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Bioactive Materials Sep 2024Cartilage defect (CD) is a common complication in osteoarthritis (OA). Impairment of chondrogenesis and cellular senescence are considered as hallmarks of OA development...
INTRODUCTION
Cartilage defect (CD) is a common complication in osteoarthritis (OA). Impairment of chondrogenesis and cellular senescence are considered as hallmarks of OA development and caused failure of cartilage repair in most clinical CD cases. Exploring markers for cellular senescence in CD patients might provide new perspectives for osteoarthritic CD patients. In the present study, we aim to explore senescent markers in CD patients with OA to fabricate a senescence-targeted SMSC organoid hydrogel for cartilage repair.
METHODS
Clinical cartilage samples from cartilage defect patients were collected. Immunofluorescence staining of senescent markers and SA-β-Gal staining were used to detect the senescence state of SMSCs and chondrocytes in cartilage defect and OA patients. MicroRNA expression profiles of SMSC organoids and H2O2-treated SMSC organoids were analyzed and compared with high-throughput microRNA sequencing. Fluorescent in situ hybridization of miRNA were used to determine the expression level of miR-24 in SMSC organoids and cartilage samples. Interaction between miR-24 and its downstream target was analyzed via qRT-PCR, immunofluorescence and luciferase assay. Senescence-targeted miR-24 μS/SMSC organoid hydrogel (MSOH) was constructed for cartilage repair. Anti-senescence properties and chondrogenesis were determined in vitro for MSOH. Rats were used to evaluate the cartilage repair capacity of the MSOH hydrogel in vivo.
RESULTS
In this study, we found Osteoarthritic cartilage defect patients demonstrated upregulated cellular senescence in joint cartilage. MicroRNA sequencing demonstrated senescence marker miR-24 was negatively associated with cartilage impairment and cellular senescence in osteoarthritic CD patients. Moreover, miR-24 mimics alleviates cellular senescence to promote chondrogenesis by targeting downstream TAOK1. Also, miR-24 downregulated TAOK1 expression and promoted chondrogenesis in SMSC organoids. Senescence-targeted miR-24 μS/SMSC organoid hydrogel (MSOH) was constructed and demonstrated superior chondrogenesis in vitro. Animal experiments demonstrated that MSOH hydrogel showed better cartilage repairing effects and better maintained joint function at 24 weeks with low intra-articular inflammatory response after transplantation in rat joint. Single-cell RNA-seq of generated cartilage indicated that implanted MSOH could affect chondrocyte homeostatic state and alter the chondrocyte cluster frequency by regulating cellular glycolysis and OXPHOS, impacting cell cycle and ferroptosis to alleviate cellular senescence and prevent joint degeneration.
CONCLUSION
Osteoarthritic cartilage defect patients demonstrated upregulated cellular senescence in joint cartilage. Senescence marker miR-24 was negatively associated with cartilage impairment in osteoarthritic CD patients. miR-24 attenuates chondrocytes senescence and promotes chondrogenesis in SMSC organoids through targeting TAOK1. Senescence-targeted miR-24 microsphere/SMSC organoid composite hydrogel could successfully repair cartilage defect in osteoarthritic microenvironment via enhanced miR-24/TAOK1 signaling pathway, suggesting MSOH might be a novel therapy for cartilage repair in osteoarthritic CD patients.
PubMed: 38855061
DOI: 10.1016/j.bioactmat.2024.05.036 -
Heliyon Jun 2024Plastic pollution is a worldwide problem especially in the marine environment. Plastic items once fragmented into microplastics (MPs), can be captured by different...
Plastic pollution is a worldwide problem especially in the marine environment. Plastic items once fragmented into microplastics (MPs), can be captured by different marine species. Benthic filter feeders like sponges and polychaetas, due to their trophic strategy, are highly exposed to MPs pollution. Herein a simple but effective method to digest the fan worm and the calcareous sponge is presented: a solution with KOH and HO was able to remove quantitatively (more than 98 %) the organic matter in 3 h while an acid treatment dissolved most of spicules and chaetes in less than 30 min. MPs were easily identified both microscopically and spectroscopically on filters. Quantification in animals collected from the same environment showed that, on average, sponges accumulate fewer MPs than polychaetes (66 ± 31 and 117 ± 46 particles/g dry weight, respectively). The plastic recovery of the method was validated using three different approaches (spiking of standard PS microspheres, of common-use plastic objects, and of microplastics already weathered in marine environment). This procedure can make it easier and cost-effective to process biota in monitoring studies, providing information about bioindicator/bioremediation species.
PubMed: 38845917
DOI: 10.1016/j.heliyon.2024.e31796 -
Frontiers in Immunology 2024In a cooperative study of the University Hospital Leipzig, University of Leipzig, and the Charité Berlin on kidney transplant patients, we analysed the occurrence of...
OBJECTIVE
In a cooperative study of the University Hospital Leipzig, University of Leipzig, and the Charité Berlin on kidney transplant patients, we analysed the occurrence of HLA-specific antibodies with respect to the HLA setup of the patients. We aimed at the definition of specific HLA antigens towards which the patients produced these antibodies.
METHODS
Patients were typed for the relevant HLA determinants using mainly the next-generation technology. Antibody screening was performed by the state-of-the-art multiplex-based technology using microspheres coupled with the respective HLA alleles of HLA class I and II determinants.
RESULTS
Patients homozygous for *********, and * in the class I group and **********, and * in the class II group were found to have a significant higher antibody production compared to the heterozygous ones. In general, all HLA determinants are affected. Remarkably, * homozygous patients can produce antibodies towards all HLA-A determinants, while * homozygous ones make antibodies towards all HLA-B and selected HLA-A and C antigens, and are associated with an elevation of , and * seems to increase the risk for antibody responses against most of the HLA class I antigens (HLA-A, HLA-B, and HLA-C) in contrast to * where a lower risk towards few HLA-A and HLA-B alleles is found. The widely observed differential antibody response is therefore to be accounted to the patient's HLA type.
CONCLUSION
Homozygous patients are at risk of producing HLA-specific antibodies hampering the outcome of transplantation. Including this information on the allocation procedure might reduce antibody-mediated immune reactivity and prevent graft loss in a patient at risk, increasing the life span of the transplanted organ.
Topics: Humans; Kidney Transplantation; Homozygote; Risk Factors; HLA Antigens; Isoantibodies; Histocompatibility Testing; Alleles; Antibody Formation; Male; Female
PubMed: 38840925
DOI: 10.3389/fimmu.2024.1384823 -
Frontiers in Veterinary Science 2024Avian leukosis, a viral disease affecting birds such as chickens, presents significant challenges in poultry farming due to tumor formation, decreased egg production,...
INTRODUCTION
Avian leukosis, a viral disease affecting birds such as chickens, presents significant challenges in poultry farming due to tumor formation, decreased egg production, and increased mortality. Despite the absence of a commercial vaccine, avian leukosis virus (ALV) infections have been extensively documented, resulting in substantial economic losses in the poultry industry. This study aimed to develop alginate-chitosan composite microspheres loaded with ALV-J Gp85 protein (referred to as aCHP-gp85) as a potential vaccine candidate.
METHODS
Sodium alginate and chitosan were utilized as encapsulating materials, with the ALV-J Gp85 protein serving as the active ingredient. The study involved 45 specific pathogen-free (SPF) chickens to evaluate the immunological effectiveness of aCHP-gp85 compared to a traditional Freund adjuvant-gp85 vaccine (Freund-gp85). Two rounds of vaccination were administered, and antibody levels, mRNA expression of immune markers, splenic lymphocyte proliferation, and immune response were assessed. An animal challenge experiment was conducted to evaluate the vaccine's efficacy in reducing ALV-J virus presence and improving clinical conditions.
RESULTS
The results demonstrated that aCHP-gp85 induced a significant and sustained increase in antibody levels compared to Freund-gp85, with the elevated response lasting for 84 days. Furthermore, aCHP-gp85 significantly upregulated mRNA expression levels of key immune markers, notably TNF-α and IFN-γ. The application of ALV-J Gp85 protein within the aCHP-gp85 group led to a significant increase in splenic lymphocyte proliferation and immune response. In the animal challenge experiment, aCHP-gp85 effectively reduced ALV-J virus presence and improved clinical conditions compared to other groups, with no significant pathological changes observed.
DISCUSSION
The findings suggest that aCHP-gp85 elicits a strong and prolonged immune response compared to Freund-gp85, indicating its potential as an innovative ALV-J vaccine candidate. These results provide valuable insights for addressing avian leukosis in the poultry industry, both academically and practically.
PubMed: 38840641
DOI: 10.3389/fvets.2024.1374923 -
Langmuir : the ACS Journal of Surfaces... Jun 2024Actin, found in all eukaryotic cells as globular (G) or filamentous (F) actin, undergoes polymerization, with G-actin units changing shape to become F-actin. Thermal...
Actin, found in all eukaryotic cells as globular (G) or filamentous (F) actin, undergoes polymerization, with G-actin units changing shape to become F-actin. Thermal proteins, or proteinoids, are created by heating amino acids (160-200 °C), forming polymeric chains. These proteinoids can swell in an aqueous solution at around 50 °C, producing hollow microspheres filled with a solution, exhibiting voltage spikes. Our research explores the signaling properties of proteinoids, actin filaments, and hybrid networks combining actin and proteinoids. Proteinoids replicate brain excitation dynamics despite lacking specific membranes or ion channels. We investigate enhancing conductivity and spiking by using pure actin, yielding improved coordination in networks compared with individual filaments or proteinoids. Temperature changes (20 short-peptide supramolecular C to 80 °C) regulate conduction states, demonstrating external control over emergent excitability in protobrain systems. Adding actin to proteinoids reduces spike timing variability, providing a more uniform feature distribution. These findings support theoretical models proposing cytoskeletal matrices for functional specification in synthetic protocell brains, promoting stable interaction complexity. The study concludes that life-like signal encoding can emerge spontaneously within biological polymer scaffolds, incorporating abiotic chemistry.
Topics: Actin Cytoskeleton; Microspheres; Actins; Temperature; Animals
PubMed: 38837748
DOI: 10.1021/acs.langmuir.4c01107 -
Physica Medica : PM : An International... Jun 2024The dosimetry evaluation for the selective internal radiation therapy is currently performed assuming a uniform activity distribution, which is in contrast with...
UNLABELLED
The dosimetry evaluation for the selective internal radiation therapy is currently performed assuming a uniform activity distribution, which is in contrast with literature findings. A 2D microscopic model of the perfused liver was developed to evaluate the effect of two different Y microspheres distributions: i) homogeneous partitioning with the microspheres equally distributed in the perfused liver, and ii) tumor-clustered partitioning where the microspheres distribution is inferred from the patient specific images.
METHODS
Two subjects diagnosed with liver cancer were included in this study. For each subject, abdominal CT scans acquired prior to the SIRT and post-treatment Y positron emission tomography were considered. Two microspheres partitionings were simulated namely homogeneous and tumor-clustered partitioning. The homogeneous and tumor-clustered partitionings were derived starting from CT images. The microspheres radiation is simulated by means of Russell's law.
RESULTS
In homogenous simulations, the dose delivery is uniform in the whole liver while in the tumor-clustered simulations a heterogeneous distribution of the delivered dose is visible with higher values in the tumor regions. In addition, in the tumor-clustered simulation, the delivered dose is higher in the viable tumor than in the necrotic tumor, for all patients. In the tumor-clustered case, the dose delivered in the non-tumoral tissue (NTT) was considerably lower than in the perfused liver.
CONCLUSIONS
The model proposed here represents a proof-of-concept for personalized dosimetry assessment based on preoperative CT images.
Topics: Liver Neoplasms; Carcinoma, Hepatocellular; Humans; Microspheres; Radiotherapy Dosage; Yttrium Radioisotopes; Models, Biological; Tomography, X-Ray Computed; Radiation Dosage; Microscopy
PubMed: 38824827
DOI: 10.1016/j.ejmp.2024.103384 -
Stem Cell Research & Therapy Jun 2024Nerve guide conduits are a promising strategy for reconstructing peripheral nerve defects. Improving the survival rate of seed cells in nerve conduits is still a...
BACKGROUND
Nerve guide conduits are a promising strategy for reconstructing peripheral nerve defects. Improving the survival rate of seed cells in nerve conduits is still a challenge and microcarriers are an excellent three-dimensional (3D) culture scaffold. Here, we investigate the effect of the 3D culture of microcarriers on the biological characteristics of adipose mesenchymal stem cells (ADSCs) and to evaluate the efficacy of chitosan nerve conduits filled with microcarriers loaded with ADSCs in repairing nerve defects.
METHODS
In vitro, we prepared porous chitosan microspheres by a modified emulsion cross-linking method for loading ADSCs and evaluated the growth status and function of ADSCs. In vivo, ADSCs-loaded microcarriers were injected into chitosan nerve conduits to repair a 12 mm sciatic nerve defect in rats.
RESULTS
Compared to the conventional two-dimensional (2D) culture, the prepared microcarriers were more conducive to the proliferation, migration, and secretion of trophic factors of ADSCs. In addition, gait analysis, neuro-electrophysiology, and histological evaluation of nerves and muscles showed that the ADSC microcarrier-loaded nerve conduits were more effective in improving nerve regeneration.
CONCLUSIONS
The ADSCs-loaded chitosan porous microcarrier prepared in this study has a high cell engraftment rate and good potential for peripheral nerve repair.
Topics: Chitosan; Nerve Regeneration; Animals; Microspheres; Rats; Mesenchymal Stem Cells; Adipose Tissue; Rats, Sprague-Dawley; Sciatic Nerve; Porosity; Tissue Scaffolds; Male; Mesenchymal Stem Cell Transplantation; Cell Proliferation; Cells, Cultured
PubMed: 38824568
DOI: 10.1186/s13287-024-03753-w -
Ecotoxicology and Environmental Safety Jul 2024Microplastics (MP) can influence a plethora of fungal species within the rhizosphere. Nevertheless, there are few studies on the direct impacts of MPs on soil fungi and...
Microplastics (MP) can influence a plethora of fungal species within the rhizosphere. Nevertheless, there are few studies on the direct impacts of MPs on soil fungi and their intricate interplay with plants. Here, we investigated the impact of polyethylene microspheres (PEMS) on the ecological interactions between Fusarium solani, a plant pathogenic fungus, and Trichoderma viride, a fungal plant growth promotor, within the rhizosphere of Solanum lycopersicum (tomato). Spores of F. solani and T. viride were pre-incubated with PEMS at two concentrations, 100 and 1000 mg L. Mycelium growth, sporulation, spore germination, and elongation were evaluated. Tomato seeds were exposed to fungal spore suspensions treated with PEMS, and plant development was subsequently assessed after 4 days. The results showed that PEMS significantly enhanced the sporulation (106.0 % and 70.1 %) but compromised the spore germination (up to 27.3 % and 32.2 %) and radial growth (up to -5.2% and -21.7 %) of F. solani and T. viride, respectively. Furthermore, the 100 and 1000 mg L concentrations of PEMS significantly (p<0.05) enhanced the mycelium density of T. viride (9.74 % and 22.30 %, respectively), and impaired the germ-tube elongation of F. solani after 4 h (16.16 % and 11.85 %, respectively) and 8 h (4 % and 17.10 %, respectively). In addition, PEMS amplified the pathogenicity of F. solani and boosted the bio-enhancement effect of T. viride on tomato root growth. Further, PEMS enhanced the bio-fungicidal effect of T. viride toward F. solani (p<0.05). In summary, PEMS had varying effects on F. solani and T. viride, impacting their interactions and influencing their relationship with tomato plants. It intensified the beneficial effects of T. viride and increased the aggressiveness of F. solani. This study highlights concerns regarding the effects of MPs on fungal interactions in the rhizosphere, which are essential for crop soil colonization and resource utilization.
Topics: Solanum lycopersicum; Fusarium; Spores, Fungal; Microplastics; Rhizosphere; Soil Microbiology; Soil Pollutants; Polyethylene; Hypocreales; Microspheres; Plant Roots
PubMed: 38820874
DOI: 10.1016/j.ecoenv.2024.116518 -
Nanoscale Advances May 2024Expression of concern for 'Acceleration of ammonium phosphate hydrolysis using TiO microspheres as a catalyst for hydrogen production' by Ayman H. Zaki , , 2020, ,...
Expression of concern for 'Acceleration of ammonium phosphate hydrolysis using TiO microspheres as a catalyst for hydrogen production' by Ayman H. Zaki , , 2020, , 2080-2086, https://doi.org/10.1039/D0NA00204F.
PubMed: 38817440
DOI: 10.1039/d4na90040e -
Journal of Nanobiotechnology May 2024Extracellular vesicles (EVs) derived from human adipose-derived mesenchymal stem cells (hADSCs) have shown great therapeutic potential in plastic and reconstructive...
A quick and innovative pipeline for producing chondrocyte-homing peptide-modified extracellular vesicles by three-dimensional dynamic culture of hADSCs spheroids to modulate the fate of remaining ear chondrocytes in the M1 macrophage-infiltrated microenvironment.
BACKGROUND
Extracellular vesicles (EVs) derived from human adipose-derived mesenchymal stem cells (hADSCs) have shown great therapeutic potential in plastic and reconstructive surgery. However, the limited production and functional molecule loading of EVs hinder their clinical translation. Traditional two-dimensional culture of hADSCs results in stemness loss and cellular senescence, which is unfavorable for the production and functional molecule loading of EVs. Recent advances in regenerative medicine advocate for the use of three-dimensional culture of hADSCs to produce EVs, as it more accurately simulates their physiological state. Moreover, the successful application of EVs in tissue engineering relies on the targeted delivery of EVs to cells within biomaterial scaffolds.
METHODS AND RESULTS
The hADSCs spheroids and hADSCs gelatin methacrylate (GelMA) microspheres are utilized to produce three-dimensional cultured EVs, corresponding to hADSCs spheroids-EVs and hADSCs microspheres-EVs respectively. hADSCs spheroids-EVs demonstrate excellent production and functional molecule loading compared with hADSCs microspheres-EVs. The upregulation of eight miRNAs (i.e. hsa-miR-486-5p, hsa-miR-423-5p, hsa-miR-92a-3p, hsa-miR-122-5p, hsa-miR-223-3p, hsa-miR-320a, hsa-miR-126-3p, and hsa-miR-25-3p) and the downregulation of hsa-miR-146b-5p within hADSCs spheroids-EVs show the potential of improving the fate of remaining ear chondrocytes and promoting cartilage formation probably through integrated regulatory mechanisms. Additionally, a quick and innovative pipeline is developed for isolating chondrocyte homing peptide-modified EVs (CHP-EVs) from three-dimensional dynamic cultures of hADSCs spheroids. CHP-EVs are produced by genetically fusing a CHP at the N-terminus of the exosomal surface protein LAMP2B. The CHP + LAMP2B-transfected hADSCs spheroids were cultured with wave motion to promote the secretion of CHP-EVs. A harvesting method is used to enable the time-dependent collection of CHP-EVs. The pipeline is easy to set up and quick to use for the isolation of CHP-EVs. Compared with nontagged EVs, CHP-EVs penetrate the biomaterial scaffolds and specifically deliver the therapeutic miRNAs to the remaining ear chondrocytes. Functionally, CHP-EVs show a major effect on promoting cell proliferation, reducing cell apoptosis and enhancing cartilage formation in remaining ear chondrocytes in the M1 macrophage-infiltrated microenvironment.
CONCLUSIONS
In summary, an innovative pipeline is developed to obtain CHP-EVs from three-dimensional dynamic culture of hADSCs spheroids. This pipeline can be customized to increase EVs production and functional molecule loading, which meets the requirements for regulating remaining ear chondrocyte fate in the M1 macrophage-infiltrated microenvironment.
Topics: Humans; Chondrocytes; Extracellular Vesicles; Spheroids, Cellular; Mesenchymal Stem Cells; Peptides; MicroRNAs; Macrophages; Cells, Cultured; Microspheres; Tissue Engineering; Cell Culture Techniques, Three Dimensional; Cellular Microenvironment; Ear Cartilage; Adipose Tissue; Cell Differentiation
PubMed: 38816719
DOI: 10.1186/s12951-024-02567-5