Development and preclinical evaluation of virus-like particle vaccine against COVID-19 infection.

BACKGROUND: Vaccines that incorporate multiple SARS-CoV-2 antigens can further broaden the breadth of virus-specific cellular and humoral immunity. This study describes the development and immunogenicity of SARS-CoV-2 VLP vaccine that incorporates the four structural proteins of SARS-CoV-2. METHODS: VLPs were generated in transiently transfected HEK293 cells, purified by multimodal chromatography, and characterized by tunable-resistive pulse sensing, AFM, SEM, and TEM. Immunoblotting studies verified the protein identities of VLPs. Cellular and humoral immune responses of immunized animals demonstrated the immune potency of the formulated VLP vaccine. RESULTS: Transiently transfected HEK293 cells reproducibly generated vesicular VLPs that were similar in size to and expressing all four structural proteins of SARS-CoV-2. Alum adsorbed, K3-CpG ODN-adjuvanted VLPs elicited high titer anti-S, anti-RBD, anti-N IgG, triggered multifunctional Th1-biased T-cell responses, reduced virus load, and prevented lung pathology upon live virus challenge in vaccinated animals. CONCLUSION: These data suggest that VLPs expressing all four structural protein antigens of SARS-CoV-2 are immunogenic and can protect animals from developing COVID-19 infection following vaccination.

Circulating extracellular vesicles of patients with steroid-sensitive nephrotic syndrome have higher RAC1 and induce recapitulation of nephrotic syndrome phenotype in podocytes.

Since previous research suggests a role of a circulating factor in the pathogenesis of steroid-sensitive nephrotic syndrome (NS), we speculated that circulating plasma extracellular vesicles (EVs) are a candidate source of such a soluble mediator. Here, we aimed to characterize and try to delineate the effects of these EVs in vitro. Plasma EVs from 20 children with steroid-sensitive NS in relapse and remission, 10 healthy controls, and 6 disease controls were obtained by serial ultracentrifugation. Characterization of these EVs was performed by electron microscopy, flow cytometry, and Western blot analysis. Major proteins from plasma EVs were identified via mass spectrometry. Gene Ontology classification analysis and Ingenuity Pathway Analysis were performed on selectively expressed EV proteins during relapse. Immortalized human podocyte culture was used to detect the effects of EVs on podocytes. The protein content and particle number of plasma EVs were significantly increased during NS relapse. Relapse NS EVs selectively expressed proteins that involved actin cytoskeleton rearrangement. Among these, the level of RAC-GTP was significantly increased in relapse EVs compared with remission and disease control EVs. Relapse EVs were efficiently internalized by podocytes and induced significantly enhanced motility and albumin permeability. Moreover, relapse EVs induced significantly higher levels of RAC-GTP and phospho-p38 and decreased the levels of synaptopodin in podocytes. Circulating relapse EVs are biologically active molecules that carry active RAC1 as cargo and induce recapitulation of the NS phenotype in podocytes in vitro.NEW & NOTEWORTHY Up to now, the role of extracellular vesicles (EVs) in the pathogenesis of steroid-sensitive nephrotic syndrome (NS) has not been studied. Here, we found that relapse NS EVs contain significantly increased active RAC1, induce enhanced podocyte motility, and increase expression of RAC-GTP and phospho-p38 expression in vitro. These results suggest that plasma EVs are biologically active molecules in the pathogenesis of NS.

Stem Cell and Advanced Nano Bioceramic Interactions.

Bioceramics are type of biomaterials generally used for orthopaedic applications due to their similar structure with bone. Especially regarding to their osteoinductivity and osteoconductivity, they are used as biodegradable scaffolds for bone regeneration along with mesenchymal stem cells. Since chemical properties of bioceramics are important for regeneration of tissue, physical properties are also important for cell proliferation. In this respect, several different manufacturing methods are used for manufacturing nano scale bioceramics. These nano scale bioceramics are used for regeneration of bone and cartilage both alone or with other types of biomaterials. They can also act as carrier for the delivery of drugs in musculoskeletal infections without causing any systemic toxicity.

Boron Nano-hydroxyapatite Composite Increases the Bone Regeneration of Ovariectomized Rabbit Femurs.

Osteoporosis is a systemic metabolic disease defined by a decreased bone mineral density, microarchitectural deterioration, and an increased incidence of fragility fractures that may lead to morbidity and mortality. Boron may stimulate new bone formation and regeneration, when combined with nano-hydroxyapatite. We questioned whether injecting boron-containing nano-hydroxyapatite composites with hyaluronan increased the bone mineral density and new bone formation in osteoporotic rabbit femurs. The regenerative effects of injectable boron-containing nano-hydroxyapatite composites from 6 to 12 weeks, which may prevent osteoporotic femoral fractures, were assessed. Boron-containing (10 µg/ml) nano-hydroxyapatite composites were injected into the intramedullary femoral cavity with hyaluronan. These significantly increased the histomorphometric new bone surface to the total bone surface ratio at 6 and 9 weeks. The micro-tomographic bone volume to the total volume ratio and bone mineral density in osteoporotic rabbit femurs increased when compared to the hyaluronan (p = 0.004, p = 0.004, p = 0.004, p = 0.01, respectively) and the sham-control (p = 0.01, p = 0.004, p = 0.01, p = 0.037, respectively) groups. The boron-containing group had a higher bone mineralization and new bone formation compared to the nano-hydroxyapatite group, although the difference was not statistically significant. These findings reveal that intramedullary injection of boron-containing nano-hydroxyapatite with hyaluronan increases new bone formation and mineralization in ovariectomized rabbit femurs. Boron-containing nano-hydroxyapatite composites are promising tissue engineering biomaterials that may have regenerative potential in preventing primary and/or secondary femoral fractures in osteoporosis patients.