RIP140 regulates transcription factor HES1 oscillatory expression and mitogenic activity in colon cancer cells.

The transcription factor receptor-interacting protein 140 (RIP140) regulates intestinal homeostasis and tumorigenesis through Wnt signaling. In this study, we investigated its effect on the Notch/HES1 signaling pathway. In colorectal cancer (CRC) cell lines, RIP140 positively regulated HES1 gene expression at the transcriptional level via a recombining binding protein suppressor of hairless (RBPJ)/neurogenic locus notch homolog protein 1 (NICD)-mediated mechanism. In support of these in vitro data, RIP140 and HES1 expression significantly correlated in mouse intestine and in a cohort of CRC samples, thus supporting the positive regulation of HES1 gene expression by RIP140. Interestingly, when the Notch pathway is fully activated, RIP140 exerted a strong inhibition of HES1 gene transcription controlled by the level of HES1 itself. Moreover, RIP140 directly interacts with HES1 and reversed its mitogenic activity in human CRC cells. In line with this observation, HES1 levels were associated with a better patient survival only when tumors expressed high levels of RIP140. Our data identify RIP140 as a key regulator of the Notch/HES1 signaling pathway, with a dual effect on HES1 gene expression at the transcriptional level and a strong impact on colon cancer cell proliferation.

Gelatin methacrylate hydrogel with drug-loaded polymer microspheres as a new bioink for 3D bioprinting.

3D bioprinted hydrogel constructs are advanced systems of a great drug delivery application potential. One of the bioinks that has recently gained a lot of attention is gelatin methacrylate (GelMA) hydrogel exhibiting specific properties, including UV cross-linking possibility. The present study aimed to develop a new bioink composed of GelMA and gelatin modified by addition of polymer (polycaprolactone or polyethersulfone) microspheres serving as bioactive substance carriers. The prepared microspheres suspension in GelMA/gelatin bioink was successfully bioprinted and subjected to various tests, which showed that the addition of microspheres and their type affects the physicochemical properties of the printouts. The hydrogel stability and structure was examined using scanning electron and optical microscopy, its thermal properties with differential scanning calorimetry and thermogravimetric analysis and its biocompatibility on HaCaT cells using viability assay and electron microscopy. Analyses also included tests of hydrogel equilibrium swelling ratio and release of marker substance. Subsequently, the matrices were loaded with ampicillin and the antibiotic release was validated by monitoring the antibacterial activity on Staphylococcus aureus and Escherichia coli. It was concluded that GelMA/gelatin bioink is a good and satisfying material for potential medical use. Depending on the polymer used, the addition of microspheres improves its structure, thermal and drug delivery properties.

3D printing of cellulose nanocrystals based composites to build robust biomimetic scaffolds for bone tissue engineering.

Cellulose nanocrystals (CNC) are drawing increasing attention in the fields of biomedicine and healthcare owing to their durability, biocompatibility, biodegradability and excellent mechanical properties. Herein, we fabricated using fused deposition modelling technology 3D composite scaffolds from polylactic acid (PLA) and CNC extracted from Ficus thonningii. Scanning electron microscopy revealed that the printed scaffolds exhibit interconnected pores with an estimated average pore size of approximately 400 µm. Incorporating 3% (w/w) of CNC into the composite improved PLA mechanical properties (Young's modulus increased by ~ 30%) and wettability (water contact angle decreased by ~ 17%). The mineralization process of printed scaffolds using simulated body fluid was validated and nucleation of hydroxyapatite confirmed. Additionally, cytocompatibility tests revealed that PLA and CNC-based PLA scaffolds are non-toxic and compatible with bone cells. Our design, based on rapid 3D printing of PLA/CNC composites, combines the ability to control the architecture and provide improved mechanical and biological properties of the scaffolds, which opens perspectives for applications in bone tissue engineering and in regenerative medicine.

Fabrication of Radio-Opaque and Macroporous Injectable Calcium Phosphate Cement.

The aim of this work was the development of injectable radio-opaque and macroporous calcium phosphate cement (CPC) to be used as a bone substitute for the treatment of pathologic vertebral fractures. A CPC was first rendered radio-opaque by the incorporation of zirconium dioxide (ZrO2). In order to create macroporosity, poly lactic-co-glycolic acid (PLGA) microspheres around 100 µm were homogeneously incorporated into the CPC as observed by scanning electron microscopy. Physicochemical analyses by X-ray diffraction and Fourier transform infrared spectroscopy confirmed the brushite phase of the cement. The mechanical properties of the CPC/PLGA cement containing 30% PLGA (wt/wt) were characterized by a compressive strength of 2 MPa and a Young's modulus of 1 GPa. The CPC/PLGA exhibited initial and final setting times of 7 and 12 min, respectively. Although the incorporation of PLGA microspheres increased the force necessary to inject the cement and decreased the percentage of injected mass as a function of time, the CPC/PLGA appeared fully injectable at 4 min. Moreover, in comparison with CPC, CPC/PLGA showed a full degradation in 6 weeks (with 100% mass loss), and this was associated with an acidification of the medium containing the CPC/PLGA sample (pH of 3.5 after 6 weeks). A cell viability test validated CPC/PLGA biocompatibility, and in vivo analyses using a bone defect assay in the caudal vertebrae of Wistar rats showed the good opacity of the CPC through the tail and a significant increased degradation of the CPC/PLGA cement a month after implantation. In conclusion, this injectable CPC scaffold appears to be an interesting material for bone substitution.

Multifunctional manganese-doped Prussian blue nanoparticles for two-photon photothermal therapy and magnetic resonance imaging.

Here we demonstrate for the first time that Mn2+-doped Prussian blue nanoparticles of c.a. 70 nm act as effective agents for photothermal therapy under two-photon excitation with an almost total eradication of malignant cells (97 and 98%) at a concentration of 100 µg mL-1 24 h after NIR excitation. This effect combined with interesting longitudinal NMR relaxivity values offer new perspectives for effective imaging and cancer treatment.