A 4D printed self-assembling PEGDA microscaffold fabricated by digital light processing for arthroscopic articular cartilage tissue engineering.

Articular cartilage in synovial joints such as the knee has limited capability to regenerate independently, and most clinical options for focal cartilage repair merely delay total joint replacement. Tissue engineering presents a repair strategy in which an injectable cell-laden scaffold material is used to reconstruct the joint in situ through mechanical stabilisation and cell-mediated regeneration. In this study, we designed and 3D-printed millimetre-scale micro-patterned PEGDA biomaterial microscaffolds which self-assemble through tessellation at a scale relevant for applications in osteochondral cartilage reconstruction. Using simulated chondral lesions in an in vitro model, a series of scaffold designs and viscous delivery solutions were assessed. Hexagonal microscaffolds (750 µm x 300 µm) demonstrated the best coverage of a model cartilage lesion (at 73.3%) when injected with a 1% methyl cellulose solution. When chondrocytes were introduced to the biomaterial via a collagen hydrogel, they successfully engrafted with the printed microscaffolds and survived for at least 14 days in vitro, showing the feasibility of reconstructing stratified cartilaginous tissue using this strategy. Our study demonstrates a promising application of this 4D-printed injectable technique for future clinical applications in osteochondral tissue engineering. Supplementary Information: The online version contains supplementary material available at 10.1007/s40964-022-00360-0.

The rare circulating tumor microemboli as a biomarker contributes to predicting early colorectal cancer recurrences after medical treatment.

Early prognosis of cancer recurrence remains difficult partially due to insufficient and ineffective screening biomarkers or regimes. This study evaluated the rare circulating tumor microemboli (CTM) from liquid biopsy individually and together with circulating tumor cells (CTCs) and serum CEA/CA19-9 in a panel, on early prediction of colorectal cancer (CRC) recurrence. Stained CTCs/CTM were detected by a microfluidic chip-based automatic rare-cell imaging platform. ROC, AUC, Kaplan-Meier survival, and Cox proportional hazard models regarding 4 selected biomarkers were analyzed. The relative risk, odds ratio, predictive accuracy, and positive/negative predictive value of biomarkers individually and in combination, to predict CRC recurrence were assessed and preliminarily validated. The EpCAM+Hochest+CD45- CTCs/CTM could be found in all cancer stages, where more recurrences were observed in late-stage cases. Significant correlations between CTCs/CTM with metastatic stages and clinical treatment were illustrated. CA19-9 and CTM could be seen as independent risk factors in patient survivals, while stratified patients by grouped biomarkers on the Kaplan-Meier analyses presented more significant differences in predicting CRC recurrences. By monitoring the panel of selected biomarkers, disease progressions of 4 CRC patients during follow-up visits after first treatments within 3 years were predicted successfully. This study unveiled the value of rare CTM on clinical studies and a panel of selected biomarkers on predicting CRC recurrences in patients at the early time after medical treatment, in which the CTM and serum CA19-9 could be applied in clinical surveillance and CRC management to improve the accuracy.

Optimizing sludge dewatering efficiency with ultrasonic Treatment: Insights into Parameters, Effects, and microstructural changes.

Sludge dewatering plays a critical role in the efficient and cost-effective management of wastewater treatment plants. Ultrasonic treatment has emerged as a promising technique for improving dewatering processes. This study aims to evaluate the impact of ultrasonic treatment on sludge dewatering characteristics. A series of experiments were conducted to evaluate the dewatering characteristics of sludge under ultrasonic treatment. Experimental data was collected, and the effects of ultrasonic parameters on dewatering efficiency were analyzed. Ultrasound has the capacity to disintegrate sludge flocs, liberate tightly bound water, and enhance sludge dewatering capabilities. The application of ultrasound leads to the breakdown of sludge flocs, which facilitates a substantial amount of organic acids or carbonates. This, in turn, modifies the pH value of the sludge. Additionally, ultrasound induces instantaneous high temperature and pressure within the liquid phase, consequently elevating the temperature of the sludge slurry. Optimum ultrasound energy density and duration of ultrasound treatment exist. For the sludge samples analyzed in this investigation, it was determined that the optimal ultrasonic energy density is 9.8 W, while the optimal duration of ultrasound treatment is 30 s. Excessively escalating the sound energy density or prolonging the duration of ultrasound may yield unfavorable outcomes in terms of sludge dewatering effectiveness. To enhance sludge dewatering, it is crucial to select appropriate ultrasonic energy density and duration of ultrasonic treatment. This study demonstrates the positive impact of ultrasonic treatment on the dewatering characteristics of sludge. The findings provide valuable insights into the potential of ultrasonic technology for enhancing sludge dewatering.

Attachment of facile synthesized NaCo2O4 nanodots to SiO2 nanoflakes for sodium-rich boosted Pt-dominated ambient HCHO oxidation.

Surface alkali metal ions are typically utilized as available promoters for ambient HCHO oxidation. In this study, NaCo2O4 nanodots with two different preferential crystallographic orientations are synthesized by facile attachment to SiO2 nanoflakes with varying degrees of lattice defects. A unique Na-rich environment is established through interlayer Na+ diffusion based on the small size effect. The optimized catalyst Pt/HNaCo2O4/T2 can deal with HCHO below 5 ppm in the static measurement system with a sustained release background and produces approximately 40 ppm of CO2 in 2 h. Combining the experimental analyses with density functional theory (DFT) calculations, the possible catalytic enhancing mechanism is proposed from the support promotion perspective, and the positive synergistic effect of Na-rich, oxygen vacancies and optimized facets for Pt-dominant ambient HCHO oxidation via both kinetic and thermodynamic processes is confirmed.

Potential Values of Circulating microRNA-21 to Predict Early Recurrence in Patients with Colorectal Cancer after Treatments.

Insufficient prognosis of local recurrence contributes to the poor progression-free survival rate and death in colorectal cancer (CRC) patients. Various biomarkers have been explored in predicting CRC recurrence. This study investigated the expressions of plasma/exosomal microRNA-21 (miR-21) in 113 CRC patients by qPCR, their values of predicting CRC recurrence, and the possibility to improve the prognostic efficacy in early CRC recurrence in stratified patients by combined biomarkers including circulating miR-21s, circulating tumour cells/microemboli (CTCs/CTM), and serum carcinoembryonic antigen (CEA)/carbohydrate antigen 19-9 (CA19-9). Expressions of plasma and exosomal miR-21s were significantly correlated (p < 0.0001) in all and late-stage patients, presenting similar correlations with other biomarkers. However, stage IV patients stratified by a high level of exosomal miR-21 and stage I to III patients stratified by a high level of plasma miR-21 displayed significantly worse survival outcomes in predicting CRC recurrence, suggesting their different values to predict CRC recurrence in stratified patients. Comparable and even better performances in predicting CRC recurrence in late-stage patients were found by CTCs/CTM from our blood samples as sensitive biomarkers. Improved prognosing efficacy in CRC recurrence and better outcomes to significantly differentiate recurrence in stratified patients could be obtained by analysing combined biomarkers.

Carcinoma-specific expression of P2Y11 receptor and its contribution in ATP-induced purinergic signalling and cell migration in human hepatocellular carcinoma cells.

Extracellular ATP-induced Ca2+ signalling is critical in regulating diverse physiological and disease processes. Emerging evidence suggests high concentrations of extracellular ATP in tumour tissues. In this study, we examined the P2 receptor for ATP-induced Ca2+ signalling in human hepatocellular carcinoma (HCC) cells. Fura-2-based measurements of the intracellular Ca2+ concentration ([Ca2+]i) showed that extracellular ATP induced an increase in the [Ca2+]i in human HCC Huh-7 and HepG2 cells. NF546, a P2Y11 receptor agonist was equally effective in inducing an increase in the [Ca2+]i. In contrast, agonists for the P2X receptors (αßmeATP and BzATP), P2Y1 receptor (MRS2365) or P2Y2 receptor (MRS2768) were ineffective. In addition, ATP/NF546-induced increases in the [Ca2+]i were strongly inhibited by treatment with NF340, a P2Y11 receptor antagonist. Immunofluorescent confocal imaging and western blotting analysis consistently demonstrated the P2Y11 receptor expression in Huh-7 and HepG2 cells. Transfection with P2Y11-specific siRNA attenuated the P2Y11 receptor protein expression level and also reduced NF546-induced increase in the [Ca2+]i. Importantly, immunohistochemistry revealed that the P2Y11 receptor was expressed at very high level in human HCC tissues and, by contrast, it was barely detected in normal liver tissues. Trans-well cell migration assay demonstrated that ATP and NF546 induced concentration-dependent stimulation of Huh-7 cell migration. Treatment with NF340 prevented ATP-induced stimulation of cell migration. Taken together, our results show carcinoma-specific expression of the P2Y11 receptor and its critical role in mediating ATP-inducing Ca2+ signalling and regulating cell migration in human HCC cells.

Expression of P2 Purinergic Receptors in Mesenchymal Stem Cells and Their Roles in Extracellular Nucleotide Regulation of Cell Functions.

Extracellular ATP and other nucleotides induce autocrine and/or paracrine purinergic signalling via activation of the P2 receptors on the cell surface, which represents one of the most common signalling mechanisms. Mesenchymal stem cells (MSC) are a type of multipotent adult stem cells that have many promising applications in regenerative medicine. There is increasing evidence to show that extracellular nucleotides regulate MSC functions and P2 receptor-mediated purinergic signalling plays an important role in such functional regulation. P2 receptors comprise ligand-gated ion channel P2X receptors and G-protein-coupled P2Y receptors. In this review, we provide an overview of the current understanding with respect to expression of the P2X and P2Y receptors in MSC and their roles in mediating extracellular nucleotide regulation of MSC proliferation, migration and differentiation. J. Cell. Physiol. 232: 287-297, 2017. © 2016 Wiley Periodicals, Inc.

Purinergic and Store-Operated Ca(2+) Signaling Mechanisms in Mesenchymal Stem Cells and Their Roles in ATP-Induced Stimulation of Cell Migration.

ATP is an extrinsic signal that can induce an increase in the cytosolic Ca(2+) level ([Ca(2+) ]c ) in mesenchymal stem cells (MSCs). However, the cognate intrinsic mechanisms underlying ATP-induced Ca(2+) signaling in MSCs is still contentious, and their importance in MSC migration remains unknown. In this study, we investigated the molecular mechanisms underlying ATP-induced Ca(2+) signaling and their roles in the regulation of cell migration in human dental pulp MSCs (hDP-MSCs). RT-PCR analysis of mRNA transcripts and interrogation of agonist-induced increases in the [Ca(2+) ]c support that P2X7, P2Y1 , and P2Y11 receptors participate in ATP-induced Ca(2+) signaling. In addition, following P2Y receptor activation, Ca(2+) release-activated Ca(2+) Orai1/Stim1 channel as a downstream mechanism also plays a significant role in ATP-induced Ca(2+) signaling. ATP concentration-dependently stimulates hDP-MSC migration. Pharmacological and genetic interventions of the expression or function of the P2X7, P2Y1 and P2Y11 receptors, and Orai1/Stim1 channel support critical involvement of these Ca(2+) signaling mechanisms in ATP-induced stimulation of hDP-MSC migration. Taken together, this study provide evidence to show that purinergic P2X7, P2Y1 , and P2Y11 receptors and store-operated Orai1/Stim1 channel represent important molecular mechanisms responsible for ATP-induced Ca(2+) signaling in hDP-MSCs and activation of these mechanisms stimulates hDP-MSC migration. Such information is useful in building a mechanistic understanding of MSC homing in tissue homeostasis and developing more efficient MSC-based therapeutic applications. Stem Cells 2016;34:2102-2114.