the entrapment of porous silk fibroin into the PCL cage decreased the high compressive modulus in a favorable matter in the wet state thanks to the silk fibroin's high swelling properties. The in vitro studies with human stem cells or meniscocytes seeded constructs, demonstrated that the EiC scaffolds had superior cell adhesion, metabolic activity, and proliferation compared to the PCL cage scaffolds. Upon subcutaneous implantation of scaffolds in nude mice, all groups were free of adverse incidents, and mildly invaded by inflammatory cells with neovascularization, while the EiC scaffolds showed better tissue infiltration. The results of this work indicated that the EiC scaffolds of PCL and silk fibroin are favorable for meniscus tissue engineering, and the findings are encouraging for further studies using a larger animal model.

Tuning Enzymatically Crosslinked Silk Fibroin Hydrogel Properties for the Development of a Colorectal Cancer Extravasation 3D Model on a Chip.

Microfluidic devices are now the most promising tool to mimic in vivo like scenarios such as tumorigenesis and metastasis due to its ability to more closely mimic cell's natural microenvironment (such as 3D environment and continuous perfusion of nutrients). In this study, the ability of 2% and 3% enzymatically crosslinked silk fibroin hydrogels with different mechanical properties are tested in terms of colorectal cancer cell migration, under different microenvironments in a 3D dynamic model. Matrigel is used as control. Moreover, a comprehensive comparison between the traditional Boyden chamber assay and the 3D dynamic microfluidic model in terms of colorectal cancer cell migration is presented. The results show profound differences between the two used biomaterials and the two migration models, which are explored in terms of mechanical properties of the hydrogels as well as the intrinsic characteristics of the models. Moreover, the developed 3D dynamic model is validated by demonstrating that hVCAM-1 plays a major role in the extravasation process, influencing extravasation rate and traveled distance. Furthermore, the developed model enables precise visualization of cancer cell migration within a 3D matrix in response to microenvironmental cues, shedding light on the importance of biophysical properties in cell behavior.