The IPGA1-ANGUSTIFOLIA module regulates microtubule organisation and pavement cell shape in Arabidopsis.

Plant cells continuously experience mechanical stress resulting from the cell wall that bears internal turgor pressure. Cortical microtubules align with the predicted maximal tensile stress direction to guide cellulose biosynthesis and therefore results in cell wall reinforcement. We have previously identified Increased Petal Growth Anisotropy (IPGA1) as a putative microtubule-associated protein in Arabidopsis, but the function of IPGA1 remains unclear. Here, using the Arabidopsis cotyledon pavement cell as a model, we demonstrated that IPGA1 forms protein granules and interacts with ANGUSTIFOLIA (AN) to cooperatively regulate microtubule organisation in response to stress. Application of mechanical perturbations, such as cell ablation, led to microtubule reorganisation into aligned arrays in wild-type cells. This microtubule response to stress was enhanced in the IPGA1 loss-of-function mutant. Mechanical perturbations promoted the formation of IPGA1 granules on microtubules. We further showed that IPGA1 physically interacted with AN both in vitro and on microtubules. The ipga1 mutant alleles exhibited reduced interdigitated growth of pavement cells, with smooth shape. IPGA1 and AN had a genetic interaction in regulating pavement cell shape. Furthermore, IPGA1 genetically and physically interacted with the microtubule-severing enzyme KATANIN. We propose that the IPGA1-AN module regulates microtubule organisation and pavement cell shape.

WIF1 enhanced dentinogenic differentiation in stem cells from apical papilla.

BACKGROUND: Odontogenic mesenchymal stem cells (MSCs) isolated from tooth tissues are a reliable resource that can be utilized for dental tissue regeneration. Exploration of the mechanisms underlying the regulation of their differentiation may be helpful for investigating potential clinical applications. The stem cell niche plays an important role in maintaining cell functioning. Previous studies found that Wnt inhibitory factor 1 (WIF1) is more highly expressed in apical papilla tissues than in stem cells from apical papilla (SCAPs) using microarray analysis. However, the function of WIF1 in SCAPs remains unclear. In the present study, we investigated the function of WIF1 during dentinogenic differentiation in SCAPs. METHODS: A retrovirus containing HA-WIF1 was used to overexpress WIF1 in SCAPs. Using Western blot analysis, we verified the expression of HA-WIF1. Alkaline phosphatase (ALP) activity assays, Alizarin Red staining and quantitative calcium analysis were performed to investigate the in vitro potential for dentinogenic differentiation in SCAPs. The expression of dentinogenesis-associated genes DSPP, DMP1, Runx2 and OSX were assayed using real-time RT-PCR. Transplantation experiments were used to measure dentinogenesis potential in vivo. RESULTS: The real time RT-PCR results showed that WIF1 was more highly expressed in apical papilla tissues than in SCAPs, and its expression was increased during the process of dentinogenic differentiation. Overexpression of WIF1 enhanced ALP activity and mineralization in vitro, as well as the expression of DSPP, DMP1 and OSX in SCAPs. Moreover, in vivo transplantation experiments revealed that dentinogenesis in SCAPs was enhanced by WIF1 overexpression. CONCLUSION: These results suggest that WIF1 may enhance dentinogenic differentiation potential in dental MSCs via its regulation of OSX and identified potential target genes that could be useful for improving dental tissue regeneration.

LIM homeobox 8 reduced apoptosis and promoted periodontal tissue regeneration function of dental pulp stem cells.

Stem cell-mediated tissue regeneration is a promising strategy for repairing tissue defects and functional reconstruction in periodontitis, a common disease that leads to the loss of alveolar bone and teeth. However, stem cell apoptosis, widely observed during tissue regeneration, impairs its efficiency. Therefore, the regulation of stem cell apoptosis is critical for improving regeneration efficiency. The LIM homeobox 8 gene LHX8, belongs to the LIM homeobox family, which was involved in tooth morphogenesis. Here, we found that LHX8 was significantly expressed in dental pulp. LHX8 knockdown significantly increased dental pulp mesenchymal stem cells (DPSCs) apoptosis, as confirmed by RT-PCR, western blotting, flow cytometry, and transmission electron microscopy. Additionally, LHX8 overexpression inhibited apoptosis and enhanced the osteo/odontogenic differentiation potential of hDPSCs in vitro. Furthermore, LHX8-overexpression could enhance the periodontal tissue regeneration efficiency of hDPSCs in mice with periodontitis. In conclusion, the present study indicates that LHX8 inhibits stem cell apoptosis and promotes functional tissue formation in stem cell-based tissue regeneration engineering, suggesting a new therapeutic target to increase the efficacy of periodontal tissue regeneration.

Facile engineered macrophages-derived exosomes-functionalized PLGA nanocarrier for targeted delivery of dual drug formulation against neuroinflammation by modulation of microglial polarization in a post-stroke depression rat model.

Post-stroke depression (POSD) is a common difficulty and most predominant emotional syndrome after stroke often consequences in poor outcomes. In the present investigation, we have designed and studied the neurologically active celastrol/minocycline encapsulated with macrophages-derived exosomes functionalized PLGA nanoformulations (CMC-EXPL) to achieve enhanced anti-inflammatory behaviour and anti-depressant like activity in a Rat model of POSD. The animal model of POSD was established through stimulating process with chronic unpredictable mild stress (CUM) stimulations after procedure of middle cerebral artery occlusion (MCAO). Neuronal functions and Anti-inflammation behaviours were observed by histopathological (H&E) examination and Elisa analyses, respectively. The anti-depressive activity of the nanoformulations treated Rat models were evaluated by open-field and sucrose preference test methods. Microglial polarization was evaluated via flow-cytometry and qRT-PCR observations. The observed results exhibited that prepared nanoformulations reduced the POSD-stimulated depressive-like activities in rat models as well alleviated the neuronal damages and inflammatory responses in the cerebral hippocampus. Importantly, prepared CMC-EXPL nanoformulation effectively prevented the M1 pro-inflammatory polarization and indorsed M2 anti-inflammatory polarization, which indicates iNOS and CD86 levels significantly decreased and upsurged Arg-1 and CD206 levels. CMC-EXPL nanoformulation suggestively augmented anti-depressive activities and functional capability and also alleviated brain inflammation in POSD rats, demonstrating its therapeutic potential for POSD therapy.

Circularly polarized coherent anti-Stokes Raman scattering microscopy.

We demonstrate circularly polarized coherent anti-Stokes Raman scattering (CP-CARS) microscopy that significantly suppresses the nonresonant background for high-contrast vibrational imaging. Circularly polarized pump and Stokes fields with opposite handedness are used to excite CARS signal. In this case, theoretically the nonresonant CARS signal and resonant CARS signal from isotropic media will completely vanish, while the resonant CARS signal from anisotropic structures can still exist. This allows CARS imaging of anisotropic samples with enhanced resonant contrast. Furthermore, we performed CP-CARS imaging on fibroin fibers from silkworm silk, and the results confirmed its effectiveness in background suppression.

Nanoscale hematoporphrin-based frameworks for photo-sono synergistic cancer therapy via utilizing Al(III) as metal nodes rather than heavy metals.

Nanoscale metal-organic frameworks composed of heavy metal ions (such as Fe3+ and Cu2+) as metal nodes have been utilized for cancer therapy, but they suffer from serious quenching in fluorescence and photo-sono sensitization due to their paramagnetism and unsaturated 3d orbitals. To solve these problems, we synthesize nanoscale hematoporphrin-based frameworks with Al3+ ions as metal nodes (AlHFs) rather than heavy metals and achieve enhanced photo-sono therapy of malignant tumors. The hydrophilic AlHFs are prepared by first assembling hematoporphrin molecules and Al(III) trimers via covalent coordination and then surface-modifying them with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(polyethylene glycol) (DSPE-PEG) molecules. Under excitation with 660 nm light or ultrasound, AlHFs-PEG can produce 3.6-fold or 2.8-fold more 1O2 species than the as-synthesized nanoscale Fe-hematoporphrin frameworks (FeHFs) because the Al3+ ions without 3d orbitals are not beneficial for energy transfer, while Fe3+ ions with unsaturated 3d orbitals and paramagnetism can cause significant energy transfer. AlHFs-PEG exhibits high biocompatibility and can be engulfed by cells to produce intracellular 1O2 for efficient destruction of cells. With the high biosafety and the photo-sono sensitization, the growth rate of tumors in mice with the AlHFs-PEG injection is significantly inhibited upon exposure to both light and ultrasound, showing higher therapeutic efficacy than photodynamic therapy or sonodynamic therapy alone. Therefore, the present work not only presents the preparation of AlHFs-PEG for tumor photo-sono therapy but also provides some insights for developing nanoscale frameworks with light metal ions as metal nodes.

Recent Advances in Cell and Functional Biomaterial Treatment for Spinal Cord Injury.

Spinal cord injury (SCI) is a devastating central nervous system disease caused by accidental events, resulting in loss of sensory and motor function. Considering the multiple effects of primary and secondary injuries after spinal cord injury, including oxidative stress, tissue apoptosis, inflammatory response, and neuronal autophagy, it is crucial to understand the underlying pathophysiological mechanisms, local microenvironment changes, and neural tissue functional recovery for preparing novel treatment strategies. Treatment based on cell transplantation has become the forefront of spinal cord injury therapy. The transplanted cells provide physical and nutritional support for the damaged tissue. At the same time, the implantation of biomaterials with specific biological functions at the site of the SCI has also been proved to improve the local inhibitory microenvironment and promote axonal regeneration, etc. The combined transplantation of cells and functional biomaterials for SCI treatment can result in greater neuroprotective and regenerative effects by regulating cell differentiation, enhancing cell survival, and providing physical and directional support for axon regeneration and neural circuit remodeling. This article reviews the pathophysiology of the spinal cord, changes in the microenvironment after injury, and the mechanisms and strategies for spinal cord regeneration and repair. The article will focus on summarizing and discussing the latest intervention models based on cell and functional biomaterial transplantation and the latest progress in combinational therapies in SCI repair. Finally, we propose the future prospects and challenges of current treatment regimens for SCI repair, to provide references for scientists and clinicians to seek better SCI repair strategies in the future.

Development of Polymer-Ceramic-Metal Graded Acoustic Matching Layers via Cold Sintering.

A family of three phase, polymer-ceramic-metal (Poly-cer-met) electrically conducting composites was developed via cold sintering for acoustic matching application in medical ultrasound transducers. A range of acoustic impedance ( Z ) between MRayl with low attenuation (<3.5 dB/mm, measured at 10 MHz) was achieved in composites of zinc oxide, silver, and in thermoplastic polymers like Ultem polyetherimide (PEI) or polytetrafluoroethylene (PTFE) at sintering pressure less than 50 MPa and temperature of 150 °C. Densities exceeding 95% were achieved, with resistivities less than 1 Ω -cm. The acoustic velocity was homogeneous across the part (variations <5%). The acoustic velocities exceeded 2500 m/s for Z above 12 MRayl. The experimentally measured acoustic impedance of ZnO/Ag/PEI composites was observed to be in close agreement with the theoretical logarithmic model developed for different volume fractions of individual phases at the percolation limit for Ag. Thus, the acoustic properties of this family of matching layers (MLs) can be predicted to a good approximation before experimental realization. Additionally, a non-conducting low Z (5 MRayl MRayl) with acoustic velocities exceeding 2000 m/s was achieved using hydrozincite as the ceramic component. Scaling of the composites to 2'' diameter was demonstrated. A -6 dB bandwidth greater than 85% was measured for a three ML ultrasound transducer, fabricated using a single cold sintered layer ( Z = 19 MRayl) and two other commercial layers in the stack. Finally, a co-cold sintered graded prototype consisting of three tape-casted formulations corresponding to Z = 5 , 9, and 19 MRayl, while still retaining the correct distributions of the components was demonstrated.

Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro.

A major challenge in coronavirus vaccination and treatment is to counteract rapid viral evolution and mutations. Here we demonstrate that CRISPR-Cas13d offers a broad-spectrum antiviral (BSA) to inhibit many SARS-CoV-2 variants and diverse human coronavirus strains with >99% reduction of the viral titer. We show that Cas13d-mediated coronavirus inhibition is dependent on the crRNA cellular spatial colocalization with Cas13d and target viral RNA. Cas13d can significantly enhance the therapeutic effects of diverse small molecule drugs against coronaviruses for prophylaxis or treatment purposes, and the best combination reduced viral titer by over four orders of magnitude. Using lipid nanoparticle-mediated RNA delivery, we demonstrate that the Cas13d system can effectively treat infection from multiple variants of coronavirus, including Omicron SARS-CoV-2, in human primary airway epithelium air-liquid interface (ALI) cultures. Our study establishes CRISPR-Cas13 as a BSA which is highly complementary to existing vaccination and antiviral treatment strategies.

Introducing RGD peptides on PHBV films through PEG-containing cross-linkers to improve the biocompatibility.

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), a biodegradable polyester, has been a good candidate of biomaterial employed in tissue engineering. However, the PHBV film is hydrophobic and has no recognition sites for cell attachment. In this study, PHBV films are activated by ammonia plasma treatment to produce amino groups on the surface, followed by sequential reactions with a heterobifunctional cross-linker containing a segment of poly(ethylene glycol) (PEG) and further with RGD-containing peptides. XPS analyses of modified surfaces after each reaction step reveal that the RGD-containing peptides have been covalently grafted onto PHBV films. The result of cell viability assay indicates that the RGD-modified PHBV films exhibit a distinctly improved cellular compatibility. Moreover, according to the results of serum adsorption tests by optical waveguide lightmode spectroscopy (OWLS) and fibrinogen adsorption tests by enzyme-linked immunosorbent assay (ELISA) on unmodified and modified PHBV surfaces, the introduced PEG chains can significantly decrease the nonspecific adsorption of proteins from serum and fibrinogen from plasma, thus decreasing the risk of thrombus formation and improving the blood compatibility of implanted materials.