Exosomal miR-673-5p from fibroblasts promotes Schwann cell-mediated peripheral neuron myelination by targeting the TSC2/mTORC1/SREBP2 axis.

Peripheral myelination is a complicated process, wherein Schwann cells (SCs) promote the formation of the myelin sheath around the axons of peripheral neurons. Fibroblasts are the second resident cells in the peripheral nerves; however, the precise function of fibroblasts in SC-mediated myelination has rarely been examined. Here, we show that exosomes derived from fibroblasts boost myelination-related gene expression in SCs. We used exosome sequencing, together with bioinformatic analysis, to demonstrate that exosomal microRNA miR-673-5p is capable of stimulating myelin gene expression in SCs. Subsequent functional studies revealed that miR-673-5p targets the regulator of mechanistic target of the rapamycin (mTOR) complex 1 (mTORC1) tuberous sclerosis complex 2 in SCs, leading to the activation of downstream signaling pathways including mTORC1 and sterol-regulatory element binding protein 2. In vivo experiments further confirmed that miR-673-5p activates the tuberous sclerosis complex 2/mTORC1/sterol-regulatory element binding protein 2 axis, thus promoting the synthesis of cholesterol and related lipids and subsequently accelerating myelin sheath maturation in peripheral nerves. Overall, our findings revealed exosome-mediated cross talk between fibroblasts and SCs that plays a pivotal role in peripheral myelination. We propose that exosomes derived from fibroblasts and miR-673-5p might be useful for promoting peripheral myelination in translational medicine.

Metabolic analysis of the antidepressive effects of Yangxinshi Tablet in a vascular depression model in mice.

In recent years, vascular depression has become the focus of international attention. Yangxinshi Tablet (YXST) is usually used in cthe linic for the treatment of arrhythmia and heart failure, but we found that it also has antidepressive effects. The objective of the study was to identify biomarkers related to vascular depression in hippocampus and explore the antidepressive effects of YXST on the mouse model. Untargeted metabolomics based on UHPLC-Q-TOF/MS was applied to identify significantly differential biomarkers between the model group and control group. Unsupervised principal component analysis (PCA) was used to scan the tendency of groups and partial least squares-discriminant analysis (PLS-DA) to distinguish between the vascular depressive mice and the sham. PCA stores showed clear differences in metabolism between the vascular depressive mice and sham groups. The PLS-DA model exhibited 38 metabolites as the biomarkers to distinguish the vascular depressive mice and the sham. Further, YXST significantly regulated 22 metabolites to normal levels. The results suggested that YXST has a comprehensive antidepressive effect on vascular depression via regulation of multiple metabolic pathways including amino acid, the tricarboxylic acid cycle and phosphoglyceride metabolisms. These findings provide insight into the pathophysiological mechanism underlying vascular depression and the mechanism of YXST.

Cell culture on suspended fiber for tissue regeneration: A review.

Cell culturing is a cornerstone of tissue engineering, playing a crucial role in tissue regeneration, drug screening, and the study of disease mechanisms. Among various culturing techniques, 3D culture systems, particularly those utilizing suspended fiber scaffolds, offer a more physiologically relevant environment than traditional 2D monolayer cultures. These 3D scaffolds enhance cell growth, differentiation, and proliferation by mimicking the in vivo cellular milieu. This review focuses on the critical role of suspended fiber scaffolds in tissue engineering. We compare the effectiveness of 3D suspended fiber scaffolds with 2D culture systems, discussing their respective benefits and limitations in the context of tissue regeneration. Furthermore, we explore the preparation methods of suspended fiber scaffolds and their potential applications. The review concludes by considering future research directions for optimizing suspended fiber scaffolds to address specific challenges in tissue regeneration, underscoring their significant promise in advancing tissue engineering and regenerative medicine.

Multifunctional double-network hydrogel with antibacterial and anti-inflammatory synergistic effects contributes to wound healing of bacterial infection.

Wound infection not only hinders the time sequence of tissue repair, but also may lead to serious complications. Multifunctional wound dressings with biocompatibility, excellent mechanical properties and antibacterial properties can promote wound healing during skin infection and reduce the use of antibiotics. In this study, a multifunctional dual-network antibacterial hydrogel was constructed based on the electrostatic interaction of two polyelectrolytes, hydroxypropyl trimethyl ammonium chloride chitosan (HACC) and sodium alginate (SA). Attributing to the suitable physical crosslinking between HACC and SA, the hydrogel not only has good biocompatibility, mechanical property, but also has broad-spectrum antibacterial properties. In vivo results showed that the hydrogel could regulate M2 polarization, promote early vascular regeneration, and create a good microenvironment for wound healing. Therefore, this hydrogel is an effective multifunctional wound dressing. Consequently, we propose a novel hydrogel with combined elements to expedite the intricate repair of wound infection.

Correction: Innovative wound management: creating dynamic Alg-Mg/SF hydrogels for controlled Mg2+ release in wound healing.

[This corrects the article DOI: 10.1039/D4RA00793J.].

Innovative wound management: creating dynamic Alg-Mg/SF hydrogels for controlled Mg2+ release in wound healing.

Antibacterial hydrogels have gained considerable attention for soft tissue repair, particularly in preventing infections associated with wound healing. However, developing an antibacterial hydrogel that simultaneously possesses excellent cell affinity and controlled release of metal ions remains challenging. This study introduces an antibacterial hydrogel based on alginate modified with bisphosphonate, forming a coordination complex with magnesium ions. The hydrogel, through an interpenetrating network with silk fibroin, effectively controls the release of magnesium ions and enhances strain resistance. The Alg-Mg/SF hydrogel not only demonstrates outstanding biocompatibility and broad-spectrum antibacterial properties but also stimulates macrophages to secrete anti-inflammatory factors. This advanced Alg-Mg/SF hydrogel provides a convenient therapeutic approach for chronic wound management, showcasing its potential applications in wound healing and other relevant biomedical fields.

Skin derived precursors induced Schwann cells mediated tissue engineering-aided neuroregeneration across sciatic nerve defect.

A central question in neural tissue engineering is how the tissue-engineered nerve (TEN) translates detailed transcriptional signals associated with peripheral nerve regeneration into meaningful biological processes. Here, we report a skin-derived precursor-induced Schwann cell (SKP-SC)-mediated chitosan/silk fibroin-fabricated tissue-engineered nerve graft (SKP-SCs-TEN) that can promote sciatic nerve regeneration and functional restoration nearly to the levels achieved by autologous nerve grafts according to behavioral, histological, and electrophysiological evidence. For achieving better effect of neuroregeneration, this is the first time to jointly apply a dynamic perfusion bioreactor and the ascorbic acid to stimulate the SKP-SCs secretion of extracellular matrix (ECM). To overcome the limitation of traditional tissue-engineered nerve grafts, jointly utilizing SKP-SCs and their ECM components were motivated by the thought of prolongating the effect of support cells and their bioactive cues that promote peripheral nerve regeneration. To further explore the regulatory model of gene expression and the related molecular mechanisms involved in tissue engineering-aided peripheral nerve regeneration, we performed a cDNA microarray analysis of gene expression profiling, a comprehensive bioinformatics analysis and a validation study on the grafted segments and dorsal root ganglia tissues. A wealth of transcriptomic and bioinformatics data has revealed complex molecular networks and orchestrated functional regulation that may be responsible for the effects of SKP-SCs-TEN on promoting peripheral nerve regeneration. Our work provides new insights into transcriptomic features and patterns of molecular regulation in nerve functional recovery aided by SKP-SCs-TEN that sheds light on the broader possibilities for novel repair strategies of peripheral nerve injury.

Synthesis and neuroprotective effects of the fluorine substituted salidroside analogues in the PC12 cell model exposed to hypoglycemia and serum limitation.

Salidroside (Sal) is a natural antioxidant extracted from the root of Rhodiola rosea L., a traditional Chinese medicinal plant, which elicits neuroprotective effects in the treatment of ischemic stroke. In an attempt to improve its neuroprotective effects, fluorine substituted Sal analogues were synthesized and their neuroprotective activities against the hypoglycemia and serum limitation induced cell injury in differentiated PC12 cells were evaluated. The target compounds displayed strong protective effects on the cell viability against the damage caused by hypoglycemia and serum limitation, especially for D1, which had a great potency superior to Sal and efficiently inhibited hypoglycemia and serum limitation induced cell nuclear morphologic changes and the increased apoptotic rate in a dose-dependent manner. These new findings may provide potentially important information for further development of Sal analogues and lay the basis for further studies on the cerebral ischemic stroke and neurodegenerative diseases for human clinical treatment.

Synthesis and protective effects of novel salidroside analogues on glucose and serum depletion induced apoptosis in PC12 cells.

Salidroside is a natural product isolated from Rhodiola rosea L. which possesses a wide range of biological activities, especially neuroprotective effects in the treatment of ischemic stroke. In an attempt to improve its neuroprotective effects, a series of novel salidroside analogues were synthesized and their neuroprotective activities were evaluated against the glucose and serum depletion-induced cell death in differentiated PC12 cells. Most target compounds displayed protective effects on the cell viability, especially for compound 6, which had a great potency superior to salidroside. MTT assay and Hoechst 33342 staining collectively showed that pretreatment with 6 attenuated cell viability loss and reduced apoptotic death in cultured PC12 cells with glucose and serum depletion. And its neuroprotective effects might be associated with the increase of the apoptosis-related protein Bcl-2/Bax expression ratio, and also with the inhibition of caspase-3 activation. Therefore, our new findings may provide potentially important information for further development of salidroside analogues and lay the basis for further studies on the cerebral ischemic stroke and neurodegenerative diseases for human clinical treatment.

Leveraging artificial intelligence to identify high-risk patients for postoperative sore throat: An observational study.

Postoperative sore throat (POST) is a prevalent complication after general anesthesia and targeting high-risk patients help in its prevention. This study developed and validated a machine learning model to predict POST. A total number of 834 patients who underwent general anesthesia with endotracheal intubation were included in this study. Data from a cohort of 685 patients was used for model development and validation, whilst a cohort of 149 patients served for external validation. The prediction performance of random forest (RF), neural network (NN), and XGBoost models was compared using comprehensive performance metrics. The Local Interpretable Model-Agnostic Explanations (LIME) methods elucidated the best-performing model. POST incidences across training, validation, and testing cohorts were 41.7%, 38.4%, and 36.2%, respectively. Five predictors were age, sex, endotracheal tube cuff pressure, endotracheal tube insertion depth, and the time interval between extubation and the first drinking of water after extubation. After incorporating these variables, the NN model demonstrated superior generalization capabilities in predicting POST when compared to the XGBoost and RF models in external validation, achieving an area under the receiver operating characteristic curve (AUROC) of 0.81 (95% CI 0.74-0.89) and a precision-recall curve (AUPRC) of 0.77 (95% CI 0.66-0.86). The model also showed good calibration and clinical usage values. The NN model outperforms the XGBoost and RF models in predicting POST, with potential applications in the healthcare industry for reducing the incidence of this common postoperative complication.

Fibroblast exosomal TFAP2C induced by chitosan oligosaccharides promotes peripheral axon regeneration via the miR-132-5p/CAMKK1 axis.

Chitosan and its degradation product, oligosaccharides, have been shown to facilitate peripheral nerve regeneration. However, the underlying mechanisms are not well understood. In this study, we analyzed the protein expression profiles in sciatic nerves after injury using proteomics. A group of proteins related to exosome packaging and transport is up-regulated by chitosan oligosaccharides (COS), implying that exosomes are involved in COS-induced peripheral nerve regeneration. In fact, exosomes derived from fibroblasts (f-EXOs) treated with COS significantly promoted axon extension and regeneration. Exosomal protein identification and functional studies, revealed that TFAP2C is a key factor in neurite outgrowth induced by COS-f-EXOs. Furthermore, we showed that TFAP2C targets the pri-miRNA-132 gene and represses miR-132-5p expression in dorsal root ganglion neurons. Camkk1 is a downstream substrate of miR-132-5p that positively affects axon extension. In rats, miR-132-5p antagomir stimulates CAMKK1 expression and improves axon regeneration and functional recovery in sciatic nerves after injury. Our data reveal the mechanism for COS in axon regeneration, that is COS induce fibroblasts to produce TFAP2C-enriched EXOs, which are then transferred into axons to promote axon regeneration via miR-132-5p/CAMKK1. Moreover, these results show a new facet of fibroblasts in axon regeneration in peripheral nerves.

Matrix stiffness triggers chemoresistance through elevated autophagy in pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) has a signature of extremely high matrix stiffness caused by a special desmoplastic reaction, which dynamically stiffens along with the pathological process. The poor prognosis and low five-year survival rate of PDAC are partly owing to chemoresistance triggered by substrate stiffness. Understanding the potential mechanisms of matrix stiffness causing PDAC chemoresistance is of great significance. In this study, methacrylated gelatin hydrogel was used as platform for PANC-1 and MIA-PaCa2 cell culture. The results indicated that compared to soft substrate, stiff substrate distinctively reduced the gemcitabine sensitivity of pancreatic cancer. Intriguingly, transmission electron microscopy, immunofluorescence staining, western blot and qRT-PCR assay showcased that the number of autophagosomes and the expression of LC3 were elevated. The observations indicate that matrix stiffness may regulate the autophagy level, which plays a vital role during chemoresistance. In brief, soft substrate exhibited low autophagy level, while the counterpart displayed elevated autophagy level. In order to elucidate the underlying interaction between matrix stiffness-mediated cell autophagy and chemoresistance, rescue experiments with rapamycin and chloroquine were conducted. We found that inhibiting cell autophagy dramatically increased the sensitivity of pancreatic cancer cells to gemcitabine in the stiff group, while promoting autophagy-driven chemoresistance in the soft group, demonstrating that matrix stiffness modulated chemoresistance via autophagy. Furthermore, RNA-seq results showed that miR-1972 may regulate autophagy level in response to matrix stiffness. Overall, our research shed light on the synergistic therapy of PDAC combined with gemcitabine and chloroquine, which is conducive to promoting a therapeutic effect.

Progress in methods for evaluating Schwann cell myelination and axonal growth in peripheral nerve regeneration via scaffolds.

Peripheral nerve injury (PNI) is a neurological disorder caused by trauma that is frequently induced by accidents, war, and surgical complications, which is of global significance. The severity of the injury determines the potential for lifelong disability in patients. Artificial nerve scaffolds have been investigated as a powerful tool for promoting optimal regeneration of nerve defects. Over the past few decades, bionic scaffolds have been successfully developed to provide guidance and biological cues to facilitate Schwann cell myelination and orientated axonal growth. Numerous assessment techniques have been employed to investigate the therapeutic efficacy of nerve scaffolds in promoting the growth of Schwann cells and axons upon the bioactivities of distinct scaffolds, which have encouraged a greater understanding of the biological mechanisms involved in peripheral nerve development and regeneration. However, it is still difficult to compare the results from different labs due to the diversity of protocols and the availability of innovative technologies when evaluating the effectiveness of novel artificial scaffolds. Meanwhile, due to the complicated process of peripheral nerve regeneration, several evaluation methods are usually combined in studies on peripheral nerve repair. Herein, we have provided an overview of the evaluation methods used to study the outcomes of scaffold-based therapies for PNI in experimental animal models and especially focus on Schwann cell functions and axonal growth within the regenerated nerve.

Experimental Investigation on the Diffusion Law of Polymer Slurry Grouted in Sand.

Polymer slurry is widely used in underground engineering treatment, but due to the concealed nature of underground projects, the diffusion pattern of slurry in the sand has been little studied. In this study, the basic physical properties of water-based polyurethane, oil-based polyurethane, and acrylate and epoxy resin were compared, and the performance of chemical grouted sands with different polymer slurry/sand mass ratios (PS/S) was tested. The higher the PS/S, the better the mechanical and impermeability properties of the chemical grouted sands. In this paper, water-based polyurethane was selected to carry out orthogonal tests on the diffusivity of slurry in sands. This experiment investigated the degree of influence of sand quality, grouting pressure and sand compactness on the diffusion of slurry in sands. The test results show that, in terms of factors affecting the final pressure of grouting, the sand density has the greatest influence, followed by the sand quality, and the grouting pressure is the smallest. In terms of slurry diffusibility, grouting pressure has the greatest influence, followed by sand compactness, and the sand quality is the smallest. The diffusion mechanism of slurry in the sand was deduced from the morphology of chemical grouted sands. Water-based polyurethane showed splitting-compression-penetration diffusion in sands of different grades, and the diffusion pattern of the slurry was not the same in low-pressure (1-1.5 MPa) grouting and high-pressure (2 MPa) grouting, and lateral splitting occurred in the case of high-pressure grouting diffusion.

Study of the Structural Mechanical Properties of Drainage Canals Rehabilitated by Spraying Method.

A large number of drainage pipes and canals in China have been in disrepair for a long time and there have been problems such as leakage and corrosion. In response to these problems, this paper studies a non-excavation technology for repairing the arched canal structure-the in-situ spraying method. To study the influence of the original canal structure on the mechanical characteristics of the lining structure by in-situ spraying and the restraint effect on the lining structure, a field model test with a similar ratio of 1:2 was conducted in the field test pit. By conducting four stages of three-point concentrated load loading tests, the mechanical characteristics of the lining structure were investigated to reveal the influence of the canal structure on the force of the lining structure. The test results show that: the maximum crack width of the newly added lining structure is 0.27 mm and the normal service ultimate bearing capacity of the arched structure repaired by H-70 reaches 150 kN; comparing the loading test and the numerical simulation results, the difference between the two vault displacement results is 4.65% and the results are relatively consistent. The displacement of the bottom of the lining structure is small and the participation of the bottom plate is small, while the displacement of the upper arch structure of the lining is significantly larger than the lateral displacement, indicating that the canal structure can effectively limit the lateral displacement of the newly added lining and that the canal structure is greatly reduced. The bending moment of the lining structure is improved and the restraint effect on the arch foot is more obvious. This paper proposes the use of H-70 to repair arched canal structures by the in-situ spraying method and seeks to prove the feasibility of this method and fill the gap of research in this area. This paper provides the structural design basis and experimental knowledge for the construction of the repair method, which has important practical significance for the pipeline repair project in the future.

Conductive biocomposite hydrogels with multiple biophysical cues regulate schwann cell behaviors.

Peripheral nerve injuries are serious clinical events, and surgical treatment has certain limitations. Conductive hydrogels are promising biomaterials for neural tissue engineering, as they can enhance the functionality of neurons and Schwann cells (SCs) by mimicking the biophysical and biochemical cues existing in the natural extracellular matrix. It remains unexplored, however, whether there is a connection between the effects of different cues, such as hydrogel elasticity and conductivity, on SC fate. In the present work, we fabricated a series of conductive biocomposite hydrogels with the combination of silk fibroin (SF) and graphene oxide (GO) nanosheets and demonstrated an approach to control hydrogel electrical conductivity, independent of matrix elasticity and polymer concentration. Our results indicated that the soft substrates play a more critical role in SC survival, proliferation, spreading, and gene expression of neurotrophic factors, while the increased conductivity may also be beneficial to SC functional behaviors. These findings may promote the understanding of cell-matrix interactions and provide new insights for the design of neural tissue engineering scaffolds.

Synthesis and protective effects of aralkyl alcoholic 2-acetamido-2-deoxy-ß-D-pyranosides on hypoglycemia and serum limitation induced apoptosis in PC12 cell.

Neuroprotective agents have been in the focus of attention in the treatment of ischemic stroke. Salidroside, a phenylpropanoid glycoside isolated from Rhodiola rosea L., possessed a wide range of biological activities, especially neuroprotection. In an attempt to improve neuroprotective effects of new salidroside analogs for ischemic stroke, a series of novel aralkyl alcoholic 2-acetamido-2-deoxy-ß-d-pyranosides were synthesized and their protective activities against the hypoglycemia and serum limitation induced cell death in rat pheochromocytoma cells (PC12 cells) were studied. Most compounds showed strong neuroprotective effects, especially for 4g and 4h, which exhibited a great potency superior to salidroside. MTT assay, Hoechst 33342 staining, and flow cytometry with annexin V/PI staining collectively showed that pretreatment with 4g and 4h attenuated cell viability loss and apoptotic cell death in cultured PC12 cells. Caspase-3 colorimetric assay and Rhodamine 123 staining revealed the changes in expression levels of caspase-3 and mitochondrial membrane potential in PC12 cells on exposure to hypoglycemia and serum limitation with and without 4g and 4h pretreatment, respectively. All the results suggested that 4g and 4h protects the PC12 cells against hypoglycemia and serum limitation induced apoptosis possibly by modulation of apoptosis-related gene expression and restoration of the mitochondrial membrane potential. Therefore, these novel findings may provided a new framework for the design of new aralkyl alcoholic 2-acetamido-2-deoxy-ß-d-pyranosides as neuroprotective agents for treating cerebral ischemic stroke and neurodegenerative diseases.

Synthesis and biological evaluation of two salidroside analogues in the PC12 cell model exposed to hypoglycemia and serum limitation.

Salidroside is a phenylpropanoid glycoside isolated from Rhodiola rosea L., a traditional Chinese medicinal plant, and has displayed a broad spectrum of pharmacological properties. In this paper, two analogues were prepared with the glucosamine and N-acetylglucosamine as glycosyl donor, 2-(4-hydroxyphenyl)ethanol as glycosyl acceptor. The effects of them over PC12 cell model exposed to hypoglycemia and serum limitation were assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Flow Cytometry and Western blot analysis.

Durability Study on High-Performance Fiber-Reinforced Mortar under Simulated Wastewater Pipeline Environment.

The acid-alkaline-inducd corrosive environments inside wastewater concrete pipelines cause concrete structural deterioration and substantial economic losses all over the world. High-performance concrete/mortar (HPC) was designed to have better resistance to corrosive environments, with enhanced service life. However, the durability of HPC in wastewater pipeline environments has rarely been studied. A high-performance mortar mixture (M) reinforced by supplemental materials (including fly ash and silica fume) and polyvinyl alcohol (PVA) fibers, together with a mortar mixture (P) consisting of cement, sand and water with similar mechanical performance, were both designed and exposed to simulated wastewater pipeline environments. The visual appearance, dimensional variation, mass loss, mechanical properties, permeable pore volume, and microstructure of the specimens were measured during the corrosion cycles. More severe deterioration was observed when the alkaline environment was introduced into the corrosion cycles. Test results showed that the M specimens had less permeable pore volume, better dimensional stability, and denser microstructure than the P specimens under acid-alkaline-induced corrosive environments. The mass-loss rates of the M specimens were 66.1-77.2% of the P specimens after 12 corrosion cycles. The compressive strength of the M specimens was 25.5-37.3% higher than the P specimens after 12 cycles under corrosive environments. Hence, the high-performance mortar examined in this study was considered superior to traditional cementitious materials for wastewater pipeline construction and rehabilitation.

Biomimetic hybrid scaffold of electrospun silk fibroin and pancreatic decellularized extracellular matrix for islet survival.

Islet transplantation is considered as one of the promising treatment options for curing diabetes. However, the extracellular matrix (ECM) is destroyed during the process of islet isolation and extraction, which leads to decreased islet activity in vitro. ECM-based biomaterials which used to reconstruct the microenvironment of cells have been applied in various fields. In this study, an electrospinning hybrid scaffolds with silk fibroin (SF) and pig pancreatic decellularized extracellular matrix (P-dECM) have been prepared to mimic the islet ECM in vivo. Furthermore, the activity and function of islet were evaluated in vitro. The microstructures, hydrophilia and the main components of scaffolds were characterized by SEM, contact angle analysis and immunohistochemical experiment. The toxicity of stents was assessed by MTT assay. Cell activity and function were estimated by the live-dead cell staining, immunofluorescence, glucose-stimulated insulin secretion assay and q-PCR. A nanofiber scaffold with good hydrophilicity, non-toxic and retention of key ECM components has been obtained, which can improve the survival and promote and function of islets. This scaffold can be a promising candidate for pancreatic tissue engineering and provides a new strategy for islet transplantation.