Background. Effective biomarkers are needed to enable personalized medicine for pancreatic cancer patients. This study analyzes the prognostic value, in early pancreatic cancer, of single circulating tumor cell (CTC) and CTC clusters from the central venous catheter (CVC) and portal blood (PV). Methods. In total, 7 mL of PV and CVC blood from 35 patients with early pancreatic cancer were analyzed. CTC were isolated using a positive immunomagnetic selection. The detection and identification of CTC were performed by immunocytochemistry (ICC) and were analyzed by Epi-fluorescence and confocal microscopy. Results. CTC and the clusters were detected both in PV and CVC. In both samples, the CTC number per cluster was higher in patients with grade three or poorly differentiated tumors (G3) than in patients with well (G1) or moderately (G2) differentiated. Patients with fewer than 185 CTC in PV exhibited a longer OS than patients with more than 185 CTC (24.5 vs. 10.0 months; p = 0.018). Similarly, patients with fewer than 15 clusters in PV showed a longer OS than patients with more than 15 clusters (19 vs. 10 months; p = 0.004). These significant correlations were not observed in CVC analyses. Conclusions. CTC presence in PV could be an important prognostic factor to predict poor prognosis in early pancreatic cancer. In addition, the number of clustered-CTC correlate to a tumor negative differentiation degree and, therefore, could be used as a diagnostic biomarker for pancreatic cancer.

The current treatments to restore skeletal muscle defects present several injuries. The creation of scaffolds and implant that allow the regeneration of this tissue is a solution that is reaching the researchers' interest. To achieve this, electrospinning is a useful technique to manufacture scaffolds with nanofibers with different orientation. In this work, polycaprolactone and gelatin solutions were tested to fabricate electrospun scaffolds with two degrees of alignment between their fibers: random and aligned. These scaffolds can be seeded with myoblast C2C12 and then stimulated with a mechanical bioreactor that mimics the physiological conditions of the tissue. Cell viability as well as cytoskeletal morphology and functionality was measured. Myotubes in aligned scaffolds (9.84 ± 1.15 μm) were thinner than in random scaffolds (11.55 ± 3.39 μm; P = 0.001). Mechanical stimulation increased the width of myotubes (12.92 ± 3.29 μm; P < 0.001), nuclear fusion (95.73 ± 1.05%; P = 0.004), and actin density (80.13 ± 13.52%; P = 0.017) in aligned scaffolds regarding the control. Moreover, both scaffolds showed high myotube contractility, which was increased in mechanically stimulated aligned scaffolds. These scaffolds were also electrostimulated at different frequencies and they showed promising results. In general, mechanically stimulated aligned scaffolds allow the regeneration of skeletal muscle, increasing viability, fiber thickness, alignment, nuclear fusion, nuclear differentiation, and functionality.

Circulating Tumor Cells (CTCs) are considered a prognostic marker in pancreatic cancer. In this study we present a new approach for counting CTCs and CTC clusters in patients with pancreatic cancer using the Isoflux^TM System with the Hough transform algorithm (Hough-Isoflux^TM). The Hough-Isoflux^TM approach is based on the counting of an array of pixels with a nucleus and cytokeratin expression excluding the CD45 signal. Total CTCs including free and CTC clusters were evaluated in healthy donor samples mixed with pancreatic cancer cells (PCCs) and in samples from patients with pancreatic ductal adenocarcinoma (PDAC). The Isoflux^TM System with manual counting was used in a blinded manner by three technicians who used Manual-Isoflux^TM as a reference. The accuracy of the Hough-Isoflux^TM approach for detecting PCC based on counted events was 91.00% [84.50, 93.50] with a PCC recovery rate of 80.75 ± 16.41%. A high correlation between the Hough-Isoflux^TM and Manual-Isoflux^TM was observed for both free CTCs and for clusters in experimental PCC (R² = 0.993 and R² = 0.902 respectively). However, the correlation rate was better for free CTCs than for clusters in PDAC patient samples (R² = 0.974 and R² = 0.790 respectively). In conclusion, the Hough-Isoflux^TM approach showed high accuracy for the detection of circulating pancreatic cancer cells. A better correlation rate was observed between Hough-Isoflux^TM approach and with the Manual-Isoflux^TM for isolated CTCs than for clusters in PDAC patient samples.

Additive manufacturing techniques allow the customized design of medical devices according to the patient's requirements. Enteroatmospheric fistula is a pathology that benefits from this personalization due to its extensive clinical variability since the size and morphology of the wound differ extensively among patients. Standard prosthetics do not achieve proper isolation of the wound, leading to a higher risk of infections. Currently, no effective personalized technique to isolate it has been described. In this work, we present the workflow for the design and manufacture of customized devices adapted to the fistula characteristics as it evolves and changes during the treatment with Negative Pressure Wound Therapy (NPWT). For each case, a device was designed with dimensions and morphology depending on each patient's requirements using white light scanning, CAD design, and additive manufacturing. The design and manufacture of the devices were performed in 230.50 min (184.00–304.75). After the placement of the device, the wound was successfully isolated from the intestinal content for 48–72 h. The therapy was applied for 27.71 ± 13.74 days, and the device was redesigned to adapt to the wound when geometrical evolutionary changes occur during the therapy. It was observed a decrease in weekly cures from 23.63 ± 10.54 to 2.69 ± 0.65 (p = 0.001). The fistulose size was reduced longitudinal and transversally by 3.25 ± 2.56 cm and 6.06 ± 3.14 cm, respectively. The wound depth also decreased by 1.94 ± 1.08 cm. In conclusion, customization through additive manufacturing is feasible and offers promising results in the generation of personalized devices for the treatment of enteroatmospheric fistula.

Introduction: A positive surgical margin (R1 resection) is a relevant risk factor for local recurrence in patients with pancreatic ductal adenocarcinoma of the pancreas (PDAC). An intraoperative liquid biopsy (ILB) based on tumor cell mobilization could help to detect R1 resection intraoperatively. Objective: To evaluate the potential role of the intraoperative circulating tumor cells (CTCs) and cluster mobilization on the R0/R1 detection. Methods: Sixty-three patients with resectable PDAC of the head of the pancreas were prospective enrolled under the CETUPANC trial. Open pancreaticoduodenectomy (PD) was done in all patients. Intraoperative CTCs and clusters were determined during PD. Results: The overall rate of R1 resection was 34.9% (22/63 patients). Multivariate analysis showed that factors associated with R1 resection (AUC=0.920) were the presence of undifferentiated G3 tumor (P=0.017), microscopic vascular invasion (P=0.016), and the intraoperative increase of both free CTCs and clusters in portal vein determination from the beginning to the end of the surgery (P=0.002 and P=0.005, respectively). A specific logistic regression model, including delta end to baseline CTCs and cluster mobilization to achieve a combined cut-off to detect R1 detection was calculated (AUC=0.799). The obtained R1-index based on ILB had 84% of sensitivity and 68% of specificity to detect R1 resection. Conclusions: The ILB based on the intraoperative mobilization of CTCs and clusters from the beginning to the end of the PD was a predictive factor to detect R1 resection in patients with PDAC.