Open-source 3D printers mean objects can be quickly and efficiently produced. However, design and fabrication parameters need to be optimized to set up the correct printing procedure; a procedure in which the characteristics of the printing materials selected for use can also influence the process. This work focuses on optimizing the printing process of the open-source 3D extruder machine RepRap, which is used to manufacture poly(ε-caprolactone) (PCL) scaffolds for cell culture applications. PCL is a biocompatible polymer that is free of toxic dye and has been used to fabricate scaffolds, i.e., solid structures suitable for 3D cancer cell cultures. Scaffold cell culture has been described as enhancing cancer stem cell (CSC) populations related to tumor chemoresistance and/or their recurrence after chemotherapy. A RepRap BCN3D+ printer and 3 mm PCL wire were used to fabricate circular scaffolds. Design and fabrication parameters were first determined with SolidWorks and Slic3r software and subsequently optimized following a novel sequential flowchart. In the flowchart described here, the parameters were gradually optimized step by step, by taking several measurable variables of the resulting scaffolds into consideration to guarantee high-quality printing. Three deposition angles (45°, 60° and 90°) were fabricated and tested. MCF-7 breast carcinoma cells and NIH/3T3 murine fibroblasts were used to assess scaffold adequacy for 3D cell cultures. The 60° scaffolds were found to be suitable for the purpose. Therefore, PCL scaffolds fabricated via the flowchart optimization with a RepRap 3D printer could be used for 3D cell cultures and may boost CSCs to study new therapeutic treatments for this malignant population. Moreover, the flowchart defined here could represent a standard procedure for non-engineers (i.e., mainly physicians) when manufacturing new culture systems is required.

months (p = 0,013). There were no significant differences in age, ECOG PS, histology, size, therapies or treatment line. Conclusions: The NLR remains a prognostic factor in both diseases, with or without EGFR mutation, but appear to have less impact on the outcome of EGFR mutated patients. In NSCLC EGFR mutated maybe inflammatory index could have implications on therapeutic choice, especially subsequent lines and his role deserves further study. Legal entity responsible for the study: Mario Scartozzi. Funding: Has not received any funding.

Background: Tumor metastasis is the main cause of death of cancer patients and the biggest hurdle for cancer cure. The identification of decisive drivers of metastasis is thus an urgent therapeutic need.Methods: Cancer cell spheroids, wound healing and cell aggregation assays were utilized to explore cell-cell adhesion capacity. Immunofluorescence confocal microscopy and flow cytometry analysis were utilized to quantify expression of target proteins, IHC analysis quantified the expression of target molecules in primary tumors and metastases. Pre-clinical models of orthotopic growth of colon cancer and metastatic diffusion to the liver were utilized. Xenotransplant transcriptome analysis assessed EMT determinant transcription. Patients: 24 distinct case series of breast, colon, uterus, ovary, stomach, lung, and pancreatic cancers, for a total number of 13,042 primary tumors were analyzed. KaplaneMeier plots were used to illustrate survival and metastatic relapse in investigated cohorts.Results: We identify functional inactivation of highly expressed E-cadherin as a pivotal driver of metastatic diffusion in human cancer. E-cadherin is inactivated by binding to Trop-2, which causes release from the cytoskeleton, loss of cell-cell adhesion and activation of b-catenin, while maintaining epithelial differentiation. This leads to antiapoptotic signaling, increased cell migration capacity and enhanced cancer cell survival. This global, Trop-2/E-cadherin/b-catenin-driven pro-metastatic program was recapitulated in human cancer, and was shown to profoundly impact on breast, colon, ovary, uterus, stomach cancer metastatic diffusion. Conclusions:We identify functional inactivation of E-cadherin by Trop-2 as a pivotal driver of EMT-less metastatic diffusion in human cancer. This global, Trop-2/E-cadherin/b-cateninedriven pro-metastatic program profoundly impacts on the survival of patients bearing breast, colon, uterus, ovary, stomach, lung, pancreas tumours, paving the way for novel diagnostic procedures and anti-cancer therapies.Legal entity responsible for the study: The authors.