Neuroprotection is an important approach for the treatment of spinal cord injury (SCI). Minocycline (MC), a known neuroprotective agent, has been utilized for SCI treatment, but its therapeutic effect is limited by instability and low bioavailability. Herein, we developed an innovative micellar thermosensitive hydrogel (MCPP-M-gel) that encapsulates MC in polyethylene glycol (PEG)-poly(lactide-co-glycolic acid) (PLGA) micelles to enhance its therapeutic efficacy in a rat model of SCI. The micelles were synthesized via the thin-film hydration method and characterized for encapsulation efficiency, particle size, zeta potential, and polydispersity index (PDI). MCPP-M-gel demonstrated favorable physico-mechanical properties and extended MC release over 72 hours in vitro without cytotoxic effects on neural crest-derived ectoderm mesenchymal stem cells (EMSCs). Thereafter, MC, MCPP-M, MCPP-M-gel and a blank micellar thermosensitive gel were injected into the injured site of SCI rats. Histopathological evaluation demonstrated that MCPP-M-gel could promote neuronal regeneration at the injured site of the SC after 28 days. Immunofluorescence techniques revealed that MCPP-M-gel increased the expression of neuronal class III β-tubulin (Tuj1), myelin basic protein (MBP), growth-associated protein 43 (GAP43), neurofilament protein-200 (NF-200) and nestin as well as reduced glial-fibrillary acidic protein (GFAP) expression in damaged areas of the SC. In conclusion, this study innovatively developed MCPP-M-gel based on a PEG-PLGA copolymer as a biomaterial, laying a solid foundation for further research and application of MCPP-M-gel in SCI models or other neurodegenerative diseases.

Background Spinal cord injury (SCI) remains a significant health concern, with limited available treatment options. This condition poses significant medical, economic, and social challenges. SCI is typically categorized into primary and secondary injuries. Inflammation, oxidative stress, scar formation, and the immune microenvironment impede axon regeneration and subsequent functional restoration. Numerous studies have shown that the destruction of the blood–brain barrier (BBB) and microvessels is a crucial factor in severe secondary injury. Additionally, reactive oxygen species (ROS)-induced lipid peroxidation significantly contributes to endothelial cell death. Pericytes are essential constituents of the BBB that share the basement membrane with endothelial cells and astrocytes. They play a significant role in the establishment and maintenance of BBB. Results Immunofluorescence staining at different time points revealed a consistent correlation between pericyte coverage and angiogenesis, suggesting that pericytes promote vascular repair via paracrine signaling. Pericytes undergo alterations in cellular morphology and the transcriptome when exposed to hypoxic conditions, potentially promoting angiogenesis. We simulated an early ischemia-hypoxic environment following SCI using glucose and oxygen deprivation and BBB models. Co-culturing pericytes with endothelial cells improved barrier function compared to the control group. However, this enhancement was reduced by the exosome inhibitor, GW4869. In vivo injection of exosomes improved BBB integrity and promoted motor function recovery in mice following SCI. Subsequently, we found that pericyte-derived exosomes exhibited significant miR-210-5p expression based on sequencing analysis. Therefore, we performed a series of gain- and loss-of-function experiments in vitro. Conclusion Our findings suggest that miR-210-5p regulates endothelial barrier function by inhibiting JAK1/STAT3 signaling. This process is achieved by regulating lipid peroxidation levels and improving mitochondrial function, suggesting a potential mechanism for restoration of the blood-spinal cord barrier (BSCB) after SCI.

Disruption of the blood-spinal cord barrier (BSCB) leads to inflammatory cell infiltration and neural cell death, thus, contributing to poor functional recovery after spinal cord injury (SCI). Previous studies have suggested that Sirtuin 1 (SIRT1), an NAD+-dependent class III histone deacetylase, is abundantly expressed in endothelial cells and promotes endothelial homeostasis. However, the role of SIRT1 in BSCB function after SCI remains poorly defined. Here, we report that SIRT1 is highly expressed in spinal cord endothelial cells, and its expression significantly decreases after SCI. Using endothelial cell-specific SIRT1 knockout mice, we observed that endothelial cell-specific knockout of SIRT1 aggravated BSCB disruption, thus, resulting in widespread inflammation, neural cell death and poor functional recovery after SCI. In contrast, activation of SIRT1 by the agonist SRT1720 had beneficial effects. In vitro, knockdown of SIRT1 exacerbated IL-1β-induced endothelial barrier disruption in bEnd.3 cells, whereas overexpression of SIRT1 was protective. Using RNA-seq and IP/MS analysis, we identified p66Shc, a redox protein, as the potential target of SIRT1. Further studies demonstrated that SIRT1 interacts with and deacetylates p66Shc, thereby attenuating oxidative stress and protecting endothelial barrier function. Overall, our results indicate that SIRT1 decreases endothelial ROS production and attenuates BSCB disruption by deacetylating p66Shc after SCI, and suggest that SIRT1 activation has potential as a therapeutic approach to promote functional recovery against BSCB disruption following SCI.

Spinal cord injury is a serious disabling disease that not only causes physical defects, but also causes psychological problems for patients. In the current study, a novel multi-dimensional functional composite nanotube was prepared by simultaneously riveting nerve growth factor (NGF)-loaded PLGA microspheres and tissue-engineered cells (BMSCs) into the PLA fiber nanotube by a designed thermosensitive hydrogel to improve spinal cord injury (SCI) repair. The PLGA microspheres allow the protection and long-term controlled release of NGF to guarantee enough time and drug concentration to promote the growth and differentiation of cells into axons and promote axonal regeneration. Thermosensitive hydrogels are served not only to provide a tunable physiologically simulated microenvironment, but also as scaffolds to provide a suitable matrix for cells and drugs, and PLA nanotube ensures the seal of the released NGF in the nanotube to improve its drug bioavailability, as well as the supporting and guiding of the nerve growth. The physicochemical properties of the composite nanotube are investigated and the repair efficacy is evaluated according to the functional recovery, histological, immunohistochemistry, immunofluorescence examinations as well as the westernblot analysis. The administration of the composite nanotube in the SCI rat models obviously improves the regeneration of the neuron, axon, myelin sheath, neural stem cell and nissnei body but decreases the astrocytes formation, as well as enhances the integrity of the spinal cord tissue and the recovery of locomotor functions. This present work provides a future perspective for NGF application and a potential strategy for SCI repair.

The nonunion following a fracture is associated with severe patient morbidity and economic consequences. Currently, accumulating studies are focusing on the importance of macrophages during fracture repair. However, details regarding the process by which macrophages facilitate endochondral ossification (EO) are largely unknown. In this study, we present evidence that apoptotic chondrocytes (ACs) are not inert corpses awaiting removal, but positively modulate the osteoinductive ability of macrophages. In vivo experiments revealed that fatty acid (FA) metabolic processes up-regulated following EO. In vitro studies further uncovered that FAs derived from ACs are taken up by macrophages mainly through macrophage scavenger receptor 1 (MSR1). Then, our functional experiments confirmed that these exogenous FAs subsequently activate peroxisome proliferator-activated receptor α (PPARα), which further facilitates lipid droplets generation and fatty acid oxidation (FAO). Mechanistically, elevated FAO is involved in up-regulating the osteoinductive effect by generating BMP7 and NAD+/SIRT1/EZH2 axis epigenetically controls BMP7 expression in macrophages cultured with ACs culture medium. Our findings advanced the concept that ACs could promote bone regeneration by regulating metabolic and function reprogram in macrophages and identified macrophage MSR1 represents a valuable target for fracture treatments.

Neuroinflammation is an important cause of poor prognosis in patients with spinal cord injury. pyroptosis is a new type of inflammatory cell death. Treg cells has been shown to play an anti-inflammatory role in a variety of inflammatory diseases, including inflammatory bowel disease, amyotrophic lateral sclerosis, and arthritis. However, little is known about Treg cells' potential role in pyroptosis following spinal cord injury. The aim of this research was to look into the effect of Treg cells to motor function recovery, pyroptosis and the mechanism behind it after SCI. Here, we found that pyroptosis mainly occurred in microglia on the seventh day after spinal cord injury. Konckout Treg cells resulted in widely pyroptosis and poor motor recovery after SCI. In conversely, over-infiltration of Treg cell in mice by tail vein injection had beneficial effects following SCI.Treg cell-derived exosomes promote functional recovery by inhibiting microglia pyroptosis in vivo. Bioinformatic analysis revealed that miRNA-709 was significantly enriched in Treg cells and Treg cell-secreted exosomes. NKAP has been identified as a miRNA-709 target gene. Moreover, experiments confirmed that Treg cells targeted the NKAP via exosomal miR-709 to reduce microglia pyroptosis and promote motor function recovery after SCI. More importantly, The miR-709 overexpressed exosomes we constructed significantly reduced the inflammatory response and improved motor recovery after spinal cord injury. In brief, our findings indicate a possible mechanism for communication between Treg cells and microglia, which opens up a new perspective for alleviating neuroinflammation after SCI.

As a worldwide medical problem, spinal cord injury has no clear and effective treatment to improve its prognosis. Hence, new treatment strategies for spinal cord injury with good therapeutic efficacy have been actively pursued. As a new drug loading system, acetal dextran nanoparticles (SAD) have good biocompatibility and biodegradability. Therefore, we designed spermine-functionalized acetal-dextran (SAD) nanoparticles and encapsulated paclitaxel (PCL) into them. This design can ensure the sustained release of paclitaxel in the injured area for 4 days and promote the extension of nerve processes in vitro. In our experiment, we found that paclitaxel-loaded SAD nanoparticles (PCL@SAD) decreased the level of chondroitin sulfate proteoglycan in the rat spinal cord injury model, which reduced the scar repair of the injured site and changed the inhibitory environment after spinal cord injury. This reveals that PCL@SAD can effectively protect the injured spinal cord and ultimately improve the functional recovery of the injured spinal cord. One single injection of PCL@SAD shows better therapeutic effect than that of PCL. This study opens an exciting perspective toward the application of neuroprotective PCL@SAD for the treatment of severe neurological diseases.

The objective was to design a scaffold that could continuously deliver nerve growth factor (NGF) combined with neurally differentiated bone marrow mesenchymal stem cells (BMSCs) to promote better recovery of spinal cord injury (SCI) in rats. BMSCs were induced to differentiate into neurons for 6 days in vitro, and then seeded on a NGF persistent delivery scaffold, both were transplanted to SCI rats in combination. Relevant extensive tests were conducted 1, 4 and 8 weeks after transplantation. The results showed that the scaffold had a stable ability to continuously release NGF and that the BMSCs on the scaffold could successfully differentiate into nerve cells. The results of Bacco, Beattie and Bresnahan (BBB) scores, inclined plane tests and electrophysiological investigations revealed that the rats in the combined regimen had better locomotor functional recovery. The results of H&E/Nissl staining, Golgi staining and immunofluorescence showed that the rats in the combined regimen retained the most neurons and had the least cavities and more formations of dendritic spines. Similarly, the positive rate was high for MAP2, NeuN and MBP, and low for GFAP. The graft of the NGF persistent delivery scaffold seeded with neurally differentiated BMSCs significantly reduced the formation of cavities and glial scars at the SCI sites and promoted neuronal survival, axonal regeneration and locomotor function recovery. Compared with the single graft of NGF persistent delivery scaffold or the single graft of neurally differentiated BMSCs, this combined scheme had a better effect in promoting the recovery of SCI.

A sandwich structured composite membrane for longtime controlled release of nerve growth factor (NGF) to repair spinal cord injury (SCI) is prepared through electrospray. In this system, PLA film is used as the sealing layer to prevent drug diffusion and provide mechanical support, PLGA microspheres as the sandwich layer to load and controlled release NGF, and chitosan (CS) film as the planting layer to seed bone marrow mesenchymal stem cells (BMSCs). The composite membrane has good biocompatibility and can effectively promote PC-12 cells to differentiate into neurons. In addition, the composite membrane can be directly applied to the injured areas without further damage. The longtime sustained release of NGF guaranteed enough requirement time for SCI repair, which will decrease the administration frequency and improve patient compliance. The administration of BMSCs coupled with the sandwich composite membrane effectively relieves SCI, decreases cavity formation, enhances neuronal regeneration and tissue repair, as well as improves the recovery of locomotor functions. Overall, this present work provides a future perspective for the treatment of SCI by the NGF-loaded sandwich composite membrane with prolonged drug release function.

The clearance of myelin debris is a critical step in the functional recovery following spinal cord injury (SCI). As phagocytes do, microvascular endothelial cells (MECs) participate in myelin debris clearance at the injury site within one week. Our group has verified that G protein-coupled receptor kinase 2 interacting protein-1 (GIT1) is essential in autophagy and angiogenesis, both of which are tightly related to the uptake and degradation of myelin debris by MECs. Here, we analyzed the performance and mechanism of GIT1 in myelin debris clearance after SCI. The SCI contusion model was established and in vitro MECs were treated with myelin debris. Better recovery from traumatic SCI was observed in the GIT1 WT mice than in the GIT1 KO mice. More importantly, we found that GIT1 prompted MECs to clear myelin debris and further enhanced MECs angiogenesis in vivo and in vitro. Mechanistically, GIT1-mediated autophagy contributed to the clearance of myelin debris by MECs. In this study, we demonstrated that GIT1 may prompt MECs to clear myelin debris via autophagy and further stimulate MECs angiogenesis via upregulating VEGF. Our results indicate that GITI may serve as a promising target for accelerating myelin debris clearance and improving SCI recovery.

Nanoscale bioactive glass particles have greater bioactivity than microscale bioactive glass particles, due to their high-specific surface area and fast ion release rate in body fluid. However, preparation of bioactive glass nanoparticles (BGNPs) is difficult since calcium is not easy to be highly doped into the forming silica atom network, leading to an uneven distribution and a low content of calcium. In addition, BGNPs are usually prepared in a dilute solution to avoid agglomeration of the nanoparticles, which decreases the production efficiency and increases the cost. In this work, BGNPs are prepared by a method of the reactive flash nanoprecipitation (RFNP) as well as a traditional sol–gel method. The results indicate that the BGNPs by the RFNP present a smaller size, narrower size distribution, more uniform composition, and better bioactivity than those by the traditional sol–gel method. The obtained BGNPs have uniform compositions close to the feed values. The high and even doping of calcium in the BGNPs is achieved. This successful doping of calcium into nanoparticles by the RFNP demonstrates a promising way to effectively generate high-quality BGNPs for bone repairs.

The interactions of ions and molecules with material surface are highly dependent on the surface properties of the material. Therefore, the distribution of ions or molecules near the material surface may be affected by the surface properties. This phenomenon can be significant enough for controlling the structure of a material synthesized in the sub-micrometer scale confinement space of a template. This work confirms that inverse opals are perfect templates for offering confinement space, while their different surface properties can strongly affect ion and block copolymer distribution in the confinement space. This surface effect principle can be used for the controlled synthesis of colloids with complex composition. As an example, four kinds of mesoporous magnetic bioactive glass colloids with ordered mesopores, core-shell structure, open surface pores, or disordered mesopores are prepared by using polystyrene and carbon inverse opal templates. This work reveals that inverse opal templates possess great advantage in controlled synthesizing colloidal structures due to their surface effect on ions and molecules and confinement space.

Arsenic trioxide (As2O3), an effective drug for leukemia, is limited to be used for solid tumor treatment due to its high side effects. In this study, polyethylene glycol (PEG) and lactobionic acid (LA) modified chitosan (PLC) was synthesized and was used to coat poly(lactide-co-glycolide) (PLGA) nanoparticles for encapsulation and targeted release of As2O3 in liver cancer treatment. The As2O3-loaded PLGA/PLC nanoparticles (As2O3-PLGA/PLC NPs) were fabricated through double emulsion-solvent evaporation method and were optimized by orthogonal tests. As2O3-PLGA/PLC NPs presented suitable physical stability, positive charge, high encapsulation efficiency and drug loading, and good biocompatibility. As expected, the NPs can quickly release enough dose of As2O3 in a short time and then sustain the drug concentration. The As2O3-PLGA/PLC NPs showed effective inhibition of SMMC-7721 cells while having lower cytotoxicity against normal human liver cells (LO2 cells). Furthermore, In vivo study showed that the NPs did not present toxic effects on kidney and liver, but showed relatively high growth inhibition effect on liver tumor. Therefore, this PLGA/PLC NPs could be an effective and safe drug delivery system for liver cancer chemotherapy.

Background: Peripheral nerves can self-regenerate after traumatic injury, although their self-regeneration ability is limited after severe nerve injury. After peripheral nerve injury, the distal nerve stumps undergo Wallerian degeneration while the proximal nerve stumps undergo a regeneration process. Methods: Here, to decipher genetic changes and involved biological processes in the proximal nerve stumps after peripheral nerve injury, microarray data (GSE30165) were analyzed. Differentially expressed genes in the proximal nerve stumps at 0.5 h, 1 h, 3 h, 6 h, 9 h, 1 d, 4 d, 7 d, and 14 d after rat sciatic nerve transection were subjected to Ingenuity pathway analysis (IPA) bioinformatic analysis. Results: Cytokine signaling, cellular immune response, nuclear receptor signaling, disease-specific pathways, and organismal growth and development were significantly activated in the proximal nerve stumps after nerve transection. Organ development, inflammation and immune response, diseases and organ abnormalities, and cellular behavior-related biological functions were highly involved. Conclusions: The expression levels of differentially expressed genes in biological function "Organismal Injury and Abnormalities" were displayed and validated. Our current study helps to obtain a better understanding of the biological processes of peripheral nerve regeneration, especially the regeneration process in the proximal nerve stumps, and thus may help to discover new therapeutic methods that can promote nerve regeneration.

Purpose of the study: Edaravone is an oxygen free radical scavenger that is widely used to treat ischemic injury to the nervous system. This study investigated the effect of edaravone pretreatment on neurons subjected to oxygen-glucose deprivation/recovery (OGD/R) injury. Materials and methods: Common neurons were subjected to oxygen and glucose deprivation for 1 h, followed by oxygen and glucose recovery for 0.5, 2, 6 and 12 h to establish the OGD/R model. Autophagy was assessed by electron microscope observation of autophagosomes, cell immunofluorescence, mRFP-GFP-LC3 virus cell fluorescence and western blotting analyses of the autophagy-related proteins. The findings showed that at OGD/R 2 h autophagy was high. Next, neurons were pretreated with different concentrations of edaravone (0, 5, 10, 25, 50 and 100 mu M) before establishing the OGD/R model. Western blotting was used to analyze the expression of autophagy-related proteins. The CCK-8 assay was used to analyze cell viability after pretreatment with different concentrations of edaravone. Optimal inhibition of autophagy was achieved with the concentration of edaravone 50 mu M. Neurons pretreated with 50 mu M edaravone and established OGD/R model were analyzed for autophagy levels. Results: At every OGD/R time point autophagy was lower in neurons pretreated with edaravone than in those not pretreated with the drug. The difference was statistically significant without OGD/R 12 h. Conclusions: Pretreatment with edaravone may reduce the level of autophagy in neurons subjected to OGD/R injury.

A polylactic acid-polyethylene glycol block copolymer (PLA-b-PEG) was used as an additive to prepare gelatin/PLA-b-PEG blend films for the first time. The PEG molecule block enhanced the compatibility of the PLA molecule block with gelatin, which greatly improved the excellent mechanical and gas barrier properties of the gelatin film. The film contained 5 wt% PLA-b-PEG possessed the highest tensile strength and the highest elastic modulus. When the PLA-b-PEG content further increased to 20 wt%, the tensile strength, elastic modulus and elongation at the break of the blend film were all higher than pure gelatin film, suggesting that the gelatin/PLA-b-PEG blend film was pliable and tough. The blend film possessed not only excellent oxygen barrier property, but also a much-improved water barrier property. The degradation rate of the blend film was elongated controllably by regulating the content of the PLA-b-PEG copolymer. The blend film showed great potential in the application of food packaging.